US 2012.0034613A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2012/0034613 A1 Gopaul et al. (43) Pub. Date: Feb. 9, 2012

(54) APPARATUS AND METHOD FOR TESTING Related U.S. Application Data RELATIONSHIPS BETWEEN EXPRESSION AND PHYSICAL APPEARANCE (60) Provisional application No. 61/370,190, filed on Aug. OFSKN 3, 2010. Publication Classification (75) Inventors: Remona Gopaul, Spanish Fork, UT (US); Dale G. Kern, Hyde Park, (51) Int. Cl. UT (US); Janet F. Lephart, Orem, CI2O I/68 (2006.01) UT (US) CI2M I/34 (2006.01) (52) U.S. Cl...... 435/6.12: 435/6.13:435/287.2 (73) Assignee: Nst Products, Inc., Provo, UT (57) ABSTRACT The disclosure is directed to apparatus and methods for test (21) Appl. No.: 13/196,288 ing relationships between gene expression and physical appearance of skin and methods of assessing the efficacy of (22) Filed: Aug. 2, 2011 skin anti-aging agents. Patent Application Publication Feb. 9, 2012 Sheet 1 of 18 US 2012/0034613 A1

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APPARATUS AND METHOD FOR TESTING 0008. The present disclosure has been developed against RELATIONSHIPS BETWEEN GENE this backdrop. EXPRESSION AND PHYSICAL APPEARANCE OFSKN SUMMARY 0009. In a first aspect, the present disclosure is directed to CROSS-REFERENCE TO RELATED a method of testing to identify associated with one or APPLICATIONS more physical attributes of skin aging. The methods comprise exposing a first sample of skin cells or tissue to an 0001. This application claims priority to U.S. Provisional agent, determining a first set of expression levels of a plurality Application No. 61/370,190 filed Aug. 3, 2010, entitled of genes in the first sample of human skin, comparing the first Apparatus and Method for Testing Relationships Between set of expression levels to a second set of expression levels, Gene Expression and Physical Appearance of Skin, the the second set of expression levels corresponding to expres sion levels of human skin tissue not exposed to the agent, to entire content of which is hereby incorporated herein by identify a first Subset of genes having a fold change difference reference in its entirety. in expression level between the exposed and unexposed samples that meets a first, selected biological relevance level. FIELD selecting from the first Subset of genes a second Subset of genes, each gene being associated with a biochemical path 0002 The present disclosure relates to apparatus and way associated with physical appearance of skin aging, methods for testing to identify and select genes associated selecting from the second Subset of genes, at least one skin with certain physical attributes of skin and methods of assess attribute Subset of genes, each gene in the skin attribute Subset ing the efficacy of a skin anti-aging agent. being associated with a biochemical pathway relating to the skin attribute that is shown in the comparing step to have been regulated in a more youthful direction for that biochemical BACKGROUND pathway and skin attribute, exposing a second sample of human skin tissue to the agent, determining the levels of 0003. The sequencing of the and the con expression for the at least one skin attribute Subset of genes in tinual development of analytical methods to quickly and inex the second sample of human skin tissue using a method for pensively measure whole-genome gene expression changes determining expression levels that is different than that used has created an overload of information and a need for meth for the first sample of human skin tissue, and selecting a third ods to organize, focus and reduce the data resulting from gene subset of genes from the at least one skin attribute subset of level analyses of biological processes. genes whose expression levels in the second sample of human 0004 Analysis of the biological process of aging may be skin tissue meet a second, selected biological relevance level performed at the genomic level. Skin is the largest organ of and whose direction of regulation conforms to the more the human body. The skin aging process is influenced by youthful direction used in selecting the at least one skin many different extrinsic (e.g., environmental) or intrinsic (in attribute Subset of genes. cluding genetic and biochemical pathway) factors. 0010. In some embodiments the biochemical pathway associated with the physical appearance of skin aging com 0005. However, examining all the changes at the human prises at least one of skin structural synthesis, skin genome or genomic product level can be overwhelming, par structural degradation and maintenance, extracellular matrix ticularly when Strategies of anti-aging are examined. In skin, assembly, cellular differentiation, skin barrier component there are thousands of changes in gene expression with chro synthesis, skin barrier integrity, water regulation, or regula nological aging and photo-aging (Robinson et al., Genomic tion of melanin production and control. driven insights into changes in aging skin, J. Drugs Derma 0011. In some embodiments, the skin attribute for the at tol., 2009, 8(7 Suppl):s8-11). Not only does genome-wide least one skin attribute Subset of genes is skin structure, skin testing produce massive data sets, the literature reporting on pigmentation, skin hydration or turnover. genetic research is also constantly growing, making it diffi 0012. In another embodiment, the first, selected biological cult to bring together all desired knowledge and test results. relevance level is about a two fold difference between the 0006 Focusing research resources on a smaller group of exposed and unexposed samples. genes and/or associated , and the biochemical path 0013. In some embodiments, the human skin tissue com ways associated with the groups of genes and/or proteins may prises skin cells comprising at least one of keratinocytes, be used to reduce confusion and distraction from the many fibroblasts, adipocytes, melanocytes or combinations thereof. unknown or irrelevant factors and variables. In addition, there 0014. In another embodiment, the first set of expression is a cost to including genes in an analysis. If fewer genes are levels of a plurality of gene comprises expression levels for analyzed due to a better focus, the costs of research are essentially the full human genome. reduced. 0015. In other embodiments, the method of determining 0007. On the other hand, for an organ as complex as skin expression levels that is different than that used for the first and a problem Such as aging that has multiple dimensions, too sample of human tissue is a method using an RNA quantifi narrow a focus also may be problematic, by excluding genetic cation metric. pathways that play a role in skin aging. An approach includ 0016. In some embodiments, in the step of selecting from ing an overall strategy to slow down physical manifestations the second Subset of genes, at least one skin attribute Subset of of the aging process in skin by examining at the gene level genes, each gene in the skin attribute Subset being associated several mechanisms of aging simultaneously may be used, with a biochemical pathway relating to the skin attribute that instead of in-depth analysis of each individual gene. is shown in the comparing step to have been regulated in a US 2012/0034613 A1 Feb. 9, 2012 more youthful direction for that biochemical pathway and skin structure, skin pigmentation, skin hydration or cell turn skin attribute comprising performing this step for a plurality over. In another embodiment in the system, the first, selected of skin attribute subsets of genes, and the step selecting a third biological relevance level is about a two fold difference Subset of genes from the at least one skin attribute subset of between the exposed and unexposed samples. genes whose expression levels in the second sample of human 0021. In other embodiments in the system, the human skin skin tissue meet a second, selected biological relevance level tissue comprises skin cells comprising at least one of kerati and whose direction of regulation conforms to the more nocytes, fibroblasts, adipocytes, melanocytes or combina youthful direction used in selecting the at least one skin tions thereof. attribute Subset of genes comprising performing this step for I0022. In some embodiments in the system the first set of a plurality of skin attribute subsets of genes. expression levels of a plurality of genes comprises expression 0017. In another embodiment, the plurality of skin levels for essentially the full human genome. attribute subsets of genes are two or more skin attribute subset 0023. In another embodiment in the system, the second of genes selected from the group consisting of skin structure, instrument for determining expression levels that is different skin pigmentation, skin hydration and cell turnover. than that used for the first sample of human tissue is an 0018. In other embodiments, the method further com instrument using an RNA quantification metric. prises determining the levels of expression for additional 0024. In a third aspect, methods of assessing the efficacy genes associated with a biochemical pathway associated with of a skin anti-aging agent are disclosed. The methods com skin aging in the second sample of human skin tissue using a prise exposing a first sample of human skin tissue to an agent, method for determining expression levels that is different determining a first set of expression levels of a plurality of than that used for the first sample of human tissue, and select genes in the first sample of human skin, comparing the first set ing for the third subset of genes those genes from the addi of expression levels to a second set of expression levels, the tional genes associated with a biochemical pathway associ Second set of expression levels corresponding to expression ated with skin aging whose expression levels in the second levels of human skin tissue not exposed to the agent, to sample of human skin tissue meet a second, selected biologi identify a first Subset of genes having a fold change difference cal relevance level and whose direction of regulation con in expression level between the exposed and unexposed forms to the more youthful direction of regulation of the samples that meets a first, selected biological relevance level, associated biochemical pathway. Selecting from the first subset of genes a second subset of 0019. In a second aspect, a computer based system of genes, each gene being associated with a biochemical path testing to identify genes associated with one or more physical Way associated with physical appearance of skin aging, attributes of skin aging comprises a first instrument for expos Selecting from the second subset of genes, at least one skin ing a first sample of human skin tissue to an agent and deter attribute subset of genes, each gene in the skin attribute subset mining a first set of expression levels of a plurality of genes in being associated with a biochemical pathway relating to the the first sample of human skin, a computer module for com skin attribute that is shown in the comparing step to have been paring the first set of expression levels to a second set of regulated in a more youthful direction for that biochemical expression levels, the second set of expression levels corre pathway and skin attribute, exposing a second sample of sponding to expression levels of human skin tissue not human skin tissue to the agent, determining the levels of exposed to the agent to identify a first subset of genes having expression for the at least one skin attribute subset of genes in a fold change difference in expression level between the the second sample of human skin tissue using a method for exposed and unexposed samples that meet a first, selected determining expression levels that is different than that used biological relevance level, a computer module for accessing a for the first sample of human skin tissue, selecting a third Stored data set identifying genes, each gene being associated Subset of genes from the at least one skin attribute subset of with a biochemical pathway associated with physical appear genes whose expression levels in the second sample of human ance of skin aging and for selecting from the first subset a skin tissue meet a second, selected biological relevance level Second subset comprising those genes also in the second and whose direction of regulation conforms to the more Subset, a computer module for selecting from the second youthful direction used in selecting the at least one skin Subset of genes, at least one skin attribute subset of genes, attribute subset of genes, and comparing the third subset of each gene in the skin attribute subset being associated with a genes to a previously determined third subset of genes for a biochemical pathway relating to the skin attribute that is second agent, thereby showing the efficacy of the skin anti shown in the comparing step to have been regulated in a more aging agent. youthful direction for that biochemical pathway and skin 0025. In some embodiments of the method, the skin attribute, a second instrument for exposing a second sample attribute for the at least one skin attribute subset of genes is of human skin tissue to the agent and for determining the skin structure, skin pigmentation, skin hydration or cell turn levels of expression for the at least one skin attribute subset of OVer. genes in the second sample of human skin tissue using a 0026. In another embodiment, the method for determining method for determining expression levels that is different expression levels that is different than that used for the first than that used for the first sample of human tissue, a computer sample of human tissue is a method using an RNA quantifi module for selecting a third subset of genes from the at least cation metric. one skin attribute subset of genes whose expression levels in the second sample of human skin tissue meet a second, BRIEF DESCRIPTION OF THE DRAWINGS selected biological relevance level and whose direction of regulation conforms to the more youthful direction used in 0027. Those skilled in the art will understand that the Selecting the at least one skin attribute subset of genes. drawings, described herein, are for illustration purposes only. 0020. In some embodiments in the system, the skin The drawings are not intended to limit the scope of the present attribute for the at least one skin attribute subset of genes is disclosure. US 2012/0034613 A1 Feb. 9, 2012

0028 FIGS. 1A-1K are a flowchart schematically show illary fibroblasts but not reticular fibroblasts. A new view of ing, in a simplified example, steps in a process for choosing skin morphogenesis and aging. PLoS ONE 3(12):e4066, genes for inclusion in a functional youth gene assembly. 2008). 0029 FIG. 2 depicts percent of change over time for dif 0044) If one first examines genes associated with specific ferent Sub-categories of wrinkles, including crow's feet, mechanisms of aging, then by grouping genes from several under eye, and cheek areas. mechanisms (associated with attributes of aging or preserv 0030 FIG.3 depicts percent change over time for a variety ing youthfulness) one can compile a functional youth gene of facial attributes. profile expression or a functional youth gene assembly that is 0031 FIG. 4 depicts percent change over time for corne associated with a specific tissue, such as the skin, and specific ometer measurements of skin hydration. attributes of it. 0032 FIG. 5 depicts percent change over time for cutom eter measurements of skin extensibility. Agents 0033 FIG. 6 depicts percent change over time for ultra 0045 An agent generally refers to a substance that causes Sound measurements of skin density. a change in tissue observed. An agent is chosen based on the 0034 FIG. 7 shows a schematic diagram for a system, ability (or expected ability) of the agent to affect signs of including gene expression level testing devices and data aging, presumably by reason of an effect on expression levels processing components for carrying out the method dis of genes or gene products, including epigenetic effects. closed, and data sets developed and used as the method pro 0046 Salicin is an agent that has shown effects on multiple ceeds. signs of skin aging. See Applicant's co-pending patent appli 0035 FIG. 8 shows the relationships of various data sets cation Ser. No. 12/058,201, publication number US 2009/ and Subsets developed leading to a confirmed skin attribute 0246152 A1, hereby incorporated by reference in its entirety. Subset defining a functional youth gene assembly. 0047. In some embodiments, the skin anti-aging agent chosen for the experimental testing is salicin. Salicin DETAILED DESCRIPTION (CHsO4) or 2-(Hydroxymethyl)phenyl f-D-glucospyra Definitions noside) is an alcoholic beta-glycoside that contains D-glu cose. Salicin is obtained from several species of the white 0036. The term “functional youth gene assembly” refers to willow bark tree. Salicin is commercially available as a white, groups of genes encompassing one or more biochemical path crystalline, water soluble powder from, for example, Sigma ways or mechanisms of aging, addressable for functional Aldrich (St. Louis, Mo.). restoration or stabilization of a more youthful state in the skin. 0048. Other agents may be chosen based on the ability of 0037. The term “skin attributes' refers to characteristics or the Substance to bring about a biological or chemical effect in qualities of human skin. tissue to reflect a more youthful state of the tissue. Choice of 0038. The term “biochemical pathway associated with an agent is dependent upon the objective one is trying to skin’ refers to a sequence of reactions and interactions among obtain. An agent could be chosen for its apparent wide-spec genes/proteins leading to a specific biochemical end product trum anti-aging benefits, including effect on skin hydration, relevant to at least one skin biological processes. skin structure, skin pigmentation and skin cell turnover, oran 0039. The term “biochemical pathway associated with agent could be chosen specifically for a single objective, e.g., physical appearance of skin aging refers to a biochemical hydration of the skin, associated with a more youthful state of pathway that leads to biochemical end products that cause a epidermal tissue. For example, ingredients such as retinoids, less youthful state in the skin. niacinamide, N-acetylglucosamine have a wide spectrum of 0040. The term “skin anti-aging agent” refers to a sub anti-aging benefits. stance that causes a biological or chemical change in the skin to reflect a more youthful state in the skin. DNA Microarray Technology 0041 Reference is now made in detail to certain embodi ments of systems and methods. The disclosed embodiments Data Processing Overview are not intended to be limiting of the claims. To the contrary, 0049. In various aspects, the present disclosure relates to the claims are intended to cover all alternatives, modifica the analysis of skin tissue samples by microarray-based tech tions, and equivalents. nology and transforming the microarray output data into use 0042. The present disclosure provides a system and ful Subsets of data identifying particular genes of interest. method in which the genes expressed in human derived skin 0050. In some embodiments, the methods of the present cells in a human equivalent skin tissue model are tested for disclosure comprise analyzing at least one test sample of a linkage to the physical appearance of human facial skin skin anti-aging agent on human or human-derived skin tissue, aging. Human derived skin cells include, for example, fibro by using microarray-based technology to obtain information blasts, keratinocytes, adipocytes and melanocytes. relating to changes in expression levels, if any. 0.043 Gene expression profiling using high-throughput 0051. A reference sample is a sample that lacks the pres methodologies such as DNA microarrays has proven to be a ence of a skin anti-aging agent. Test and reference samples powerful approach in exploring the complex processes of may be obtained from a biological Source comprising human aging, which involve many genetic pathways. This technique or human-derived skin cells or human or human-equivalent allows researchers to Scan essentially the entire genome for tissue, by any Suitable method of nucleic acid isolation and/or age-related changes in gene expression, or changes in gene extraction. In various aspects, the test sample and the refer expression as a result of anti-aging strategies. Such analyses ence sample are extracted RNA. have demonstrated an association between aging and wide 0.052 Array hybridization experiments allow the analysis spread gene expression. (Solene M. Fortunel NO, Pageon H, of thousands of genes in one experiment. Microarrays are Asselineau D, Aging alters fitnctionally human dermal pap Solid Supports made of either nylon or silicon which house US 2012/0034613 A1 Feb. 9, 2012 thousands of transcripts at fixed locations. The DNA is example, P, S, H, C, I, I, and others; fluorescent printed, spotted or synthesized on the Support. This method is dyes; chemiluminescent agents such as, for example, acri based on hybridization probing which uses fluorescently dinium esters, stabilized dioxetanes, and others; micropar labeled nucleic acids as probes to identify complementary ticles such as, for example, quantum dots, nanocrystals, phos sequences. Single stranded DNA is made up of 4 different phors and others; enzymes such as, for example, those used in nucleotides, adenine (A), thymine (T), guanine (G), and an ELISA, horseradish peroxidase, beta-galactosidase, cytosine (C). Adenine pairs with thymine and guanine pairs luciferase, alkaline phosphatase and others; colorimetric with cytosine. Hybridization occurs when a group of nucle labels such as, for example, dyes, colloidal gold and others; otides finds their complementary partners. Microarray magnetic labels such as, for example, DynabeadsTM particles: experiments measure the level of hybridization of each DNA and biotin, dioxigenin or otherhaptens and proteins for which on the Support via fluorescently labeled tags. antisera or monoclonal antibodies are available. 0053. There are three different types of probes that are 0057 The microarray or chip used for testing is then incu commonly used in hybridization experiments: genomic DNA bated overnight with both reference and experimental probes, cDNA probes and oligonucleotide probes. This pro cDNAs. Certain cDNA will hybridize with the complemen vides the different terms namely “DNA array”, “cDNA array” tary Strands from its gene that is covalently bound to a grid or "oligonucleotide array' depending on what type of probe is spot on the chip. The chips are then washed to remove any used. (Kumar, A., Goel, G., Fehrenbach, E., Puniya, K. A., unbound cDNAs. Two computerized images are then pro Singh, K. Microarrays: The Technology, Analysis and Appli duced by Scanning first to detect the grid spots containing cation, Engineering in Life Sciences 5(3), 215-222 (2005); cDNAs labeled with green dye, and second to detect the spots Jiang, N. et al. Methods for evaluating gene expression from containing the red-labeled cDNAs. The computer also pro Affymetrix microarray datasets, BMC Bioinformatics 9,284 duces a combination of the two images showing a yellow spot (2008); Auer, H., Newsom, D. L., Kornacker, K. Expression for grids spots containing both red and green labeled cDNAs. Profiling. Using Affymetrix GeneChip Microarrays, Methods These yellow spots represent transcripts that are expressed Mol Biol. 509, 35-46 (2009)). under both sets of conditions. 0054. In conducting a DNA microarray experiment, total 0058. In addition to producing images, microarray experi RNA is extracted from the samples to be tested. The purified ments yields quantitative data for each spot on the chip, RNA is then analyzed for quality and quantity (>1 micro resulting in large datasets where bioinformatics tools are grams). Reverse transcriptase is then used to transcribe the needed for complete analysis. Parametric t-test with a Ben mRNA into cDNA. The nucleotides used to synthesize the jamini and Hochberg false discovery rate correction is the cDNA are labeled with either agreen or reddye, one color for most common statistical parameter used for microarrays to reference conditions or the other color for experimental con identify genes with a statistically significant p value equal to ditions. The test samples and the reference samples may be or less than 0.05 and with a fold change of 2.0 and greater (or differentially labeled with any detectable substance or moi other suitable expression level criterion, expressed as a fold eties. The detectable substances or moieties may be selected change threshold or otherwise). Genes are then either Such that they generate signals that can be readily measured grouped by biological function or relation to a particular and Such that the intensity of the signals is proportional to the disease, depending on the objectives of the study. amount of labeled nucleic acids present in the sample. The 0059. These array-based methods of genetic analyses for detectable Substances or moieties may also be selected Such skin cell samples allow the research analyst to develop data that they generate localized signals, thereby allowing resolu for essentially the entire human genome as an initial step, but tion of the signals from each spot on an array. economics and the need to limit focus make it desirable to 0055 Methods for labeling nucleic acids are well-known analyze this data with a goal of limiting the number of genes in the art. For exemplary reviews of labeling protocols, label addressed in later analysis steps. Techniques to permit focus detection techniques and recent developments in the field, ing of resources on particular conditions, mechanisms and (see e.g., Kricka, Ann, Clin. Biochem. (2002), 39: 114-129; interventions can save time and cost. van Gijlswijk et al., Expert Rev. Mol. Diagn. (2001), 1:81-91; 0060. The array-based data is of sufficient volume that it is and Joos et al., J. Biotechnol. (1994), 35: 135-153). Standard desirable (and likely necessary) to carry out the analysis that nucleic acid labeling methods include: incorporation of transforms a genome-wide set of data from microarray equip radioactive agents, direct attachment of fluorescent dyes or of ment into Smaller, focused sets of data using a computer enzymes, chemical modification of nucleic acids to make based system. FIG. 7 shows a schematic diagram for a system them detectable immunochemically or by other affinity reac 700, including data-processing components, for carrying out tions, and enzyme-mediated labeling methods including, the method disclosed, including data sets developed and pro without limitation, random priming, nick translation, PCR cessed as the method proceeds. System 700 includes a data and tailing with terminal transferase. Other suitable labeling processing system 710 that may be implemented with a desk methods include psoralen-biotin, photoreactive azido deriva top computer, a cluster of computers, a group of computer tives, and DNA alkylating agents. In various embodiments, resources in a cloud, a Supercomputer or any other configu test sample and reference sample nucleic acids are labeled by ration of at least one CPU, memory and an operating system Universal Linkage System, which is based on the reaction of 712 that permits data from a full genome microarray 780 to be monoreactive cisplatin derivatives with the N7 position of received in a database 730. The system 700 further includes guanine moieties in DNA (see, e.g., Heetebrijet al., Cytoge various process application modules 720, including statistics net. Cell. Genet. (1999), 87: 47-52). modules 722 and other data processing modules and param 0056. Any of a wide variety of detectable substances or eters 724 for performing steps outlined in the flowchart of moieties can be used to label test and/or reference samples. FIGS. 1A-1K that carry out the data processing described in Suitable detectable substances or moieties include, but are not greater detail below. The database 730 receives from the limited to: Various ligands; radioneuclides such as, for microarray 780 raw or semiprocessed data that are produced US 2012/0034613 A1 Feb. 9, 2012 in one or more runs of the microarray. This data may include pathway that either leads to more or less youthful appearance. data comprising a full genome data set 750a derived from a To the extent a goal is to identify agents to influence biologi sample of agent-exposed tissue 782 and from a full genome cal pathways that enhance youthfulness, it is significant to control data set 750b derived from a sample of tissue not identify both the genes that can be up-regulated to cause a agent-exposed 784. These data will be subject to computa more youthful state, as well as the genes that can be down tions that produce a further data set 750c, with computed regulated to reduce action of a pathway that leads to less ratios by gene of levels of gene expression in exposed and youthful state. Thus, the skin attribute data sets 756 are coded unexposed tissue (782/784) as detected by microarray 780. to identify for each gene, the up or down regulation of a The computations further include applying to this data set of pathway that either leads to more or less youthful appearance ratios 750c a criterion for whether the level of gene expression as to the particular skin attribute involved. associated with the agent, is significant, in either an up-regu 0063. The results embodied in a skin attribute intersection lated or a down-regulated direction. The criterion may be a data set 758 may be considered preliminary and will be con selected and stored parameter in the applications modules sidered more reliable if they can in some way be confirmed, or 720, specifically in the data processing modules and param refined. The present system in one embodiment develops and eters 724. Application of a stored “fold’ criterion to the ratio analyzes further data to provide possible confirmation and data set 750c for the full genome yields a fold criterion data refining. As seen in FIG. 7, a second instrument for determin set 752, i.e., a data set listing genes found by testing to have a ing levels of gene expression, such as an instrument 790 that level of expression in the agent-exposed tissue that meet the relies on PCR techniques for determining expression levels fold criterion. may be employed. The candidate genes whose expression 0061 Also in the database 730 are data sets developed level is the focus for testing in this instrument are selected from literature 786 on the biological pathways that have been based on one or more of the skin attribute data sets 758. In one reported as associated with various genes. One data set 754 embodiment, the candidate genes identified in one of these identifies genes reported as having biological pathways that sets 758 may be supplemented for second instrument testing are significant for skin. (The data set 754 may be derived from with other candidate genes of interest, based on secondary literature by automated keyword and/or metadata analysis of research or simply because they are useful to provide refer the text of Scientific journals, patents or other sources report ence values. In one embodiment, the other candidate genes of ing on activity of particular genes, including non-published interest may be selected by searching in literature and in fold studies, or may be built by the input of one or more scientific criterion result data 752 for genes associated with aging (al experts. For this purpose, it may be helpful to build a database though not specifically with skin aging) and showing up of sources annotated with metadata 787 that permit ready regulation of an anti-aging biochemical pathway. Thus, a identification of each gene associated with skin (or other PCR Candidate data set 760 is assembled to define the ana organs) and, for genes associated with skin, as addressing lytical focus of gene expression level testing in the PCR biochemical pathways associated with particular skin Testing instrument 790. attributes.) This data set may be used in an intersection analy 0064. The test result data from the PCR Testing instrument sis to identify the genes in a larger test result data set, for 790 show levels of gene expression for the selected candidate example the full genome microarray data sets 750a, 750b or genes when the same agent used to develop data in the upper the fold criterion result data set 752, that are related to skin portion of FIG. 7 is exposed to a new tissue sample. In one and also meet the specified fold criterion that led to the fold embodiment, this is the same type of skin tissue sample as criterion result data set 752, forming a new skin pathway used to develop data in microarray 780 in the upper portion of intersection data set 755. Thus, the literature data set 754 may FIG. 7. The level of gene expression data, e.g., PCRAC, data be used as a filter for the test data to provide a focus on genes 762, are developed for each selected candidate gene. These for which data from the literature data set supports a pathway data are statistically cleaned and processed, then correlated of interest in skin. with a particular skin attribute intersection data set 758 hav 0062. The data sets derived from literature can also have a ing the same genes or with data for corresponding genes narrower focus within the broader area of skin. For example, found in data sets 752 or 755. In some instances, for a par a skin attribute data set 756 may be derived that identifies ticular candidate gene, a high level of expression in the same genes reported as having biological pathways that are signifi direction as was associated with more youthful appearance in cant for a particular attribute of skin, Such as skin structure or the intersection data set 758 will confirm that the gene and its skin pigmentation. Such a data set may be used in an inter associated biological pathway(s) appears to be capable of section analysis to identify the genes in a larger test result data agent stimulation for expression that leads to more youthful set, for example the skin pathway intersection data set 755, to appearance as to aparticular skin attribute. A set of such genes identify genes that are related to a specific skin structure and then may become part of a final functional youth gene assem also meet the specified fold criterion that led to the fold bly 770 for the skin attribute. A different set of candidate data criterion data set 752, forming a new skin attribute intersec 760 based on a different skin attribute intersection data set tion data set 758. The skin attribute intersection data set 758 758, and the set of PCRAC, test data 762 developed for those is then focused on genes related to one specific skin structure; candidates may become part of a final functional youth gene an attribute data set 756 for another skin attribute, e.g., skin assembly 772 for a different skin attribute. Each such assem pigmentation, can lead to a different skin attribute intersec bly would appear to provide a more useful and economic tion data set 758. At this level of focus, assuming the goal of focus for further study than any whole genome study. The focus is to identify genes associated with more youthful processes leading the various data sets identified in FIG. 7 are manifestations of a skin attribute, the skin attribute data sets discussed in greater detail below. also include up or down regulation coding. That is, if a gene 0065. The discussion below proceeds at two levels. At one is associated with a particular skin attribute, it may be level, with reference to FIGS. 1A-1K., a highly simplified reported as involved in either the up or down regulation of a example assuming a genome with only genes a-hand with US 2012/0034613 A1 Feb. 9, 2012

entirely hypothetical data is used to show the process steps. At water. An agent that has some known effects on skin aging a second level, with reference to Tables 1-6, the same process may be useful for revealing gene-based effects, but other is described, but Tables 1-6 show actual test data from lab agents may be selected. testing using actual skin tissue samples and equipment that 0073. The first samples of the human equivalent skin tis finds expression levels for actual genes that are included in the Sue are exposed to the skin anti-aging agent 104. Untreated human genome and the literature data sets (754, 756) are cultures, or human skin tissues not exposed to the agent, serve derived from actual gene literature. as controls or reference samples. (See FIG. 7 at 782, 784) Global Gene Expression Levels 0074 RNA is extracted from each of the human skin cells, 0066. In the present method and system, expression pro or cultures, using an RNeasy R. Fibrous kit (Qiagen, Valencia, filing for thousands of genes, Substantially all of the genes of Calif.) following the manufacturer's protocol. (RNeasy(R) the human genome, is performed to determine a first set of Fibrous Tissue Handbook, November 2006). cDNA is syn expression levels of a plurality of genes in a first sample of thesized from 100 ng of total RNA, and then converted to agent-exposed human or human derived skin. biotin-labeled amplified RNA (aRNA) using an Affymetrix 0067 Genome refers to all nucleic acid sequences, coding GeneChip(R3' IVT Express kit, according to the manufactur and non-coding, present in each cell type of a subject. The er's instructions. (Affymetrix User Manual GeneChip(R) 3' term also includes all naturally occurring or induced varia IVT Express Kit (2008)). tions of these sequences that may be present in a mutant or 0075. In some embodiments, the samples are hybridized disease variant of any cell type, including, for example, tumor to Affymetrix GeneChip(R) HG U133 Plus 2.0 microarrays, cells. Genomic DNA and genomic nucleic acids are thus washed, stained and scanned according to Affymetrix proto nucleic acids isolated from a nucleus of one or more cells, and cols. The microarray laser Scanner measures fluorescence include nucleic acids derived from, isolated from, amplified intensities of all of the transcripts on the gene chip; the fluo from, or cloned from genomic DNA, as well as synthetic rescence of each transcript is compared among each of the versions of all or any part of a genome. samples. (See FIG. 7 at 780). 0068 For example, the human genome consists of 0076 Results from these experiments reveal that the skin approximately 3.0x10 base pairs of DNA organized into 46 anti-aging agent selected for exploration may have influenced distinct . The genome of a normal human dip the activity of any of more than 15,000 genes in the human loid somatic cell consists of 22 pairs of autosomes (chromo genome. somes 1 to 22) and either chromosomes X and Y (male) or a pair of Xs (female) for a total of 46 chromo 0077. The activity is measured by determining expression SOS. levels for genes in the first exposed tissue sample (test 0069. In some embodiments, Affymetrix R DNA microar sample) 106. This involves reading the array that has a human ray technology is used for measuring global gene expression equivalent tissue model exposed to the agent. This results in a in human in vitro skin cultures. Microarrays are ideal for set of data that is stored in database 730 of a data processing simultaneously measuring the effects of a test compound on system 710 for implementing the method described herein, the activity of thousands of genes in the human genome. (See FIG. 7 at 750a). (Microchip Methods in Diagnostics, vol. 509, chapter 3, 0078. To give a basis for comparison, the method uses a Expression Profiling. Using Affymetrix GeneChip Microar reference tissue that has not been exposed to the agent. The rays, Auer et al. (2009)). same skin model is used to provide data on expression levels 0070 For one embodiment, EpidermFTTM Skin Model when the agent is not present. The reference level is measured (EFT-400) full thickness skin cultures (Mattek Corp., Ash by determining expression levels for genes in the first unex land, Mass.) is used as a skin model. These cultures contain posed sample (reference sample) 108. Again, this results in a normal, human-derived epidermal keratinocytes from neona set of data that is stored in database 730 of a data processing tal foreskin tissue and normal human-derived dermal fibro system 710 for implementing the method described herein. blasts, from mammary tissue. These cells are cultured to form (See FIG. 7 at 750b). a multilayered, highly differentiated model of the human 007.9 The agent-exposed and reference data are analyzed dermis and epidermis. The model parallels human skin and is to determine the affects of agent exposure. The expression useful for in vitro testing, where a microarray is used to level data for each gene of the test sample are compared with develop and collect data. Skin models typically contain the corresponding data of the reference sample to obtain a human derived skin cells cultured to form a model of skin ratio of the data 110. (See FIG. 7 at 750c). At least part of the tissue. Generally, these skin models are referred to as “human gathering, storing and comparing of the expression level data equivalent skin tissue' or “human derived skin tissue'. Skin is performed by a computer, such as the data processing cells include keratinocytes, fibroblasts, adipocytes and mel system 710, with CPU 712 (see FIG. 7) for performing data anocytes. access and storage and various computations specified by (0071 FIGS. 1A-1K show steps in a flowchart 100 with Software modules corresponding to the functions occurring at steps for choosing from the simplified hypothetical genes a-h. various described steps of this method. genes for a functional youth gene assembly. Reference 0080 Abioinformatics statistical analysis is conducted on numerals associated with method steps and tabular data on data 112 to identify and characterize candidate genes. A Para flowchart 100 appear in the description of the method below. metric t-test with a Benjamini and Hochberg false discovery 0072 The steps begin with selecting an agent that is a rate correction is the most common statistical parameter used candidate to help skin appearance 102 (or explore effects of for microarrays to identify genes with a statistically signifi skin aging). In some embodiments, the agent tested is salicin cant p value equal to or less than 0.05 and with a selected at a concentration of 0.5% salicin, available from Symrise biological relevance level, e.g., a fold change of 2.0 and Corporation (Teterborro, N.J.). The salicin is dissolved in greater. To help organize the results, genes are either grouped US 2012/0034613 A1 Feb. 9, 2012 by biological function or relation to a particular disease, I0086. As shown in row 206, example genes b, c, e,f, g, and depending on the objectives of the study. h have ratios of 3, 2.5, 4, 0.2, 3.33 and 12, respectively. Thus, with a fold change threshold selected at 2, the table in FIG. 1B Selecting First Subset of Genes shows the example genes b, c, e, f, g, and h, meet the fold 0081 Flowchart 100 in FIG. 1A has an illustration of a change criterion. This is the first subset as described for the table of the extremely simplified hypothetical example set of hypothetical example. genes (for simplicity identified as “a-h') with rows showing I0087. In the test data from skin model experiments, fold expression levels in the exposed sample 202, expression lev changes of at least about 2.0, about 2.0, of at least about 3.0, els in unexposed sample 204, and the ratio 206 of row 204 between about 2.0 and about 4.0, between about 2.0 and about compared to row 202 (row 202-row 204-row 206). (The data 7.0, and even between 2.0 and about 200.0 may be selected as in rows 202 and 204 are entirely hypothetical, for purposes of biologically relevant. Fold change significance may vary an example, and row 206 shows a computation from the based on the instrument used for testing, tissue sample and hypothetical data.) other factors. For Affymetrix DNA microarray data, using the 0082 FIG. 1B shows an expanded table of the example set specific statistically scientific parameter, a fold change of 2 or of genes. Row 208 shows the ratios of row 206 as fold changes, reflecting that genes may be up-regulated or down more is biologically significant. (The fold change criterion is regulated. a selectable parameter 724 in process application modules 0083. The raw dataset from the first subset of genes, row 720.) 208, is then filtered, using bioinformatics methods, to identify 0088. As shown on row 206 of the table on FIG. 1B, and characterize genes. In particular, the data is filtered to example genes a and d have ratios of 1.625 (up), and 0.75 focus on genes with biologically relevant fold change values. (down), respectively, or fold changes of 1.625 and 1.33, In data from a typical microarray device, e.g., data from the respectively, both below a fold change selection criterion of 2 Affymetrix testing, the up or down regulation of the gene is for up-regulation or down-regulation. By comparison, for also identifiable. The row 210 shows the regulation direction gene e, the expression level was higher in the exposed test of the hypothetical gene. sample than in the reference, so in comparison to the refer 0084. In some embodiments at least part of the analysis of ence, this gene was up-regulated. The column “f value would data is performed by a computer. Statistical data analysis may be a "down” regulation gene situation, with a fold change of be carried out using GeneSpring GX software (version 10). A 5 (a ratio of 2/10-0.2). Thus, gene fin the example also meets parametric t-test with a Benjamini and Hochberg false dis the fold change criterion. Genes not meeting the fold change covery rate correction is performed to identify genes with a criterion and not chosen for the first Subset of genes (a and d. statistically significant p value equal to or less than 0.05. in the example) may be considered for additional research (Statistical analysis may be performed by one or more of the based on secondary research factors 116. (Secondary process application modules 720. (See FIG. 7). In one set of research factors are discussed below Table 6.) data collected, the 0.05% salicin treated cultures (N=7) are I0089 Correspondingly, as will be seen below and in Table compared with untreated control cultures (N=4) after 24 1, the actual skin model data test results from the microarray hours of stimulation. instrument 780 were subjected to the same fold change crite 0085. Returning to the simplified example, at step 114, the rion as in the simplified example; thus genes with fold system selects a first Subset of genes (b, c, e, f, g, and h) with changes reported below 2.0 were not included in any of the biologically relevant fold changes in the level of expression, tables for skin model actual experimental results. for example, those calculated to be at least a two fold change. 0090 Table 1 below shows experimental data from a (See row 208). As noted with reference to rows 202,204,206, microarray for 0.05% salicin treated cultures (N=7) com the ratio is measured from the first exposed sample (test pared with untreated control cultures (N=4) corresponding to sample), as compared to the first unexposed sample (refer the simplified hypothetical data in the table of FIG. 1B. (See ence sample). The method also involves identifying which also, FIG. 7 at 752). This is a list of more than 2,300 genes genes are regulated in a particular direction (up or down tested by the Affymetrix microarray test methods with filter regulated). (See row 210). In one embodiment, a ratio of data ing of data as described in the example to focus on genes with in row 202 to data in row 204, shows up-regulation if the ratio biologically relevant fold change values. Table 1 thus shows is greater than 1 and down-regulation is the ratio is less that 1. the first subset for the microarray data.

TABLE 1.

Fold Probe Set ID change Direction Gene Symbol Gene Title 1556.410 a. at 24.649403 up KRTAP19-1 keratin associated protein 19-1 210229 s at 16.55O27 up CSF2 colony stimulating factor 2 (granulocyte-macrophage) 205114 S at 15.552O67 up CCL3 chemokine (C-C motif) ligand 3 fif CCL3L1 if chemokine (C-C motif) ligand 3-like 1 CCL3L3 if fit chemokine (C-C motif) ligand 3-like LOCA28830 3 similar to C-C motif chemokine 3 like 1 precursor (Small-inducible cytokine A3-like 1) (Tonsillar lymphocyte LD78 beta protein) (LD78-beta(1-70)) (G0/G1 switch US 2012/0034613 A1 Feb. 9, 2012 8

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title regulatory protein 19-2) (GOS19-2 protein) (PAT 464.2) 213418 a 14.241181 l HSPA6 heat shock 70 kDa protein 6 (HSP70B) 205943 a 12.26O297 l TDO2 tryptophan 2,3-dioxygenase 214038 a 11.567539 l CCL8 chemokine (C-C motif) ligand 8 207442 a. 11.091.33 l CSF3 colony stimulating factor 3 (granulocyte) 241031 a 10.97.546S l FAM148A family with sequence similarity 148, member A 225207 a 10.136719 l PDK4 pyruvate dehydrogenase kinase, isozyme 4 205931 s at 9.909108 l CREBS cAMP responsive element binding protein 5 229228 a 9.767824 l CREBS cAMP responsive element binding protein 5 206924 a 9.694227 l IL11 interleukin 11 207526 S. at 9.089992 l IL1RL1 interleukin 1 receptor-like 1 206569 a 8.736957 l IL24 interleukin 24 207850 a 8.702696 l CXCL3 chemokine (C X—C motif) ligand 3 206926 s at 7.970876 l IL1 interleukin 11 206176 a 7.765054 l BMP6 bone morphogenetic protein 6 216248 s at 7.5919886 l NR4A2 nuclear receptor Subfamily 4, group A, member 2 204622 X at 7.5477886 l NR4A2 nuclear receptor Subfamily 4, group A, member 2 209774 X at 7.546OO76 l CXCL2 chemokine (C X—C motif) ligand 2 233011 at 7.322324 l ANXA1 Annexin A1, mRNA (cDNA clone MGC: 32774 IMAGE:4662939) 21738.8 s at 7.2951.15 l KYNU kynureninase (L-kynurenine hydrolase) 216979 at 7.2O3941 l NR4A3 nuclear receptor Subfamily 4, group A, member 3 204621 s at 7.06227 l NR4A2 nuclear receptor Subfamily 4, group A, member 2 1554685 a. at 7.039142 l KIAA1199 KIAA11.99 228501 at 6.957408 l GALNTL2 UDP-N-acetyl-alpha-D- galactosamine:polypeptide N acetylgalactosaminyltransferase-like 2 235004 at 6.917818 l RBM24 RNA binding motif protein 24 202643 s at 6.915339 l TNFAIP3 tumor necrosis factor, alpha-induced protein 3 235086 at 6.8579264 l THEBS1 (clone lambda-TS-33) thrombospondin (THBS) mRNA, 5' end 206382 s at 6.826926 l BDNF brain-derived neurotrophic factor 1554997 a. at 6.812711 l PTGS2 prostaglandin-endoperoxide synthase 2 (prostaglandin GH synthase and cyclooxygenase) 237411 at 6.7217474 l ADAMTS6 ADAM metallopeptidase with hrombospondin type 1 motif, 6 21294.2 s at 6.522843 l KIAA1199 KIAA11.99 23.0748 at 6.413 O154 l SLC16A6 solute carrier family 16, member 6 (monocarboxylic acid transporter 7) 2393.67 at 6.3149343 l BDNF brain-derived neurotrophic factor 204932 at 6.287326 l TNFRSF11B tumor necrosis factor receptor Superfamily, member 11b 204933 s at 6.249607 l TNFRSF11B tumor necrosis factor receptor Superfamily, member 11b 236361 at 6.1742O6 l GALNTL2 UDP-N-acetyl-alpha-D- galactosamine:polypeptide N acetylgalactosaminyltransferase-like 2 208075 S. at 6.120267 l CCL7 chemokine (C-C motif) ligand 7 226614 S at 6.01704O7 l FAM167A amily with sequence similarity 167, member A 206137 at S.972.218 l RIMS2 regulating synaptic membrane exocytosis 2 210133 at S.958449 l CCL11 chemokine (C-C motif) ligand 11 1557257 at 5.733714 l BCL10 CDNA FLJ25924 fis, clone CBR05109 215078 at 5.7122235 l SOD2 Superoxide dismutase 2, mitochondrial 202644 S at 5.705O176 l TNFAIP3 tumor necrosis factor, alpha-induced protein 3 221577 x at 5.573193 l GDF15 growth differentiation factor 15 2064.07 s at S.S16O13 l CCL13 chemokine (C-C motif) ligand 13 216598 s at 54O7172 l CCL2 chemokine (C-C motif) ligand 2 203708 at 5.313648 l PDE4B phosphodiesterase 4B, cAMP-specific US 2012/0034613 A1 Feb. 9, 2012 9

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title (phosphodiesterase E4 dunce homolog, Drosophila) 242809 at S.274104 l L1RL1 ST2 protein 20984.0 s at S.160858 l LRRN3 leucine rich repeat neuronal 3 204748 at S.148792 l PTGS2 prostaglandin-endoperoxide synthase 2 (prostaglandin GH synthase and cyclooxygenase) 203.645 S. at S.145445 l CD163 CD163 molecule 20984.1 s at S.O964932 l LRRN3 leucine rich repeat neuronal 3 207038 at 5.0897655 l SLC16A6 solute carrier family 16, member 6 (monocarboxylic acid transporter 7) 211302 s at S.OOO998 l PDE4B phosphodiesterase 4B, cAMP-specific (phosphodiesterase E4 dunce homolog, Drosophila) 210663 s at 4.955.7395 l KYNU kynureninase (L-kynurenine hydrolase) 215506 s at 4.9070177 l DIRAS3 DIRAS family, GTP-binding RAS-like 3 220817 at 4.90531 l TRPC4 transient receptor potential cation channel, Subfamily C, member 4 215501 s at 4.86OO24 l DUSP10 dual specificity phosphatase 10 209990 s. at 4.761S633 l GABBR2 gamma-aminobutyric acid (GABA) B receptor, 2 203680 a 4.729812 l PRKAR2B protein kinase, cAMP-dependent, regulatory, type II, beta 207815 a. 4.719714 l PF4V1 platelet factor 4 variant 1 206167 s at 4.712O314 l ARHGAP6 Rho GTPase activating protein 6 21.0997 a 4.71.1954 l HGF hepatocyte growth factor (hepapoietin A: scatter factor) 213524 s at 4.703OSO6 l GOS2 GOG1switch 2 206950 a 4.6845O2 l SCN9A Sodium channel, voltage-gated, type X, alpha Subunit 215049 X at 4.532594 l CD163 CD163 molecule 224071 a 4.500.9127 l IL20 interleukin 20 232017 a 4.494.8945 l TP2 ight junction protein 2 (Zona occludens 2) 239461 a 4.4746847 l GALNTL2 UDP-N-acetyl-alpha-D- galactosamine:polypeptide N acetylgalactosaminyltransferase-like 2 205681 a 4.4743O4 l BCL2A1 BCL2-related protein A1 201107 s at 4472S666 l THEBS1 hrombospondin 1 203895 a 4.425,0426 l PLCB4 phospholipase C, beta 4 212353 a 4.4241323 l SULF1 Sulfatase 1 1569020 at 4.418171 l NEDD9 neural precursor cell expressed, developmentally down-regulated 9 242329 a 4.370621 l LOC4O1317 hypothetical LOC4O1317 217999 s at 4.368414 l PHILDA1 pleckstrin -like domain, amily A, member 1 220088 a 4.3576527 l CSAR1 complement component 5a receptor 1 213921 a 4.35561 l SST Somatostatin 210998 s at 4.350414 l HGF hepatocyte growth factor (hepapoietin A: scatter factor) 205992 S at 4.32588 l L15 interleukin 15 242767 a 4.3130975 l LMCD1 CDNA FLJ52480 complete cols, highly similar to LIM and cysteine-rich domains protein 1 203817 a 4.3O71494 l GUCY1B3 guanylate cyclase 1, Soluble, beta 3 203304 a 4.288485 l BAMBI BMP and activin membrane-bound inhibitor homolog (Xenopus laevis) 237132 a. 4.2871.304 l TP2 ight junction protein 2 (Zona occludens 2) 207332 S at 4.2861 SOS l TFRC transferrin receptor (p90, CD71) 201860 s at 4.228699 l PLAT plasminogen activator, tissue 1552694 at 4.224.9846 l SLC2A13 solute carrier family 2 (facilitated glucose transporter), member 13 117 at 4.210481 l HSPA6 heat shock 70 kDa protein 6 (HSP7OB') 207978 s at 4.165331 l NR4A3 nuclear receptor Subfamily 4, group A, member 3 200664 S at 4.1382117 l DNAB1 DnaJ (Hsp40) homolog, Subfamily B, member 1 229160 at 4.138O2 l MUM1L1 melanoma associated antigen (mutated) -like 1 220818 s at 4.1121902 l TRPC4 transient receptor potential cation channel, Subfamily C, member 4 US 2012/0034613 A1 Feb. 9, 2012 10

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 210755 at 4...10634.04 l HGF hepatocyte growth factor (hepapoietin A: scatter factor) 2173.71 s at 4...106031 l L15 interleukin 15 204160 s at 4.1OO3O46 l ENPP4 ectonucleotide pyrophosphatase/phosphodiesterase 4 (putative function) 205476 at 4.091709 l CCL20 chemokine (C-C motif) ligand 20 203549 s at 4.07S3236 l LPL ipoprotein lipase 234.066 at 4.OS28307 l L1RL1 ST2 protein 203896 s at 4.0160823 l PLCB4 phospholipase C, beta 4 15561.34 a at 3.9963.338 l B3GNTS Homo sapiens, clone MAGE: 5122250, mRNA 205410 s at 3.9841.306 l ATP2B4 ATPase, Ca++ transporting, plasma membrane 4 210287 s at 3.9SO3276 l FLT1 ms-related kinase 1 (vascular endothelial growth factorivascular permeability factor receptor) 219778 at 3.9186013 l ZFPM2 Zinc finger protein, multitype 2 204273 at 3.9157705 l EDNRB endothelin receptor type B 208.691 at 3.91473O8 l TFRC transferrin receptor (p90, CD71) 1556629 a. at 3.913148 l SNAP2S HUMSNAP25B(F) 209960 at 3.912294 l HGF hepatocyte growth factor (hepapoietin A: scatter factor) 228962 at 3.9099233 l PDE4D phosphodiesterase 4D, cAMP-specific (phosphodiesterase E3 dunce homolog, Drosophila) 15531.33 at 3.861SS3 l C9orf72 chromosome 9 open reading frame 72 204421 S at 3.8445234 l FGF2 fibroblast growth factor 2 (basic) 229435 at 3.841,224 l GLIS3 GLIS family Zinc finger 3 214582 at 3.82721.83 l PDE3B phosphodiesterase 3B, c0MP-inhibited 229.199 at 3.818O137 l SCN9A Sodium channel, voltage-gated, type X, alpha Subunit 203548 s at 3.8146.515 l LPL ipoprotein lipase 22421.9 s at 3.8127234 l TRPC4 transient receptor potential cation channel, Subfamily C, member 4 202422 S at 3.78SS804 l ACSL4 acyl-CoA synthetase long-chain family member 4 204385 a 3.7706523 l KYNU kynureninase (L-kynurenine hydrolase) 206701 x at 3.767451 l EDNRB endothelin receptor type B 212099 a 3.7642O62 l RHOB ras homologgene family, member B 227176 a 3.75334.86 l SLC2A13 solute carrier family 2 (facilitated glucose transporter), member 13 20104.4 x at 3.7260537 l DUSP1 dual specificity phosphatase 1 205.266 a 3.7O18826 l LIF eukemia inhibitory factor (cholinergic differentiation factor) 221563 a 3.6977322 l DUSP10 dual specificity phosphatase 10 230258 a 3.689,173 l GLIS3 GLIS family Zinc finger 3 21591.0 s at 3.6631.63 l FNDC3A fibronectin type III domain containing 3A 215033 a 3.6403422 l TM4SF1 transmembrane 4 L six family member 1 215966 x at 3.6169035 l GK3P glycerol kinase 3 pseudogene 1552695 a. at 3.5807984 l SLC2A13 solute carrier family 2 (facilitated glucose transporter), member 13 204931 a 3.57953.79 l TCF21 21 209959 a 3.5782444 l NR4A3 nuclear receptor Subfamily 4, group A, member 3 215034 S. at 3.575,6443 l TM4SF1 transmembrane 4 L six family member 1 235591 a 3.5697513 l SSTR1 Somatostatin receptor 1 202843 a 3.56OO66S l DNATB9 DnaJ (Hsp40) homolog, Subfamily B, member 9 231367 s at 3.558.707 l LOC6471.31 hypothetical LOC647131 2025.08 s at 3.5226371 l SNAP2S synaptosomal-associated protein, 25kDa 15696.17 at 3.5221984 l OSBP2 CDNA clone IMAGE:3632045 202672 s at 3.5205257 l ATF3 activating transcription factor 3 2108.37 s at 3.S2O149 l PDE4D phosphodiesterase 4D, cAMP-specific (phosphodiesterase E3 dunce homolog, Drosophila) 218000 s at 3.S197904 l PHILDA1 pleckstrin homology-like domain, family A, member 1 214632 at 3.51.0114 l NRP2 neuropilin 2 215977 X at 3.SO9982 l GK glycerol kinase US 2012/0034613 A1 Feb. 9, 2012 11

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 219935 a. 3.4860334 l ADAMTSS ADAM metallopeptidase with thrombospondin type 1 motif, 5 209101 a 3.474422 l CTGF connective tissue growth factor 244804 a 3.4737628 l SQSTM1 Sequestosome 1 (SQSTM1), transcript variant 1 mRNA 212224 a 3.462966 l ALDH1A1 aldehyde dehydrogenase 1 family, member A1 212354 a 3.4603515 l SULF1 Sulfatase 1 204422 S at 3.455492 l FGF2 fibroblast growth factor 2 (basic) 229357 a 3.450523 l ADAMTSS ADAM metallopeptidase with thrombospondin type 1 motif, 5 209071 s at 3.4271708 l RGSS regulator of G-protein signaling 5 205088 a 3.4216924 l MAMLD1 mastermind-like domain containing 1 217998 a 3.4190629 l LOC652993 hypothetical LOC652993 ff, pleckstrin i. PHILDA1 homology-like domain, family A, member 1 227361 a 3.409933 l HS3ST3B1 heparan sulfate (glucosamine) 3-O- Sulfotransferase 3B1 217997 a 3.4O90981 l PHILDA1 pleckstrin homology-like domain, family A, member 1 218177 a 3.39026 l CHMP1B chromatin modifying protein 1B 204271 s at 3.385063 l EDNRB endothelin receptor type B 204491 a 3.38O295 l PDE4D phosphodiesterase 4D, cAMP-specific (phosphodiesterase E3 dunce homolog, Drosophila) 229273 a 3.3785.377 l SALL1 sal-like 1 (Drosophila) 206411 s at 3.3458,714 l ABL2 w-abl Abelson murine leukemia viral oncogene homolog 2 (arg, Abelson related gene) 219195 a 3.344138 l PPARGC1A peroxisome proliferator-activated receptor gamma, coactivator 1 alpha 231031 a 3.3397727 l KGFLP2 keratinocyte growth factor-like protein 2 206029 a 3.3350718 l ANKRD1 ankyrin repeat domain 1 (cardiac muscle) 211555. S. at 3.3216224 l GUCY1B3 guanylate cyclase 1, Soluble, beta 3 217167 x at 3.31938.31 l GK glycerol kinase 210511 s at 3.3132741 l INHIBA inhibin, beta A 210836 X at 3.3O34697 l PDE4D phosphodiesterase 4D, cAMP-specific (phosphodiesterase E3 dunce homolog, Drosophila) 207387 s at 3.292S398 l GK glycerol kinase 209961 s at 3.291533 l HGF hepatocyte growth factor (hepapoietin A: scatter factor) 1554779 S at 3.28,1243 l PHILDB2 pleckstrin homology-like domain, family B, member 2 243357 at 3.27.09684 l NEGR1 neuronal growth regulator 1 206893 at 3.26491.76 l SALL1 sal-like 1 (Drosophila) 1552632 a. at 3.2559857 l ARSG arylsulfatase G 21184.4 S at 3.2455873 l NRP2 neuropilin 2 212094 at 3.234917 l PEG10 paternally expressed 10 232235 at 3.2294521 l DSEL dermatan Sulfate epimerase-like 220416 at 3.2220476 l ATP8B4 ATPase, class I, type 8B, member 4 1555.257 a. at 3.2004O68 l MYO3B myosin IIIB 225566 at 3.1977339 l NRP2 neuropilin 2 205207 at 3.1970735 l IL6 interleukin 6 (interferon, beta 2) 240556 at 3.1939862 l DCN EST from clone 130486, 5' end 209387 s at 3.1869683 l TM4SF1 transmembrane 4 L six family member 1 206025 s at 3.1820982 l TNFAIP6 tumor necrosis factor, alpha-induced protein 6 233126 s at 3.1724448 l OLAH oleoyl-ACP hydrolase 209386 at 3.1714O27 l TM4SF1 transmembrane 4 L six family member 1 205782 at 3.1584346 l FGF7 fibroblast growth factor 7 (keratinocyte growth factor) 1553271 at 3.1546SO2 l DIP2B DIP2 disco-interacting protein 2 homolog B (Drosophila) 227487 s at 3.1483.386 l SERPINE2 Serpin peptidase inhibitor, clade E (nexin, plasminogen activator inhibitor type 1), member 2 (SERPINE2), transcript variant 1, mRNA 202558 s at 3.1297736 l HSPA13 heat shock protein 70 kDa family, member 13 219155 at 3.12S4O86 l PITPNC1 phosphatidylinositol transfer protein, cytoplasmic 1 US 2012/0034613 A1 Feb. 9, 2012 12

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 226814 at 3.12.1966 l ADAMTS9 ADAM metallopeptidase with hrombospondin type 1 motif, 9 225946 at 3.1174657 l RASSF8 Ras association (RalGDSAF-6) domain family (N-terminal) member 8 1554163 at 3.112223 l TWIST2 twist homolog 2 (Drosophila) 200800 s at 3.11194.42 l HSPA1A heat shock 70 kDa protein 1A i? heat HSPA1B shock 70 kDa protein 1B 201631 s at 3.109209S l IER3 immediate early response 3 202304 at 3.109106 l FNDC3A fibronectin type III domain containing 3A 223754 at 3.101.5599 l MGC13057 hypothetical protein MGC13057 219975 X at 3.09.0888S l OLAH oleoyl-ACP hydrolase 218546 at 3O864964 l C1orf115 open reading frame 115 205935 at 3.0835295 l FOXF1 orkhead box F1 229529 at 3.0813122 l TCF21 transcription factor 21 2284.90 at 3.0776792 l ABHD2 abhydrolase domain containing 2 206118 at 3.0758493 l STAT4 signal transducer and activator of transcription 4 219279 at 3.074901 l DOCK10 dedicator of cytokinesis 10 222018 at 3.0714924 l NACA nascent polypeptide-associated NACA2 complex alpha subunit i? nascent NACAP1 polypeptide-associated complex alpha Subunit 2 nascent-polypeptide associated complex alpha polypeptide pseudogene 1 225842 at 3.0654O18 l PHILDA1 pleckstrin homology-like domain, amily A, member 1 1555167 s at 3.064SO18 l NAMPT nicotinamide phosphoribosyltransferase 201502 s at 3.0644495 l NFKBIA nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha 228195 at 3.054O12 l MGC13057 hypothetical protein MGC13057 209183 s at 3.0345566 l C10orf10 chromosome 10 open reading frame 10 209833 at 3.0264297 l CRADD CASP2 and RIPK1 domain containing adaptor with death domain 204596 s at 3.016O112 l STC1 Stanniocalcin 1 204440 at 3.015682S l CD83 CD83 molecule 2294.61 x at 3.014515 l NEGR1 neuronal growth regulator 1 201041 s at 3.0053132 l DUSP1 dual specificity phosphatase 1 205356 at 3.0O39837 l USP13 ubiquitin specific peptidase 13 (isopeptidase T-3) 216316 x at 2.998.2364 l GK GK3P glycerol kinase if glycerol kinase 3 pseudogene 219257 s at 2.9839.096 l SPHK1 sphingosine kinase 1 205239 a 2.983703 l AREG If amphiregulin amphiregulin B AREGB 225847 a 2.9832149 l AADACL1 arylacetamide deacetylase-like 1 214370 a 2.98O2.194 l S100A8 Calcium-binding protein in macrophages (MRP-8) macrophage migration inhibitory factor (MIF)- related protein 206756 a 2.9774O6S l CHST7 carbohydrate (N-acetylglucosamine 6 O) sulfotransferase 7 213325 a. 2.9673648 l PVRL3 poliovirus receptor-related 3 220054 a 2.967118 l IL23A interleukin 23, alpha subunit p19 203372 s at 2.9669046 l SOCS2 Suppressor of cytokine signaling 2 231899 a 2.96.64657 l ZC3H12C Zinc finger CCCH-type containing 12C 237215 s at 2.95O1724 l TFRC transferrin receptor (p90, CD71) 227613 a 2.935803 l ZNF331 Zinc finger protein 331 231944 a 2.932O385 l ERO1LB ERO1-like beta (S. cerevisiae), mRNA (cDNA clone MGC: 26065 MAGE: 4829502) 243296 a 2.9062977 l NAMPT GOS9 mRNA, instability elements 207 754 a 2.90.39783 l RASSF8 Ras association (RalGDSAF-6) domain family (N-terminal) member 8 214681 a 2.89692.62 l GK glycerol kinase 225606 a 2.894.8586 l BCL2L11 BCL2-like 11 (apoptosis facilitator) 229225 a. 2.893.143 l NRP2 neuropilin 2 208962 s at 2.88.9333 l FADS1 atty acid desaturase 1 219926 a 2.8884964 l POPDC3 popeye domain containing 3 241611 s at 2.88.746 l FNDC3A fibronectin type III domain containing US 2012/0034613 A1 Feb. 9, 2012 13

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 227235 a. 2.8838294 l GUCY1A3 guanylate cyclase 1, Soluble, alpha 3 24.1762 a 2.878.3326 l FBXO32 F-box protein 32 (FBXO32), transcript variant 2 mRNA 236645 a. 2.86956.79 l HBP1 HMG-box transcription factor 1 206618 a 2.864942 l IL18R1 interleukin 18 receptor 1 223510 a 2.864709 l NRP2 neuropilin 2 208591 s at 2.851,169 l PDE3B phosphodiesterase 3B, c0MP-inhibited 213103 a 2.8436816 l STARD13 StAR-related lipid transfer (START) domain containing 13 224.942 a. 2.8339565 l PAPPA pregnancy-associated plasma protein A, pappalysin 1 24-1763 s at 2.832837 l FBXO32 F-box protein 32 (FBXO32), transcript variant 2 mRNA 229.088 a 2.831386 l ENPP1 ectonucleotide pyrophosphatase/phosphodiesterase 1 202779 s at 2.83OS142 l LOCA31049 similar to Ubiquitin-conjugating if UBE2S enzyme E2S (Ubiquitin-conjugating enzyme E2-24 kDa) (Ubiquitin-protein igase) (Ubiquitin carrier protein) (E2 EPF5) i? ubiquitin-conjugating enzyme E2S 225142 at 2.82S2O53 up JHDM1D jumonji C domain containing histone demethylase 1 homolog D (S. cerevisiae) 232825 s at 2.82O7173 up DSEL dermatan Sulfate epimerase-like 212092 at 2.812921S up PEG10 paternally expressed 10 1552508 at 2.8024O6S up KCNE4 potassium voltage-gated channel, Isk related family, member 4 213931 at 2.8021426 up ID2 ID2B inhibitor of DNA binding 2, dominant negative helix-loop-helix protein if inhibitor of DNA binding 2B, dominant negative helix-loop-helix protein (pseudogene) 235996 at 2.7879438 l RASSF8 Ras association (RalGDSAF-6) domain family (N-terminal) member 8 204457 s at 2.7829573 l GAS1 growth arrest-specific 1 207386 at 2.7815282 l CYP7B1 cytochrome P450, family 7, Subfamily B, polypeptide 1 2094.06 at 2.773616 l BAG2 BCL2-associated athanogene 2 20738.8 s at 2.7678.204 l PTGES prostaglandin E synthase 204818 at 2.7658327 l HSD17B2 hydroxysteroid (17-beta) dehydrogenase 2 201108 s at 2.7555368 l THEBS1 hrombospondin 1 224229 s at 2.7508733 l AKT3 v-akt murine thymoma viral oncogene homolog 3 (protein kinase B, gamma) 235368 a 2.7506857 l ADAMTSS ADAM metallopeptidase with hrombospondin type 1 motif, 5 203827 a 2.7443364 l WIPI1 WD repeat domain, phosphoinositide interacting 1 237867 s at 2.7432.609 l PID1 phosphotyrosine interaction domain containing 1 206806 a 2.7395847 l DGKI diacylglycerol kinase, iota 224.941 a 2.7371583 l PAPPA pregnancy-associated plasma protein A, pappalysin 1 231779 a 2.7335684 l RAK2 interleukin-1 receptor-associated kinase 2 204038 s at 2.7315595 l LPAR1 ySophosphatidic acid receptor 1 201981 a 2.7272985 l PAPPA pregnancy-associated plasma protein A, pappalysin 1 212374 a 2.726O473 l FEM1B em-1 homolog b (C. elegans) 21.6005 a 2.7193272 l TNC Tenascin 229310 a 2.71912S l KLHL29 kelch-like 29 (Drosophila) 211958 a 2.71.18793 l GFBP5 insulin-like growth factor binding protein 5 214702 a 2.70994.62 l FN1 fibronectin 1 224220 X at 2.7084796 l TRPC4 transient receptor potential cation channel, Subfamily C, member 4 235723 a 2.7O6666 l BNC2 basonuclin 2 211965 a 2.7050216 l ZFP36L1 Zinc finger protein 36, C3H type-like 1 225337 a 2.704214 l ABHD2 abhydrolase domain containing 2 209070 s at 2.700427 l RGSS regulator of G-protein signaling 5 1562606 a. at 2.7OOO384 l LOC440O28 hypothetical gene Supported by BCO40853 210764 S at 2.6922395 l CYR61 cysteine-rich, angiogenic inducer, 61 US 2012/0034613 A1 Feb. 9, 2012 14

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 222379 at 2.6855981 l KCNE4 potassium voltage-gated channel, Isk related family, member 4 205794 S at 2.6822994 l NOVA1 neuro-oncological ventral antigen 1 207535 S. at 2.67OOOS8 l NFKB2 nuclear factor of kappa light polypeptide gene enhancer in B-cells 2 (p49, p100) 204015 s at 2.66827 S4 l DUSP4 dual specificity phosphatase 4 217738 a 2.6675475 l NAMPT nicotinamide phosphoribosyltransferase 221569 a 2.6674O23 l AHI1 Abelson helper integration site 1 236140 a 2.666789S l GCLM glutamate-cysteine ligase, modifier Subunit 1554.462 a. at 2.66640O2 l DNATB9 DnaJ (Hsp40) homolog, Subfamily B, member 9 212344 a 2.6652708 l SULF1 Sulfatase 1 202557 a 2.6646934 l HSPA13 heat shock protein 70 kDa family, member 13 22494.0 s at 2.6638696 l PAPPA pregnancy-associated plasma protein A, pappalysin 1 220655 a. 2.6540391 l TNIP3 TNFAIP3 interacting protein 3 216199 s at 2.654O227 l MAP3K4 mitogen-activated protein kinase kinase kinase 4 230206 a 2.6534097 l DOCKS Dedicator of cytokinesis 5, mRNA (cDNA clone IMAGE: 3347029) 204557 s at 2.6526,718 l DZIP1 DAZ interacting protein 1 206390 x at 2.65.15956 l PF4 platelet factor 4 226322 a. 2.6468,246 l TMTC1 transmembrane and tetratricopeptide repeat containing 1 203925 a. 2.6462543 l GCLM glutamate-cysteine ligase, modifier Subunit 220955 X at 2.64.4316 l RAB23 RAB23, member RAS oncogene amily 208322 s at 2.64381.SS l ST3GAL1 ST3 beta-galactoside alpha-2,3- sialyltransferase 1 225,033 a 2.640394 l ST3GAL1 ST3 beta-galactoside alpha-2,3- sialyltransferase 1 208964 S at 2.6379926 l FADS1 atty acid desaturase 1 219134 a 2.6371877 l ELTD1 EGF, latrophilin and seven transmembrane domain containing 1 232504 a 2.6357627 l LOC285628 hypothetical protein LOC285628 2391.63 a 2.63.56966 l UBE2B ubiquitin-conjugating enzyme E2B (RAD6 homolog) 205924 a 2.6338.334 l RAB3B RAB3B, member RAS oncogene amily 225532 a 2.63293SS l CABLES1 CdkS and Ablenzyme substrate 1 228785 a 2.6269226 l ZNF281 Full length insert cDNA clone ZEO9A11 1554980 a. at 2.623143 l ATF3 activating transcription factor 3 212607 a 2.6.224685 l AKT3 v-akt murine thymoma viral oncogene homolog 3 (protein kinase B, gamma) 227.058 a 2.6.2094O2 l C13orf53 chromosome 13 open reading frame 33 238649 a 2.62O7957 l PITPNC1 phosphatidylinositol transfer protein, cytoplasmic 1 242163 a 2.6.196032 l THRAP3 hyroid hormone receptor associated protein 3 229947 a 2.6.18586 l PI15 peptidase inhibitor 15 223264 a 2.6175916 l MESDC1 mesoderm development candidate 1 242283 a 2.6143296 l C1orf67 chromosome 1 open reading frame 67 DNAH14 if dynein, axonemal, heavy chain 14 202842 s at 2.612975 l DNAB9 DnaJ (Hsp40) homolog, Subfamily B, member 9 214472 a. 2.6077855 l HIST1H2AD histone cluster 1, H2ad histone cluster 1, H3a histone cluster 1, H3b HIST1H3A f/f histone cluster 1, H3c histone if HIST1H3B cluster 1, H3d if histone cluster 1, H3e if HIST1H3C f/f histone cluster 1, H3f histone cluster 1, H3 gift histone cluster 1, H3h HIST1H3D f/f histone cluster 1, H3i histone if HIST1H3E cluster 1, H3 if HIST1H3F

HIST1H3G US 2012/0034613 A1 Feb. 9, 2012 15

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title HIST1H3H if HIST1H3I if HIST1H3J 218980 a 2.60654.04 l FHOD3 ormin homology 2 domain containing 3 217590 s. at 2.6O19518 l TRPA1 transient receptor potential cation channel, Subfamily A, member 1 211756 a 2.5990396 l PTHLH parathyroid hormone-like hormone 227080 a 2.59890S6 l ZNF697 Zinc finger protein 697 220346 a 2.5971677 l MTHFD2L methylenetetrahydrofolate dehydrogenase (NADP+ dependent) 2 ike 221942 s at 2.5942.783 l GUCY1A3 guanylate cyclase 1, Soluble, alpha 3 205825 a. 2.5902O19 l PCSK1 proprotein convertase Subtilisinkexin type 1 222945 X at 2.5855827 l OLAH oleoyl-ACP hydrolase 226931 a 2.583O119 l TMTC1 transmembrane and tetratricopeptide repeat containing 1 24-1765 a 2.5736842 l CPM carboxypeptidase M 230237 a 2.5733.738 l ADCYAP1 adenylate cyclase activating polypeptide 1 (pituitary) 209808 X at 2.5724.812 l ING1 inhibitor of growth family, member 1 227945 a. 2.5660486 l TBC1D1 TBC1 (tre-2/USP6, BUB2, ccdc16) domain family, member 1 214446 a 2.5570273 l ELL2 elongation factor, RNA polymerase II, 2 221782 a 2.5525873 l DNAJC10 DnaJ (Hsp40) homolog, subfamily C, member 10 223.058 a 2.5435784 l FAM107B family with sequence similarity 107, member B 202213 s at 2.5383.074 l CUL4B cullin 4B 237106 a 2.5334878 l SLC11A2 NRAMP2 235927 a 2.5293667 l XPO1 exportin 1 (CRM1 homolog, yeast) 230494 a 2.526OO12 l SLC20A1 Solute carrier family 20 (phosphate transporter), member 1, mRNA (cDNA clone MGC: 8767 IMAGE: 3918690) 204897 a 2.52S4738 l PTGER4 prostaglandin E receptor 4 (Subtype EP4) 214469 a 2.5204768 l HIST1H2AB histone cluster 1, H2ab histone cluster 1, H2ae HIST1H2AE 204472 a. 2.5198975 l GEM GTP binding protein overexpressed in skeletal muscle 206026 s at 2.519742 l TNFAIP6 tumor necrosis factor, alpha-induced protein 6 210367 s at 2.5187645 l PTGES prostaglandin E synthase 217996 at 2.5148838 l PHILDA1 pleckstrin homology-like domain, amily A, member 1 227123 at 2.5144148 l RAB3B Small GTP binding protein RAB3B (RAB3B) 208885 at 2.5127752 l LCP1 ymphocyte cytosolic protein 1 (L- plastin) 214254 at 2.5125573 l MAGEA4 melanoma antigen family A, 4 226886 at 2.512313 l GFPT1 -fructose-6-phosphate transaminase 1 205566 at 2.5082593 l ABHD2 abhydrolase domain containing 2 221781 s at 2.5082314 l DNAJC10 DnaJ (Hsp40) homolog, Subfamily C, member 10 1554741 s at 2.5060341 l FGF7 fibroblast growth factor 7 (keratinocyte KGFLP1 if growth factor) if keratinocyte growth KGFLP2 actor-like protein 1 / keratinocyte growth factor-like protein 2 206300 s at 2.5052757 l PTHLH parathyroid hormone-like hormone 206805 at 2.SO4S469 l SEMA3A Sema domain, immunoglobulin domain (Ig), short basic domain, Secreted, (semaphorin) 3A 202052 s at 2. SO331.93 l RAI14 retinoic acid induced 14 213993 at 2.4992S3 l SPON1 spondin 1, extracellular matrix protein 209897 s at 2.4991436 l SLIT2 slithomolog 2 (Drosophila) 229530 at 2.498.387 l GUCY1A3 guanylate cyclase 1, Soluble, alpha 3 215285 S at 2.4979303 l PHTF1 putative homeodomain transcription factor 1 US 2012/0034613 A1 Feb. 9, 2012 16

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 218615 s at 2.49741.63 l TMEM39A transmembrane protein 39A 220615 s at 2.4965284 l FAR2 fatty acyl CoA reductase 2 205303 a 2.492.71 l KCNJ8 potassium inwardly-rectifying channel, Subfamily J, member 8 222735 a. 2.4891634 l TMEM38B transmembrane protein 38B 2095O1 a 2.487 S13 l CDR2 cerebellar degeneration-related protein 2, 62 kDa 205767 a 2.486.593 l EREG epiregulin 205830 a. 2.48.58525 l CLGN calmegin 228128 x at 2.48174OS l PAPPA pregnancy-associated plasma protein A, pappalysin 1 229256 a 2.4780662 l PGM2L1 phosphoglucomutase 2-like 1 22305.9 s at 2.4761326 l FAM107B family with sequence similarity 107, member B 228551 a 2.473978 l DENNDSB DENN/MADD domain containing 5B 205098 a 2.47395.16 l CCR1 chemokine (C-C motif) receptor 1 220272 a 2.47 10832 l BNC2 basonuclin 2 212975 a 2.4708278 l DENND3 DENN/MADD domain containing 3 222690 s. at 2.469726 l TMEM39A transmembrane protein 39A 203239 s at 2.46571.9S l CNOT3 CCR4-NOT transcription complex, subunit 3 203823 a 2.4619107 l RGS3 regulator of G-protein signaling 3 203373 a 2.46O2883 l SOCS2 Suppressor of cytokine signaling 2 210692 s at 2.456732 l SLC43 A3 solute carrier family 43, member 3 205139 s at 2.4541962 l UST uronyl-2-sulfotransferase 220145 a. 2.4529 l MAP9 microtubule-associated protein 9 226743 a 2.4S1173 l SLFN11 schlafen family member 11 219093 a 2.4472542 l PID1 phosphotyrosine interaction domain containing 1 213112 S at 2.4471.107 l SQSTM1 sequestosome 1 219228 a 2.4418485 l ZNF331 Zinc finger protein 331 1554110 at 2.44O1536 l CDCP1 CUB domain containing protein 1 217649 a 2.4384716 l ZFANDS Zinc finger, AN1-type domain 5 2268.04 a 2.4369S45 l FAM2OA family with sequence similarity 20, member A 243403 X at 2.4360526 l CPM carboxypeptidase M 214701 s at 2.4357622 l FN1 fibronectin 1 207522 s at 2.434122 l ATP2A3 ATPase, Ca++ transporting, ubiquitous 219117 s at 2.431.4268 l FKBP11 FK506 binding protein 11, 19 kDa 21184.0 s at 24287648 l PDE4D phosphodiesterase 4D, cAMP-specific (phosphodiesterase E3 dunce homolog, Drosophila) 239001 a 2.428.258 l MGST1 Microsomal glutathione S-transferase 1 (MGST1), transcript variant 1d, mRNA 219427 a 2.4277742 l FAT4 FAT tumor Suppressor homolog 4 (Drosophila) 228.540 a 2.426083 l QKI quaking homolog, KH domain RNA binding (mouse) 205547 s at 2.4258394 l TAGLN transgelin 214375 a. 2.425571 l LOC729222 similar to mKIAA1230 protein fif PPFIBP1 PTPRF interacting protein, binding protein 1 (liprin beta 1) 204037 a 2.4254246 l LPAR1 ySophosphatidic acid receptor 1 213704 a 2.4229991 l RABGGTB Rab geranylgeranyltransferase, beta Subunit 219118 a 2.422199 l FKBP11 FK506 binding protein 11, 19 kDa 225582 a 2.4213138 l TPRIP inositol 1,4,5-triphosphate receptor interacting protein 229.307 a 2.42O8326 l ANKRD28 ankyrin repeat domain 28 214290 s. at 2.42O4948 l HIST2H2AA3 histone cluster 2, H2aa3 fff histone cluster 2, H2aa4 HIST2H2AA4 201739 a 2.4191535 l SGK1 serum glucocorticoid regulated kinase 1 239336 a 2.4168572 l THEBS1 (clone lambda-TS-33) thrombospondin (THBS) mRNA, 5' end 1559400 S. at 2.4151716 l PAPPA pregnancy-associated plasma protein A, pappalysin 1 205066 s at 2.4135945 l ENPP1 ectonucleotide pyrophosphatase/phosphodiesterase 1 220034 at 2.41 13884 l RAK3 interleukin-1 receptor-associated kinase 3 208963 X at 2.4111784 l FADS1 fatty acid desaturase 1 US 2012/0034613 A1 Feb. 9, 2012 17

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 213496 a 2.4059057 l LPPR4 plasticity related gene 1 205453 a 2.4039452 l HOXB2 homeobox B2 205119 s at 2.4022236 l FPR1 formyl peptide receptor 1 232224 a 2.401911S l MASP1 mannan-binding lectin peptidase 1 (C4C2 activating component of Ra reactive factor) 202392 s at 2.4O164 l PISD phosphatidylserine decarboxylase 212906 a 2.4009886 l GRAMID1B GRAM domain containing 1B, mRNA (cDNA clone IMAGE: 3854666) 218801 a 2.4OOO664 l UGCGL2 UDP-glucose ceramide glucosyltransferase-like 2 212122 a. 2.39834.67 l RHOQ ras homologgene family, member Q 22O153 a 2.394.9509 l ENTPD7 ectonucleoside triphosphate diphosphohydrolase 7 204036 a 2.390SS94 l LPAR1 lysophosphatidic acid receptor 1 228461 a 2.3892233 l SH3RF3 SH3 domain containing ring finger 3 205304 is at 2.3868387 l KCNJ8 potassium inwardly-rectifying channel, Subfamily J, member 8 212350 a 2.386301 l TBC1D1 TBC1 (tre-2/USP6, BUB2, ccdc16) domain family, member 1 205003 a 2.3832471 l DOCK4 dedicator of cytokinesis 4 235339 a 2.376645 l SETDB2 SET domain, bifurcated 2 220012 a 2.3763833 l ERO1LB ERO1-like beta (S. cerevisiae) 44783 s at 2.372O322 l HE hairy fenhancer-of-split related with YRPW motif 1 1554290 at 2.3719432 l HERC3 hect domain and RLD 3 1570351 at 2.370291 l ADAMTS6 ADAM metallopeptidase with thrombospondin type 1 motif, 6 204222 S at 2.366,1144 l GLIPR1 GLI pathogenesis-related 1 222880 at 2.364.9035 l AKT3 V-akt murine thymoma viral oncogene homolog 3 (protein kinase B, gamma) 235593 at 2.3646,154 l LOC100128821 hypothetical protein LOC100128821 /// F. ZEB2 Zinc finger E-box binding homeobox2 232263 at 2.3646133 l SLC6A15 Solute carrier family 6 (neutral transporter), member 15 1552701 a. at 2.3634586 l CARD16 caspase recruitment domain family, member 16 224807 at 2.3633232 l GRAMD1A GRAM domain containing 1A 204470 at 2.362897 l CXCL1 chemokine (C X—C motif) ligand 1 (melanoma growth stimulating activity, alpha) 230263 s at 2.3622723 l DOCKS Dedicator of cytokinesis 5, mRNA (cDNA clone IMAGE: 3347029) 213035 at 2.3611116 l ANKRD28 ankyrin repeat domain 28 202986 at 2.3609338 l ARNT2 aryl-hydrocarbon receptor nuclear translocator 2 209437 s at 2.36OO132 l SPON1 spondin 1, extracellular matrix protein 206613 s at 2.3585851 l TAF1A TATA box binding protein (TBP)- associated factor, RNA polymerase I, A, 48 kDa 24.2079 at 2.357697 l RGS12 regulator of G-protein signaling 12 211924 S at 2.356,7264 l PLAUR plasminogen activator, urokinase receptor 203836 s at 2.3556855 l MAP3KS mitogen-activated protein kinase kinase kinase 5 1555471 a. at 2.354568 l FMN2 ormin 2 201042 at 2.3514817 l TGM2 transglutaminase 2 (C polypeptide, protein-glutamine-gamma glutamyltransferase) 223217 s at 2.3498836 l NFKBIZ. nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, Zeta 1555103 s at 2.349.5677 l FGF7 fibroblast growth factor 7 (keratinocyte growth factor) 221815 at 2.3482256 l ABHD2 abhydrolase domain containing 2 1558143 a at 2.346621 l BCL2L11 BCL2-like 11 (apoptosis facilitator) 223774 at 2.3389.745 l SNEHG12 Small nucleolar RNA host gene 12 (non-protein coding) 1553194 at 2.3387334 l NEGR1 neuronal growth regulator 1 231015 at 2.3381798 l KLF15 Kruppel-like factor 15 1552578 a. at 2.337757 l MYO3B myosin IIIB 202340 x at 2.33773 l NR4A1 nuclear receptor Subfamily 4, group A, member 1 US 2012/0034613 A1 Feb. 9, 2012 18

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 202609 at 2.334.194 up EPS8 epidermal growth factor receptor pathway substrate 8 210941 at 2.333396 up PCDHT protocadherin 7 203921 at 2.331509 up CHST2 carbohydrate (N-acetylglucosamine-6- O) sulfotransferase 2 1562102 at 2.3314924 up AKR1C1 Aldo-keto reductase family 1, member C1 (dihydrodiol dehydrogenase 1; 20 alpha (3-alpha)-hydroxysteroid dehydrogenase), mRNA (cDNA clone MGC: 42600 IMAGE: 4825338) 222305 at 2.3291554 l HK2 hexokinase 2 21731.0 s at 2.3269384 l FOXJ3 forkhead box J3 205128 x at 2.326936 l PTGS1 prostaglandin-endoperoxide synthase 1 (prostaglandin GH synthase and cyclooxygenase) 238989 a 2.326773 l C1GALT1C1 C1GALT1-specific chaperone 1 235770 a 2.3237767 l MASP1 mannan-binding lectin serine peptidase 1 (C4C2 activating component of Ra reactive factor) 226103 a 2.3234518 l NEXN nexilin (Factin binding protein) 220841 s at 2.3223.662 l AHI1 Abelson helper integration site 1 228817 a 2.32098.72 l ALG9 asparagine-linked 9, alpha-1,2-mannosyltransferase homolog (S. cerevisiae) 227688 a 2.318O34 l LRCH2 leucine-rich repeats and calponin homology (CH) domain containing 2 236290 a 2.3161294 l DOK6 docking protein 6 222736 s at 2.3157086 l TMEM38B transmembrane protein 38B 229584 a 2.31394.08 l LRRK2 leucine-rich repeat kinase 2 203414 a 2.313451 l MMD monocyte to macrophage differentiation-associated 207570 a 2.312389 l SHOX short stature homeobox 205.214 a 2.3090763 l STK17B serine/ kinase 17b 207630 s at 2.3O87645 l CREM cAMP responsive element modulator 207237 a 2.3O84.404 l KCNA3 potassium voltage-gated channel, shaker-related subfamily, member 3 228699 a 2.306360S l NRP2 Vascular endothelial cell growth factor 65 receptor 2 (VEGF165R2) 215813 s at 2.3061.419 l PTGS1 prostaglandin-endoperoxide synthase 1 (prostaglandin GH synthase and cyclooxygenase) 209180 at 2.3054507 l RABGGTB Rab geranylgeranyltransferase, beta Subunit 218696 at 2.3052793 l EIF2AK3 eukaryotic translation initiation factor 2-alpha kinase 3 223370 at 2.302342 l PLEKHA3 pleckstrin homology domain containing, family A (phosphoinositide binding specific) member 3 205100 at 2.2997909 l GFPT2 glutamine-fructose-6-phosphate transaminase 2 212609 s at 2.2997367 l AKT3 Clones 23920 and 2392.1 mRNA Sequence 1555.281 X at 2.2991943 l ARMC8 armadillo repeat containing 8 204944 a 2.2989373 l PTPRG protein tyrosine phosphatase, receptor type, G 229438 a 2.2988OSS l LOC100132244 CDNA: FLJ22487 fis, clone HRC10931 212902 a 2.298.442 l SEC24A SEC24 family, member A (S. cerevisiae) 224826 a 2.298.3568 l RP5- hypothetical protein KIAA1434 1022P6.2 218772 X at 2.2961-67 l TMEM38B transmembrane protein 38B 241372 a. 2.29.598.62 l ZC3H6 Zinc finger CCCH-type containing 6 229555 a. 2.295903 l GALNTS UDP-N-acetyl-alpha-D- galactosamine:polypeptide N acetylgalactosaminyltransferase 5 (GalNAc-T5) 202375 a. 2.295754 l SEC24D SEC24 family, member D (S. cerevisiae) 1559582 at 2.2931254 l RHOQ ras homologgene family, member Q 1555279 at 2.29071 l ARMC8 armadillo repeat containing 8 203475 a. 2.2897353 l CYP19A1 cytochrome P450, family 19, Subfamily A, polypeptide 1 204984 a 2.2885683 l GPC4 glypican 4 1560007 at 2.28589 l LOC645984 hypothetical LOC645984 US 2012/0034613 A1 Feb. 9, 2012 19

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 23.5236 at 2.284.801S l LOC100131897 Uncharacterized protein LOC100131897 (LOC100131897), mRNA 1559060 a. at 2.284.3337 l FNIP1 MRNA, cDNA DKFZp451AO64 (from clone DKFZp451AO64) 204089 x at 2.2831.46 l MAP3K4 mitogen-activated protein kinase kinase kinase 4 227621 at 2.2817085 l WTAP Wilms tumor 1 associated protein (WTAP), transcript variant 2, mRNA 203498 at 2.28O8936 l RCAN2 regulator of calcineurin 2 230207 s at 2.27943O4 l DOCKS Dedicator of cytokinesis 5, mRNA (cDNA clone IMAGE: 3347029) 213425 at 2.2791493 l WNTSA wingless-type MMTV integration site family, member 5A 220745 at 2.2776432 l L19 interleukin 19 235338 s at 2.2773378 l SETDB2 SET domain, bifurcated 2 200790 at 2.27561.64 l ODC1 ornithine decarboxylase 1 218280 x at 2.2754815 l HIST2H2AA3 histone cluster 2, H2aa3 histone cluster 2, H2aa4 HIST2H2AA4 20472.0 s at 2.2753987 l DNAJC6 DnaJ (Hsp40) homolog, subfamily C, member 6 202820 a 2.272O664 l AHR aryl hydrocarbon receptor 228423 a 2.270422 l MAP9 microtubule-associated protein 9 203424 is at 2.2697344 l GFBP5 insulin-like growth factor binding protein 5 221918 a 2.26818O8 l PCTK2 PCTAIRE protein kinase 2 238877 a 2.26654.46 l EYA4 eyes absent homolog 4 (Drosophila) 202721 s at 2.2655363 l GFPT1 glutamine-fructose-6-phosphate transaminase 1 213159 a 2.265123 l PCNX pecanex homolog (Drosophila) 203837 a 2.261806S l MAP3KS mitogen-activated protein kinase kinase kinase 5 244808 a 2.261.4517 l GRAMD1A GRAM domain containing 1A, mRNA (cDNA clone IMAGE: 5921205) 222802 a 2.2602103 l EDN1 endothelin 1 20272.2 s at 2.2597966 l GFPT1 glutamine-fructose-6-phosphate transaminase 1 228368 a 2.2592928 l ARHGAP2O Rho GTPase activating protein 20 228653 a 2.25894.45 l SAMDS sterile alpha motif domain containing 5 2244.55 s at 2.2587.087 l ADPGK ADP-dependent glucokinase 1570515 a. at 2.258047 l FILIP1 filamin A interacting protein 1 44790 s. at 2.257748 l C13orf18 chromosome 13 open reading frame 18 LOC72897O // hypothetical LOC728970 213836 s at 2.2555509 l WIPI1 WD repeat domain, phosphoinositide interacting 1 225589 a 2.254314 l SH3RF1 SH3 domain containing ring finger 1 209545 S. at 2.2526133 l RIPK2 receptor-interacting serine-threonine kinase 2 1556113 at 2.2524776 l DKFZp451A211 DKFZp451A211 protein 206249 a 2.2523835 l MAP3K13 mitogen-activated protein kinase kinase kinase 13 244852 a 2.2S14307 l DSEL dermatan Sulfate epimerase-like 203980 a 2.2SO6835 l FABP4 atty acid binding protein 4, adipocyte 212585 a. 2.25O1442 l OSBPL8 oxysterol binding protein-like 8 210845 s at 2.249447 l PLAUR plasminogen activator, urokinase receptor 223463 a 2.2488134 l RAB23 RAB23, member RAS oncogene amily 209204 a 2.24691.99 l LMO4 LIM domain only 4 227445 a. 2.242928S l ZNF689 Zinc finger protein 689 223222 a. 2.2426856 l SLC25A19 solute carrier family 25 (mitochondrial hiamine pyrophosphate carrier), member 19 209277 a 2.2425933 l TFPI2 issue factor pathway inhibitor 2 205138 s at 2241066 l UST uronyl-2-sulfotransferase 219390 a 2.240858 l FKBP14 FK506 binding protein 14, 22 kDa 223315 a. 2.24O703 l NTN4 netrin 4 213309 a 2.24O6566 l PLCL2 phospholipase C-like 2 218541 s at 2.2391.245 l C8orfA. chromosome 8 open reading frame 4 210510 s at 2.238.3678 l NRP1 neuropilin 1 212915 a. 2.2363226 l PDZRN3 PDZ domain containing ring finger 3 226890 a 2.2358947 l WDR35 WD repeat domain 35 US 2012/0034613 A1 Feb. 9, 2012 20

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 224.523 s at 2.234SS38 l C3orf26 chromosome 3 open reading frame 26 213029 at 2.234S178 l NFIB nuclear factor IB 204597 X at 2.2341,132 l STC1 Stanniocalcin 1 204595 S. at 2.2336535 l STC1 Stanniocalcin 1 210875 s at 2.233O45 l ZEB1 Zinc finger E-box binding homeobox 1 210257 X at 2.231 l CUL4B cullin 4B 2301.23 a 2.2295592 l NECAP2 NECAP endocytosis associated 2 2295.88 a 2.229439 l DNAJC10 DnaJ (Hsp40) homolog, subfamily C, member 10 220092 s at 2.2293727 l ANTXR1 anthrax toxin receptor 1 213417 a 2.228S94 l TBX2 T-box2 228986 a 2.2255878 l OSBPL8 oxysterol binding protein-like 8 201531 a 2.22S1139 l ZFP36 Zinc finger protein 36, C3H type, homolog (mouse) 210896 s at 2.224665 l ASPH aspartate beta-hydroxylase 222619 a 2.222882S l ZNF281 Zinc finger protein 281 220254 a 2.2226748 l LRP12 low density lipoprotein-related protein 12 219910 a 2.220998 l FICD FIC domain containing 227351 a 2.22O7675 l C16orf52 chromosome 16 open reading frame 52 218401 s at 2.2176473 l ZNF281 Zinc finger protein 281 201289 a 2.2144566 l CYR61 cysteine-rich, angiogenic inducer, 61 202543 s at 2.2127845 l GMFB glia maturation factor, beta 213113 s at 2.21 1486 l SLC43 A3 solute carrier family 43, member 3 226337 a 2.210516S l SCYL1BP1 SCY1-like 1 binding protein 1 221958 s at 2.209569 l GPR177 G protein-coupled receptor 177 219985 a 2.2090566 l HS3ST3A1 heparan sulfate (glucosamine) 3-O- Sulfotransferase 3A1 223866 a 2.2079794 l ARMC2 armadillo repeat containing 2 221978 a 2.2O7626 l HLA-F major histocompatibility complex, class I, F 240728 a 2.20726 l PLCB4 Phospholipase C beta 4 (PLCB4) 213338 a 2.20540O2 l TMEM158 transmembrane protein 158 204151 X at 2.205384 l AKR1C1 aldo-keto reductase family 1, member C1 (dihydrodiol dehydrogenase 1; 20 alpha (3-alpha)-hydroxysteroid dehydrogenase) 1554414 a at 2.205.3685 l OSGIN2 oxidative stress induced growth inhibitor family member 2 209708 at 2.2048678 l MOXD1 monooxygenase, DBH-like 1 2298.68 s at 2.2048671 l GDF15 Macrophage inhibitory cytokine-1 (MIC-1) 230O31 at 2.2040954 l HSPAS heat shock 70 kDa protein 5 (glucose regulated protein, 78 kDa) 203857 s at 2.2O38O1 l PDIAS protein disulfide isomerase family A, member 5 239286 at 2.2O35995 l CDH11 cadherin 11, type 2, OB-cadherin (osteoblast) 225174 at 2.2O341O1 l DNAJC10 DnaJ (Hsp40) homolog, subfamily C, member 10 235706 at 2.202396 l CPM carboxypeptidase M 205495 S. at 2.2022398 l GNLY granulysin 235550 at 2.201.9966 l MAP9 microtubule-associated protein 9 210275 s at 2.2013817 l ZFANDS Zinc finger, AN1-type domain 5 216218 s at 2.2008579 l PLCL2 phospholipase C-like 2 202888 s at 2.2007616 l ANPEP alanyl (membrane) aminopeptidase 210191 s at 2.1.997S6 l PHTF1 putative homeodomain transcription factor 1 209781 s at 2.199483 l KHDRBS3 KH domain containing, RNA binding, signal transduction associated 3 1554036 at 2.1.96S 137 l ZBTB24 Zinc finger and BTB domain containing 24 238669 at 2.1943119 l PTGS1 prostaglandin-endoperoxide synthase 1 (prostaglandin GH synthase and cyclooxygenase) 21303.6 X at 2.19.1737 l ATP2A3 ATPase, Ca++ transporting, ubiquitous 227220 at 2.1902707 l NFXL1 nuclear transcription factor, X-box binding-like 1 236129 at 2.1839883 l GALNTS CDNA FLJ75131 complete cols, highly similar to Homo sapiens UDP-N- acetyl-alpha-D- galactosamine:polypeptide N US 2012/0034613 A1 Feb. 9, 2012 21

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title acetylgalactosaminyltransferase 5 (GalNAc-T5) (GALNT5), mRNA 1555724 S at 2.1835961 l TAGLN transgelin 208510 s at 2.1819134 l PPARG peroxisome proliferator-activated receptor gamma 229271 x at 2.1817298 l COL11A1 collagen, type XI, alpha 1 219631 at 2.18O8488 l LRP12 low density lipoprotein-related protein 12 218193 s at 2.1798851 l GOLT1B golgi transport 1 homolog B (S. cerevisiae) 227660 at 2.1793392 l ANTXR1 anthrax toxin receptor 1 1552309 a at 2.176896 l NEXN nexilin (Factin binding protein) 214061 at 2.1762.218 l WDR67 WD repeat domain 67 227294 at 2.1758094 l ZNF689 Zinc finger protein 689 223296 at 2.175538 l SLC25A33 solute carrier family 25, member 33 203425 s at 2.1755157 l GFBP5 insulin-like growth factor binding protein 5 226001 at 2.1747112 l KLHLS kelch-like 5 (Drosophila) 206445 s at 2.1745844 l PRMT1 protein methyltransferase 1 213988 s at 2.1742477 l SAT1 spermidine?spermine N1 acetyltransferase 1 214657 s at 2.1736887 l NCRNA00O84 non-protein coding RNA 84 210350 x at 2.1736152 l NG1 inhibitor of growth family, member 1 204556 s at 2.1734612 l DZIP1 DAZ interacting protein 1 207345 at 2.170217 l FST ollistatin 220301 at 2.16967 S4 l CCDC102B coiled-coil domain containing 102B 203574 at 2.169319 l NFIL3 nuclear factor, interleukin 3 regulated 215058 at 2.1687279 l DENNDSB DENN/MADD domain containing 5B 238497 at 2.1683OO)4 l TMEM136 transmembrane protein 136 2394.68 at 2.1671 67 l MKX mohawk homeobox 1554474 a at 2.1654644 l MOXD1 monooxygenase, DBH-like 1 205991 s a 2.1652021 l PRRX1 paired related homeobox 1 219500 at 2.1651778 l CLCF1 cardiotrophin-like cytokine factor 1 207829 s a 2.1641598 l BNIP1 BCL2 adenovirus E1B 19 kDa interacting protein 37226 at 2.1634657 l BNIP1 BCL2 adenovirus E1B 19 kDa interacting protein 20985.0 s a 2.160621 l CDC42EP2 CDC42 effector protein (Rho GTPase binding) 2 217741 s a 2.1604116 l ZFANDS Zinc finger, AN1-type domain 5 1554140 at 2.1590114 l WDR78 WD repeat domain 78 205110 s a 2.1575923 l FGF13 fibroblast growth factor 13 63825 at 2.157092 l ABHD2 abhydrolase domain containing 2 228702 at 2.1557388 l FLJ43663 hypothetical LOC378805 226800 at 2.1555.407 l EFCAB7 EF-hand calcium binding domain 7 202952 s a 2.155327 l ADAM12 ADAM metallopeptidase domain 12 217739 s a 2.155086 l NAMPT nicotinamide phosphoribosyltransferase 2251.85 at 2.1548986 l MRAS muscle RAS oncogene homolog 214211 at 2.1543927 l FTH1 erritin, heavy polypeptide 1 235311 at 2.15348 l FKBP14 FK506 binding protein 14, 22 kDa, mRNA (cDNA clone MGC: 12218 MAGE: 4042173) 225871 at 2.153166 l STEAP2 six transmembrane epithelial antigen of he prostate 2 1561042 at 2.1507823 l ITGB1 integrin, beta 1 (fibronectin receptor, beta polypeptide, antigen CD29 includes MDF2, MSK12) 204790 at 2.1506758 l SMAD7 SMAD family member 7 203518 at 2.1503856 l LYST ysosomal trafficking regulator 208415 X at 2.1496897 l ING1 inhibitor of growth family, member 1 230291 s at 2.149512S l NFIB HMGIC/NFIB fusion protein (HMGIC/NFIB) 213032 at 2.146494 l NFIB nuclear factor IB 214913 at 2.1450849 l ADAMTS3 ADAM metallopeptidase with hrombospondin type 1 motif, 3 209436 at 2.1450522 l SPON1 spondin 1, extracellular matrix protein 210355 at 2.14395O2 l PTHLH parathyroid hormone-like hormone 225436 at 2.1422665 l FAM108C1 amily with sequence similarity 108, member C1 1554.602 at 2.1414428 l RBM8A RNA binding motif protein 8A 204420 at 2.1413243 l FOSL1 FOS-like antigen 1 204742 s at 2.1387715 l PDSSB PDS5, regulator of cohesion maintenance, homolog B (S. cerevisiae) US 2012/0034613 A1 Feb. 9, 2012 22

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 235.182 at 2.138126 l ISM1 isthmin 1 homolog () 203740 at 2.1379848 l MPHOSPH6 M-phase phosphoprotein 6 215997 s at 2.1378837 l CUL4B cullin 4B 219558 at 2.13.78727 l ATP13A3 ATPase type 13A3 223758 s at 2.137832 l GTF2H2 general transcription factor IIH, polypeptide 2, 44 kDa 24-1986 at 2.1363626 l BMPER BMP binding endothelial regulator 232267 at 2.135431 l GPR133 G protein-coupled receptor 133 210841 s at 2.132714 l NRP2 neuropilin 2 229.623 at 2.13186O7 l FLJ12993 Hypothetical LOC441027 (FLJ12993), mRNA 224,461 s at 2.13164 l AIFM2 apoptosis-inducing factor, mitochondrion-associated, 2 219774 at 2.13 OOSO7 l CCDC93 coiled-coil domain containing 93 229306 at 2.12945S l FAM148B Family with sequence similarity 148, member B (FAM148B), mRNA 226142 at 2.1290789 l GLIPR1 GLI pathogenesis-related 1 210424 S at 2.1265538 l GOLGA8A golgi autoantigen, golgin Subfamily a, 8A golgi autoantigen, golgin GOLGA8B Subfamilya, 8B 232004 at 2.1262782 l HNRNPR heterogeneous nuclear ribonucleoprotein R 228950 S at 2.1261652 l GPR177 G protein-coupled receptor 177 23.9415 at 2.1261606 l MAP9 microtubule-associated protein 9 213093 at 2.12S2O6 l PRKCA protein kinase C, alpha 213033 s at 2.1251779 l NFIB nuclear factor IB 218574 s at 2.1227338 l LMCD1 LIM and cysteine-rich domains 1 201467 s at 2.12 1963S l NQO1 NAD(P)H dehydrogenase, quinone 1 224978 s at 2.1217666 l USP36 ubiquitin specific peptidase 36 232311 at 2.1214871 l B2M Beta 2-mu = beta 2-microglobulin human, SK-MEL-33 cells, mRNA Mutant, 433 nt 211981 at 2.1.21121 l COL4A1 collagen, type IV, alpha 1 219471 at 2.11986O2 l C13orf18 chromosome 13 open reading frame 18 LOCA2897O // hypothetical LOC728970 214014 at 2.1.19122 l CDC42EP2 CDC42 effector protein (Rho GTPase binding) 2 235O19 at 2.11691.9S l CPM carboxypeptidase M 218810 at 2.1168468 l ZC3H12A Zinc finger CCCH-type containing 12A 210519 s at 2.116195 l NQO1 NAD(P)H dehydrogenase, quinone 1 219790 s. at 2.11SO982 l NPR3 natriuretic peptide receptor Ciguanylate cyclase C (atrionatriuretic peptide receptor C) 215O12 a 2.1141691 l ZNF451 Zinc finger protein 451 203853 S at 2.114138 l GAB2 GRB2-associated binding protein 2 229553 a 2.1131918 l PGM2L1 phosphoglucomutase 2-like 1 203066 a 2.112S245 l GALNAC4S- B cell RAG associated protein 6ST 221881 s at 2.1089115 l CLIC4 chloride intracellular channel 4 229414 a 2.108.2094 l PITPNC1 phosphatidylinositol transfer protein, cytoplasmic 1 228082 a 2.108O198 l ASAM adipocyte-specific adhesion molecule 225212 a 2.1078928 l SLC25A2S solute carrier family 25 (mitochondrial carrier; phosphate carrier), member 25 204115 a. 2.10781OS l GNG11 guanine nucleotide binding protein (G protein), gamma 11 229.194 a 2.1072176 l PCGF5 polycomb group ring finger 5 219283 a 2.1071.42 l C1GALT1C1 C1GALT1-specific chaperone 1 215785 S at 2.1066854 l CYFIP2 cytoplasmic FMR1 interacting protein 2 217202 s at 2.1064858 l GLUL glutamate-ammonia ligase (glutamine synthetase) 228949 a 2.1053102 l GPR177 G protein-coupled receptor 177 226136 a 2.10407 l GLIPR1 GLI pathogenesis-related 1 225590 a 2.1038868 l SH3RF1 SH3 domain containing ring finger 1 226160 a 2.1O3OSO7 l H6PD hexose-6-phosphate dehydrogenase (glucose 1-dehydrogenase) 236154 a 2.1026752 l QKI CDNA FLJ39382 fis, clone PERIC2OOO473 225,034 a 2.099384 l ST3GAL1 ST3 beta-galactoside alpha-2,3- sialyltransferase 1 202724 S at 2.0982,356 l FOXO1 forkhead box O1 1559477 s at 2.098.0742 l MEIS1 Meis homeobox 1 US 2012/0034613 A1 Feb. 9, 2012 23

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 226731 at 2.0977373 l PELO Pelota major mRNA, complete cols: alternatively spliced 220253 s at 2.0964284 l LRP12 low density lipoprotein-related protein 12 1555270 a. at 2.0956323 l WFS1 Wolfram syndrome 1 (wolframin) 219054 a 2.0956144 l CSOrf23 chromosome 5 open reading frame 23 155396.2 s at 2.094OS28 l RHOB ras homologgene family, member B 2014.68 s at 2.092926S l NQO1 NAD(P)H dehydrogenase, quinone 1 211725 s at 2.0922117 l BID BH3 interacting domain death agonist 210665 a 2.091 109 l TFPI tissue factor pathway inhibitor (lipoprotein-associated coagulation inhibitor) 243797 a 2.089624 l STK17B serine/threonine kinase 17b 20445.6 s at 2.0887516 l GAS1 growth arrest-specific 1 243631 a 2.0873.725 l LOC727815 hypothetical LOC727815 204396 s at 2.0871058 l GRKS G protein-coupled receptor kinase 5 213994 S at 2.0860326 l SPON1 spondin 1, extracellular matrix protein 213790 a 2.0849094 l ADAM12 ADAM metallopeptidase domain 12 244752 a 2.083O863 l ZNF438 Zinc finger protein 438 212554 a 2.0824976 l CAP2 CAP, adenylate cyclase-associated protein, 2 (yeast) 239108 a 2.08.1294 l FAR2 Fatty acyl CoA reductase 2, mRNA (cDNA clone MGC: 22328 MAGE: 4732586) 219284 a 2.0798855 l HSPBAP1 HSPB (heat shock 27 kDa) associated protein 1 221586 s at 2.0788653 l E2F5 E2F transcription factor 5, p130 binding 218665 a 2.0779874 l FZD4 rizzled homolog 4 (Drosophila) 240395 a 2.077514 l LOC100128727 hypothetical LOC100128727 224,583 a 2.0774055 l COTL1 coactosin-like 1 (Dictyostelium) 203426 s at 2.07494.54 l GFBP5 insulin-like growth factor binding protein 5 238.893 a 2.0748.096 l LOC338.758 hypothetical protein LOC338758 212298 a 2.072682 l NRP1 neuropilin 1 210145 a. 2.071591 l PLA2G4A phospholipase A2, group IVA (cytosolic, calcium-dependent) 2294.04 a 2.0715733 l TWIST2 twist homolog 2 (Drosophila) 205659 a 2.071.4095 l HDAC9 histone deacetylase 9 213173 a 2.071243 l PCNX pecanex homolog (Drosophila) 229287 a 2.0710356 l PCNX pecanex homolog (Drosophila) 227027 a 2.0672464 l GFPT1 glutamine-fructose-6-phosphate transaminase 1 205290 s. at 2.064931 l BMP2 bone morphogenetic protein 2 227658 s at 2.0645.716 l PLEKHA3 pleckstrin homology domain containing, family A (phosphoinositide binding specific) member 3 209324 S at 2.06432S3 l RGS16 regulator of G-protein signaling 16 209699 X at 2.0639524 l AKR1C2 aldo-keto reductase family 1, member C2 (dihydrodiol dehydrogenase 2; bile acid binding protein; 3-alpha hydroxysteroid dehydrogenase, type III) 22105.9 s at 2.063764 l COTL1 coactosin-like 1 (Dictyostelium) 219374 s at 2.06233SS l ALG9 asparagine-linked glycosylation 9, alpha-1,2-mannosyltransferase homolog (S. cerevisiae) 229942 at 2.0615158 l BNC2 basonuclin 2 212120 at 2.0612128 l RHOQ ras homologgene family, member Q 242281 at 2.060927 l GLUL glutamate-ammonia ligase (glutamine synthetase) 202214 S at 2.05838.68 l CUL4B cullin 4B 201150 s at 2.0579739 l TIMP3 TIMP metallopeptidase inhibitor 3 203927 at 2.0S68886 l NFKBIE nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, epsilon 220980 s at 2.0566983 l ADPGK ADP-dependent glucokinase 212454 x at 2.OS3964 l HNRPDL HiRNPJKTBP 235391 at 2.OS37112 l FAM92A1 family with sequence similarity 92, member A1 202784 S at 2.OS3589 l NNT nicotinamide nucleotide transhydrogenase US 2012/0034613 A1 Feb. 9, 2012 24

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 203810 at 2.0534089 l DNAB4 DnaJ (Hsp40) homolog, subfamily B, member 4 222731 at 2.0529802 l ZDHHC2 Zinc finger, DHHC-type containing 2 219682 s at 2.OS1939 l TBX3 T-box 3 200755 s at 2.OS17964 l CALU calumenin 210198 s at 2.OS1592 l PLP1 proteolipid protein 1 203294 is at 2.0499084 l LMAN1 lectin, mannose-binding, 1 210243 s at 2.0492911 l B4GALT3 UDP-Gal:betaGlcNAc beta 14 galactosyltransferase, polypeptide 3 201772 a 2.0489492 l AZIN1 antizyme inhibitor 1 218513 a 2.046744 l C4orfA3 chromosome 4 open reading frame 43 212110 a 2.0458658 l SLC39A14 solute carrier family 39 (zinc transporter), member 14 244246 a 2.04551.89 l MIPOL1 mirror-image polydactyly 1 235359 a 2.0454898 l LRRC33 eucine rich repeat containing 33 211952 a. 2.0442.946 l IPO5 importin 5 2392.61 s at 2.0437918 l CORIN corin, serine peptidase 40560 at 2.0437784 l TBX2 T-box2 231824 a 2.0437.446 l LARP2 La ribonucleoprotein domain family, member 2 238417 a 2.0436635 l PGM2L1 phosphoglucomutase 2-like 1 235061 a 2.0422926 l PPM1 K protein phosphatase 1 K (PP2C domain containing) 214077 x at 2.0414855 l MEIS3P1 Meis homeobox 3 pseudogene 1 212977 a 2.039824 l CXCR7 chemokine (C X-C motif) receptor 7 218178 s at 2.0365124 l CHMP1B chromatin modifying protein 1B 202581 a 2.0352576 l HSPA1A heat shock 70 kDa protein 1A i? heat HSPA1B shock 70 kDa protein 1B 239781 a 2.034794 l hCG 1815504 CG1815504 204845 S. at 2.03438.78 l ENPEP glutamyl aminopeptidase (aminopeptidase A) 221207 s at 2.033731 l NBEA neurobeachin 242814 a 2.033107 l SERPINB9 Serpin peptidase inhibitor, clade B (ovalbumin), member 9 225793 a 2.032O432 l LIX1L, Lix1 homolog (mouse)-like 211467 s at 2.0315952 l NFIB nuclear factor IB 205499 a 2.0314283 l SRPX2 Sushi-repeat-containing protein, X linked 2 202906 s at 2.03.13315 l NBN nibrin 222343 a 2.0312388 l BCL2L11 BCL2-like 11 (apoptosis facilitator) 204719 a 2O3O84 l ABCA8 ATP-binding cassette, Sub-family A (ABC1), member 8 244128 x at 2.03.01714 l GLIS1 GLIS family Zinc finger 1 202014 a 2.03OO674 l PPP1R15A protein phosphatase 1, regulatory (inhibitor) subunit 15A 210001 s at 2.028.387 l SOCS1 Suppressor of cytokine signaling 1 203184 a 2.0282688 l FBN2 fibrillin 2 203835 a. 2.0257106 l LRRC32 leucine rich repeat containing 32 213435 a. 2.0254.192 l SATB2 SATB homeobox2 203001 s at 2.O2S328 l STMN2 stathmin-like 2 216594 x at 2.0242531 l AKR1C1 aldo-keto reductase family 1, member C1 (dihydrodiol dehydrogenase 1; 20 alpha (3-alpha)-hydroxysteroid dehydrogenase) 221062 at 2.0242138 l HS3ST3B1 heparan sulfate (glucosamine) 3-O- Sulfotransferase 3B1 221685 s at 2.0240426 l CCDC99 coiled-coil domain containing 99 210007 s at 2.023.8538 l GPD2 glycerol-3-phosphate dehydrogenase 2 (mitochondrial) 209676 at 2.0226,731 l TFPI tissue factor pathway inhibitor (lipoprotein-associated coagulation inhibitor) 219921 s at 2.02.20945 l DOCKS dedicator of cytokinesis 5 212558 at 2.0209737 l SPRY1 sprouty homolog 1, antagonist of FGF signaling (Drosophila) 228728 at 2O184631 l C7orf58 chromosome 7 open reading frame 58 1557258 a. at 2.O1812S l BCL10 CDNA FLJ25924 fis, clone CBR05109 213302 at 2.017492 l PFAS phosphoribosylformylglycinamidine synthase 209289 at 2.01588.18 l NFIB nuclear factor IB 230559 X at 20148904 l FGD4 FYVE, RhoGEF and PH domain containing 4 220093 at 20146642 l ANTXR1 anthrax toxin receptor 1 US 2012/0034613 A1 Feb. 9, 2012 25

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 214587 a 2014571.4 l COL8A1 collagen, type VIII, alpha 1 213359 a 2.013772 l HNRNPD HnRNP-C like protein 200648 s at 2.01.20223 l GLUL glutamate-ammonia ligase (glutamine synthetase) 241902 a 2.0113673 l MKX mohawk homeobox 204114 a 2.0113 l NID2 nidogen 2 (Osteonidogen) 225385 S at 20110688 l HNRPLL heterogeneous nuclear ribonucleoprotein L-like 227983 a 2.0109632 l RILPL2 Rab interacting lysosomal protein-like 2 203194 s at 2O106804 l NUP98 nucleoporin 98 kDa 1567107 s at 2009 OO67 l TPM4 tropomyosin 4 201941 a 2.0O86033 l CPD carboxypeptidase D 202710 a 2.OO79546 l BET1 blocked early in transport 1 homolog (S. cerevisiae) 202908 a 2.OO73936 l WFS1 Wolfram syndrome 1 (wolframin) 202302 s at 2007O374 l RSRC2 arginine serine-rich coiled-coil 2 210839 s at 2.0057895 l ENPP2 ectonucleotide pyrophosphatase/phosphodiesterase 2 216235 s at 2.OOS2087 l EDNRA endothelin receptor type A 218995 s at 20046716 l EDN1 endothelin 1 238.049 at 2.0043075 l GRAMD3 GRAM domain containing 3 1554334 a at 2.OO3O234 l DNAJA4 DnaJ (Hsp40) homolog, Subfamily A, member 4 202804 at 2.OO30096 l ABCC1 ATP-binding cassette, Sub-family C (CFTR/MRP), member 1 219872 at 2.0O28145 l C4orf18 chromosome 4 open reading frame 18 237056 at 2.OO2S263 l NSC inscuteable homolog (Drosophila) 209290 s. at 2.OO2O652 l NFIB nuclear factor IB 203404 at 2.OO2O292 l ARMCX2 armadillo repeat containing, X-linked 2 200799 at 2.OO19038 l HSPA1A heat shock 70 kDa protein 1A heat HSPA1B shock 70 kDa protein 1B 226025 at 2.OO13952 l ANKRD28 ankyrin repeat domain 28 207710 at 2OOSO969 OW LCE2B ate cornified envelope 2B 221470 s at 100.546O4 OW L1F7 interleukin 1 family, member 7 (Zeta) 211548 s at 92.36748 OW HPGD hydroxyprostaglandin dehydrogenase 5-(NAD) 1553.081 at 88.25469 OW WFDC12 WAP four-disulfide core domain 12 203914 x at 78.771.54 OW HPGD hydroxyprostaglandin dehydrogenase 5-(NAD) 231930 a. 71.26495 OW ELMOD1 ELMOCED-12 domain containing 1 1560531 at 71.235 OW LCE1B ate cornified envelope 1B 203913 s at 67.8O365 OW HPGD hydroxyprostaglandin dehydrogenase 5-(NAD) 206643 a SS867226 OW HAL histidine ammonia-lyase 211549 s at 53.149216 OW HPGD hydroxyprostaglandin dehydrogenase 5-(NAD) 240420 a 52.46811 OW AADACL2 arylacetamide deacetylase-like 2 209309 a 51.06968 OW AZGP1 alpha-2-glycoprotein 1, zinc-binding 1569410 at 45.9146S OW FLG2 filaggrin family member 2 239787 a 41.795918 OW KCTD4 potassium channel tetramerisation domain containing 4 216935 a. 40.454136 OW C1orf246 chromosome 1 open reading frame 46 1553602 at 33.973.278 OW MUCL1 mucin-like 1 207 720 a 32.353355 OW LOR oricrin 220625 s at 31.788977 OW ELF5 E74-like factor 5 (ets domain transcription factor) 223720 a 25.761408 OW SPINK7 serine peptidase inhibitor, Kazal type 7 (putative) 241412 a 25479097 OW BTC betacellulin 224,555 x at 21.945213 OW L1F7 interleukin 1 family, member 7 (Zeta) 235.514 a 21.287827 OW ASPRV1 aspartic peptidase, retroviral-like 1 237974 a 21.200142 OW ABHD12B abhydrolase domain containing 12B 1552532 a. at 20.6454.66 OW ATP6V1C2 ATPase, H+ transporting, lysosomal 42 kDa, V1 subunit C2 226188 a 20.39768 OW HSPC159 galectin-related protein 202688 a 20.340603 OW TNFSF10 tumor necrosis factor (ligand) Superfamily, member 10 202687 s at 20.3O895 OW TNFSF10 tumor necrosis factor (ligand) Superfamily, member 10 205159 a 20.243301 OW CSF2RB colony stimulating factor 2 receptor, beta, low-affinity (granulocyte macrophage) US 2012/0034613 A1 Feb. 9, 2012 26

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 1553534 at 2O.OOO898 OW NLRP10 NLR family, pyrin domain containing 10 211712 s at 9.846176 OW ANXA9 annexin A9 228766 at 9.823524 OW CD36 CD36 molecule (thrombospondin receptor) 219476 at 9.656.67 OW C1orf116 chromosome 1 open reading frame 116 206295 at 9.309084 OW IL18 interleukin 18 (interferon-gamma inducing factor) 220635 at 9.078548 OW PSORS1C2 psoriasis Susceptibility 1 candidate 2 223816 at 8.854.237 OW SLC46A2 solute carrier family 46, member 2 240512 X at 8.449383 OW KCTD4 potassium channel tetramerisation domain containing 4 1552544 at 8.410263 OW SERPINA12 Serpin peptidase inhibitor, clade A (alpha-1 antiproteinase, antitrypsin), member 12 207908 at 8.13667 OW KRT2 keratin 2 202454 s at 7.877632 OW ERBB3 v-erb-b2 erythroblastic leukemia viral oncogene homolog 3 (avian) 233488 at 7.474O66 OW RNASE7 ribonuclease, RNase A family, 7 2256.67 s at 7.434992 OW FAM84A amily with sequence similarity 84, member A 234331 s at 7.273813 OW FAM84A amily with sequence similarity 84, member A 22.7717 at 7.084187 OW FLJ41603 FLJ41603 protein 203608 at 7.O13868 OW ALDHSA1 aldehyde dehydrogenase 5 family, member A1 227241 at 6.848,194 OW MUC15 mucin 15, cell Surface associated 209604 S at 6.352.425 OW GATA3 GATA binding protein 3 204733 at 6.282O7 OW KLK6 kallikrein-related peptidase 6 1564307 a at 5.90927 OW A2ML.1 alpha-2-macroglobulin-like 1 2096O2 s at 5.703371 OW GATA3 GATA binding protein 3 1554195 a. at 5.667729 OW C5orf16 chromosome 5 open reading frame 46 222484 S at S.34712 OW CXCL14 chemokine (C X—C motif) ligand 14 1554252 a. at S.34O231 OW LASS3 LAG1 homolog, ceramide synthase 3 203798. S. at 5.19176 OW VSNL1 visinin-like 1 232170 a 5.098.177 OW S100A7A S100 calcium binding protein A7A 209493 a SO42625 OW PDZD2 PDZ domain containing 2 219795 a S.O10469 OW SLC6A14 Solute carrier family 6 (amino acid transporter), member 14 213135 a. 4.684513 OW TIAM1 T-cell lymphoma invasion and metastasis 1 237120 a 4.594.361 OW KRTTT keratin 77 226789 a 4S18254 OW LOC6471-21 embigin homolog (mouse) pseudogene 1555773 at 3.957752 OW BPIL2 bactericidal permeability-increasing protein-like 2 205439 a 3.7O6697 OW GSTT2 glutathione S-transferase theta 2 219695 a 3.657985 OW SMPD3 sphingomyelin phosphodiesterase 3, neutral membrane (neutral sphingomyelinase II) 213933 a 3.S83514 OW PTGER3 prostaglandin E receptor 3 (Subtype EP3) 214329 X at 3.483.85 OW TNFSF10 tumor necrosis factor (ligand) Superfamily, member 10 218454 a 3.372534 OW FLJ22662 hypothetical protein FLJ22662 213780 a 3.321 OW TCHH trichohyalin 231867 a 3.271721 OW ODZ2 odz, Odd Oziten-m homolog 2 (Drosophila) 227238 a 3.252493 OW MUC15 mucin 15, cell Surface associated 243386 a 3.0263.48 OW CASZ1 castor Zinc finger 1 207324 S at 2.947083 OW DSC1 desmocollin 1 232165 a. 2.4O15O2 OW EPPK1 epiplakin 1 204393 s at 2.384473 OW ACPP acid phosphatase, prostate 1552797 s at 2.1677S4 OW PROM2 prominin 2 219681 s at 2.068543 OW RAB11FIP1 RAB11 family interacting protein 1 (class I) 219115 s at 2.066238 OW IL2ORA interleukin 20 receptor, alpha 203797 at 2.020528 OW VSNL1 visinin-like 1 1569886 a. at 2.018516 OW GLB1L3 galactosidase, beta 1-like 3 212531 at 20141.98 OW LCN2 lipocalin 2 200965 s at 187853 OW ABLIM1 actin binding LIM protein 1 206177 s at 1.859509 OW ARG1 arginase, liver 227449 at 1.78.4953 OW EPHA4 EPH receptor A4 US 2012/0034613 A1 Feb. 9, 2012 27

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 205108 s at 751713 OW APOB apolipoprotein B (including Ag(x) antigen) 210096 at 683115 OW CYP4B1 cytochrome P450, family 4, Subfamily B, polypeptide 1 217014 S at 667914 OW AZGP1 alpha-2-glycoprotein 1, zinc-binding AZGP1P1 alpha-2-glycoprotein 1, zinc-binding pseudogene 1 1564333 a at 665463 OW PSAPL1 prosaposin-like 1 222891 s at 499.077 OW BCL11A B-cell CLL/lymphoma 11A (zinc finger protein) 224262 a 446743 OW IL1E10 interleukin 1 family, member 10 (theta) 238778 a 35283 OW MPP7 membrane protein, palmitoylated 7 (MAGUK p55 subfamily member 7) 219895 a 33746 OW FAM7OA family with sequence similarity 70, member A 210020 x at 268426 OW CALML3 calmodulin-like 3 210085 s at 268161. OW ANXA9 annexin A9 207367 a 168736 OW ATP12A ATPase, H+/K+ transporting, nongastric, alpha polypeptide 207008 a 107841 OW IL8RB interleukin 8 receptor, beta 206193 s at 105947 OW CDSN corneodesmosin 218963 s at O944303 OW KRT23 keratin 23 (histone deacetylase inducible) 206192 a. O.920915 OW CDSN corneodesmosin 207254 a O.88.806 OW SLC15A1 Solute carrier family 15 (oligopeptide transporter), member 1 220724 a O.7S6084 OW FL21511 hypothetical protein FLJ21511 219858 s at O.733388 OW MFSD6 major facilitator Superfamily domain containing 6 236119 s at 0.709725 OW SPRR2G Small proline-rich protein 2G 232164 s at 0.52053 OW EPPK1 epiplakin 1 207381 a O421.057 OW ALOX12B arachidonate 12-lipoxygenase, 12R type 229764 a O4O72S4 OW TPRG1 tumor protein p53 regulated 1 218002 s at O403229 OW CXCL14 chemokine (C X—C motif) ligand 14 227752 a. O.369038 OW SPTLC3 serine palmitoyltransferase, long chain base subunit 3 206385 S at 0.252723 OW ANK3 ankyrin 3, node of Ranvier (ankyrin G) 236471 a O.205697 OW NFE2L3 nuclear factor (erythroid-derived 2)- ike 3 205513 a O.141289 OW TCN1 transcobalamin I (vitamin B12 binding protein, R binder family) 219313 a O.O72841 OW GRAMD1C GRAM domain containing 1C 228575 a. O.O42443 OW IL2ORB interleukin 20 receptor beta 227177 a 9.99791OS OW CORO2A coronin, actin binding protein, 2A 220090 a 9.935171 OW CRNN cornulin 1556793 a at 9.737699 OW FAM83C amily with sequence similarity 83, member C 222242 S at 9.652O21 OW KLKS kallikrein-related peptidase 5 242204 a 94978.31 OW WFDC5 WAP four-disulfide core domain 5 228523 a 9.446722 OW NANOS1 nanos homolog 1 (Drosophila) 2261.85 a 9.266244 OW CDS1 CDP-diacylglycerol synthase (phosphatidate cytidylyltransferase) 1 237690 a 9.185.177 OW GPR115 G protein-coupled receptor 115 214071 a 9.122895 OW MPPE1 MRNA, cDNA DKFZp686K2379 (from clone DKFZp686K2379) 225540 a 9.112212 OW MAP2 microtubule-associated protein 2 227209 a 9.101.238 OW CNTN1 Contactin 2 precursor (CNTN1) 220624 S at 9.078715 OW ELF5 E74-like factor 5 (ets domain transcription factor) 243871 a 9.058688 OW LOC100.130476 PREDICTED: Homo sapiens similar to hCG2036711 (LOC100.130476), mRNA 242998 a 8.8784485 OW RDH12 retinol dehydrogenase 12 (all-transf9 cis 11-cis) 219995 s at 8.8615S OW ZNF750 Zinc finger protein 750 225846 a 8.854407 OW RBM3SA RNA binding motif protein 35A 227747 a 8.839586 OW MPZL3 myelin protein zero-like 3 213056 a 8.79SO81 OW FRMD4B FERM domain containing 4B 204702 s at 8.784606 OW NFE2L3 nuclear factor (erythroid-derived 2)- like 3 238017 a 8.7703905 OW SDR16CS short chain dehydrogenase/reductase family 16C, member 5 US 2012/0034613 A1 Feb. 9, 2012 28

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 220414 a 8.748902 OW CALMLS calmodulin-like 5 225792 a 8.72326 OW HOOK1 hook homolog 1 (Drosophila) 209442 x at 8.710819 OW ANK3 ankyrin 3, node of Ranvier (ankyrin G) 204469 a 8.7OO343 OW PTPRZ1 protein tyrosine phosphatase, receptor type, Z polypeptide 1 226213 a 8.644O65 OW ERBB3 v-erb-b2 erythroblastic leukemia viral oncogene homolog 3 (avian) 205220 a 8.635616 OW GPR109B G protein-coupled receptor 109B 21936.9 s at 8.634883 OW OTUB2 OTU domain, ubiquitin aldehyde binding 2 206595 a 8.616346 OW CST6 cystatin E/M 1556361 s at 8.6023O3S OW ANKRD13C ankyrin repeat domain 13C 1559226 X at 8.588SS1 OW LCE1E late cornified envelope 1E 243582 a 8.550055 OW SH3RF2 SH3 domain containing ring finger 2 221666 s at 8.532O53 OW PYCARD PYD and CARD domain containing 204455 a. 8.512845 OW DST dystonin 201348 a 8. SO4907 OW GPX3 glutathione peroxidase 3 (plasma) 212538 a 8.472947 OW DOCK9 dedicator of cytokinesis 9 211361 s at 8.422554 OW SERPINB13 Serpin peptidase inhibitor, clade B (ovalbumin), member 13 205969 a 8.273929 OW AADAC arylacetamide deacetylase (esterase) 220013 a 8.225659 OW ABHD9 abhydrolase domain containing 9 219756 S at 8.1738O3 OW POF1B premature ovarian failure, 1B 23O349 a 8.170966 OW XKRX XK, Kell group complex subunit-related, X-linked 238654 a 8.165287 OW LOC147645 hypothetical protein LOC147645 220723 s at 8.1235 OW FL21511 hypothetical protein FLJ21511 225325 a. 8.10OSO3 OW MFSD6 major facilitator Superfamily domain containing 6 236514 a 8.0631.93 OW ACOT8 HIV-Nefassociated acyl CoA hioesterase (hNAACTE) 224650 a 7.978321 OW MAL2 mal, T-cell differentiation protein 2 205709 S at 7.964628 OW CDS1 CDP-diacylglycerol synthase (phosphatidate cytidylyltransferase) 1 215704 a 7.9460135 OW FLG PLAC2 placenta-specific 2 (non protein coding) 223544 a 7.94.1029 OW TMEM19 transmembrane protein 79 243722 a. 7.872192 OW PYDC1 PYD (pyrin domain) containing 1 206642 a. 7.8642S35 OW DSG1 desmoglein 1 204952 a 7.828.9723 OW LYPD3 LY6/PLAUR domain containing 3 217087 a 7.8114066 OW C1orf68 chromosome 1 open reading frame 68 222549 a 7.7580996 OW CLDN1 claudin 1 229385 s at 7.6575327 OW PLAC2 placenta-specific 2 (non-protein coding) 219267 a 7.655961 OW GLTP glycolipid transfer protein 217272 s at 7.6130595 OW SERPINB13 Serpin peptidase inhibitor, clade B (ovalbumin), member 13 1555383 a at 7.5732O36 OW POF1B premature ovarian failure, 1B 238028 a 7.5382996 OW LOC100128918 hypothetical protein LOC100128918 15525.02 s at 7.5355606 OW RHBDL2 rhomboid, veinlet-like 2 (Drosophila) 205568 a 7.4951296 OW AQP9 aquaporin 9 205442 a. 7.491568 OW MFAP3L microfibrillar-associated protein 3-like 227955 S. at 7.4095197 OW EFNAS ephrin-A5 21.0833 a 7.343155 OW PTGER3 prostaglandin E receptor 3 (Subtype EP3) 212543 a 7.342191 OW AIM1 absent in melanoma 1 218186 a 7.331.6717 OW RAB2S RAB25, member RAS oncogene family 219403 s at 7.320557 OW HPSE heparanase 210074 a 7.306036 OW CTSL2 cathepsin L2 21985.0 s at 7.2735214 OW EHF ets homologous factor 228708 a 7.271517 OW RAB27B Small GTP-binding protein Rab27b 227202 a 7.2712536 OW CNTN1 Contactin 2 precursor (CNTN1) 228538 a 7.258556 OW ZNF662 Zinc finger protein 662 232158 x at 7.256812 OW NPAL1 NIPA-like domain containing 1 226803 a 7.2564917 OW CHMP4C chromatin modifying protein 4C 20648.8 s at 7.236786 OW CD36 CD36 molecule (thrombospondin receptor) 218035 s at 7.19059 OW RBM47 RNA binding motif protein 47 205185 a. 7.1835594 OW SPINKS serine peptidase inhibitor, Kazal type 5 206115 a. 7.160374 OW EGR3 early growth response 3 221854 a 7.152312 OW PKP1 plakophilin 1 (ectodermal dysplasiaskin fragility syndrome) US 2012/0034613 A1 Feb. 9, 2012 29

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 23977O a 7.1368814 OW FAM62C family with sequence similarity 62 (C2 domain containing), member C 214091 s at 7.1271 777 OW GPX3 glutathione peroxidase 3 (plasma) 218764 a 7.12O812 OW PRKCH protein kinase C, eta 214536 a 7.1027665 OW SLURP1 secreted LY6 PLAUR domain containing 1 222496 s at 7.0O82264 OW RBM47 RNA binding motif protein 47 232056 a 7.0050O87 OW SCEL Sciellin 217496 s at 6.9SOO76 OW DE insulin-degrading enzyme 215465 a 6.94.4839S OW ABCA12 ATP-binding cassette, Sub-family A (ABC1), member 12 22907O a 6.942199 OW C6orf1 OS chromosome 6 open reading frame 105 208937 S at 6.8969717 OW D1 inhibitor of DNA binding 1, dominant negative helix-loop-helix protein 219630 a. 6.89.24527 OW PDZK1P1 PDZK1 interacting protein 1 1553454 at 6.8833647 OW RPTN repetin 1553589 a. at 6.8793.49 OW PDZK1P1 PDZK1 interacting protein 1 219388 a 6.834852 OW GRHL2 grainyhead-like 2 (Drosophila) 207326 a 6.8310184 OW BTC betacellulin 228038 a 6.828.2814 OW SOX2 SRY (sex determining region Y)-box 2 36499 at 6.82723 OW CELSR2 cadherin, EGF LAG seven-pass G-type receptor 2 (flamingo homolog, Drosophila) 231148 a 6.815531 OW IGFL2 IGF-like family member 2 213611 a 6.770,561 OW AQP5 aquaporin 5 231849 a 6.6379747 OW KRT8O keratin 80 226535 a. 6.6363 OW ITGB6 integrin, beta 6 204942 s at 6.6221.83 OW ALDH3B2 aldehyde dehydrogenase 3 family, member B2 203178 a 6.61S657 OW GATM amidinotransferase (L- arginine:glycine amidinotransferase) 1557136 at 6.6O1717 OW ATP13A4 ATPase type 13A4 213927 a 6.5928116 OW MAP3K9 mitogen-activated protein kinase kinase kinase 9 220945 X at 6.560972 OW MANSC1 MANSC domain containing 1 206125 s at 6.5SOS314 OW KLK8 kallikrein-related peptidase 8 202712 s at 6.507372 OW CKMT1A, if creatine kinase, mitochondrial 1A fif CKMT1B creatine kinase, mitochondrial 1B LOC100.133623 similar to Creatine kinase, ubiquitous mitochondrial precursor (U-MtCK) (Mia-CK) (Acidic-type mitochondrial creatine kinase) 1561225 at 6.498.2934 OW LOC338.579 hypothetical protein LOC338579 224189 x at 6.491583 OW EHF ets homologous factor 211788 s at 6.48531.63 OW TREX2 three prime repair exonuclease 2 203180 at 6.4791 126 OW ALDH1A3 aldehyde dehydrogenase 1 family, member A3 1555890 at 6.4574604 OW OR2A2OP olfactory receptor, family 2, Subfamily OR2A9P A, member 20 pseudogene if olfactory receptor, family 2, Subfamily A, member 9 pseudogene 226482 s at 6.4535813 OW hCG 20857 hiosulfate sulfurtransferase KAT, J. RP11- putative KAT protein 544M22.4 208156 x at 6.4456OS3 OW EPPK1 epiplakin 1 228948 a 6.4166746 OW EPHA4 EPH receptor A4 223658 a 6.4O2442 OW KCNK6 potassium channel, Subfamily K, member 6 230179 a 6.3794336 OW LOC285812 hypothetical protein LOC285812 1554253 a at 6.3690333 OW LASS3 LAG1 homolog, ceramide synthase 3 240304 S at 6.356037 OW TMC5 transmembrane channel-like 5 207109 a 6.272226 OW POU2F3 POU class 2 homeobox 3 206114 a 6.2673.96S OW EPHA4 EPH receptor A4 219232 s at 6.265491 OW EGLN3 egl nine homolog 3 (C. elegans) 41469 at 6.2596.33 OW PI3 peptidase inhibitor 3, skin-derived 238710 a 6.2384377 OW TMEM86A transmembrane protein 86A 202193 a 6.232698 OW LIMK2 LIM domain kinase 2 220664 a 6.2243,595 OW SPRR2C Small proline-rich protein 2C (pseudogene) 203021 a. 6.1999.464 OW SLPI Secretory leukocyte peptidase inhibitor 210461 s at 6.193O823 OW ABLIM1 actin binding LIM protein 1 210015 s at 6.1631.96 OW MAP2 microtubule-associated protein 2 US 2012/0034613 A1 Feb. 9, 2012 30

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 206392 s at 6.1626806 down RARRES1 retinoic acid receptor responder (tazaroteine induced) 1 212570 at 6.1608458 down ENDOD1 endonuclease domain containing 1 214070 s at 6.1544147 down ATP1 OB ATPase, class V, type 10B 226863 at 6.136124 down FAM110C amily with sequence similarity 110, member C 207192 at 6.115O15S down DNASE1L2 deoxyribonuclease I-like 2 215125 s at 6.1119647 down UGT1A1 UDPglucuronosyltransferase 1 family, UGT1A10 polypeptide A1 / UDP UGT1A3 glucuronosyltransferase 1 family, UGT1 A4 polypeptide A10 / UDP UGT1AS glucuronosyltransferase 1 family, UGT1A6 polypeptide A3 // UDP UGT1A7 glucuronosyltransferase 1 family, UGT1A8 polypeptide A4 i? UDP UGT1A9 glucuronosyltransferase 1 family, polypeptide A5 // UDP glucuronosyltransferase 1 family, polypeptide A6/// UDP glucuronosyltransferase 1 family, polypeptide A7 /?/ UDP glucuronosyltransferase 1 family, polypeptide A8 // UDP glucuronosyltransferase 1 family, polypeptide A9 1554921 a. at 6.0999765 OW SCEL Sciellin 225645 at 6.08.21557 OW EHF Ets homologous factor, mRNA (cDNA clone MGC:47678 IMAGE: 6055934) 219936 s at 6.08.06403 OW GPR87 G protein-coupled receptor 87 219532 at 6.073317 OW ELOVL4 elongation of very long chain fatty acids (FEN1/Elo2, SUR4/Elo3, yeast)- ike 4 216258 s at 6.0713954 OW SERPINB13 Serpin peptidase inhibitor, clade B (ovalbumin), member 13 203328 x at 6.0695624 OW IDE insulin-degrading enzyme 23.0323 s at 6.0668OO6 OW TMEM4SB transmembrane protein 45B 227180 a 6.0664496 OW ELOVL7 ELOVL family member 7, elongation of long chain fatty acids (yeast) 219497 s at 6.0622134 OW BCL11A B-cell CLL/lymphoma 11A (zinc finger protein) 212560 a 6.057562 OW SORL sortilin-related receptor, L(DLR class) A repeats-containing 216074 X at 6.0346746 OW WWC WW and C2 domain containing 1 21.0117 a 6.022336 OW SPAG1 sperm associated antigen 1 221872 a. 5.9994.154 OW RARRES1 retinoic acid receptor responder (tazaroteine induced) 1 238.096 a 5.9605427 OW LOC284O23 hypothetical protein LOC284023 209603 a S.944.856 OW GATA3 GATA binding protein 3 215.554 a S.90897.04 OW GPLD glycosylphosphatidylinositol specific phospholipase D1 203691 a 5.895547 OW PI3 peptidase inhibitor 3, skin-derived 21O130 s at 5.8894.96 OW TM7SF2 transmembrane 7 Superfamily member 2 1553989 a. at 5.889157 OW ATP6V1C2 ATPase, H+ transporting, lysosomal 42 kDa, V1 subunit C2 235230 a. S. 864316 OW PLCXD2 phosphatidylinositol-specific phospholipase C, X domain containing 2 1554897 s at S.809435 OW RHBDL2 rhomboid, veinlet-like 2 (Drosophila) 236172 a 5.80707 OW LTB4R eukotriene B4 receptor 228865 a S.8040857 OW C1orf116 chromosome 1 open reading frame 116 223659 a S.8040533 OW TMPRSS13 transmembrane protease, serine 13 205778 a S.80O858 OW KLK7 kallikrein-related peptidase 7 219529 a 5.7376733 OW CLIC3 chloride intracellular channel 3 216615 s at S. 662SO1 OW HTR3A 5-hydroxytryptamine (serotonin) receptor 3A 238567 a 5.6408434 OW SGPP2 sphingosine-1-phosphate phosphotase 2 206165 s at 5.62O2765 OW CLCA2 chloride channel regulator 2 206008 a S. 6143503 OW TGM1 transglutaminase 1 (Kpolypeptide epidermal type I, protein-glutamine gamma-glutamyltransferase) 211906 s at S. 612926 OW SERPINB4 Serpin peptidase inhibitor, clade B (ovalbumin), member 4 US 2012/0034613 A1 Feb. 9, 2012 31

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 235.955 at S. 6074624 OW MARVELD2 MARVEL domain containing 2 206214 at S.S946O4 OW PLA2G7 phospholipase A2, group VII (platelet activating factor acetylhydrolase, plasma) 202286 s at S.S906916 OW TACSTD2 tumor-associated calcium signal transducer 2 206164 a S.S694.885 OW CLCA2 chloride channel regulator 2 239381 a S.S62S143 OW KLK7 kallikrein-related peptidase 7 227014 a 5.5577726 OW ASPHD2 aspartate beta-hydroxylase domain containing 2 208539 X at 5.553559 OW SPRR2B Small proline-rich protein 2B 222847 s at 5.53052 OW EGLN3 egl nine homolog 3 (C. elegans) 227450 a 5.5204797 OW ERP27 endoplasmic reticulum protein 27 kDa 225615 a. 5.4997 22 OW FFO2 intermediate filament family orphan 2 219461 a S.4821987 OW PAK6 p21 protein (Cdc42, Rac)-activated kinase 6 206683 a S.4811287 OW ZNF165 Zinc finger protein 165 1555.310 a. at S.4699.154 OW PAK6 p21 protein (Cdc42, Rac)-activated kinase 6 242828 a 5.4434 OW FIGN idgetin 224329 s at 5.4422174 OW CNF cornifelin 219998 a 5.4138575 OW HSPC159 galectin-related protein 33767 at S.4136043 OW NEF neurofilament, heavy polypeptide 205363 a S.4128237 OW BBOX1 butyrobetaine (gamma), 2-oxoglutarate dioxygenase (gamma-butyrobetaine hydroxylase) 1 211597 s at S.409704 OW HOPX HOP homeobox 203962 s at 5.383551 OW NEBL nebullette 206032 a 5.376.959 OW DSC3 desmocollin 3 227570 a 5.374O797 OW TMEM86A transmembrane protein 86A 226926 a 5.3653264 OW DMKN dermokine 202191 s at 5.363796 OW GAS7 growth arrest-specific 7 206482 a 5.34657 OW PTK6 PTK6 protein tyrosine kinase 6 223611 s at S.3417473 OW LNX1 igand of numb-protein X1 204379 s at S.287966 OW FGFR3 fibroblast growth factor receptor 3 212992 a 5.2794547 OW AHNAK2 AHNAK nucleoprotein 2 217528 a S.258966 OW CLCA2 chloride channel regulator 2 2365.34 a 5.2476907 OW BNIPL BCL2 adenovirus E1B 19 kD interacting protein like 201131 s at 5.239927 OW CDH1 cadherin 1, type 1, E-cadherin (epithelial) 235099 a S.228119 OW CMTM8 CKLF-like MARVEL transmembrane domain containing 8 226755 a. S.2196O16 OW LOC642587 CDNA FLJ33794 fis, clone CTONG1OOOOO9 235146 a S.2194705 OW TMCC3 transmembrane and coiled-coil domain family 3 1554593 s at 5.2057705 OW SLC1A6 solute carrier family 1 (high affinity aspartate? glutamate transporter), member 6 205832 at 5.1992.483 OW CPA4 carboxypeptidase A4 213085 s at 5.183778 OW WWC1 WW and C2 domain containing 1 229518 at 5.1793.13 OW FAM46B family with sequence similarity 46, member B 226177 at S.172866 OW GLTP glycolipid transfer protein 200862 at 5.17O659 OW DHCR 24 24-dehydrocholesterol reductase 212242 at S.147954 OW TUBA4A tubulin, alpha 4a 24.2103 at S.142739 OW TMEM86A transmembrane protein 86A 206166 s at 5.137547 OW CLCA2 chloride channel regulator 2 210715 s at S.117244 OW SPINT2 serine peptidase inhibitor, Kunitz type, 2 1554912 at S.1171923 OW FAM62C family with sequence similarity 62 (C2 domain containing), member C 1553.077 at S.116311 OW SDR9C7 short chain dehydrogenase/reductase family 9C, member 7 206628 at S.108023 OW SLC5A1 Solute carrier family 5 (sodium glucose cotransporter), member 1 228698 at S.O9854 OW SOX7 SRY (sex determining region Y)-box 7 22O161 s at S.O8.9994 OW EPB41L4B erythrocyte membrane protein band 4.1 like 4B 202421 at 5.0736.53 OW IGSF3 immunoglobulin Superfamily, member 3 238909 at 5.072O587 OW S100A10 Calpactin I light chain, 5'UTR region 225299 at S.O482273 OW MYOSB myosin VB 235141 at 5.0444136 OW MARVELD2 MARVEL domain containing 2 US 2012/0034613 A1 Feb. 9, 2012 32

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 201236 s at S.O438776 OW BTG2 BTG family, member 2 1555382 at S.O42S2OS OW POF1B premature ovarian failure, 1B 1557094 at S.O291085 OW LOC653110 hypothetical LOC653110 201243 s at S.O24576 OW ATP1B1 ATPase, Na+/K+ transporting, beta 1 polypeptide 207114 at 5.O127378 OW LY6G6C lymphocyte antigen 6 complex, locus G6C 227676 at 4.999608 OW FAM3D family with sequence similarity 3, member D 2196.80 at 4.9981O1 OW NLRX1 NLR family member X1 202295 s at 4.97334.86 OW CTSH cathepsin H 206561 s at 4.97.185 OW AKR1B10 aldo-keto reductase family 1, member B10 (aldose reductase) 218717 s at 4.965496 OW LEPREL1 leprecan-like 1 20921.2 s at 4.963OS27 OW KLFS Kruppel-like factor 5 (intestinal) 20741.4 S at 4.9624557 OW PCSK6 proprotein convertase Subtilisinkexin type 6 242271 at 4.962O56 OW SLC26A9 solute carrier family 26, member 9 21691.8 s at 4.960929 OW DST dystonin 204855 at 4.9595795 OW SERPINB5 Serpin peptidase inhibitor, clade B (ovalbumin), member 5 1553929 at 4.9470873 OW ACER1 alkaline ceramidase 1 203961 at 4.92486 OW NEBL nebullette 205807 s at 4.9016848 OW TUFT1 tuftelin 1 203453 at 4.897337 OW SCNN1A Sodium channel, nonvoltage-gated 1 alpha 222383 s at 4.875127 OW ALOXE3 arachidonate lipoxygenase 3 1552319 a. at 4.8667984 OW KLK8 kallikrein-related peptidase 8 216733 s at 4.83671.43 OW GATM glycine amidinotransferase (L- arginine:glycine amidinotransferase) 208153 s at 4819246 OW FAT2 FAT tumor Suppressor homolog 2 (Drosophila) 1559224 at 4.81139 OW LCE1E ate cornified envelope 1E 222892 s at 4.811261 OW TMEM40 transmembrane protein 40 213992 at 4.80O3573 OW COL4A6 collagen, type IV, alpha 6 206023 at 4.788.6114 OW NMU neuromedin U 214734 at 47853S13 OW EXPHS exophilin 5 60474 at 4785.0847 OW FERMT1 ermitin family homolog 1 (Drosophila) 219498 s at 4.7693.014 OW BCL11A B-cell CLL/lymphoma 11A (zinc finger protein) 59625 at 4.763774 OW NOL3 nucleolar protein 3 (apoptosis repressor with CARD domain) 202179 at 4.752.7394 OW BLMH bleomycin hydrolase 236128 at 4.73937 OW ZNF91 Zinc finger protein 91 203642 s at 4.73S424 OW COBLL1 COBL-like 1 205011 at 4.726828 OW WWASA von Willebrand factor A domain containing 5A 203407 at 4.7218986 OW PPL periplakin 205590 at 4.7O12725 OW RASGRP1 RAS guanyl releasing protein 1 (calcium and DAG-regulated) 222603 at 4.700272 OW ERMP1 endoplasmic reticulum metallopeptidase 1 203779 S at 4.6950703 OW MPZL2 myelin protein zero-like 2 210834 S. at 4.694923 OW PTGER3 prostaglandin E receptor 3 (Subtype EP3) 223484 at 4.6944O65 OW C15Crfa3 chromosome 15 open reading frame 48 206033 s at 4.676169 OW DSC3 desmocollin 3 2323.06 at 4.6732864 OW CDH26 cadherin-like 26 215808 at 4.67278 OW KLK10 kallikrein-related peptidase 10 202411 at 4.671116 OW IFI27 interferon, alpha-inducible protein 27 207802 at 4.665757 OW CRISP3 cysteine-rich secretory protein 3 221667 s at 4.658692 OW HSPB8 heat shock 22 kDa protein 8 219087 at 4.6529336 OW ASPN asporin 218741 at 4.6508865 OW CENPM centromere protein M 203741 s at 4.6428866 OW ADCY7 adenylate cyclase 7 203074 at 4.63983 OW ANXA8 annexin A8 annexin A8-like 1 ANXA8L1 annexin A8-like 2 ANXA8L2 201286 at 4.6295.676 OW SDC1 Syndecan 1 231733 at 4.617893 OW CARD18 caspase recruitment domain family, member 18 2098.73 s at 4.6149898 OW PKP3 plakophilin 3 US 2012/0034613 A1 Feb. 9, 2012 33

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 212573 at 4.6O98948 OW ENDOD1 endonuclease domain containing 1 244780 at 4.6O221 OW SGPP2 sphingosine-1-phosphate phosphotase 2 225177 at 4.5943675 OW RAB11FIP1 RAB11 family interacting protein 1 (class I) 223322 at 4.591433 OW RASSFS Ras association (RalGDSAF-6) domain family member 5 227309 at 4.5890346 OW YOD1 YOD1 OTU deubiquinating enzyme 1 homolog (S. cerevisiae) 206515 at 4.58716S OW CYP4F3 cytochrome P450, family 4, Subfamily F, polypeptide 3 204995 at 4.58.68692 OW CDK5R1 cyclin-dependent kinase 5, regulatory subunit 1 (p35) 202826 at 4.585347 OW SPINT1 serine peptidase inhibitor, Kunitz type 1 205651 X at 4.5728 OW RAPGEF4 Rap guanine nucleotide exchange actor (GEF) 4 22274.6 s at 4.57OO636 OW BSPRY B-box and SPRY domain containing 219722 s at 4555332 OW GDPD3 glycerophosphodiester phosphodiesterase domain containing 3 205538 at 4.5494-61 OW CORO2A coronin, actin binding protein, 2A 210619 s at 4.5378,346 OW HYAL1 hyaluronoglucosaminidase 1 204990 s. at 4.5270753 OW ITGB4 integrin, beta 4 227204 at 4.515133 OW PARD6G par-6 partitioning defective 6 homolog gamma (C. elegans) 238,063 at 4.513,157 OW TMEM154 transmembrane protein 154 21935.8 s at 4.5108666 OW ADAP2 Arf6AP with dual PH domains 2 206276 at 4.498863 OW LY6D ymphocyte antigen 6 complex, locus D 209792 S at 4.4899607 OW KLK10 kallikrein-related peptidase 10 208892 s at 4.48.9426 OW DUSP6 dual specificity phosphatase 6 206265 s at 4.488534 OW GPLD1 glycosylphosphatidylinositol specific phospholipase D1 235852 a 4.484227 OW STON2 CDNA FLJ37480 fis, clone BRAWH2013866, highly similar to Homo sapiens stonin 2 mRNA 221107 a 4.484O99 OW CHRNA9 cholinergic receptor, nicotinic, alpha 9 2196.91 a 4.478.258 OW SAMD9 sterile alpha motif domain containing 9 226499 a 4.476OSS OW NRARP MRNA full length insert cDNA clone EUROIMAGE 1499812 227556 a 4.47O7904 OW NME7 non-metastatic cells 7, protein expressed in (nucleoside-diphosphate kinase) 1558846 at 4.46SS1 OW PNLIPRP3 pancreatic lipase-related protein 3 215425 a. 4.46542 OW BTG3 BTG family, member 3 226226 a 4.4539366 OW TMEM4SB transmembrane protein 45B 204204 a 4.4521403 OW SLC31A2 Solute carrier family 31 (copper transporters), member 2 208893 s at 4.4509206 OW DUSP6 dual specificity phosphatase 6 204029 a 4.4487 OW CELSR2 cadherin, EGF LAG seven-pass G-type receptor 2 (flamingo homolog, Drosophila) 201656 a 4.4418707 OW TGA6 integrin, alpha 6 206034 a 4.426019 OW SERPINB8 Serpin peptidase inhibitor, clade B (ovalbumin), member 8 206391 a 4.4257607 OW RARRES1 retinoic acid receptor responder (tazaroteine induced) 1 228531 a 4.4111724 OW SAMD9 sterile alpha motif domain containing 9 203638 s at 4.4092S6S OW FGFR2 fibroblast growth factor receptor 2 206264 a 4.40854O7 OW GPLD1 glycosylphosphatidylinositol specific phospholipase D1 201015 s at 4.4O7748 OW UP junction plakoglobin 226029 a 4.401417 OW WANGL2 vang-like 2 (van gogh, Drosophila) 202053 s at 4.3994246 OW ALDH3A2 aldehyde dehydrogenase 3 family, member A2 205675 a. 4.38894.13 OW MTTP microsomal triglyceride transfer protein 203917 a 4.387866S OW CXADR coxsackie virus and adenovirus receptor 1560250 S at 4.384614 OW LOC284242 hypothetical protein LOC284242 201242 S at 4.3839946 OW ATP1B1 ATPase, Na+/K+ transporting, beta 1 polypeptide 203126 at 4.3744597 OW IMPA2 inositol(myo)-1 (or 4)- monophosphatase 2 219412 at 4.3728795 OW RAB38 RAB38, member RAS oncogene family US 2012/0034613 A1 Feb. 9, 2012 34

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 22O124 a 4.371851 OW GAN gigaxonin 211067 s at 4.34.44204 OW GAS7 growth arrest-specific 7 220066 a 4.3412647 OW NOD2 nucleotide-binding oligomerization domain containing 2 230188 a 4.31929S4 OW ICHTHYIN ichthyin protein 2025.04 a 4.3078O84 OW TRIM29 tripartite motif-containing 29 1569144 a at 4.3075566 OW C9orf169 chromosome 9 open reading frame 169 LOC100.130547 /?/ hypothetical protein LOC100.130547 226733 a 4.2923.603 OW PFKFB2 6-phosphofructo-2-kinase/fructose-2,6- biphosphatase 2 222829 s at 4.2921.16 OW IL2ORA interleukin 20 receptor, alpha 229720 a 4.2816586 OW BAG1 BCL2-associated athanogene 219395 a 4.2743 106 OW RBM3SB RNA binding motif protein 35B 208891 a 4.263SO26 OW DUSP6 dual specificity phosphatase 6 203585 a. 4.2597046 OW ZNF185 Zinc finger protein 185 (LIM domain) 221 690 s. at 4.2S36693 OW NLRP2 NLR family, pyrin domain containing 2 231270 a 4.2472186 OW CA13 carbonic anhydrase XIII 244692 a 4.2340446 OW CYP4F22 cytochrome P450, family 4, Subfamily F. polypeptide 22 220413 a 4.2331395 OW SLC39A2 solute carrier family 39 (zinc transporter), member 2 202289 s at 4.229875 OW TACC2 transforming, acidic coiled-coil containing protein 2 211362 s at 4.22O6O2 OW SERPINB13 Serpin peptidase inhibitor, clade B (ovalbumin), member 13 203327 a 4.2173615 OW DE insulin-degrading enzyme 1555.173 at 4.212S28 OW STX19 Syntaxin 19 213924 a 4.212SOS OW MPPE1 MRNA, cDNA DKFZp686K2379 (from clone DKFZp686K2379) 204484 a 4.199939 OW PIK3C2B phosphoinositide-3-kinase, class 2, beta polypeptide 226907 a 4.199478 OW PPP1R14C protein phosphatase 1, regulatory (inhibitor) subunit 14C 205863 a 4.1991963 OW S100A12 S100 calcium binding protein A12 222881 a 4.1936674 OW HPSE heparanase 201884 a 4.1872177 OW CEACAMS carcinoembryonic antigen-related cell adhesion molecule 5 228570 a 4.181567 OW BTBD11 BTB (POZ) domain containing 11 210138 a 4.1791.59 OW RGS2O regulator of G-protein signaling 20 228O10 a 4.178918 OW PPP2R2C protein phosphatase 2 (formerly 2A), regulatory subunit B, gamma isoform 232181 a 4.1646137 OW LOC153346 hypothetical protein LOC153346 41660 at 4.1577396 OW CELSR1 cadherin, EGF LAG seven-pass G-type receptor 1 (flamingo homolog, Drosophila) 227197 a 4.152967 OW SGEF Src homology 3 domain-containing guanine nucleotide exchange factor 225822 a. 4.1515694 OW TMEM12S transmembrane protein 125 219474 a 4.142456 OW C3orf52 chromosome 3 open reading frame 52 205900 a 4.1411896 OW KRT1 keratin 1 203367 a 4.1374893 OW DUSP14 dual specificity phosphatase 14 225671 a 4.136382 OW SPNS2 spinster homolog 2 (Drosophila) 55081 at 4.132S61 OW MICALL1 MICAL-like 1 223832 s at 4.1.23751 OW CAPNS2 calpain, Small subunit 2 214490 a 4.1.185994 OW ARSF arylsulfatase F 210372 s at 4.1101165 OW TPD52L1 tumor protein D52-like 1 209863 s at 4.1084O65 OW TP63 tumor protein p63 210297 s at 4.10O8377 OW MSMB microseminoprotein, beta 224210 s at 4.098.7024 OW PXMP4 peroxisomal membrane protein 4, 24 kDa 221245 s at 4.0908217 OW FZD5 rizzled homolog 5 (Drosophila) 205977 s at 4087086 OW EPHA1 EPH receptor A1 230563 at 4.084.1923 OW RASGEF1A RasGEF domain family, member 1A 226272 at 4.082O107 OW RCAN3 RCAN family member 3 204004 at 4.077613 OW PAWR PRKC, apoptosis, WT1, regulator 2055.34 at 4O76567 OW PCDHT protocadherin 7 239272 at 4.O74428 OW MMP28 matrix metallopeptidase 28 204300 at 4.0636954 OW PET112L PET112-like (yeast) 235879 at 4.043492 OW MBNL1 MBNL protein 1554.179 s at 4.0318265 OW LYNX1 Ly6/neurotoxin 1 235272 at 4.0244374 OW SBSN Suprabasin 235085 at 4.018871 OW PRAGMIN homolog of rat pragma of Rind2 US 2012/0034613 A1 Feb. 9, 2012 35

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 213954 at 4O13676 down FAM169A family with sequence similarity 169, member A 226490 at 4O12788 down NHSL1 NHS-like 1 211002 s at 4.012248 down TRIM29 tripartite motif-containing 29 212706 at 3.99.73824 down LOC100132214 similar to HSPC047 protein / similar to RAS p21 protein activator 4 if LOC100.133005 similar to HSPC047 protein / RAS p21 protein activator 4 LOC10O134722 RASA4 225354 s at 3.99.22047 OW SH3BGRL2 SH3 domain binding rich protein like 2 205190 at 3.9871655 OW PLS1 plastin 1 (I isoform) 202054 S at 3.9822264 OW ALDH3A2 aldehyde dehydrogenase 3 family, member A2 232449 at 3.9744756 OW BCO2 beta-caroteine oxygenase 2 218342 s at 3.969063S OW ERMP1 endoplasmic reticulum metallopeptidase 1 206043 s at 3.963122 OW ATP2C2 ATPase, Ca++ transporting, type 2C, member 2 206284 X at 3.962128 OW CLTB clathrin, light chain (Lcb) 209126 X at 3.952012 OW KRT6B keratin 6B 1553213 a at 3.939.0984 OW KRT78 keratin 78 210413 x at 3.9384.09 OW SERPINB3 Serpin peptidase inhibitor, clade B SERPINB4 (ovalbumin), member 3 i? serpin peptidase inhibitor, clade B (ovalbumin), member 4 223895 s at 3.9289596 OW EPN3 epsin 3 202546 at 3.9224277 OW WAMP8 vesicle-associated membrane protein 8 (endobrevin) 206453 s at 3.92.1748 OW NDRG2 NDRG family member 2 205020 s at 3.9133503 OW ARLAA ADP-ribosylation factor-like 4A 213279 at 3.907437 OW DHRS1 dehydrogenase/reductase (SDR family) member 1 214838 at 3.9040148 OW SFT2D2 SFT2 domain containing 2 210102 at 3.8968651 OW WWASA von Willebrand factor A domain containing 5A 225687 at 3.8950646 OW FAM83D family with sequence similarity 83, member D 1553695 a. at 3.8867338 OW NLRX1 NLR family member X1 226064 s at 3.882SO26 OW DGAT2 diacylglycerol O-acyltransferase homolog 2 (mouse) 205773 at 3.876O667 OW CPEB3 cytoplasmic polyadenylation element binding protein 3 223232 s at 3.8725953 OW CGN cingulin 209570 S. at 3.870752 OW D4S234E DNA segment on chromosome 4 (unique) 234 expressed sequence 224806 at 3.8689046 OW TRIM2S tripartite motif-containing 25 1554.648 a. at 38646846 OW DUOXA1 dual oxidase maturation factor 1 213501 at 3.8544102 OW ACOX1 acyl-Coenzyme A oxidase 1, palmitoyl 220786 s at 3.844398 OW SLC38A4 solute carrier family 38, member 4 216060 S at 3.84O2324 OW DAAM1 dishevelled associated activator of morphogenesis 1 224435 at 3.8286.057 OW C10orf57 chromosome 10 open reading frame 57 C10orf58 if chromosome 10 open reading frame 58 2024.89 s at 3.826976 OW FXYD3 FXYD domain containing ion transport regulator 3 228124 at 3.8211079 OW ABHD12 abhydrolase domain containing 12 223748 at 3.8204598 OW SLC4A11 Solute carrier family 4, Sodium borate transporter, member 11 225301 s at 3.8147264 OW MYOSB myosin VB 205030 at 3.8O3794 OW FABP7 atty acid binding protein 7, brain 203997 at 3.7991052 OW PTPN3 protein tyrosine phosphatase, non receptor type 3 206409 at 3.792147 OW TIAM1 T-cell lymphoma invasion and metastasis 1 230464 at 3.7890499 OW S1PR5 sphingosine-1-phosphate receptor 5 21708.0 s at 3.788767 OW HOMER2 homer homolog 2 (Drosophila) 228596 at 3.78S9466 OW LOC728377 similar to rho guanine nucleotide exchange factor 5 219121 s at 3.7856553 OW RBM3SA RNA binding motif protein 35A 206605 at 3.7852893 OW P11 26 serine protease US 2012/0034613 A1 Feb. 9, 2012 36

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 218796 at 3.784O903 OW FERMT1 ermitin family homolog 1 (Drosophila) 206004 at 3.78.30596 OW TGM3 transglutaminase 3 (Epolypeptide, protein-glutamine-gamma glutamyltransferase) 204519 s at 3.7812307 OW PLLP plasma membrane proteolipid (plasmolipin) 203128 at 3.7796.383 OW SPTLC2 serine palmitoyltransferase, long chain base subunit 2 231875 at 3.7795398 OW KIF21A kinesin family member 21A 212096 s at 3.7734342 OW MTUS1 mitochondrial tumor suppressor 1 226003 at 3.77.00498 OW KIF21A kinesin family member 21A 200832 s at 3.7699654 OW SCD stearoyl-CoA desaturase (delta-9- desaturase) 208.614 S at 3.7652352 OW FLNB filamin B, beta (actin binding protein 278) 35820 at 3.763104 OW GM2A GM2 ganglioside activator 235048 at 3.7585585 OW FAM169A amily with sequence similarity 169, member A 202540 S at 3.7533727 OW HMGCR 3-hydroxy-3-methylglutaryl-Coenzyme A reductase 210375 at 3.74.46036 OW PTGER3 prostaglandin E receptor 3 (Subtype EP3) 219010 at 3.74.44217 OW C1orf106 chromosome 1 open reading frame 106 226129 at 3.7394128 OW FAM83H amily with sequence similarity 83, member H 232151 at 3.7381117 OW MACC1 metastasis associated in colon cancer 1 236313 at 3.736902 OW CDKN2B cyclin-dependent kinase inhibitor 2B (p15, inhibits CDK4) 227962 at 3.7365305 OW ACOX1 acyl-Coenzyme A oxidase 1, palmitoyl 235626 at 3.7264879 OW CAMK1D calcium calmodulin-dependent protein kinase ID 1559190 S. at 3.7257981 OW RDH13 CDNA PSEC0082 fis, clone NT2RP2004966, highly similar to Retinol dehydrogenase 13 (EC 1.1.1.—) 227735 S. at 3.7186708 OW C10orf 9 chromosome 10 open reading frame 99 227109 at 3.7156994 OW CYP2R1 cytochrome P450, family 2, Subfamily R, polypeptide 1 242773 at 3.714043 OW SLC5A1 Solute carrier family 5 (sodium glucose cotransporter), member 1 223541 at 3.713O34 OW HAS3 hyaluronan synthase 3 1553057 at 3.7106702 OW SERPINB12 Serpin peptidase inhibitor, clade B (ovalbumin), member 12 214279 s at 3.7052615 OW NDRG2 NDRG family member 2 221566 s at 3.7041054 OW NOL3 nucleolar protein 3 (apoptosis repressor with CARD domain) 219496 at 3.6988SS4 OW ANKRDS7 ankyrin repeat domain 57 225806 at 3.698.343S OW JUB jub, ajuba homolog (Xenopus laevis) 204765 at 3.6892798 OW ARHGEFS Rho guanine nucleotide exchange factor (GEF) 5 207430s at 3.6888688 OW MSMB microseminoprotein, beta 226382 at 3.6853971 OW LOC283070 hypothetical protein LOC283070 201693 s at 3.6807284 OW EGR1 early growth response 1 206277 at 3.679081 OW P2RY2 purinergic receptor P2Y, G-protein coupled, 2 2198.21 s at 3.678.7474 OW GFOD1 glucose-fructose oxidoreductase domain containing 1 212737 at 3.6714 OW GM2A GM2 ganglioside activator 209719 X at 3.6645396 OW SERPINB3 Serpin peptidase inhibitor, clade B (ovalbumin), member 3 205172 x at 36626048 OW CLTB clathrin, light chain (Lcb) 1553764 a at 3.662437 OW JUB jub, ajuba homolog (Xenopus laevis) 204351 at 3.660O852 OW S1 OOP S100 calcium binding protein P 203148 s at 3.6579587 OW TRIM14 tripartite motif-containing 14 209000 s at 3.6571388 OW 40064 septin 8 205786 s at 3.6454937 OW ITGAM integrin, alpha M (complement component 3 receptor 3 subunit) 223298 s at 3.64094-54 OW NTSC3 5'-nucleotidase, cytosolic III 1568868 at 3.6400533 OW CYP27C1 cytochrome P450, family 27, Subfamily C, polypeptide 1 210544 s at 3.63294.89 OW ALDH3A2 aldehyde dehydrogenase 3 family, member A2 US 2012/0034613 A1 Feb. 9, 2012 37

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title

212314 at 3.628.236 OW KIAAO746 KIAA0746 protein fit serine SERINC2 incorporator 2 206.421 s at 3.622729 OW SERPINB7 Serpin peptidase inhibitor, clade B (ovalbumin), member 7 202539 s at 3.6214926 OW HMGCR 3-hydroxy-3-methylglutaryl-Coenzyme A reductase 1553211 at 3.6197479 OW ANKFN1 ankyrin-repeat and fibronectin type III domain containing 1 20276.1 s at 3.61974OS OW SYNE2 spectrin repeat containing, nuclear envelope 2 205783 a 3.6192052 OW KLK13 kallikrein-related peptidase 13 225502 a 3.607785 OW DOCK8 dedicator of cytokinesis 8 216905 s at 3.6036117 OW ST14 Suppression of tumorigenicity 14 (colon carcinoma) 221779 a 3.6O17182 OW MICALL1 MICAL-like 1 204734 a 3.6002238 OW KRT1S keratin 15 231732 a 3.594498 OW SMPD3 sphingomyelin phosphodiesterase 3, neutral membrane (neutral sphingomyelinase II) 220030 a. 3.58.35576 OW STYK1 serine/threoninetyrosine kinase 1 39248 at 3.5794623 OW AQP3 aquaporin 3 (Gill blood group) 222876 s at 3.5791473 OW ADAP2 Arf6AP with dual PH domains 2 204636 a 3.5764592 OW COL17A1 collagen, type XVII, alpha 1 210553 x at 3.5725062 OW PCSK6 proprotein convertase Subtilisinkexin type 6 225864 a 3.5664208 OW FAM84B amily with sequence similarity 84, member B 209211 a 3.56306OS OW KLFS Kruppel-like factor 5 (intestinal) 1552777 a. at 3.5581756 OW RAET1E retinoic acid early transcript 1E 239547 a 3.5570898 OW HS3ST6 heparan sulfate (glucosamine) 3-O- Sulfotransferase 6 224839 s at 3.SS47O16 OW GPT2 glutamic pyruvate transaminase (alanine aminotransferase) 2 1553364 at 3.SS30663 OW PNPLA1 patatin-like phospholipase domain containing 1 235117 at 3.55O1385 OW CHAC2 ChaC, cation transport regulator homolog 2 (E. coli) 222354 at 3.540919:S OW F11R F11 receptor 219976 at 3.54039 OW HOOK1 hook homolog 1 (Drosophila) 211372 s at 3.5362O17 OW L1R2 interleukin 1 receptor, type II 219316 s at 3.5322564 OW FLVCR2 eline leukemia virus Subgroup C cellular receptor family, member 2 224646 x at 3.5317452 OW H19 H19, imprinted maternally expressed transcript (non-protein coding) 65438 at 3.5268037 OW KIAA1609 KIAA1609 226666 at 3.51850.58 OW DAAM1 dishevelled associated activator of morphogenesis 1 219597 s at 3.S163934 OW dual oxidase 1 205421 at 3.5158527 OW Solute carrier family 22 (extraneuronal monoamine transporter), member 3 206714 at 3.51.17545 OW ALOX1SB arachidonate 15-lipoxygenase, type B 219752 at 3.5069795 OW RASAL1 RAS protein activator like 1 (GAP1 ike) 203256 at 3.5057578 OW CDH3 cadherin 3, type 1, P-cadherin (placental) 207558 s at 3.SO35906 OW PITX2 paired-like homeodomain 2 231969 at 3.49967SS OW STOX2 storkhead box2 205249 at 3.4971406 OW EGR2 early growth response 2 (Krox-20 homolog, Drosophila) 1563900 at 3.4969769 OW FAM83B amily with sequence similarity 83, member B 210026 s at 3.4745O2 OW CARD10 caspase recruitment domain family, member 10 225.095 at 3.4744895 OW SPTLC2 KIAAO526 protein 1553333 at 3.4741.37 OW C1orf161 chromosome 1 open reading frame 161 239710 at 3.472O347 OW FIGN idgetin 203780 at 3.4676228 OW MPZL2 myelin protein zero-like 2 221843 s at 3.461.0984 OW KIAA1609 KIAA1609 208191 X at 3.4576283 OW PSG4 pregnancy specific beta-1-glycoprotein 4 201428 at 3.45399.02 OW CLDN4 claudin 4 229296 at 3.4521377 OW LOC10O128SO1 hypothetical protein LOC100128501 211043 s at 3.446346 OW CLTB clathrin, light chain (Lcb) 218432 at 3.4458OOS OW F-box protein 3 US 2012/0034613 A1 Feb. 9, 2012 38

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 201005 a. 3.4436.064 OW CD9 CD9 molecule 210868 s at 3.4410589 OW ELOVL6 ELOVL family member 6, elongation of long chain fatty acids (FEN1/Elo2, SUR4/Elo3-like, yeast) 227 276 a 3.43982OS OW PLXDC2 plexin domain containing 2 213820 s at 3.4357908 OW STARDS StAR-related lipid transfer (START) domain containing 5 209301 a 3.4329O16 OW CA2 carbonic anhydrase II 204941 s at 3.42871.07 OW ALDH3B2 aldehyde dehydrogenase 3 family, member B2 223199 a 3.4269447 OW MKNK2 MAP kinase interacting serine/threonine kinase 2 224327 s at 3.4269319 OW DGAT2 diacylglycerol O-acyltransferase homolog 2 (mouse) 235678 a 3.42678O2 OW GM2A GM2 ganglioside activator 236225 a. 3.426428 OW GGT6 gamma-glutamyltransferase 6 221567 a 3.4161S6S OW NOL3 nucleolar protein 3 (apoptosis repressor with CARD domain) 204546 a 3.4134994 OW KIAAOS13 KIAAOS13 204058 a 3.4133024 OW ME1 malic enzyme 1, NADP(+)-dependent, cytosolic 210065 s at 3.41.22272 OW UPK1B uroplakin 1B 1557165 S at 3.4083939 OW KLHL18 kelch-like 18 (Drosophila) 205014 a 340602 OW FGFBP1 fibroblast growth factor binding protein 1 221669 s at 3.4037082 OW ACAD8 acyl-Coenzyme A dehydrogenase amily, member 8 225525 a. 3.4O10828 OW CTA- KIAA1671 protein i? hypothetical 221G9.4 protein LOC100131004 LOC100131004 231118 a 3.3923123 OW ANKRD35 ankyrin repeat domain 35 229337 a 3.39.13348 OW USP2 ubiquitin specific peptidase 2 219670 a 3.381.4268 OW BENDS BEN domain containing 5 20504.8 s a 3.38.12659 OW PSPH phosphoserine phosphatase 1554.062 at 3.3779898 OW XG Xgblood group 2228.66 s a 3.372543 OW FLVCR2 eline leukemia virus Subgroup C cellular receptor family, member 2 219429 a 3.369738 OW FA2H atty acid 2-hydroxylase 232500 a 3.36646 OW C20orf74 chromosome 20 open reading frame 74 209600 s a 3.3606262 OW ACOX1 acyl-Coenzyme A oxidase 1, palmitoyl 236213 a 3.3596435 OW LOC100.130885 hypothetical protein LOC10013O885 225536 a 3.354181 OW TMEMS4 transmembrane protein 54 201534 s a 3.353898 OW UBL3 ubiquitin-like 3 228729 a 3.34.9102 OW CCNB1 cyclin B1 223839 s a 3.3401856 OW SCD if stearoyl-CoA desaturase (delta-9- SCDOS desaturase) i? stearoyl-CoA desaturase opposite strand 243611 a 3.33S2149 OW MICALCL MICAL C-terminal like 211382 s a 3.33S143 OW TACC2 transforming, acidic coiled-coil containing protein 2 22O149 a 3.3329418 OW C2Orfs4 chromosome 2 open reading frame 54 227856 a 3.3276808 OW C4orf32 chromosome 4 open reading frame 32 228469 a 3.3271294 OW PPID Cyclophilin-40 230518 a 3.325.8517 OW MPZL2 myelin protein zero-like 2 209558 s at 3.322O49 OW HIP1R huntingtin interacting protein 1 related 227.015 a. 3.3209336 OW ASPHD2 aspartate beta-hydroxylase domain containing 2 201287 s at 3.32O23 OW SDC1 Syndecan 1 205470 s at 3.3177328 OW KLK11 kallikrein-related peptidase 11 209569 X at 3.3130386 OW D4S234E DNA segment on chromosome 4 (unique) 234 expressed sequence 228.640 at 3.3072002 OW PCDHT protocadherin 7 225.001 at 3.305868 OW RAB3D RAB3D, member RAS oncogene family 232082 X at 3.3027992 OW SPRR3 Small proline-rich protein 3 223694 at 3.3023326 OW TRIM7 tripartite motif-containing 7 215393 s at 3.301.1584 OW COBLL1 COBL-like 1 229114 at 3.30.10995 OW GAB1 GRB2-associated binding protein 1 2354.05 at 3.2997503 OW GSTA4 glutathione S-transferase alpha 4 232090 at 3.2993267 OW LOC100128178 similar to hCG2041313 22825.6 s at 3.298O163 OW EPB41L4A erythrocyte membrane protein band 4.1 like 4A US 2012/0034613 A1 Feb. 9, 2012 39

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 227134 a 3.2938037 OW SYTL1 synaptotagmin-like 1 218150 a 3.2934453 OW ARLSA ADP-ribosylation factor-like 5A 205403 a 3.2927.191 OW IL1R2 interleukin 1 receptor, type II 231771 a 3.2887797 OW GJB6 gap junction protein, beta 6, 30 kDa 227782 a. 3.2884O97 OW ZBTB7C Zinc finger and BTB domain containing 7C 227461 a 3.287732 OW STON2 stonin 2 203509 a 3.2846527 OW SORL1 sortilin-related receptor, L(DLR class) A repeats-containing 223168 a 3.283O467 OW RHOU ras homologgene family, member U. 209631 s at 3.26491.26 OW GPR37 G protein-coupled receptor 37 (endothelin receptor type B-like) 205029 s at 3.2629297 OW FABP7 fatty acid binding protein 7, brain 226649 a 3.2610345 OW PANK1 pantothenate kinase 1 227889 a 3.25648 OW LPCAT2 lysophosphatidylcholine acyltransferase 2 213094 a 3.2563322 OW GPR126 G protein-coupled receptor 126 244261 a 3.2S21746 OW L28RA interleukin 28 receptor, alpha (interferon, lambda receptor) 201340 S at 3.2456722 OW ENC1 ectodermal-neural cortex (with BTB ike domain) 217974 a 3.2413647 OW TM7SF3 transmembrane 7 Superfamily member 3 223631 s at 3.2411804 OW C19Crf33 chromosome 19 open reading frame 33 230765 a. 3.238S905 OW KIAA1239 KIAA1239 1552566 at 3.232O094 OW BTBD16 BTB (POZ) domain containing 16 225613 a 3.23.06.297 OW LOC100128443 hypothetical protein LOC100128443/// A. MAST4 microtubule associated serine/threonine kinase family member 4 205776 a 3.222692 OW FMO5 flavin containing monooxygenase 5 203007 X at 3.2214603 OW LY PLA1 ySophospholipase I 2291.03 a 3.2176034 OW WNT3 wingless-type MMTV integration site amily, member 3 224367 a 3.2115495 OW BEX2 brain expressed X-linked 2 225834 a 3.21 13602 OW FAM72A, . amily with sequence similarity 72, FAM72B member A family with sequence GCUD2 similarity 72, member B if gastric cancer up-regulated-2 230266 a 3.2071729 OW RAB7B RAB7B, member RAS oncogene amily 206059 a 3.2045915 OW ZNF91 Zinc finger protein 91 225807 a 3.198O147 OW UB jub, ajuba homolog (Xenopus laevis) 203786 s at 3.196283 OW TPD52L1 tumor protein D52-like 1 1554895 a. at 3.193OOO3 OW RHBDL2 rhomboid, veinlet-like 2 (Drosophila) 203665 a 3.1928,537 OW HMOX1 heme oxygenase (decycling) 1 204363 a 3.19.15643 OW F3 coagulation factor III (thromboplastin, issue factor) 238.164 a 3.190472 OW USP6NL USP6 N-terminal like 212322 a. 3.1838899 OW SGPL1 sphingosine-1-phosphate lyase 1 1566766 a. at 3.1814OOS OW MACC1 metastasis associated in colon cancer 1 207126 X at 3.1803102 OW UGT1A1 UDPglucuronosyltransferase 1 family, UGT1A10 polypeptide A1 / UDP UGT1 A4 glucuronosyltransferase 1 family, UGT1A6 polypeptide A10 / UDP UGT1A8 glucuronosyltransferase 1 family, UGT1A9 polypeptide A4 i? UDP glucuronosyltransferase 1 family, polypeptide A6/// UDP glucuronosyltransferase 1 family, polypeptide A8 // UDP glucuronosyltransferase 1 family, polypeptide A9 222223 s at 3.176874 down IL1R5 interleukin 1 family, member 5 (delta) 239694 at 3.174275 down TRIM7 tripartite motif-containing 7 206969 at 3.1713967 down KRT34 keratin 34 207540 S at 3.1631026 down SYK spleen tyrosine kinase 235857 at 3.1571.46 down KCTD11 potassium channel tetramerisation domain containing 11 205829 at 3.1498.919 down HSD17B1 hydroxysteroid (17-beta) dehydrogenase 1 20972O is at 3.1471493 down SERPINB3 Serpin peptidase inhibitor, clade B (ovalbumin), member 3 226698 at 3.1410942 down FCHSD1 FCH and double SH3 domains 1 203147 s at 3.140271 down TRIM14 tripartite motif-containing 14 US 2012/0034613 A1 Feb. 9, 2012 40

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 214036 at 3.1385825 OW EFNAS ephrin-A5 1558378 a. at 3.1375O17 OW AHNAK2 AHNAK nucleoprotein 2 203887 s at 3.1370428 OW THEBD hrombomodulin 205015 S. at 3.13355O2 OW TGFA transforming growth factor, alpha 218756 s at 3.1297085 OW DHRS11 dehydrogenase/reductase (SDR family) member 11 204256 at 3.1214519 OW ELOVL6 ELOVL family member 6, elongation of long chain fatty acids (FEN1/Elo2, SUR4/Elo3-like, yeast) 206912 at 3.120S63 OW FOXE1 orkhead box E1 (thyroid transcription actor 2) 209885 at 3.12O1258 OW RHOD ras homologgene family, member D 229638 at 3.1183026 OW IRX3 iroquois homeobox 3 220249 at 3.116939 OW HYAL4 hyaluronoglucosaminidase 4 210347 s at 3.11.198O2 OW BCL11A B-cell CLL/lymphoma 11A (zinc finger protein) 224480 s at 3.1057024 OW AGPAT9 -acylglycerol-3-phosphate O acyltransferase 9 236359 at 3.10485 OW SCN4B Sodium channel, voltage-gated, type V, beta 229374 at 3.10OS 714 OW EPHA4 EPH receptor A4 226617 at 3.0999246 OW ARLSA ADP-ribosylation factor-like 5A 221856 s at 3.0998528 OW FAM63A amily with sequence similarity 63, member A 238720 at 3.099.1333 OW OMG OMGP mRNA for oligodendrocyte myelin glycoprotein 214295 at 3.0939,116 OW KIAAO485 hypothetical LOC57235 21895.1 s at 3.0916,145 OW PLCXD1 phosphatidylinositol-specific phospholipase C, X domain containing 1 223574 X at 3.0913527 OW PPP2R2C protein phosphatase 2 (formerly 2A), regulatory subunit B, gamma isoform 224685 a. 3.0898316 OW MLLT4 myeloid lymphoid or mixed-lineage eukemia (trithorax homolog, Drosophila); translocated to, 4 231778 a 3.089387 OW DLX3 distal-less homeobox 3 229290 a 3.0886714 OW DAPL1 death associated protein-like 1 21882.9 s at 3.0885O17 OW CHD7 chromodomain helicase DNA binding protein 7 223894 s at 3.0799034 OW AKTIP AKT interacting protein 2261.87 a 3.0795898 OW CDS1 CDP-diacylglycerol synthase (phosphatidate cytidylyltransferase) 1 227944 a 3.077466 OW PTPN3 protein tyrosine phosphatase, non receptor type 3 241813 a 3.0757666 OW MBD1 methyl-CpG binding domain protein 1 203287 a 3.07.20277 OW LAD1 adinin 1 229546 a 3.0575104 OW LOC6536O2 hypothetical LOC653602 218849 s at 3.0525377 OW PPP1R13L protein phosphatase 1, regulatory (inhibitor) subunit 13 like 1558281 a. at 3.0518956 OW TMEM184A transmembrane protein 184A 224.579 a 3.04471.47 OW SLC38A1 solute carrier family 38, member 1 222830 a. 3.04.40314 OW GRHL1 grainyhead-like 1 (Drosophila) 209605 a 3.037887 OW TST hiosulfate sulfurtransferase (rhodanese) 206632 s at 3.0369 OW APOBEC3B apolipoprotein B mRNA editing enzyme, catalytic polypeptide-like 3B 21303.0 s at 3.0361433 OW PLXNA2 plexin A2 213787 s at 3.0355 OW EBP emopamil binding protein (sterol isomerase) 212095 s at 3.0337477 OW MTUS1 mitochondrial tumor suppressor 1 1564308 a. at 3.033596S OW MPP7 membrane protein, palmitoylated 7 (MAGUK p55 subfamily member 7) 204059 s at 3.028911 OW ME1 malic enzyme 1, NADP(+)-dependent, cytosolic 218922 s at 3.02.78475 OW LASS4 LAG1 homolog, ceramide synthase 4 219990 at 3.02.1536 OW E2F8 E2F transcription factor 8 244758 at 3.021281 OW SCAND3 SCAN domain containing 3 201427 s at 3.01.1953 OW SEPP1 Selenoprotein P. plasma, 1 237899 at 3.01176OS OW LOCA29994 hypothetical LOC72.9994 233814 at 3.OOS24O7 OW EFNAS Receptor tyrosine kinase ligand LERK 7 precursor (EPLG7) 226829 at 3.0O2S802 OW AFAP1L2 actin filament associated protein 1-like 2 203397 s at 3.002098 OW GALNT3 UDP-N-acetyl-alpha-D- galactosamine:polypeptide N US 2012/0034613 A1 Feb. 9, 2012 41

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title acetylgalactosaminyltransferase 3 (GalNAc-T3) 2228.09 X at 3.OO19972 OW C14Orf65 chromosome 14 open reading frame 65 234725 s at 3.0007038 OW SEMA4B Sema domain, immunoglobulin domain (Ig), transmembrane domain (TM) and short cytoplasmic domain, (semaphorin) 4B 203560 at 2.9994.648 OW GGEH gamma-glutamyl hydrolase (conjugase, olylpolygammaglutamyl hydrolase) 233565 s at 2.9988334 OW SDCBP2 Syndecan binding protein (Syntenin) 2 1553212 at 2.99641 O1 OW KRT78 keratin 78 204135 at 2.9959981 OW FILIP1L, filamin A interacting protein 1-like 223278 at 2.995849 OW GJB2 gap junction protein, beta 2, 26 kDa 227385 at 2.995O788 OW PPAPDC2 phosphatidic acid phosphatase type 2 domain containing 2 216347 s at 2.9949014 OW PPP1R13B protein phosphatase 1, regulatory (inhibitor) subunit 13B 203359 is at 2.993.3915 OW MYCBP c-myc binding protein 242064 at 2.99.17743 OW SDK2 sidekick homolog 2 (chicken) 1553114 a at 2.987O312 OW PTK6 PTK6 protein tyrosine kinase 6 236266 at 2.98SSO3 OW RORA Hypothetical protein LOC283666, mRNA (cDNA clone MAGE: 4750925) 242317 at 2.9845624 OW HIGD1A HIG1 domain family, member 1A 209203 s at 2.98.4112 OW BICD2 bicaudal Dhomolog 2 (Drosophila) 226245 at 2.98O1052 OW KCTD1 potassium channel tetramerisation domain containing 1 214765 s at 2.9758816 OW NAAA N-acylethanolamine acid amidase 223216 x at 2.97O3996 OW ZNF395 Zinc finger protein 395 221215 S. at 2.97.02475 OW RIPK4 receptor-interacting serine-threonine kinase 4 221081 s at 2.9699,113 OW DENND2D DENN/MADD domain containing 2D 202154 x at 2.96.69414 OW TUBB3 tubulin, beta 3 208126 s at 2.9666116 OW CYP2C18 cytochrome P450, family 2, Subfamily C, polypeptide 18 213476 x at 2.96.6195 OW TUBB3 tubulin, beta 3 203722 at 2.9630923 OW ALDH4A1 aldehyde dehydrogenase 4 family, member A1 227135 at 2.9602783 OW NAAA N-acylethanolamine acid amidase 214705 at 2.9595 757 OW INADL naD-like (Drosophila) 211985 S at 2.958S447 OW CALM1 calmodulin 1 (phosphorylase kinase, CALM2 delta) i? calmodulin 2 (phosphorylase CALM3 kinase, delta) i? calmodulin 3 (phosphorylase kinase, delta) 238,513 at 2.94.96446 OW PRRG4 Proline rich Gla (G-carboxyglutamic acid) 4 (transmembrane), mRNA (cDNA clone MGC: 19793 MAGE: 3841745) 243837 X at 2.947061 OW LOC100128500 hypothetical protein LOC100128500 218677 at 2.946.3606 OW S100A14 S100 calcium binding protein A14 218900 at 2.9451632 OW CNNM4 cyclin M4 220266 s at 2.9415443 OW KLF4 Kruppel-like factor 4 (gut) 204341 at 2.94O9094 OW TRIM16 tripartite motif-containing 16 230769 at 2.939519 OW DENND2C DENN/MADD domain containing 2C 232693 s at 2.9363637 OW FBXO16 F-box protein 16 fit zinc finger protein ZNF395 395 226246 at 2.93593.93 OW KCTD1 potassium channel tetramerisation domain containing 1 226421 at 2.933277 OW AMMECR1 Alport syndrome, mental retardation, midface hypoplasia and elliptocytosis chromosomal region gene 1 225655 at 2.92.76872 OW UHRF1 ubiquitin-like with PHD and ring finger domains 1 222717 at 2.92.61.97 OW SDPR. serum deprivation response (phosphatidylserine binding protein) 235871 at 2.92S3323 OW LIPEH lipase, member H 223339 at 2.92S2O36 OW ATPIF1 ATPase inhibitory factor 1 223233 s at 2.9242868 OW CGN cingulin 225051 at 2.9183056 OW EPB41 erythrocyte membrane protein band 4.1 (elliptocytosis 1, RH-linked) 209372 x at 2.91.4044 OW TUBB2A, . tubulin, beta 2A tubulin, beta 2B TUBB2B US 2012/0034613 A1 Feb. 9, 2012 42

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 224495 at 2.91.37444 down TMEM107 transmembrane protein 107 218722 s at 2.9108763 down CCDC51 coiled-coil domain containing 51 228221 at 2.9098802 down SLC44A3 solute carrier family 44, member 3 207955 at 2.907911 down CCL27 chemokine (C-C motif) ligand 27 206094 x at 2.9065409 down UGT1A1 if UDPglucuronosyltransferase 1 family, UGT1A10 polypeptide A1 / UDP UGT1A3 glucuronosyltransferase 1 family, UGT1 A4 if polypeptide A10 / UDP UGT1AS glucuronosyltransferase 1 family, UGT1A6 polypeptide A3 // UDP UGT1A7 glucuronosyltransferase 1 family, UGT1A8 polypeptide A4 i? UDP UGT1A9 glucuronosyltransferase 1 family, polypeptide A5 // UDP glucuronosyltransferase 1 family, polypeptide A6/// UDP glucuronosyltransferase 1 family, polypeptide A7 /?/ UDP glucuronosyltransferase 1 family, polypeptide A8 // UDP glucuronosyltransferase 1 family, polypeptide A9 231830 x at 2.902842 OW RAB11FIP1 RAB11 family interacting protein 1 (class I) 1554966 a. at 2.9024658 OW FILIP1L, filamin A interacting protein 1-like 22.6959 a 2.9023726 OW LOC283070 CDNA FLJ40058 fis, clone TCOLN100018O 206400 a 2.9014132 OW LGALS7 ectin, galactoside-binding, soluble, 7 LGALS7B if lectin, galactoside-binding, soluble, 7B 1554246 at 2.89799.17 OW C1orf210 chromosome 1 open reading frame 210 227736 a 2.8947847 OW C10orf 9 chromosome 10 open reading frame 99 221123 x at 2.8909688 OW ZNF395 Zinc finger protein 395 222890 a 2.8908694 OW CCDC113 coiled-coil domain containing 113 208190 s. at 2.89.0325 OW LSR lipolysis stimulated lipoprotein receptor 229396 a 2.8896SO6 OW OVOL1 ovo-like 1 (Drosophila) 227034 a 2.88846S OW ANKRDS7 ankyrin repeat domain 57 219648 a 2.8866937 OW MREG melanoregulin 218792 s at 2.8844924 OW BSPRY B-box and SPRY domain containing 204542 a. 2.88.17751 OW ST6GALNAC2 ST6 (alpha-N-acetyl-neuraminyl-2,3- beta-galactosyl-1,3)-N- acetylgalactosaminide alpha-2,6- sialyltransferase 2 238962 a 2.877806 OW ZNF681 Zinc finger protein 681 227964 a 2.8771.96 OW FRMD8 FERM domain containing 8 238964 a 2.8724544 OW FIGN idgetin 201564 S at 2.872O4SS OW FSCN1 ascin homolog 1, actin-bundling protein (Strongylocentrotus purpuratus) 38340 at 2.87.08196 OW HIP1R huntingtin interacting protein 1 related 218032 a 2.87O3704 OW SNN Stannin 204547 a 2.87O3O67 OW RAB4OB RAB40B, member RAS oncogene amily 213506 a 2.8534102 OW F2RL1 coagulation factor II (thrombin) receptor-like 1 235095 a 2.851771 OW CCDC64B coiled-coil domain containing 64B 202962 a 2.8498514 OW KIF13B kinesin family member 13B 200606 a 2.84.82468 OW DSP desmoplakin 220578 a 2.8452191 OW ADAMTSL4 ADAMTS-like 4 21878.9 s at 2.84.0476 OW C11orf71 chromosome 11 open reading frame 71 242093 a 2.8397858 OW SYTL5 synaptotagmin-like 5 228975 a 2.8345933 OW SP6 Sp6 transcription factor 1569555 at 2.832668 OW GDA guanine deaminase 201694 S at 2.832S536 OW EGR1 early growth response 1 21.0128 s at 2.8322942 OW LTB4R leukotriene B4 receptor 213805 a 2.8317797 OW ABHDS abhydrolase domain containing 5 238762 a 2.8312757 OW MTHFD2L methylenetetrahydrofolate dehydrogenase (NADP+ dependent) 2 like 1558111 at 2.8277514 OW MBNL1 muscleblind-like (Drosophila) 223000 s at 2.8271255 OW F11R F11 receptor 227998 at 2.82364O6 OW S100A16 S100 calcium binding protein A16 US 2012/0034613 A1 Feb. 9, 2012 43

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title

238206 a 2.822679S OW relaxininsulin-like family peptide receptor 1 1552648 a. at 2.8.2174 OW tumor necrosis factor receptor Superfamily, member 10a 235850 a 2.8.215306 OW WDRSB WD repeat domain 5B 200636 s at 2.816918 OW PTPRF protein tyrosine phosphatase, receptor type, F 218779 X at 2.81.3133 OW EPS8-like 1 220456 a 2.81.1995 OW serine palmitoyltransferase, long chain base subunit 3 210652 s at 2.8102167 OW tetratricopeptide repeat domain 39A 239853 a 2.809029 OW kinesin light chain 3 205765 a. 2.8077855 OW cytochrome P450, family 3, Subfamily A, polypeptide 5 1552620 at 2.8073O84 OW SPRR4 Small proline-rich protein 4 225779 a 2.80724OS OW SLC27A4 solute carrier family 27 (fatty acid transporter), member 4 203699 s at 2.806607 OW DIO2 deiodinase, iodothyronine, type II 231810 a 2.80636.36 OW BRI3BP Cervical cancer oncogene binding protein 221893 s at 2.7909083 OW ADCK2 aarF domain containing kinase 2 209260 a 2.79O1292 OW SFN stratifin 205054 a 2.7879531 OW NEB nebulin 213050 a 2.7861958 OW COBL cordon-bleu homolog (mouse) 218028 a 2.7856.884 OW ELOVL1 elongation of very long chain fatty acids (FEN1/Elo2, SUR4/Elo3, yeast)- like 224496 s at 2.78.19924 OW TMEM107 transmembrane protein 107 1553031 at 2.7813523 OW GPR115 G protein-coupled receptor 115 223497 at 2.7785342 OW FAM13SA family with sequence similarity 135, member A 204856 at 2.77795 74 OW B3GNT3 UDP-GlcNAc:beta Gal beta-1,3-N- acetylglucosaminyltransferase 3 220911 s at 2.7676141 OW KIAA1305 KIAA1305 219696 at 2.7632923 OW DENND1B DENN/MADD domain containing 1B 1559096 X at 2.75.8654 OW FBXO9 F-box protein 9 204976 s at 2.7562022 OW AMMECR1 Alport syndrome, mental retardation, midface hypoplasia and elliptocytosis chromosomal region gene 1 218149 s at 2.7463,727 OW ZNF395 Zinc finger protein 395 229513 at 2.74618OS OW STRBP Chromosome 9 open reading frame 45, mRNA (cDNA clone MGC:45613 MAGE: 2989018) 235605 at 2.74O469S OW CASZ1 castor Zinc finger 1 2132O1 is at 2.7397995 OW TNNT1 troponin T type 1 (skeletal, slow) 204141 at 2.738532 OW TUBB2A tubulin, beta 2A 23O398 at 2.7380996 OW TNS4 ensin 4 23.8013 at 2.7366548 OW PLEKHA2 pleckstrin homology domain containing, family A (phosphoinositide binding specific) member 2 204532 x at 2.7337,391 OW UDPglucuronosyltransferase 1 family, polypeptide A1 / UDP glucuronosyltransferase 1 family, polypeptide A10 / UDP glucuronosyltransferase 1 family, polypeptide A4 i? UDP glucuronosyltransferase 1 family, polypeptide A6/// UDP glucuronosyltransferase 1 family, polypeptide A8 // UDP glucuronosyltransferase 1 family, polypeptide A9 204554 at 2.731 1485 down PPP1R3D protein phosphatase 1, regulatory (inhibitor) subunit 3D 205552 s at 2.72798.44 down OAS1 2',5'-oligoadenylate synthetase 1, 40.46 kDa 228565 at 2.727 6409 down KIAA1804 mixed lineage kinase 4 228155 at 2.7267SO4 down C10orf57 chromosome 10 open reading frame 57 C10orf58 if chromosome 10 open reading frame 58 223427 s at 2.7255745 down erythrocyte membrane protein band 4.1 like 4B US 2012/0034613 A1 Feb. 9, 2012 44

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 204867 a 2.7246413 OW GCHFR GTP cyclohydrolase I feedback regulator 221802 s at 2.723564 OW KIAA1598 KIAA1598 231807 a 2.7220626 OW KIAA1217 KIAA1217 205818 a 2.72.16518 OW DBC1 deleted in bladder cancer 1 226106 a 2.7186909 OW RNF141 ring finger protein 141 205251 a 2.7182S6 OW PER2 period homolog 2 (Drosophila) 201249 a 2.7168.527 OW SLC2A1 solute carrier family 2 (facilitated glucose transporter), member 1 215891 s at 2.712683 OW GM2A GM2 ganglioside activator 206102 a 2.7O67106 OW GINS1 GINS complex subunit 1 (Psfl homolog) 205676 a 2.704932 OW CYP27B1 cytochrome P450, family 27, Subfamily B, polypeptide 1 230475 a 2.704732 OW C1 Sorf59 chromosome 15 open reading frame 59 1554878 a. at 2.7038OO2 OW ABCD3 ATP-binding cassette, Sub-family D (ALD), member 3 226968 a 2.7027438 OW KIF1B kinesin family member 1B 229997 a 2.70270.18 OW WANGL1 vang-like 1 (van gogh, Drosophila) 222833 a 2.696.728S OW LPCAT2 lysophosphatidylcholine acyltransferase 2 202449 s at 2.694.6986 OW RXRA retinoid X receptor, alpha 222912 a 2.6944733 OW ARRB1 arrestin, beta 1 1555292 at 2.6942291 OW FAM4OB family with sequence similarity 40, member B 240303 a 2.6937366 OW TMC5 Transmembrane channel-like 5, mRNA (cDNA clone IMAGE: 5265527) 203641 s at 2.6928S94 OW COBLL1 COBL-like 1 208999 a 2.6917293 OW 40064 septin 8 210716 s at 2.6889458 OW CLIP1 CAP-GLY domain containing linker protein 1 204503 a 2.6869638 OW EVPL envoplakin 220658 s at 2.6862872 OW ARNTL2 aryl hydrocarbon receptor nuclear translocator-like 2 57163 at 2.6833892 OW ELOVL1 elongation of very long chain fatty acids (FEN1/Elo2, SUR4/Elo3, yeast)- like 1 230630 at 2.6827O3 OW AK3L1 adenylate kinase 3-like 1 ff adenylate AK3L2 kinase 3-like 2 226690 at 2.68.1126 OW ADCYAP1R1 CDNA FLJ39226 fis, clone OCBBF2OO7232 218845 at 2.68O393 OW DUSP22 dual specificity phosphatase 22 212702 s at 2.68O374 OW BICD2 bicaudal D homolog 2 (Drosophila) 224.516 s at 2.67937 OW CXXC5 CXXC finger 5 201732 s at 2.6788O18 OW CLCN3 chloride channel 3 239273 s at 2.6781023 OW MMP28 matrix metallopeptidase 28 205.739 x at 2.67809S OW ZNF107 Zinc finger protein 107 225597 at 2.6771371 OW SLC45A4 solute carrier family 45, member 4 204430 s at 2.6757636 OW SLC2AS solute carrier family 2 (facilitated glucosef fructose transporter), member 5 212921 at 2.6731923 OW SMYD2 SET and MYND domain containing 2 24.2123 at 2.671885S OW PAQR7 progestin and adipoC receptor family member VII 227 271 at 2.6716337 OW FGF11 fibroblast growth factor 11 225987 at 2.669063 OW STEAP4 STEAP family member 4 209790 s. at 2.6684937 OW CASP6 caspase 6, apoptosis-related cysteine peptidase 222477 s at 2.6653984 OW TM7SF3 transmembrane 7 Superfamily member 3 203411 s at 2.663246 OW LMNA lamin AC 224,580 at 2.66036S OW SLC38A1 solute carrier family 38, member 1 205569 at 2.656O183 OW LAMP3 lysosomal-associated membrane protein 3 236496 at 2.653787 OW DEGS2 degenerative spermatocyte homolog2, lipid desaturase (Drosophila) 231775 at 2.652O64 OW TNFRSF1 OA tumor necrosis factor receptor Superfamily, member 10a 211950 at 2.6516557 OW UBR4 ubiquitin protein ligase E3 component n-recognin 4 207431 s at 2.645OOO2 OW DEGS1 degenerative spermatocyte homolog 1, lipid desaturase (Drosophila) 200635 s at 2.6411211 OW PTPRF protein tyrosine phosphatase, receptor type, F US 2012/0034613 A1 Feb. 9, 2012 45

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 230252 at 2.640716 OW LPARS lysophosphatidic acid receptor 5 226584 S at 2.64O212S OW FAM110A family with sequence similarity 110, member A 233955 X at 2.6379757 OW CXXC5 CXXC finger 5 213624 at 2.63438 OW SMPDL3A sphingomyelin phosphodiesterase, acid-like 3A 212449 s at 2.632708 OW LYPLA1 lysophospholipase I 231755 at 2.6293309 OW IL1E8 interleukin 1 family, member 8 (eta) 216836 s at 2.6269796 OW ERBB2 v-erb-b2 erythroblastic leukemia viral oncogene homolog2, neuroglioblastoma derived oncogene homolog (avian) 21208.6 X at 2.6258197 OW LMNA lamin AC 211984 a 2.623374 OW CALM1 calmodulin 1 (phosphorylase kinase, CALM2 delta) i? calmodulin 2 (phosphorylase CALM3 kinase, delta) i? calmodulin 3 (phosphorylase kinase, delta) 229522 a. 2.6224053 OW SDR42E1 short chain dehydrogenase/reductase family 42E, member 1 229545 a. 2.621OSS6 OW FERMT1 fermitin family homolog 1 (Drosophila) 218373 a 2.6198244 OW AKTIP AKT interacting protein 2095.02 s at 2.6195574 OW BALAP2 BAI1-associated protein 2 1554541 a. at 2.6188276 OW GPRIN2 G protein regulated inducer of neurite outgrowth 2 228067 a 2.6184916 OW C2Orfss chromosome 2 open reading frame 55 218174 s at 2.6181328 OW C10orf57 chromosome 10 open reading frame 57 234931 a 2.61595.03 OW AYP1p1 AYP1 pseudogene 1 1553906 S. at 2.6134384 OW FGD2 FYVE, RhoGEF and PH domain containing 2 232893 a 2.61261.34 OW LMBRD2 LMBR1 domain containing 2 21538.0 s at 2.6118267 OW GGCT gamma-glutamyl cyclotransferase 1552691 at 2.6064768 OW ARL11 ADP-ribosylation factor-like 11 209930 S. at 2.603701 OW NFE2 nuclear factor (erythroid-derived 2), 45kDa 225327 at 2.603O378 OW KIAA1370 KIAA1370 226198 at 2.602333 OW TOM1L2 target of myb1-like 2 (chicken) 210872 X at 2.6014338 OW GAS7 growth arrest-specific 7 200831 s at 2.6OO4403 OW SCD stearoyl-CoA desaturase (delta-9- desaturase) 227701 at 2.5987177 OW C10orf118 chromosome 10 open reading frame 118 230986 at 2.5968,971 OW KLF8 CDNA selection clone ADS40 220944 at 2.5951362 OW PGLYRP4 peptidoglycan recognition protein 4 225611 at 2.593.996S OW LOC100128443 hypothetical protein LOC100128443/// A. MAST4 microtubule associated serine/threonine kinase family member 4 237159 X at 2.5931 S63 OW AP1 S3 adaptor-related protein complex 1, sigma 3 subunit 208977 X at 2.5907562 OW TUBB2C tubulin, beta 2C 202735 at 2.5905764 OW EBP emopamil binding protein (sterol isomerase) 203535 at 2.590224 OW S100A9 S100 calcium binding protein A9 221 698 s at 2.586482S OW CLEC7A C-type lectin domain family 7, member A 202596 at 2.58641.98 OW ENSA endosulfine alpha 229776 at 2.58.56552 OW SLCO3A1 Solute carrier organic anion transporter family, member 3A1 218802 at 2.5770578 OW CCDC109B coiled-coil domain containing 109B 219230 at 2.5765135 OW TMEM100 transmembrane protein 100 226769 at 2.5724.943 OW FIBIN fin bud initiation factor homolog (Zebrafish) 211661 x at 2.56998O1 OW PTAFR platelet-activating factor receptor 218556 at 2.5698657 OW ORMDL2 ORM1-like 2 (S. cerevisiae) 209218 at 2.566318S OW SQLE squalene epoxidase 1557828 a. at 2.564371 OW CSOrf28 CDNA FLJ36759 fis, clone UTERU2O18566 207169 X at 2.5612175 OW DDR1 discoidin domain receptor tyrosine kinase 1 213174 at 2.5579.307 OW TTC9 tetratricopeptide repeat domain 9 236760 at 2.SS66704 OW AMMECR1 Alport syndrome, mental retardation, midface hypoplasia and elliptocytosis chromosomal region, gene 1, mRNA (cDNA clone IMAGE: 5405764) US 2012/0034613 A1 Feb. 9, 2012 46

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 224008 s at 2.SS3O894 OW KCNK7 potassium channel, Subfamily K, member 7 203970 S. at 2.546217 OW PEX3 peroxisomal biogenesis factor 3 239533 at 2.539709 OW GPR155 G protein-coupled receptor 155 202814 s at 2.5392O75 OW HEXIM1 hexamethylene bis-acetamide inducible 1 212479 s at 2.5390 153 OW RMINDSA required for meiotic nuclear division 5 homolog A (S. cerevisiae) 208188 at 2.5387063 OW KRT9 keratin 9 218888 s at 2.537O66 OW NETO2 neuropilin (NRP) and tolloid (TLL)- ike 2 202081 at 2.536.6287 OW IER2 immediate early response 2 1569065 s at 2.5342972 OW C15Orf62 chromosome 15 open reading frame 62 238.687 X at 2.531887 OW ZNF770 Zinc finger protein 770 214639 s at 2.53O4656 OW HOXA1 homeobox A1 2008.64 s at 2.5266373 OW RAB11A RAB11A, member RAS oncogene amily 22104.4 S at 2.5264642 OW TRIM34 tripartite motif-containing 34 if TRIM6 tripartite motif-containing 6 fif TRIM6 TRIM6- TRIM34 readthrough transcript TRIM34 226002 at 252624.94 OW GAB1 GRB2-associated binding protein 1 218237 s at 2.524939S OW SLC38A1 solute carrier family 38, member 1 228320 x at 2.5236.611 OW CCDC64 coiled-coil domain containing 64 218094 S at 2.5231547 OW DBNDD2 if dysbindin (dystrobrevin binding SYS1- protein 1) domain containing 2 DBNDD2 SYS1-DBNDD2 readthrough transcript 204451 at 2.52.15912 OW FZD1 frizzled homolog 1 (Drosophila) 237030 at 2.52O7937 OW ACPP acid phosphatase, prostate 203215 s at 2.51994S1 OW MYO6 myosin VI 204334 at 2.5181212 OW KLF7 Kruppel-like factor 7 (ubiquitous) 221610 s at 2.5177839 OW STAP2 signal transducing adaptor family member 2 224414 S at 2.5176234 OW CARD6 caspase recruitment domain family, member 6 219428 s at 2.5164838 OW PXMP4 peroxisomal membrane protein 4, 24 kDa 223805 at 2.5157578 OW OSBPL6 oxysterol binding protein-like 6 208534 S. at 2.515689 OW RASA4 RAS p21 protein activator 4 RAS RASA4P p21 protein activator 4 pseudogene 222725 s at 2.5121596 OW PALMD palmdelphin 207847 s at 2.5091352 OW MUC1 mucin 1, cell Surface associated 212882 at 2.5058832 OW KLHL18 kelch-like 18 (Drosophila) 204653 at 2.5O22001 OW TFAP2A transcription factor AP-2 alpha (activating enhancer binding protein 2 alpha) 21951.8 s at 2. SOO2704 OW ELL3 elongation factor RNA polymerase II ike 3 201939 at 2.4999.027 OW PLK2 polo-like kinase 2 (Drosophila) 210582 s at 2.499.334 OW LIMK2 LIM domain kinase 2 237252 at 2.494-65O1 OW THEBD hrombomodulin 201349 at 2.490851 OW SLC9A3R1 solute carrier family 9 (sodium hydrogen exchanger), member 3 regulator 1 203216 s at 2.48899.13 OW MYO6 myosin VI 213256 at 2.488.1077 OW 39875 membrane-associated ring finger (C3HC4)3 204989 s at 2.487853S OW ITGB4 integrin, beta 4 213154 s at 2.484296 OW BICD2 bicaudal D homolog 2 (Drosophila) 214058 at 2.4831367 OW MYCL1 v-myc myelocytomatosis viral oncogene homolog 1, lung carcinoma derived (avian) 220638 s at 2.4813514 OW CBLC Cas-Br-M (murine) ecotropic retroviral transforming sequence c 218180 s at 2.4810977 OW EPS8L2 EPS8-like 2 235333 at 2.480436 OW B4GALT6 UDP-Gal:betaGlcNAc beta 14 galactosyltransferase, polypeptide 6 203632 s at 2.4795.673 OW GPRCSB G protein-coupled receptor, family C, group 5, member B 229568 at 2.478.6081 OW MOBKL2B MOB1, Mps One Binder kinase activator-like 2B (yeast) 206653 at 2.4773498 OW POLR3G polymerase (RNA) III (DNA directed) polypeptide G (32 kD) US 2012/0034613 A1 Feb. 9, 2012 47

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 201432 at 2.477O842 OW CAT catalase 224810 s at 2.4760013 OW ANKRD13A ankyrin repeat domain 13A 1555942 a. at 2.4756498 OW LOC642S87 CDNA FLJ33794 fis, clone CTONG1OOOOO9 213726 x at 2.4728134 OW TUBB2C tubulin, beta 2C 242197 X at 2.4713676 OW CD36 CD36 antigen 223394 at 2.47O1357 OW SERTAD1 SERTA domain containing 1 201641 at 2.4696527 OW BST2 bone marrow stromal cell antigen 2 213288 at 2.4695723 OW MBOAT2 membrane bound O-acyltransferase domain containing 2 221841 s at 2.467558 OW KLF4 Kruppel-like factor 4 (gut) 227970 at 2.466.2564 OW GPR157 G protein-coupled receptor 157 222996 s at 2.4653852 OW CXXC5 CXXC finger 5 209566 at 2.4633334 OW NSIG2 insulin induced gene 2 214696 at 2.46279.34 OW C17orf1 chromosome 17 open reading frame 91 214234 S at 2.4618382 OW CYP3A5 cytochrome P450, family 3, Subfamily A, polypeptide 5 223471 at 2.4607677 OW RAB3IP RAB3A interacting protein (rabin3) 234305 s at 2.4569333 OW GSDMC gasdermin C 216388 s at 2.4562252 OW LTB4R eukotriene B4 receptor 227624 at 2.4S47067 OW TET2 et oncogene family member 2 223279 s at 2.453393 OW UACA uveal autoantigen with coiled-coil domains and ankyrin repeats 40016 g at 2.452864 OW LOC100128443 hypothetical protein LOC100128443/// A. MAST4 microtubule associated serine/threonine kinase family member 4 201341 a 2.4487681 OW ENC ectodermal-neural cortex (with BTB ike domain) 242722 a. 2.4461777 OW LMO7 LIM domain 7 224301 X at 2.4447956 OW H2AF H2A histone family, member J 225521 a 2.4442914 OW ANAPC7 anaphase promoting complex subunit 7 206355 a. 2.442836 OW GNAL guanine nucleotide binding protein (G protein), alpha activating activity polypeptide, olfactory type 219296 a 2.439S428 OW ZDHHC13 Zinc finger, DHHC-type containing 13 20075.2 s at 2.4395049 OW CAPN1 calpain 1 (muI) large subunit 209031 a 2.4321.353 OW CADM1 cell adhesion molecule 1 216641 s at 2.4319324 OW LAD adinin 1 209758 s at 2.4311805 OW MFAPS microfibrillar associated protein 5 228892 a 2.4310536 OW SH3RF2 SH3 domain containing ring finger 2 223681 s at 2.4298599 OW NADL naD-like (Drosophila) 200768 s at 2.428787 OW MAT2A methionine adenosyltransferase II, alpha 207455 a. 2.42761.83 OW P2RY1 purinergic receptor P2Y, G-protein coupled, 1 220403 s at 2.42S4568 OW P53AIP1 p53-regulated apoptosis-inducing protein 1 2298.00 a 2.4252849 OW DCLK1 KIAA0369 gene 1555404 a at 2.42292OS OW DUOXA1 dual oxidase maturation factor 1 1558834 S. at 2.420979S OW C1orf62 chromosome 1 open reading frame 62 209291 a 2.4183009 OW D4 inhibitor of DNA binding 4., dominant negative helix-loop-helix protein 210831 s at 2.41 78216 OW PTGER3 prostaglandin E receptor 3 (Subtype EP3) 1559571 a. at 2.4145973 OW ATP13A4 ATPase type 13A4 231928 at 2.4141552 OW HES2 hairy and enhancer of split 2 (Drosophila) 202005 at 2.41391 OW ST14 Suppression of tumorigenicity 14 (colon carcinoma) 2124.06 s at 2.413782 OW PCMTD2 protein-L-isoaspartate (D-aspartate) O methyltransferase domain containing 2 218826 at 2.41 01262 OW SLC35F2 solute carrier family 35, member F2 212707 s at 2.41 OO726 OW LOC100.133005 similar to RAS p21 protein activator 4 // similar to HSPC047 protein / RAS LOC100134722 p21 protein activator 4 || RAS p21 RASA4 protein activator 4 pseudogene RASA4P 208779 X at 2.4082775 OW DDR1 discoidin domain receptor tyrosine kinase 1 206873 at 2.4066236 OW CA6 carbonic anhydrase VI 221750 at 2.4064143 OW HMGCS1 3-hydroxy-3-methylglutaryl-Coenzyme A synthase 1 (soluble) 212321 at 2.4OSSO9 OW SGPL1 sphingosine-1-phosphate lyase 1 US 2012/0034613 A1 Feb. 9, 2012 48

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 201535 at 2.4043798 OW UBL3 ubiquitin-like 3 231804 at 2.4037292 OW RXFP1 relaxininsulin-like family peptide receptor 1 200703 at 2.4029148 OW DYNLL1 dynein, light chain, LC8-type 1 205968 at 2.402O16 OW KCNS3 potassium voltage-gated channel, delayed-rectifier, Subfamily S, member 3 219856 at 2.4O11526 OW C1orf116 chromosome 1 open reading frame 116 217979 at 2.4OO637 OW TSPAN13 tetraspanin 13 218739 at 2.399S485 OW ABHDS abhydrolase domain containing 5 33646 g at 2.3972.998 OW GM2A GM2 ganglioside activator 211071 s at 2.3966746 OW MLLT11 myeloid lymphoid or mixed-lineage leukemia (trithorax homolog, Drosophila); translocated to, 11 1569157 s at 2.39596.68 OW ZNF846 Zinc finger protein 846 2225.36 s at 2.393.6675 OW ZNF395 Zinc finger protein 395 213051 a 2.393.18 OW ZC3HAV1 Zinc finger CCCH-type, antiviral 1 227335 a. 2.3928O32 OW DIDO1 death inducer-obliterator 1 223265 a 2.3924136 OW SH3BPSL SH3-binding domain protein 5-like 218902 a 2.390648 OW NOTCH1 Notch homolog 1, translocation associated (Drosophila) 201975 a 2.3895.5 OW CLIP1 CAP-GLY domain containing linker protein 1 203515 S. at 2.388.306 OW PMVK phosphomevalonate kinase 202016 a 2.3868265 OW MEST mesoderm specific transcript homolog (mouse) 213280 a 2.38649 OW GARNLA GTPase activating Rap/RangAP domain-like 4 209424 S at 2.381329 OW AMACR alpha-methylacyl-CoA racemase 202720 a 2.3791854 OW TES testis derived transcript (3 LIM domains) 219272 a 2.37771.65 OW TRIM62 tripartite motif-containing 62 223454 a 2.3772516 OW CXCL16 chemokine (C X—C motif) ligand 16 205073 a 2.3759983 OW CYP22 cytochrome P450, family 2, Subfamily J, polypeptide 2 227263 a 2.3751676 OW C8orf58 chromosome 8 open reading frame 58 227725 a. 2.373.7078 OW ST6GALNAC1 ST6 (alpha-N-acetyl-neuraminyl-2,3- beta-galactosyl-1,3)-N- acetylgalactosaminide alpha-2,6- sialyltransferase 1 212830 a. 2.3727896 OW MEGF9 multiple EGF-like-domains 9 201733 a 2.3707964 OW CLCN3 chloride channel 3 224836 a 2.368579 OW TP53INP2 tumor protein p53 inducible nuclear protein 2 213562 s at 2.3671 67 OW SQLE squalene epoxidase 230076 a 2.36426O2 OW PITPNM3 PITPNM family member 3 22788.0 s at 2.363816 OW LOC100.132969 hypothetical protein LOC100132969 /// transmembrane protein 185A TMEM185A 226980 a 2.362776 OW DEPDC1B DEP domain containing 1B 224622 a. 2.3615942 OW TBC1D14 TBC1 domain family, member 14 231861 a 2.3615434 OW LRP10 ow density lipoprotein receptor-related protein 10 23O864 a 2.36OS864 OW MGC421OS serine/threonine-protein kinase NIM1 227868 a 2.35883.43 OW LOC154761 hypothetical LOC154761 228647 a 2.357218 OW LOC100049716 hypothetical protein LOC100049716 223289 s at 2.356582 OW USP38 ubiquitin specific peptidase 38 218816 a 2.3564343 OW LRRC1 eucine rich repeat containing 1 235068 a 2.355547 OW ZDHHC21 Zinc finger, DHHC-type containing 21 212861 a 2.3544.407 OW MFSD5 major facilitator Superfamily domain containing 5 2274.04 s at 2.353249S OW EGR Putative zinc finger protein mRNA, 3' flank 202669 s at 2.353O884 OW EFNB2 ephrin-B2 2025.06 at 2.3506389 OW SSFA2 sperm specific antigen 2 2281.23 s at 2.34971.57 OW ABHD12 abhydrolase domain containing 12 239598 s at 2.3495939 OW LPCAT2 ySophosphatidylcholine acyltransferase 2 201734 at 2.34892S4 OW CLCN3 Chloride channel protein 3 (CLCN3) 220599 s at 2.3469906 OW CARD14 caspase recruitment domain family, member 14 202341 s at 2.346867 OW TRIM2 tripartite motif-containing 2 225,099 at 2.346,118S OW FBXO45 F-box protein 45 225634 at 2.3442137 OW ZC3HAV1 Zinc finger CCCH-type, antiviral 1 US 2012/0034613 A1 Feb. 9, 2012 49

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title

204675 at 2.341.739 OW SRDSA1 steroid-5-alpha-reductase, alpha polypeptide 1 (3-oxo-5alpha-steroid delta 4-dehydrogenase alpha 1) 222774 S at 2.3412967 OW NETO2 neuropilin (NRP) and tolloid (TLL)- ike 2 2O7708 at 2.33811.83 OW ALOXE3 arachidonate lipoxygenase 3 226599 at 2.336998.7 OW FHDC1 FH2 domain containing 1 235475 at 2.3346572 OW LOC100129.720 CDNA clone IMAGE: 5302722 231211 s at 2.3334O2 OW LOCS41469 hypothetical protein LOC541469 202950 at 2.33293.18 OW CRYZ crystallin, Zeta (quinone reductase) 223340 at 2.3317142 OW ATL1 atlastin GTPase 1 236207 at 2.33O8241 OW SSFA2 sperm specific antigen 2 225245 X at 2.328221 OW H2A histone family, member J 1554.122 a. at 2.324.8432 OW hydroxysteroid (17-beta) dehydrogenase 12 222853 at 2.3233314 OW FLRT3 fibronectin leucine rich transmembrane protein 3 227163 at 2.3232682 OW GSTO2 glutathione S-transferase omega 2 205774 at 2.317882 OW F12 coagulation factor XII (Hageman actor) 45288 at 2.31 7875 OW ABHD6 abhydrolase domain containing 6 LOC64363S similar to DEAD H (Asp-Glu-Ala Asp/His) box polypeptide 11 201371 s at 2.31381.96 OW CUL3 cullin 3 214355 X at 2.3121972 OW CTAGE4 CTAGE family, member 4 // CTAGE LOC100142659 family member i? similar to CTAGE6 LOC441294 235987 at 2.3110762 OW PRKXIP1 protein kinase, X-linked, pseudogene 1 200813 s at 2.3108816 OW PAFAH1B1 platelet-activating factor acetylhydrolase, isoform Ib, alpha subunit 45 kDa 214651 s at 2.3090641 OW HOXA9 homeobox A9 201790 s. at 2.3061028 OW DHCR7 7-dehydrocholesterol reductase 1557458 S. at 2.3O3246S OW SHB Src homology 2 domain containing adaptor protein B 202748 a 2.3O2949 OW GBP2 guanylate binding protein 2, interferon inducible 2294.92 a. 2.300059 OW WANGL1 vang-like 1 (van gogh, Drosophila) 2251.08 a 2.2977571 OW AGPS alkylglycerone phosphate synthase 2386.38 a 2.2977488 OW SLC37A2 solute carrier family 37 (glycerol-3- phosphate transporter), member 2 223594 a 2.294.716 OW TMEM117 transmembrane protein 117 225973 a 2.294342 OW TAP2 transporter 2, ATP-binding cassette, sub-family B (MDR/TAP) 217118 s at 2.292SO84 OW chromosome 22 open reading frame 9 210383 a 2.2922642 OW Sodium channel, voltage-gated, type I, alpha subunit 208866 a 2.291897 OW CSNK1A1 casein kinase 1, alpha 1 223349 s at 2.29.13735 OW BOK BCL2-related ovarian killer 207911 s at 2.287757 OW TGM5 transglutaminase 5 212989 a 2.287.0367 OW SGMS1 sphingomyelin synthase 1 225517 a 2.2865.238 OW ZNF770 Zinc finger protein 770 235606 a 2.2857234 OW LOC344595 hypothetical LOC344595 214235 a. 2.2851.86 OW CYP3A5 cytochrome P450, family 3, Subfamily A, polypeptide 5 213430 a. 2.283703 OW RUN and FYVE domain containing 3 219528 s at 2.2819934 OW B-cell CLL/lymphoma 11B (zinc finger protein) 1563805 a. at 2.281725 OW FAM83C family with sequence similarity 83, member C 213546 at 2.276899 OW hypothetical protein DKFZp586I1420 206263 at 2.274217 OW flavin containing monooxygenase 4 225954 s at 2.273436S OW MIDN midnolin 215103 at 2.2728736 OW CYP2C18 cytochrome P450, family 2, Subfamily C, polypeptide 18 227228 s at 2.2723181 OW coiled-coil domain containing 88C 220318 at 2.272O828 OW epsin 3 227981 at 2.271106 OW cytochrome b-561 domain containing 1 202562 s at 2.26963S OW chromosome 14 open reading frame 1 202788 at 2.269064 OW mitogen-activated protein kinase activated protein kinase 3 231166 at 2.265618.6 OW GPR155 G protein-coupled receptor 155 US 2012/0034613 A1 Feb. 9, 2012 50

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 1555.716 a. at 2.2642384 down CXADR coxsackie virus and adenovirus receptor 220966 X at 2.2637703 down ARPCSL actin related protein 2/3 complex, subunit 5-like 232034 at 2.2627313 down LOC2O3274 CDNA FLJ31544 fis, clone NT2RI2OOO865 226021 at 2.2623584 down RDH10 retinol dehydrogenase 10 (all-trans) 209608 s at 2.2597988 down ACAT2 acetyl-Coenzyme A acetyltransferase 2 212978 at 2.2586613 down LRRC8B eucine rich repeat containing 8 family, member B 223288 at 2.2585 614 down USP38 ubiquitin specific peptidase 38 1555905 a. at 22584996 down C3orf23 chromosome 3 open reading frame 23 205109 s at 2.258.3284 down ARHGEF4 Rho guanine nucleotide exchange actor (GEF) 4 20446.5 s at 2.255.3155 down NA internexin neuronal intermediate filament protein, alpha 209679 s at 2.2552O56 down LOCS7228 Small trans-membrane and glycosylated protein 217856 at 2.2544947 down RBM8A RNA binding motif protein 8A 218261 at 2.253576 down AP1M2 adaptor-related protein complex 1, mu 2 subunit 231990 at 2.251771 down USP15 ubiquitin specific peptidase 15 210933 s at 2.2504241 down FSCN1 ascin homolog 1, actin-bundling protein (Strongylocentrotus purpuratus) 2098.72 s at 2.249547 down PKP3 plakophilin 3 205602 X at 2.2482867 down PSG7 pregnancy specific beta-1-glycoprotein 7 22885.1 s at 2.2482166 down ENSA endosulfine alpha 238846 a 2.2471364 down TNFRSF11A tumor necrosis factor receptor Superfamily, member 11a, NFKB activator 1553059 at 2246216S OW PGLYRP3 peptidoglycan recognition protein 3 215513 a 2.2452734 down HYMA hydatidiform mole associated and imprinted (non-protein coding) 231062 a 2.2450545 down LOC100129122 Clone IMAGE: 1257951, mRNA Sequence 204761 a 2.244495 down USP6NL USP6 N-terminal like 219517 a 2.2439237 down ELL3 elongation factor RNA polymerase II ike 3 201791 s at 2.242235 down DHCR7 7-dehydrocholesterol reductase 2100.58 a 2.2416518 down MAPK13 mitogen-activated protein kinase 13 225342 a. 22410355 down AK3L1 adenylate kinase 3-like 1 ff adenylate AK3L2 kinase 3-like 2 210059 s at 2.2387981 down MAPK13 mitogen-activated protein kinase 13 206149 a 2.236605 down CHP2 calcineurin B homologous protein 2 201566 X at 2.2364604 down ID2 inhibitor of DNA binding 2, dominant negative helix-loop-helix protein 223101 s at 2.2357152 down ARPCSL actin related protein 2/3 complex, subunit 5-like 209905 a 2.2349339 down HOXA9 homeobox A9 227383 a 2.2334142 down LOCA2782O hypothetical protein LOC727820 219330 a. 2.2328.598 down WANGL1 vang-like 1 (van gogh, Drosophila) 232103 a 2.229.358 down BPNT1 3'(2), 5'-bisphosphate nucleotidase 1 225300 a 2.2289581 down C15 orf23 chromosome 15 open reading frame 23 229596 a 2.228O157 down AMDHD1 amidohydrolase domain containing 1 232138 a 2.2262597 down MBNL2 Muscleblind-like 2 (Drosophila), mRNA (cDNA clone IMAGE: 4157895) 207236 a 2.2259998 down ZNF345 Zinc finger protein 345 227224 a 2.2250893 down RALGPS2 Ral GEF with PH domain and SH3 binding motif 2 221896 s at 2.224972 OW HIGD1A HIG1 domain family, member 1A 200637 s at 2.2237906 down PTPRF protein tyrosine phosphatase, receptor type, F 214798 at 2.2234876 down ATP2C2 ATPase, Ca++ transporting, type 2C, member 2 226915 s at 2.223121 down ARPCSL actin related protein 2/3 complex, subunit 5-like 213533 at 2.219701 OW D4S234E DNA segment on chromosome 4 (unique) 234 expressed sequence 213577 at 2.218055 down SQLE squalene epoxidase 219956 at 2.2179477 down GALNT6 UDP-N-acetyl-alpha-D- galactosamine:polypeptide N US 2012/0034613 A1 Feb. 9, 2012 51

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title acetylgalactosaminyltransferase 6 (GalNAc-T6) 200632 s at 2.2170808 OW NDRG1 N-myc downstream regulated 1 2024.88 s at 2.217OOOS OW FXYD3 FXYD domain containing ion transport regulator 3 202967 at 2.21596.03 OW GSTA4 glutathione S-transferase alpha 4 209727 at 2.2159023 OW GM2A GM2 ganglioside activator 203081 at 2.2149336 OW CTNNBIP1 catenin, beta interacting protein 1 1555416 a. at 2.214861 OW ALOX1SB arachidonate 15-lipoxygenase, type B 236725 at 2.2132413 OW WWC1 WW and C2 domain containing 1 225726 s at 2.213236 OW PLEKHH1 pleckstrin homology domain containing, family H (with MyTH4 domain) member 1 203625 X at 2.211.56 OW SKP2 CDNA: FLJ22571 fis, clone HSIO2239 206417 a 2.211349 OW CNGA1 cyclic nucleotide gated channel alpha 1 2O7602 a 2.2072852 OW TMPRSS11D transmembrane protease, serine 11D 202951 a 2.2066703 OW STK38 serine/threonine kinase 38 220985 S at 2.2O64672 OW RNF170 ring finger protein 170 205092 X at 2.20531.56 OW ZBTB1 Zinc finger and BTB domain containing 1 226844 a 2.2OSO 638 OW MOBKL2B MOB1, Mps One Binder kinase activator-like 2B (yeast) 226448 a 2.204SS17 OW FAM89A family with sequence similarity 89, member A 227997 a 2.2044806 OW L17RD interleukin 17 receptor D 220225 a. 2.2O3769 OW RX4 iroquois homeobox 4 40524 at 2.2021623 OW PTPN21 protein tyrosine phosphatase, non receptor type 21 202725 a. 2.199355 OW POLR2A polymerase (RNA) II (DNA directed) polypeptide A, 220 kDa 207065 a 2.1987OS OW KRTT5 keratin 75 224746 a 2.1967.638 OW KIAA1522 KIAA1522 202330 s a 2.196532 OW UNG uracil-DNA glycosylase 201735 is a 2.1951.425 OW CLCN3 chloride channel 3 21284.8 s a 2.194746S OW C9orf3 chromosome 9 open reading frame 3 209389 X at 2.194297 OW DBI diazepam binding inhibitor (GABA receptor modulator, acyl-Coenzyme A binding protein) 21718.8 s a 2.19428 OW C14orf1 chromosome 14 open reading frame 1 232843 s a 2.193926 OW DOCK8 dedicator of cytokinesis 8 210749 X at 2.1906087 OW DDR1 discoidin domain receptor tyrosine kinase 1 1569385 S at 2.1899562 OW TET2 et oncogene family member 2 21864.0 s a 2.1887794 OW PLEKHF2 pleckstrin homology domain containing, family F (with FYVE domain) member 2 205535 s a 2.1884.425 OW PCDHT protocadherin 7 203395 s a 2.188OO76 OW HES1 hairy and enhancer of split 1, (Drosophila) 225927 a 2.1876142 OW MAP3K1 mitogen-activated protein kinase kinase kinase 1 44120 at 2.1861. 672 OW ADCK2 aarF domain containing kinase 2 22.5367 a 2.1861136 OW PGM2 phosphoglucomutase 2 202178 a 2.1859634 OW PRKCZ protein kinase C, Zeta 205964 a 2.181216S OW ZNF426 Zinc finger protein 426 224799 a 2.1811337 OW NDFIP2 Nedd4 family interacting protein 2 2091.63 a 2.1807954 OW CYB561 cytochrome b-561 228762 a. 2.180639S OW LFNG LFNG O-fucosylpeptide 3-beta-N- acetylglucosaminyltransferase 214786 a 2.180576 OW MAP3K1 mitogen-activated protein kinase kinase kinase 1 239693 a 2.180439S OW SNX24 Sorting nexin SNX24 (SNX24) 225100 a 2.176683 OW FBXO45 F-box protein 45 212175 s at 2.17664O7 OW AK2 adenylate kinase 2 217744 is at 2.1757214 OW PERP PERP, TP53 apoptosis effector 212831 a 2.1755953 OW MEGF9 multiple EGF-like-domains 9 223.075 s at 2.168681 OW AIF1L, allograft inflammatory factor 1-like 216699 s at 2.1686.67 OW KLK1 kallikrein 1 244546 a 2.1682O29 OW CYCS cytochrome c, somatic 213365 a. 2.1674378 OW ERI2 exoribonuclease 2 201215 a. 2.1666453 OW PLS3 plastin 3 (T isoform) 203881 s at 2.165027 OW DMD dystrophin US 2012/0034613 A1 Feb. 9, 2012 52

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 208854 S at 2.1649718 OW STK24 serine/threonine kinase 24 (STE20 homolog, yeast) 212944 a 2.1649513 OW SLC5A3 solute carrier family 5 (sodium/myo inositol cotransporter), member 3 205416 s at 2.1644816 OW ATXN3 ataxin 3 228378 a 2.164461 OW C12Orf29 chromosome 12 open reading frame 29 219229 a 2.162O889 OW SLCO3A1 Solute carrier organic anion transporter family, member 3A1 218657 a 2.1619468 OW RAPGEFL1 Rap guanine nucleotide exchange factor (GEF)-like 1 204005 s at 2.161023 OW PAWR PRKC, apoptosis, WT1, regulator 208345 s at 2.16O1677 OW POU3F1 POU class 3 homeobox. 1 209733 a 2.160098 OW LOC286440 hypothetical protein LOC286440 225136 a 2.1593971 OW PLEKHA2 pleckstrin homology domain containing, family A (phosphoinositide binding specific) member 2 201565 s at 2.1581 004 OW ID2 inhibitor of DNA binding 2, dominant negative helix-loop-helix protein 235148 at 2.1576118 OW KRTCAP3 keratinocyte associated protein 3 201820 at 2.1536OO7 OW KRTS keratin 5 226269 at 2.1534703 OW GDAP1 ganglioside-induced differentiation associated protein 1 218705 s at 2.152O2O2 OW SNX24 Sorting nexin 24 235896 s at 2.1518.188 OW SMCR7 Smith-Magenis syndrome chromosome region, candidate 7 1559078 at 2.1510541 OW BCL11A CDNA FLJ58516 complete cols, highly similar to B-cell lymphoma leukemia 11A 223214 S at 2.14984O6 OW ZHX1 Zinc fingers and homeoboxes 1 229515 at 2.1496.76 OW PAWR CDNA clone IMAGE: 3892.559 209815 at 2.14831 OW PTCH1 patched homolog 1 (Drosophila) 227478 at 2.1481898 OW SETBP1 SET binding protein 1 238686 at 2.1473422 OW FBXO3 F-box protein 3 202838 at 2.1467884 OW FUCA1 lucosidase, alpha-L-1, tissue 209598 at 2.1460617 OW PNMA2 paraneoplastic antigen MA2 214975 s at 2.1457.093 OW MTMR1 myotubularin related protein 1 203276 at 2.1456895 OW LMNB1 amin B1 203431 s at 2.1429577 OW RICS Rho GTPase-activating protein 1554445 at 2.1428692 OW ZNF85 Zinc finger protein 85 208051 s at 2.141477 OW PAIP1 poly(A) binding protein interacting protein 1 225282 a 2.1402.11 OW SMAP2 Small AirfAP2 2384.51 a 2.1400113 OW MPP7 membrane protein, palmitoylated 7 (MAGUK p55 subfamily member 7) 201079 a 2.1399.426 OW SYNGR2 synaptogyrin 2 225878 a 2.1396.25 OW KIF1B kinesin family member 1B 213693 s at 2.138388 OW MUC1 mucin 1, cell Surface associated 218421 a 2.13 7857 OW CERK ceramide kinase 219090 a 2.13771.68 OW SLC24A3 solute carrier family 24 (sodium potassium calcium exchanger), member 3 226071 a 2.1368.642 OW ADAMTSL4 ADAMTS-like 4 219410 a 2.1362329 OW TMEM4SA transmembrane protein 45A 235200 a 2.1350272 OW ZNF561 Zinc finger protein 561 219459 a 2.1346,684 OW POLR3B polymerase (RNA) III (DNA directed) polypeptide B 225313 a 2.1338372 OW C20orf177 chromosome 20 open reading frame 177 243963 a 2.1331.64 OW SDCCAG8 Serologically defined colon cancer antigen 8, mRNA (cDNA clone IMAGE: 53.01251) 210958 s at 2.1311.638 OW LOC100128443 hypothetical protein LOC100128443/// A. MAST4 microtubule associated serine/threonine kinase family member 4 210609 s at 2.12971.8 OW TP53I3 tumor protein p53 inducible protein 3 209949 at 2.1277685 OW NCF2 neutrophil cytosolic factor 2 224055 x at 2.1269937 OW KCNK7 potassium channel, Subfamily K, member 7 232127 at 2.126492S OW CLCNS chloride channel 5 223455 at 2.126378 OW TCHP trichoplein, keratin filament binding 209512 at 2.1262.662 OW HSDL2 hydroxysteroid dehydrogenase like 2 207206 s at 2.1244593 OW ALOX12 arachidonate 12-lipoxygenase US 2012/0034613 A1 Feb. 9, 2012 53

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 1558924 S at 2.1232O33 OW CLIP1 CAP-GLY domain containing linker protein 1 216268 s at 2.12.27787 OW JAG1 jagged 1 (Alagille syndrome) 215100 at 2.1208134 OW C6orf1 OS chromosome 6 open reading frame 105 229685 at 2.12O31.96 OW LOC100134937 hypothetical LOC100134937 202263 at 2.1.194O17 OW CYB5R1 cytochrome b5 reductase 1 212136 at 2.1178362 OW ATP2B4 ATPase, Ca++ transporting, plasma membrane 4 225.308 s at 2.11776 OW TANC1 etratricopeptide repeat, ankyrin repeat and coiled-coil containing 1 224865 at 2.1171.43 OW FAR1 atty acyl CoA reductase 1 228954 at 2.1161342 OW LYSMD4 LysM, putative peptidoglycan-binding, domain containing 4 1563.088 a. at 2.1154768 OW LOC284.837 hypothetical protein LOC284837 222849 s at 2.1153836 OW SCRN3 secernin 3 1568815 a. at 2.1149223 OW DDXSO DEAD (Asp-Glu-Ala-Asp) box polypeptide 50 1554608 at 2.1143405 OW TGOLN2 trans-golgi network protein 2 206.857 s at 2.1133657 OW FKBP1B FK506 binding protein 1B, 12.6 kDa 213764 s at 2.1121294 OW MFAPS microfibrillar associated protein 5 21855.9 s at 2.1.115992 OW MAFB V-maf musculoaponeurotic fibrosarcoma oncogene homolog B (avian) 203609 s at 2.11.06138 OW ALDHSA1 aldehyde dehydrogenase 5 family, member A1 214605 X at 2.1089497 OW GPR1 G protein-coupled receptor 1 203888 at 2.108670S OW THEBD thrombomodulin 215549 x at 2.1083808 OW CTAGE4 CTAGE family, member 4 24-1990 at 2.1072762 OW RHOV ras homologgene family, member V 229955 at 2.1066082 OW FBXO3 F-box protein 3, mRNA (cDNA clone IMAGE: 5296,662) 222396 at 2.10537 OW HN1 hematological and neurological expressed 1 211203 s at 2.1OSO36S OW CNTN1 contactin 1 209234 at 2.104905 OW KIF1B kinesin family member 1B 1007 s at 2.10379 OW DDR1 discoidin domain receptor tyrosine kinase 1 205909 at 2.103594 OW POLE2 polymerase (DNA directed), epsilon 2 (p59 subunit) 205.248 at 2.102.7164 OW DOPEY2 dopey family member 2 229958 at 2.102053 OW CLN8 ceroid-lipofuscinosis, neuronal 8 (epilepsy, progressive with mental retardation) 222256 s at 2.101.3858 OW MUD7 jumonji domain containing 7 MD7- MJD7-PLA2G4B readthrough PLA2G4B .. transcript phospholipase A2, group PLA2G4B VB (cytosolic) 212089 at 2.1OOO233 OW LMNA aminAC 206429 at 2.0988882 OW F2RL1 coagulation factor II (thrombin) receptor-like 1 1559094 at 2.098.726 OW FBXO9 F-box protein 9 238697 at 2.0983906 OW NCRNA00O86 Hypothetical protein MGC39606, mRNA (cDNA clone MGC: 33489 MAGE: 4813443) 222904 S at 2.096099 OW TMC5 transmembrane channel-like 5 225182 at 2.095S298 OW TMEMSOB transmembrane protein 50B 203575 at 2.094.8935 OW CSNK2A2 casein kinase 2, alpha prime polypeptide 211922 s at 2.094567 OW CAT catalase 1553982 a. at 2.094S106 OW RAB7B RAB7B, member RAS oncogene amily 201839 s at 2.09322O2 OW EPCAM epithelial cell adhesion molecule 47560 at 2.0923448 OW LPHN1 atrophilin 1 212186 at 2.0908222 OW ACACA acetyl-Coenzyme A carboxylase alpha 219187 at 2.09.01995 OW FKBPL FK506 binding protein like 241946 at 2.0896347 OW ZDHHC21 Zinc finger, DHHC-type containing 21 1553021 S at 2.0895314 OW BICD2 bicaudal Dhomolog 2 (Drosophila) 203002 at 2.089333S OW AMOTL2 angiomotin like 2 20386.5 s at 2.08908 OW ADARB1 adenosine deaminase, RNA-specific, B1 (RED1 homolog rat) 200696 s at 2.0890284 OW GSN gelsolin (amyloidosis, Finnish type) 204967 at 2.0882254 OW SHROOM2 shroom family member 2 202894 at 2.0872O73 OW EPHB4 EPH receptor B4 US 2012/0034613 A1 Feb. 9, 2012 54

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title 1558292 s at 2.084S475 OW PIGW phosphatidylinositol glycan anchor biosynthesis, class W 235567 a 2.0833743 OW RORA Hypothetical protein LOC283666, mRNA (cDNA clone MAGE: 4750925) 212991 a 2.083101S OW FBXO9 F-box protein 9 216718 a 2.0828SO7 OW C1orfA6 chromosome 1 open reading frame 46 1558152 at 2.081475 OW LOC100131262 hypothetical LOC100131262 225211 a 2.08O33 OW PVRL1 poliovirus receptor-related 1 (herpesvirus entry mediator C) 202987 a 2.0792933 OW TRAF3IP2 TRAF3 interacting protein 2 222236 s at 2.0789578 OW ASAP3 Arf6AP with SH3 domain, ankyrin repeat and PH domain 3 239230 a. 2.0784.454 OW HESS hairy and enhancer of split 5 (Drosophila) 2025O1 a 2.0782366 OW MAPRE2 microtubule-associated protein, RP/EB amily, member 2 242255 a. 2.078182S OW LOC100.130837 CDNA clone IMAGE: 4799914 202587 s at 2.0776887 OW AK1 adenylate kinase 1 201032 a. 2.0772772 OW BLCAP bladder cancer associated protein 202850 a 2.076881 OW ABCD3 ATP-binding cassette, Sub-family D (ALD), member 3 225618 a 2.0766106 OW ARHGAP27 Rho GTPase activating protein 27 203666 a 2.0743.63 OW CXCL12 chemokine (C X—C motif) ligand 12 (stromal cell-derived factor 1) 218148 a 2.0743272 OW CENPT centromere protein T 203777 s at 2.0739517 OW RPS6KB2 ribosomal protein S6 kinase, 70 kDa, polypeptide 2 226929 a 2.0733O16 OW MTHFR 5,10-methylenetetrahydrofolate reductase (NADPH) 225998 a 2.0714586 OW GAB1 GRB2-associated binding protein 1 222843 a 2O714424 OW FIGNL1 fidgetin-like 1 225598 a 2.0712779 OW SLC45A4 solute carrier family 45, member 4 223213 s at 2.0709321 OW ZHX1 Zinc fingers and homeoboxes 1 39729 at 2.0675848 OW PRDX2 peroxiredoxin 2 235174 s at 2.0673625 OW LOC100128822 hypothetical protein LOC100128822 21918.4 x at 2.0669382 OW TIMM22 translocase of inner mitochondrial membrane 22 homolog (yeast) 207023 x at 2.066204 OW KRT10 keratin 10 227739 at 2.0635765 OW LOC648.245 hypothetical LOC648245 218021 at 2.0624394 OW DHRS4 dehydrogenase/reductase (SDR family) DHRS4L2 member 4 if dehydrogenase/reductase (SDR family) member 4 like 2 202342 s at 2.061941 OW TRIM2 tripartite motif-containing 2 203332 S at 2.06.0985 OW NPP5D inositol polyphosphate-5-phosphatase, 45 kDa 227255 at 2.058867 OW PDIK1L PDLIM1 interacting kinase 1 like 219968 at 2.0585458 OW ZNF589 Zinc finger protein 589 210045 at 2.058517 OW DH2 isocitrate dehydrogenase 2 (NADP+), mitochondrial 211070 x at 2.OS 63624 OW DB diazepam binding inhibitor (GABA receptor modulator, acyl-Coenzyme A binding protein) 200653 s at 2.OSS8417 OW CALM1 calmodulin 1 (phosphorylase kinase, CALM2 delta) i? calmodulin 2 (phosphorylase CALM3 kinase, delta) i? calmodulin 3 (phosphorylase kinase, delta) 200884 at 2.0550563 OW CKB creatine kinase, brain 227384 S at 2.054656 OW LOCA2782O hypothetical protein LOC727820 2014O9 s at 2.OS4OSO2 OW PPP1CB protein phosphatase 1, catalytic subunit, beta isoform 205184 at 2.OS36096 OW GNG4 guanine nucleotide binding protein (G protein), gamma 4 1554600 S. at 2.0529914 OW LMNA aminAC 201474 S at 2.051571 OW ITGA3 integrin, alpha 3 (antigen CD49C, alpha 3 subunit of VLA-3 receptor) 204633 s at 2.OSOS228 OW RPS6KAS ribosomal protein S6 kinase, 90 kDa, polypeptide 5 212443 at 2.OSO3902 OW NBEAL2 neurobeachin-like 2 210608 s at 2.0497348 OW FUT2 lucosyltransferase 2 (Secretor status included) US 2012/0034613 A1 Feb. 9, 2012 55

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title

1557036 at 2.0495107 OW Zinc finger and BTB domain containing 1 (ZBTB1), transcript variant 1 mRNA 207950 S at 2.0490613 OW ANK3 ankyrin 3, node of Ranvier (ankyrin G) 41858 at 2.0485.191 OW FRAG1 FGF receptor activating protein 1 234513 at 2.0481179 OW ELOVL3 elongation of very long chain fatty acids (FEN1/Elo2, SUR4/Elo3, yeast)- like 3 223312 at 2.0474.238 OW C2Orf7 chromosome 2 open reading frame 7 204137 at 2.045931 OW GPR137B G protein-coupled receptor 137B 229732 at 2.0447SOS OW ZNF823 Zinc finger protein 823 206600 s at 2.04438.23 OW LOC10O133772 similar to MCT // solute carrier family 6, member 5 (monocarboxylic acid SLC16AS transporter 6) 209.099 X at 2.0436616 OW JAG1 jagged 1 (Alagille syndrome) 221868 a 2.043.276 OW PAIP2B poly(A) binding protein interacting protein 2B 202790 a 2.0431542 OW CLDN7 claudin 7 227 272 a 2.0418959 OW C1 Sorf52 chromosome 15 open reading frame 52 200950 a 2.041765 OW ARPC1A actin related protein 2/3 complex, subunit 1A, 41 kDa 226413 a 2.04O7355 OW LOC4OOO27 hypothetical gene Supported by BCO47417 202039 a 2.04.00221 OW MYO18A myosin XVIIIA // TGFB1-induced TIAF1 anti-apoptotic factor 1 233528 s at 2.0383992 OW LOC652968 hypothetical protein LOC652968 211993 a 2.036943 OW WNK1 WNK deficient protein kinase 1 222668 a 2.0368426 OW KCTD15 potassium channel tetramerisation domain containing 15 204718 a 2.0357015 OW EPHB6 EPH receptor B6 202192 s at 2.0352569 OW GAS7 growth arrest-specific 7 208652 a. 2.035.0847 OW PPP2CA protein phosphatase 2 (formerly 2A), catalytic Subunit, alpha isoform 226388 a 2.034O674 OW TCEA3 transcription elongation factor A (SII), 3 226104 a 2.033753 OW RNF170 ring finger protein 170 22416O is at 2.0337167 OW ACAD9 acyl-Coenzyme A dehydrogenase amily, member 9 20912S a 2.033617 OW KRT6A keratin 6A 209529 a 2.0331414 OW PPAP2C phosphatidic acid phosphatase type 2C 226860 a 2.0323,775 OW TMEM19 transmembrane protein 19 226644 a 2.0322123 OW MIB2 mindbomb homolog 2 (Drosophila) 203747 a 2.0315.192 OW AQP3 aquaporin 3 (Gill blood group) 1553960 at 2.0313072 OW SNX21 Sorting nexin family member 21 65630 at 2.0311613 OW TMEM80 transmembrane protein 80 218171 a 2.O30586 OW VPS4B vacuolar protein sorting 4 homolog B (S. cerevisiae) 236863 a 2.0298839 OW C17orf67 chromosome 17 open reading frame 67 205293 X at 2.0297394 OW BALAP2 BAI1-associated protein 2 231115 a. 2.0290804 OW POLH polymerase (DNA directed), eta 209563 X at 2.0285957 OW CALM1 calmodulin 1 (phosphorylase kinase, CALM2 delta) i? calmodulin 2 (phosphorylase CALM3 kinase, delta) i? calmodulin 3 (phosphorylase kinase, delta) 205668 at 2.0280306 OW LY75 ymphocyte antigen 75 204347 at 2.025962 OW AK3L1 adenylate kinase 3-like 1 ff adenylate AK3L2 kinase 3-like 2 222482 at OW LOC100131851 hypothetical protein LOC100131851 /// hypothetical protein LOC100134497 fif LOC10O134497 similar to single stranded DNA binding protein 3 fill hypothetical LOC646674 LOC4O10O2 fit single stranded DNA binding protein 3 LOC646674 II. SSBP3 220056 at 2.023061 down L22RA1 interleukin 22 receptor, alpha 1 211986 at 2.022835 down AHNAK AHNAK nucleoprotein 1555097 a. at 2.022212S down PTGFR prostaglandin F receptor (FP) 225.684 at 2.0216835 down FAM33A family with sequence similarity 33, member A 209426 s at 2.0213833 down AMACR alpha-methylacyl-CoA racemase 203528 at 2.02.11873 down SEMA4D Sema domain, immunoglobulin domain (Ig), transmembrane domain (TM) and US 2012/0034613 A1 Feb. 9, 2012 56

TABLE 1-continued

Fold Probe Set ID change Direction Gene Symbol Gene Title short cytoplasmic domain, (semaphorin) 4D 222895 s at 2.021075 OW BCL11B B-cell CLL/lymphoma 11B (zinc finger protein) 230836 at 2.02O9422 OW ST8SIA4 ST8 alpha-N-acetyl-neuraminide alpha-2,8-Sialyltransferase 4 224443 at 2.0197344 OW C1orf7 chromosome 1 open reading frame 97 217845 X at 2.017.0114 OW HIGD1A HIG1 domain family, member 1A 1555964 at 2.O155482 OW ARL17 ADP-ribosylation factor-like 17 fif ARL17P1 ADP-ribosylation factor-like 17 pseudogene 1 204061 a 20146167 OW PRKX protein kinase, X-linked 213572 s at 2.O136144 OW SERPINB1 Serpin peptidase inhibitor, clade B (ovalbumin), member 1 218218 a 2.0132878 OW APPL2 adaptor protein, phosphotyrosine interaction, PH domain and leucine Zipper containing 2 1554588 a. at 2.0109353 OW TTC3OB tetratricopeptide repeat domain 30B 201563 a 2.0097337 OW SORD Sorbitol dehydrogenase 39313 at 20081642 OW WNK1 KIAA0344 gene 225551 a 2.OOS9721 OW C1orf71 chromosome 1 open reading frame 71 52285 f at 2.OO37622 OW CEP76 centrosomal protein 76 kDa 207593 a 2.0035.08 OW ABCG4 ATP-binding cassette, Sub-family G (WHITE), member 4 212115 a. 2.OO3O887 OW HN1L, hematological and neurological expressed 1-like 224605 a 2.0020971 OW C4orf3 chromosome 4 open reading frame 3 15588.82 at 2002O864 OW LOC4O1233 similar to HIV TAT specific factor 1; cofactor required for Tat activation of HIV-1 transcription 21775.2 s at 2.001705 OW CNDP2 CNDP dipeptidase 2 (metallopeptidase M20 family) 204241 at 2001.2212 OW ACOX3 acyl-Coenzyme A oxidase 3, pristanoyl 219929 s at 2.0005429 OW ZFYVE21 Zinc finger, FYVE domain containing 21

Salicin 0.5% ws. Untreated Control 24 hr treatment N = 7 Salicin 0.5%, N=4 Control RMA normalization (performed on all 42 chips together) t-test with Benjamini and Hochberg FDR correction p-value cut-off; 0.05 Fold change cutoff: 2.0

Selecting Second Subset of Genes Table 1. (See also, FIG. 7 at 752). In some embodiments the biochemical pathway associated with physical appearance of 0091 Because of the large amount of data associated with skin aging comprises at least one of skin structural protein the gene expression level changes shown in Table 1, it is synthesis, degradation and maintenance, extracellular matrix desirable to provide a further focus on genes associated with assembly, cellular differentiation, skin barrier component skin and aging. In one embodiment, further data sets extracted synthesis, skin barrier integrity, water regulation, or regula from the literature identify genes associated with physical tion of melanin production and control. Structural protein skin aging attributes based on current knowledge of the bio synthesis includes, for example, elastin formation, kerati chemical pathways in skin to define functional youth gene nocyte differentiation and collagen production. Skin barrier assemblies. Such an assembly may then provide a gene component synthesis includes, for example, hyaluronic acid expression focus for future further work on the same agent synthesis and lipid synthesis. Regulation of melanin produc used to get the initial full genome data set or for other agents. tion and control includes, for example, UV induced pigmen For this next step, the method first uses a data set that identi tation and inhibition of tyrosinase. The data set on biochemi fies particular genes associated with biochemical pathways in cal pathways associated with the physical appearance of skin skin 118 (see FIG. 1C). aging known for selected genes is preferably collected and 0092 Data about the biochemical pathways of genes are stored in database 730 in a format that promotes an intersec available from many sources of scientific literature, including tion analysis with the data of Table 1. This can be done by databases of journal articles or from available unpublished building a table of all genes known to be associated with a data. To make it more useful in the present system, data biochemical pathway associated with the physical appear collected may be Supplemented with metadata classifying the ance of skin aging and finding its intersection with Table 1, or conclusions reached in terminology or coding that clearly by starting the literature search with the genes in Table 1, associates genes with skin or particular skin attributes. (See which have already met the fold change criterion. FIG. 7 at 786). In some embodiments, research is done on 0093. However approached, the intersection analysis of biochemical pathways of the skin for any of the genes from this step reduces the data of Table 1, by selecting from the first US 2012/0034613 A1 Feb. 9, 2012 57

Subset of genes a second Subset of genes associated with the tion analysis to have a biochemical pathway associated with selected, identified biochemical pathways associated with the the physical appearance of skin aging. 0094. The genes in the second subset for actual test results physical appearance of skin aging. With reference to the are categorized according to biological function or associa simplified example of FIG. 1C of the method, the method tion with a plurality of biochemical pathways associated with selects from genes (b, c, e, f, gandh) based on a biochemical the physical appearance of skin aging. For the experimental pathway data set derived from review of scientific literature data for 0.05% salicin treated cultures (N=7) compared with 120. For example, if in the biochemical pathway data set the untreated control cultures (N=4), Table 2 (see also, FIG. 7 at gene “h” has no apparent association with a biochemical 754) shows a list of genes derived from Table 1, by selecting pathway associated with the physical appearance of skin from Table 1 approximately 200 genes identified in a data set by their relationship to human skin in the scientific literature. aging, it may be excluded from the second Subset at this point. 0.095 For each gene listed in Table 2, a reference is pro The resulting genes in the second Subset in the simplified vided which discusses the mechanism of action/biochemical example would be (b. c. e. f and g) as shown at 211. These pathway of the gene. The references are incorporated herein genes correspond to hypothetical genes found in the intersec by reference.

TABLE 2

Fold Gene Probe Set ID change Direction Symbol Reference 225337 at 2.704214 up ABHD2 Genes to Cells (2009) 14,407-424 200965 S at 11.87853 down ABLIM1 Exp Gerontol. 2006 Apr; 41(4): 387-97 202422 S at 3.7855804 up ACSL4 Arch Biochem Biophys. 1987 Sep: 257(2): 302-14 214913 at 2.1450849 up ADAMTS3 Biol Chem. 2001 Aug 24; 276(34): 31502-9 202053 S at 4.3994246 down ALDH3A2 Mol Genet Metab. 2007 Jan; 90(1): 1-9. 1555416 a. at 2.214861 down ALOX1SB ournal of Investigative Dermatology (1991) 97,291-297; doi:10.1111,1523-1747 203747 at 2.0315192 down AQP3 Biol Cell. 2005 Jul: 97(7): 479-86 228082 at 2.1080198 up ASAM Am J Respir Cell Mol Biol. 2007 Aug: 37(2): 169-85 1554980 a. at 2.623143 up ATF3 Pollack BP, Exp Dermatol. Activating transcription factor 3 (ATF3) expression is increased in erythema multiforme and is regulated by IFN gamma in human keratinocytes 205410 s at 3.984.1306 up ATP2B4 (2008) Lamellar Bodies of Human Epidermis, Molecular & Cellular Proteomics 7.11, 2151-2175 1558143 a at 2.346621 up BCL2L11 Med Invest. 2008 Aug. 55 (3–4): 204-10 206176 at 7.765054 up BMP6 Exp Cell Res. 2001 Feb 5:263(2): 265-73 209563 x a 2.0285957 down CALM1 Arch Dermatol Res. CALM2 993; 285(5):310-1 CALM3 210020 x a 11.268426 down CALML3 Arch Dermatol Res. 993; 285(5):310-1 220414 at 8.748902 down CALMLS Arch Dermatol Res. 993; 285(5):310-1 209790 S a 2.6684937 down CASP6 Fayi Xue Za Zhi. 2007 Oct; 23(5): 325-7,331 211922 s a 2.094567 down CAT ournal of Investigative Dermatology (2006) 126, 182-190 2064.07 s a 5.516013 up CCL13 Allergy Clin Immunol. 2009 Oct; 124(4): 753-60.e1. 216598 s a 5.407172 up CCL2 Allergy Clin Immunol. 2009 Oct; 124(4): 753-60.e1. 206193 s a 11.105.947 down CDSN FASEB J. 1996 Jun; 10(8): 871-81. 222549 at 7.7580996 down CLDN1 Drugs Dermatol. 2007 Jun; 6(6 Suppl): S20-4. 201428 at 3.4539902 down CLDN4 Skin Pharmacol Physiol. 2006; 19(2):71-7. Epub 2006 May 9 205830 at 2.48.58525 up CLGN Cell. 2008 Apr 18; 133(2): 223-34 224329 s at 5.4422174 down CNFN Biochem Biophys Res Commun. 2004 Jun 11; 318(4): 803-13 204636 at 3.5764592 down COL17A1 http://ghr.nlm.nih.gov/gene=col17a1 211981 at 2.121121 up COL4A1 Matrix Biol. 1998 Aug. 17(4): 279-91. 213992 at 4.8003573 down COL4A6 The Journal of Cell Biology, Vol 130, 1219-1229, Copyright (C) 1995 205931 s at 9.909108 up CREBS FEBS Lett. 2002 Jul 31; 524(1-3): 193-8 US 2012/0034613 A1 Feb. 9, 2012 58

TABLE 2-continued

Fold Gene Probe Set ID change Direction Symbol Reference 229228 a 9.767824 up CREBS FEBS Lett. 2002 Jul 31; 524(1-3): 93-8 210229 s at 16.5SO27 lb CSF2 Thromb Haemost. 2004 Aug: 92(2): 262-74 207442 a. 11.091.33 up CSF3 Dermatol Sci. 2001 Apr. 25(3): 179-88 209774 X at 7.546.0076 lb CXCL2 nvest Dermatol. 2007 May: 127(5): 1264-6 210764 S at 2.6922395 up CYR61 Biomed Sci. 2002 Jan.-Feb; 9(1): 59-67. 206806 a 2.7395847 l. DGKI PNAS May 24, 2005 vol. 102 no. 21 7595-76OO 203810 a 2.0534089 up DNAJB4 Photodermatol Photoimmunol Photomed. 2004 Jun; 2003): 129-37 202843 a 3.56OO665 up DNAB9 Mol Cancer Ther. 2008 Aug. 7 (8): 2319-27. 221782 a 2.SS2S873 lb DNAJC10 Photodermatol Photoimmunol Photomed. 2004 Jun; 2003): 129-37 207324 S at 12.947083 down DSC1 FASEB J. 1996 Jun; 10(8): 871-81. 206033 s at 4.676169 down DSC3 FASEB J. 1996 Jun; 10(8): 871-81. 244852 a 2.2514307 up DSEL Apr. 10, 2009 The Journal of Biological Chemistry, 284, 9788-9795 200606 a 2.84.82468 down DSP FASEB J. 1996 Jun; 10(8): 871-81. 20104.4 x at 3.7260537 DUSP United States Patent Application 2006O1163.19 204273 a 3.91577OS EDNRB Clinics in Dermatology, Volume 23, ssue 1, January-February 2005, Pages 56-67 214446 a 2.SS70273 lb ELL2 DNA Repair (Amst). 2007 Jun ;6(6): 841-51. Epub 2007 Mar 19 234513 a 2.0481179 down ELOVL3 Biol Chem. 2004 Feb 3:279(7): 5621-9. Epub 2003 Oct 27 204256 a 3.1214519 down ELOVL6 Prog Lipid Res. 2006 May: 45(3): 237-49 201839 s at 2.0932202 down EPCAM PLoS Genet. 2009 ul; 5(7): e1000563. Epub 2009 Jul 17 232165 a 12.401502 down EPPK FASEB J. 1996 Jun; 10(8): 871-81. 202609 a 2.334194 up EPS8 Am J Pathol. 2000 Jul: 157(1): 59-68. 205767 a 2.486.593 up ERE Biol Chem. 2000 Feb 25; 275 (8): 5748-53. 206429 a 2.0988882 down F2RL Proc AssocAm Physicians. 1997 Mar: 109(2): 190-207 203980 a 2.2506835 up FABP4 FEBS Lett. 2009 Apr 7:583(8): 1319-22. 208962 s at 2.88.9333 up FADS1 ournal of Investigative Dermatology 29, 2795-2804 (December 2009) 203184 a 2.0282688 up FBN2 nvest Dermatol. 1996 May: 106(5): 1090-5 227 271 a 2.6716337 down FGF11 ournal of Endocrinology (2005) 86,273-289 DOI: 0.1677.joe.1.06055 205110 s at 2.1575923 up FGF13 nvest Dermatol. 2004 May: 122(5): 1084-90. 204421 S at 3.8445234 up FGF2 nt J Cosmet Sci. 2009 Dec. 31(6): 419-26. 1555103 s at 2.3495677 up FGF7 Dermatol Sci. 2009 May: 54(2): 106-13. 205782 at 3.1584346 up FGF7 Dermatol Sci. 2009 May: 54(2): 106-13. 1554741 s at 2.5060341 up FGF7 Huang, T.-J., Lee, C.-J., Expression of KGFLP1 i? keratinocyte growth factor (KGF) is KGFLP2 regulated by two KGF-like proteins, KGFLP1 and KGFLP2. Submitted APR-2002 205014 at 3.40602 down FGFBP1 Biol Chem. 2000 Apr 4; 275 (15): 10802-11. 204379 s at 5.287966 down FGFR3 . Clin. Invest. 116:8 doi:10.1172 JCI281 63 204135 at 2.9959981 down FILIP1L, Nature Genetics 33,487-491 (2003) Published online: March 2003: doi:10.1038/ng1119 1569410 at 45.91465 down FLG2 PLoS One. 2009;4(4): e5227. 210287 s at 3.9503276 up FLT1 J Invest Dermatol. 2007 Oct; 127(10): 2445-52. 214702 at 2.70994.62 up FN1 Forensic Science International, Volume 126, Issue 2, Page 118 21591.0 s at 3.663163 up FNDC3A FEBS Lett. 1994 Apr 18; 343(1): 47-50 US 2012/0034613 A1 Feb. 9, 2012 59

TABLE 2-continued

Fold Gene Probe Set ID change Direction Symbol Reference 202724 s at 2.0982356 up FOXO1 Investig Dermatol Symp Proc. 2009 Aug; 14(1):60-2. 2096O2 s at 15.703371 down GATA3 BMC Genomics. 2009 Sep 7: 10:417. 221577 X at 5.573193 up GDF15 Oncogene. 2008 Jan 17:27(4): 409-20. 2354.05 at 3.2997503 down GSTA4 nt J Biochem Cell Biol. 1995 Mar; 27(3): 271-7 225245 X at 2.328221 down H2AF Exp Gerontol. 1999 Sep: 34(6): 741-54 206643 at 55.867226 down HAL Dermatol Sci. 2008 Jun; 50(3): 209-15. Epub 2008 Feb 15. 223541 at 3.713034 down HAS3 Arch Dermatol Res. 2006 Nov. 298(6): 273-82 210998 s at 4.350414 up HGF Cytokine. 2000 Jun; 12(6): 780-5. 221750 at 2.4064143 down HMGCS1 Clin Chim Acta. 1988 Jan 5; 171(1): 95-101 222881 at 4.1936674 down HPSE ournal of Investigative Dermatology (2001) 117, 1266-1273 227361 at 3.409933 up HS3ST3B1 Biochem Biophys Res Commun. 2008 Aug 8: 372(4): 681-7 202558 s at 3.1297736 up HSPA13 Cell Stress Chaperones. 2009 an; 14(1): 1-21. 200800 s at 3.1119442 up HSPA1A // Cell Stress Chaperones. 2009 HSPA1B an; 14(1): 1-21. 213418 a 14.241181 up HSPA6 Cell Stress Chaperones. 2009 Oct 7. 221667 s at 4.658692 down HSPB8 Oncogene. 2007 May 24; 26(24): 3521-31 219284 a 2.0798855 up HSPBAP1 BrJ Dermatol. 1995 Aug. 133(2): 194-202 210619 s at 4.5378346 down HYAL1 ournal of Investigative Dermatology (2007) 127,512-513 220249 a 3.116939 down HYAL4 Invest Dermatol. 1994 Mar: 102(3): 385-9 202411 a 4.671116 down FI27 ournal of Investigative Dermatology (2004) 122, 717-721 211958 a 2.71 18793 up GFBP5 Biochem Soc Trans. 2009 Aug. 37(Ptá): 882-5. 206924 a 9.694227 up L11 Allergy Clin Immunol. 2003 Apr. 111(4): 875-81 205992 S at 4.32588 up L15 Immunol. 1995 Nov 1: 155(9): 4492-6. 206295 a 19.309084 down L18 Arch Dermatol Res. 2001 ul; 293(7): 325-33 206618 a 2.864942 up L18R1 Autoimmun Rev. 2009 Sep; 9(1): 45-8 220745 a. 2.2776432 up L19 Invest Dermatol. 2003 Dec. 121(6): 1306-11. 221470 s at 100.54604 down L1F7 Wound Repair Regen. 2008 Jul-Aug: 6(4): 534-41. 205403 a 3.2927191 down L1R2 ournal of Investigative Dermatology (1996) 106, 1102-1107; doi:10.1111,1523-1747 207526 S. at 9.089992 up L1RL1 Hum Mol Genet. 2005 Oct ; 14(19): 2919-27 219115 s at 12.066238 down L2ORA Genes Immun. 2008 Jul: 9(5): 445-51. 228575 at 10.042443 down L2ORB Genes Immun. 2008 Jul: 9(5): 445-51. 220056 at 2.023.061 down L22RA1 Clin Exp Immunol. 2007 December; 50(3): 407-415. doi: 0.11111.1365-2249.2007.03511.x 220054 at 2.967118 up L23A Genes Immun. 2009 Apr; 10(3): 201-9 206569 at 8.736957 up L24 Cytokine. 2008 Jan; 41(1): 16-23 205207 at 3.1970735 up L6 ournal of Investigative Dermatology (2004) 123, 124-131 207008 at 11.107841 down L8RB Lab Invest 2000, 80: 595-604 201474 s at 2.051571 down TGA3 Clin Invest. 2008 Mar: 118(3):965-74 201656 at 4.4418707 down TGA6 Cell Biol. 1996 July 2: 134(2): 559-572 1561042 at 2.1507823 up TGB1 The Journal of Immunology, Vol 54, Issue 116058-6064 204990 s. at 4.5270753 down TGB4 Cell Biol. 1996 July 2: 134(2): 559-572 231031 at 3.3397727 up KGFLP2 Huang, T.-J., Lee, C.-J., Expression of keratinocyte growth factor (KGF) is regulated by two KGF-like proteins, KGFLP1 and KGFLP2. Submitted APR-2002 US 2012/0034613 A1 Feb. 9, 2012 60

TABLE 2-continued

Fold Gene Probe Set ID change Direction Symbol Reference 231015 at 2.3381798 up KLF15 The Journal of Clinical Endocrinology & Metabolism Vol. 94, No. 72594-2601 221841 s at 2.467558 down KLF4 Acta Biochim Biophys Sin (Shanghai). 2008 Jul; 40(7): 554-64 216699 s at 2.1686.67 down KLK1 Arch Dermatol Res. 980; 267(3): 301-11 215808 at 4.67278 OW KLK10 ournal of Investigative Dermatology (2005) 125, 1182-1189; doi:10.1111.0022 2O2X.2OOS.23933.x 205470 s at 3.3177328 down KLK11 ournal of Investigative Dermatology (2005) 125, 1182-1189; doi:10.1111.0022 2O2X.2OOS.23933.x 205783 at 3.6192052 down KLK13 ournal of Investigative Dermatology (2005) 125, 1182-1189; doi:10.1111.0022

204733 at 16.282O7 OW KLK6 Biol Chem. 2007 Feb 23; 282(8): 5834-41. 1552319 a. at 4.8667984 down KLK8 ournal of Investigative Dermatology (2005) 125, 1182-1189; doi:10.1111.0022 2O2X.2OOS.23933.x 205900 at 4.1411896 down KRT1 Gene Expr Patterns. 2005 Aug. 5(6): 801-8 204734 at 3.6002238 down KRT1S Invest Dermatol. 1999 Mar: 112(3): 362-9 218963 sat 10.9443O3 down KRT23 Molecular Oncology, Volume 1, ssue 2, Pages 181-195 201820 at 2.1536.007 down KRTS Acta Derm Venereol. 2004; 84(1): 18-22 209125 at 2.033617 down KRT6A http://ghr.nlm.nih.gov 'gene=krtoa 208188 at 2.5387O63 down KRT9 ournal of Investigative Dermatology (1994) 103,474-477; doi:10.1111,1523-1747 1556410 a. at 24.649403 up KRTAP19-1 JBiol Chem. 2002 Dec 3:277(50): 48993-9002 235148 at 2.1576118 down KRTCAP3 Br J Dermatol. 2003 Apr; 148(4): 654–64. 155.4252 a. at 15.340231 down LASS3 (2003) J. Biol. Chem. 278,43452-43459 218922 s at 3.0278475 down LASS4 Molecular Endocrinology 22(11): 2407-2419 1560531 at 71.235 OW LCE1B ournal of Investigative Dermatology (2005) 124, 1062-1070 1559224 at 4.81139 OW LCE1E ournal of Investigative Dermatology (2005) 124, 1062-1070; doi:10.1111.0022 2O2X.2OOS.23699.x 207710 at 2OO.SO969 OW LCE2B ournal of Investigative Dermatology (2005) 124, 1062-1070 212531 at 12.O14198 down LCN2 ournal of Investigative Dermatology (2006) 126,510-512. doi:10.1038/sjid.5700035 205.266 at 3.7018826 up LIF nvest Dermatol. 2001 Mar: 116(3): 476-8. 207 720 at 32.3533SS down LOR nt Arch Allergy Immunol. 2010; 151(1):28-37. 204038 s at 2.7315595 up LPAR1 nvest Dermatol. 2005 Sep; 125(3): 421-7. 203549 s at 4.0753236 up LPL Diabetes. 2001 Jul; 50(7): 1643-53 216388 s at 2.4562252 down LTB4R nvest Dermatol. 2009 Dec. 129(12): 2854-60 206249 at 2.2523835 up MAP3K13 The EMBO Journal (2006) 25, 5105-5116 216199 s at 2.6540227 up MAP3K4 Biochem J. 2006 Jun 1: 396(2): 307-16. 203836 s at 2.3556855 up MAP3KS rJ Immunol. 2005 Jun; 35(6): 1886-95. 213764 s at 2.1121294 down MFAPS Biol Chem. 1996 Jan 2: 271(2): 1096-103 239273 s at 2.6781023 down MMP28 Matrix Biol. 2009 Mar; 28(2):74-83. Epub 2009 Jan 20 219648 at 2.8866937 down MREG Biol Chem. 2009 Apr 7:284(16): 10877-89 1552578 a. at 2.337757 up MYO3B ournal of Biomedical Science (2007) 14: 87-105 US 2012/0034613 A1 Feb. 9, 2012 61

TABLE 2-continued

Fold Gene Probe Set ID change Direction Symbol Reference 217738 at 2.6675475 lb NAMPT Dec. 3, 2004 The Journal of Biological Chemistry, 279, 50754-50763. 201502 s at 3.0644495 up NFKBLA Photochem Photobiol. 1995 Sep; 62(3): 463-8. 223217 s at 2.3498836 up NFKBIZ. Biol Chem. 2004 Dec 31; 279 (53): 55493-8 202340 x at 2.33773 lb NR4A1 Biol Chem. 2008 May 2: 283(18): 12564-70 216248 s at 7.5919886 NR4A2 Invest Dermatol. 2008 Feb: 128(2): 300-10 216979 at 7.2O3941 up NR4A3 Biol Chem. 2008 May 2: 283(18): 12564-70 225566 at 3.1977339 NRP2 Proc Natl Acad Sci USA. 2001 Oct 23: 98(22): 12677-82. 208591 s at 2.851,169 up PDE3B Med Hypotheses. 2009 ul; 73(1): 118-9. 203708 at 5.313648 up PDE4B nt Immunopharmacol. 2009 an; 9(1): 55-62. 217997 at 3.4O90981 up PHILDA1 Clin Invest. 2006 Jan: 116(1):249-60. 2098.73 s at 4.6149898 down PKP3 FASEB J. 1996 Jun; 10(8): 871-81. 201860 s at 4.228699 up PLAT Am Podiatr Med Assoc. 2008 Sep-Oct; 98(5):345-52 210845 s at 2.249447 up PLAUR Dev Biol. 2008 May 1; 317(1): 187-95. 203896 s at 4.0160823 up PLCB4 ournal of Investigative Dermatology (1997) 108,748-752 213093 at 2.125206 up PRKCA Exp Dermatol. 2002 Feb; 11(1): 25-33. 220635 at 19.078548 down PSORS1C2 Journal of Investigative Dermatology (2007) 127, 1605-1614 213933 at 13.58.3514 down PTGER3 Allergy Clin Immunol. 2009 Oct: 124(4): 809-18.e2. 204897 at 2.5254738 up PTGER4 Nat Med. 2003 Jun; 9 (6): 744-9 20738.8 s at 2.7678204 up PTGES Arthritis Rheum. 2007 Nov. 56(11): 3564–74. 205128 x at 2.326936 up PTGS1 www.jimmunol.org/cgi/reprint 173/21321.pdf 1554997 a. at 6.812711 up PTGS2 J. Lipid Res. 2006 May; 47(5): 921-30 204944 at 2.2989373 up PTPRG J Biol Chem. 2006 Sep 15; 281 (37): 27389-97 212099 at 3.7642062 up RHOB JBiol Chem. 2005 Dec 30; 280(52): 43257-63. 214370 at 2.9802.194 up S100A8 Arch Dermatol Res. 2009 Aug. 301 (7): 523-9. Epub 2009 May 23. 200832 s at 3.7699654 down SCD Life Sciences Volume 84, Issues 3-4, 16 Jan. 2009, Pages 119-124 1554921 a. at 6.0999765 down SCEL Journal of Investigative Dermatology (2003) 121, 781-785; doi:10.1046/j.1523 1747.2003.12539.x 229.199 at 3.818.0137 up SCN9A J Invest Dermatol. 2005 Jun; 124(6): 1333-8. 201287 s at 3.32O23 down SDC1 Arch Dermatol Res. 2008 August: 300(7): 393-395 211906 s at 5.612926 down SERPINB4 Clin Exp Allergy. 2005 Oct: 35(10): 1327-33. 227487 s at 3.1483386 up SERPINE2 Cell Sci. 2008 May 1: 121(Pt:9): 435-43. 212321 a 2.405509 down SGPL1 Cell Signal. 2007 Apr: 19(4): 748-60 238567 a 5.6408434 down SGPP2 Cell Signal. 2007 Apr: 19(4): 748-60. 230494 a 2.5260012 up SLC20A1 Comp Funct Genomics. 2003 February;4(1): 47-55. doi: 0.1002/cfg.239 223222 a. 2.2426856 up SLC25A19 Cell Mol Life Sci. 2005 Oct: 62(19-20): 2204-27 204790 a 2.1506758 up SMAD7 Developmental Cell, Volume 11, ssue 3, Pages 301-312 219695 a 13.657985 down SMPD3 Arch Dermatol Res. 2009 Sep: 301 (8): 587-94. 1553960 at 2.0313072 down SNX21 Proc Natl Acad Sci USA. 2001 Jul 7: 98(15): 8756-61 215078 a 5.7122235 up SOD2 Cell Physiol. 1995 Dec; 165(3): 576-87. 202826 a 4.585347 down SPINT1 Am J Pathol. 2008 Nov; 173(5): 1464-75. US 2012/0034613 A1 Feb. 9, 2012 62

TABLE 2-continued

Fold Gene Probe Set ID change Direction Symbol Reference 209437 s at 2.3600132 up SPON1 Nature Genetics 38, 1304-1309 (1 Nov. 2006). doi:10.1038/ng1907 208539 X at S.SS3559 down SPRR2B ournal of Investigative Dermatology (2005) 125, 1286-1301 213921 at 4.35561 l SST Acta Derm Venereol. 1994 Mar: 74(2): 106-9 213103 at 2.8436816 up STARD13 ournal of Cell Science 118, 2791-2801 (2005) 213820 s a 3.4357908 down STARDS Biol Chem. 2003 Jun 20: 278(25): 22183-6 212353 at 4.4241323 up SULF1 Development. 2007 Sep: 134(18): 3327-38 205547 s a 2.4258.394 up TAGLN Wound Repair and Regeneration, Volume 14, Issue 4 (p 463-470) 205015 s a 3.1335502 down TGFA Invest Dermatol. 1990 Jun: 94(6 Suppl): 164S-170S 201042 at 2.3514817 up TGM2 PET December 2006 vol.319 no. 3 162-1171 203887 s a 3.1370428 down THEBD BrJ Dermatol. 1996 Aug. 135(2): 187-93 201150 s a 2.0579739 lb TIMP3 Invest Dermatol. 1998 Apr: 110(4): 416-21 209387 s a 3.1869683 up TM4SF1 Cell Sci. 2008 Mar 1: 121(Pt5): 685-94. Epub 2008 Feb 12. 21.6005 a 2.7193272 lb TNC Cell Physiol. 2006 Mar: 206(3): 718-27. 202643 s a 6.915339 up TNFAIP3 FEBS Lett. 2003 Feb 11:536(1-3): 35-40 206025 s a 3.1820982 up TNFAIP6 Am J Pathol. 2009 Nov; 175(5): 1915-28. 204932 a 6.287.326 up INFRSF11B United States Patent Application 2003O175710 202687 s a 20.30895 down TNFSF10 Gene. 2004 Oct 27: 341: 199-207. 230398 a 2.7380996 down TNS4 ournal of Investigative Dermatology (2008) 128,783-790. doi:10.1038/sjid.5700969 224220 X at 2.7084796 up TRPC4 Neurosci Lett. 2003 Dec 26:353(3): 189-92. 217979 a 2.400637 down TSPAN13 Mol Cell Biol. 2004 Jul: 24(13): 5978-88 204141 a 2.738532 down TUBB2A Am J Dermatopathol. 1990 Feb; 12(1): 17-24 202154 x at 2.9669414 down TUBB3 Am J Dermatopathol. 1990 Feb; 12(1): 17-24 218810 a 2.1168468 up ZC3H12A The American Journal of Pathology, Vol. 171, No. 1, July 2007

Salicin 0.5% ws. Untreated Control 24 hr treatment N = 7 Salicin 0.5%, N=4 Control RMA normalization (performed on all 42 chips together) t-test with Benjamini and Hochberg FDR correction p-value cut-off; 0.05 Fold change cutoff: 2.0

0096 Genes not chosen for the second subset of genes (0098. Returning to the flowchart of FIGS. 1A-1K for the may nonetheless be considered for additional research based simplified example, for one or more skin attributes of interest, on secondary research factors 122. For example, a hypotheti data is derived from the literature that identifies with each cal gene may have interesting aging-related pathways, not yet skin attribute the genes and their biochemical pathways that associated with skin, or in the case of hypothetical geneh, a are recognized as affecting the skin attribute 123. Thus, FIG. gene may have a high fold value. 1D shows a step of selecting from the second subset of hypo Grouping Second Subset of Genes with Attributes of thetical genes, which have biologically relevant fold changes Skin and are identified with biochemical pathways with the physi 0097. Applying to Table 1 the data set specifying the asso cal appearance of skin aging, further Subsets or groups asso ciations with biochemical pathways with the physical appear ciated with a particular skin attribute 124. ance of skin aging is a meaningful focusing of the data, which 0099. According to the method, genes of the second subset results in the list of Table 2 (See FIG. 7 at 754). Even the are further processed into a plurality of subsets (potentially shorter list of about 200 genes in Table 2 can benefit from overlapping) within the second Subset by categorizing or further focus. In particular, it has been found useful to further associating each gene by an association with one or more skin focus on particular skin attributes that are associated with skin attribute(s). (Geneb is in the subset of skin structure and also aging. in the Subset of skin pigmentation). For example, the genes US 2012/0034613 A1 Feb. 9, 2012

from the second subset may be transformed into skin attribute attributes, skin structure, pigmentation, cell turnover and Subsets, each associated with a particular physical sign of skin hydration. By defining these attributes and developing met aging and appearance, listed as follows in FIGS. 1E and 1 F: rics for them, based where possible on instrumentation that 0100 Skin structure attribute 126. makes the metrics more objective, research on interventions 0101 Skin pigmentation attribute 128. can be given focus. For example, with a chosen attribute and 0102 Skin hydration attribute 130. one or more genes and one or more biochemical pathways (0103 Cell turnover attribute 132. associated with it, the research can focus on particular parts of 0104 Turning to the second subset as defined for the a biochemical pathway that can be enhanced to encourage the experimental data listed in Table 2, the use of skin attribute biochemical pathways that produce a more youthful version data sets is further explained. A data set identifying the rela of that attribute or on inhibiting a biochemical pathway that tionship between a particular skin attribute and particular produces a less youthful version of that attribute. It is believed genes is developed based on the literature or on available that the biochemical pathways associated with genes can be unpublished data. The data set identifies biochemical path regulated by many different factors. The focus on particular ways of the physical appearance of skin aging known to be genes, particular biochemical pathways associated with the associated with one of the genes of Table 2 and a particular genes and particular skin attributes associated with those skin attribute; it is preferably collected and stored in a format pathways may permit identifying an “intervention' where a that promotes an intersection analysis with the data of Table 2. specific technology can target the gene expression activity for This can be done by building a table of all genes known to be a particular skin attribute to reflect a more youthful gene associated with a biochemical pathway of the physical expression profile, ultimately influencing the physical appearance of skin aging and a particular skin attribute of appearance of the skin as it ages. interest and finding its intersection with Table 2. In some 0110 Functional youth gene assembly, refers to a group of embodiments, the genes involved in various biochemical genes, encompassing one or more mechanisms of aging, pathways related to the particular attribute “skin structure' addressable for functional restoration or stabilization of a are chosen for analysis. Some of the biochemical pathways of more youthful state in the skin. Each functional youth gene interest for this skin attribute include skin structural proteins assembly may focus on a particular skin attribute that has synthesis, degradation and maintenance and extracellular youthful and non-youthful states. A functional youth gene matrix assembly. However, other skin attributes and associ assembly could also apply to characteristics in other tissues ated pathways may also be of interest. and organs. 0105. The attributes skin structure, pigmentation, cell 0111. By extension, a "youth gene family” is composed of turnover and hydration are described below. One can derive a related group of functional youth gene assemblies and from the literature for each attribute its own subset of genes would address multiple (or all) the significant attributes of associated with that attribute, which permits development of aging for skin (or another tissue or organ, such as adipose a data set identifying the relationship between each of these tissue, , brain, skeletal muscle, etc.). particular skin attributes and particular genes. However, a 0.112. Once it is defined, one can examine a functional gene may be associated with more than one skin attribute. A youth gene assembly for a specific tissue associated with a brief discussion of general principles of skin aging is a useful specific function. For example, in skin, the youth gene family preface to a discussion of skin attributes. may comprise functional youth gene assemblies for skin pig 0106 The stratum corneum is the layer of the skin that mentation, structure, hydration and cell turnover. The skin is forms the top surface layer and serves to protect the skin while an easily accessible organ with easily measured or observed controlling moisture and the flow of substances in and out of aging attributes. Therefore, one can readily examine mani the skin. As this barrier function is broken down, the skin festations of changed expression levels of a functional youth Suffers damaging effects, thus further contributing to prema gene assembly for a specific attribute of skin. ture aging. These damaging effects causing premature aging 0113. This approach is part of an overall strategy to slow of the skin are a concern for many individuals wishing to down the physical manifestation of the aging process in skin maintain healthy, youthful looking and feeling skin. by developing a composition that addresses several genetic 0107 Aging can occur from biological processes or envi mechanisms of aging simultaneously, i.e., through actions ronmental factors, and in Some cases environmental factors targeted to expression levels of the members of functional that impact biological processes. These factors alone and in youth gene assemblies, instead of in-depth analysis of an combination contribute to aging appearance and are respon individual gene. The following focuses on four skin attributes sible for the decline in skin health and function. Biological for which it is useful to define a functional youth gene assem aging, which is intrinsic, is the result of changes, often geneti bly. cally determined, that occur naturally within the body. Envi 0114 A. Skin Structure ronmental aging, which is extrinsic, is the result of free radi 0115 The skin structure group has genes that have bio cal damage generated by accumulated exposure to Sunlight chemical pathways associated with the physical appearance (photoaging), pollution, or cigarette Smoke. Also, lifestyle of skinaging that include for example, skin structural proteins choices like diet, sleep, and stress can affect how quickly one synthesis, degradation and maintenance and extracellular appears to age. matrix assembly. Examples of skin structural protein synthe 0108. Whether from biological or environmental sources, sis include elastin formation, keratinocyte differentiation and the appearance of aging results from several mechanisms of collagen production. action or biochemical pathways. For example, a loss of skin 0116. Younger skin has the ability to balance damage and structure, a slowing skin cell turnover, pigmentation changes repair to collagen, a structural protein in the skin. This bal or a decrease in skin hydration. ance keeps skin looking Smooth and wrinkle free. During the 0109. One can group many of the detectable/sensible aging process, skin begins to lose this balance. Less and less changes that occur with skin aging into four major skin collagen is created and more enzymes are produced which US 2012/0034613 A1 Feb. 9, 2012 64 break down this protein resulting in lines and wrinkles. hydration and moisture levels within the skin. Ample mois Increasing the production of structural proteins promotes turization within the extracellular matrix is a factor that helps youthful looking skin, whereas inhibiting the production of maintain the strength and integrity of the structural proteins. enzymes that break down the proteins in the skin is also Many GAGs are too large to enter through the epidermis, but beneficial. there are ingredients that have been shown to increase GAG 0117 B. Pigmentation production. 0118. The pigmentation group has genes that have bio I0128. Each skin attribute may be negatively impacted by chemical pathways associated with the physical appearance one or more mechanisms that contribute to aging in the skin, of skin aging that include for example, regulation of melanin or may be positively impacted by a mechanism that preserves production and control. All normal human skin contains chro youthful appearance. If biochemical processes affecting each mophores that give the skin a characteristic coloration. The attribute and the gene expression profile driving those bio color of the skin is mostly due to melanin, eumelanin, hemo chemical pathways or processes can be addressed (up-regu globin and, to Some degree, collagen and elastin. The primary lated or down-regulated), a skin state more characteristic of a function of pigmentation is the absorption of short wave younger individual is established. length light capable of damaging structural components in the I0129 Referring to the simplified example of FIGS. 1D-1F, deeper layers of the skin and the nuclear and mitochondrial the processing of steps 124, 126, 128, 130, 132 begins by DNA of keratinocytes, melanocytes, fibroblasts, lipocytes, focusing on one or more particular skin attributes. For each Langerhans cells, other immune system cells and neural cells skin attribute, from the literature a set of genes may be found in the skin. that has a biochemical pathway relevant to the skin attribute. 0119 Aside from a sallow appearance, seen in thinner, With sufficient knowledge of the pathway, it can be deter lighter skinned individuals with poor circulation, around the mined from the literature what the function of the pathway is globe the overall color of the skin does not reflect aging. and thus, whether that pathway will assist a more youthful However, across cultures the irregular distribution of skin state of the skin attribute of interest if the pathway is up color, sometimes called dispigmentation, is a key attribute regulated or down regulated. The data on a skin attribute and that characterizes older skin and presents in the form of genes with a biochemical pathway relevant to it may be ephelides, letingines and hyperpigmented or hypopigmented assembled as a dataset for that skin attribute. FIGS. 1D-1F SCaS. show for the simplified genome (a-h) one format. For each of 0120. The biochemical changes in the skin that drive an genes a-hin the table at step 120 and for each skin attribute, a irregular pigmentation pattern can be grouped into several row of data can show which genes have association with the categories. Increased growth factor signaling by melanocyte skin attribute of interest and show a direction of regulation of stimulating hormone and related cytokines, increased tyrosi the gene expression associated with a more youthful appear nase, an enzyme converting tyrosine to DOPA and on to ance. This is shown in the table at step 123 in one row for each melanin, and increased amounts of melanin transferred from of the skin attributes. (See rows 213a-213d). As will be seen, the melanocytes to the keratinocytes. these data sets set up the intersection processing for indi 0121. During the aging process, melanocytes can cluster vidual skin attributes that leads to the tables at steps 126, 128, together. These clusters of melanocytes can then become 130, and 132. overly active resulting in areas of hyperpigmentation, known 0.130 Correspondingly, for the microarray data in Table 2, as age spots or discoloration. from the genes in Table 2 a data set can be prepared that show 0.122 C. Cell Turnover association for a chosen skin attribute of interest, which of 0123. The cell turnover group has genes that have bio these genes has a pathway relevant to the skin attribute of chemical pathways associated with the physical appearance interest and from the function of that pathway can be deter of skin aging that include for example, cellular differentia mined a direction of regulation of the gene associated with a tion. During the aging process, the outer layers of the skin do more youthful appearance. The association of a gene and its not slough off as they once did. The adhesion of these skin pathway and one or more skin attributes may come from cells result in rough skin texture and dull, lifeless looking published or private research. Table 3 shows a dataset taken skin. When cell renewal slows with age, dead skin cells build form the genes in Table 2, and identifying those genes with a up along the pores of the skin. This build up increases the pathway relevant to the skin attribute “skin structure'. For appearance of pores, making them look larger than in youth each gene in the list (of Table 3) there is a column entry in ful looking skin. which a function relative to the skin attribute appears. The 0124. By increasing the cell renewal process, younger, resulting skin attribute data set may be input into a database healthier skin cells Surface, promoting a smoother skin tex 730, by a suitable process application module that stores and ture. When skin looks smoother, it reflects light more uni accesses such data and uses it for processing as contemplated formly. Smoother skin appears more radiant and bright. by steps 124, 126, 128, 130, 132 in the hypothetical example. Increasing cell renewal also stimulates a healthy exfoliation I0131 The intersection analysis for the microarray data in that results in the appearance of tighter pores. Table 3 is similar to that shown in steps 124, 126, 128, 130, 0125 D. Hydration 132 (which deal with all four skin attributes identified above: 0126 The skin hydration group has genes that have bio for the data in Table 2, there is only one example skin attribute chemical pathways associated with the physical appearance addressed in Table 3). The skin attribute data set of Table 3 of skin aging that include for example, skin barrier compo may be processed to derive a skin attribute subset for a pre nent synthesis, skin barrier integrity and water regulation. liminary functional gene assembly. As the data of Table 2 is Skin barrier component synthesis includes, for example, processed by intersection with a skin attribute data set desig hyaluronic acid synthesis and lipid synthesis. nating genes with an association with a skin attributes of 0127. Moisture-binding glycosaminoglycans (GAG) interest, it becomes necessary to consider for each gene the found within the extracellular skin matrix play a role in the literature-reported regulation direction for the more youthful US 2012/0034613 A1 Feb. 9, 2012 65 state of the skin attribute of interest. If a gene happens to have tion with more youthful appearance as to that attribute. For an association with more than one attribute, then a regulation “skin hydration', genes c and g are shown as having a down direction for each skin attribute is identified in the particular regulation association, and gene e is shown as having an skin attribute data set up-regulation of a gene might be favor up-regulation association with more youthful appearance as able for one skin attribute and down-regulation of the gene to that attribute. For cell turnover, gene e is shown as having favorable for a different attribute. an up-regulation association and gene fis shown as having a 0132 Referring to again to FIG. 1D of the simplified down-regulation association with more youthful appearance example, the table at 123, rows 213a-d show a simplified as to that attribute. hypothetical example of how the datasets on genes of interest 0.133 As for the microarray data, Table 3, shows a data set may reflect how the hypothetical genes are associated with a comprising genes from Table 2 for which the literature shows particular skin attribute and whether a pathway identified a connection to the skin structure attribute, including a func with the gene has an up or down regulation relative to a tion with respect to skin structure which will determine the youthful direction for the particular skin attribute. Thus, for more youthful regulation direction. Once the more youthful the “skin structure' attribute, genes b and fare shown as regulation direction is specified for a gene, the data set of having an up-regulation association with a more youthful Table 3 can be used for intersection processing to find a skin appearance as to that attribute. For 'skin pigmentation'. attribute subset list for the gene assembly for the skin struc genes band c are shown as having an up-regulation associa ture attribute.

TABLE 3

Probe Fold Gene Set ID change Direction Symbol Gene Title Function Reference 213992 at 4.80O3573 down COLAA6 tumor Structural The Journal of necrosis protein in Cell Biology, Vol actor, alpha- skin 130, 1219–1229, induced Copyright (C) 1995 protein 9 201286 at 4.6295.676 down SDC1 O cell- Arch Dermatol necrosis extracellular Res. 2008 actor, alpha- matrix August: 300(7): induced interactions 393-395 protein 8 205900 at 4.141 1896 down KRT1 O Epidermal Gene Expr necrosis structural Patterns. 2005 actor, alpha- protein August; 5(6): 801-8 induced protein 7 239272 at 4.074,428 down MMP28 O Destroys Matrix Biol. 2009 necrosis Collagen March; 28(2):74-83. actor, alpha- Epub 2009 Jan. 20 induced protein 6 209126 x at 3.952012 down KRT6B O Epidermal Acta Derm necrosis structural Venereol 2004; actor, alpha- protein 84:18-22 induced protein 5 204734 at 3.736902 down KRT1S O Epidermal J Invest necrosis structural Dermatol. 1999 actor, alpha- protein March; 112(3): induced 362-9 protein 4 204636 at 3.736902 down COL17A1 tumor Structural http://ghr.nlm.nih. necrosis protein in govgene=col17a1 actor, alpha- the skin induced protein 3 204135 at 3.736902 down FILIP1L tumor Cytoskeleton Nature Genetics necrosis remodelling 33, 487-491 actor, alpha- (2003) Published induced online: March protein 2 2003; | doi:10.1038/ng1119 200606 at 3.736902 down DSP O Cellular FASEB.J. 1996 necrosis Adhesion June; 10(8): 871-81. actor, alpha- protein induced protein 1 208188 at 3.736902 down KRT9 O structural Mech Ageing necrosis protein Dew. 2008 actor, alpha- October: 129(10): induced 563-71. Epub 2008 protein O Jun. 3. US 2012/0034613 A1 Feb. 9, 2012 66

TABLE 3-continued

Probe Fold Gene Set ID change Direction Symbol Gene Title Function Reference

201820 at 3.736902 down KRTS tumor Structural necrosis protein factor, alpha October: 129(10): induced 563-71. Epub 2008 protein 1 un. 3. 211922 s at 3.736902 down CAT tumor Protects ournal of necrosis from UV nvestigative factor, alpha damage Dermatology induced (2006) 126, 182 protein 2 90 202643 s at 6.915339 up TNFAIP3 tumor Negative FEBS Lett. 2003 necrosis Feedback Feb. 11:536(1- factor, alpha of NFKA 3):135-40 induced (aging protein 3 marker) 215078 at 5.7122235 up SOD2 Superoxide Protects ournal of dismutase 2, from UV nvestigative mitochondrial damage Dermatology (1994) 102,476 480 215910s at 3.6631.63 l fibronectin Collagen FEBS Lett. 1994 type III Assembly Apr. 18; 343(1): domain 47-SO containing 3A 204422 s at 3.455492 l fibroblast Inhibits Arteriosclerosis, growth factor matrix Thrombosis, and 2 (basic) collagen Vascular Biology. synthesis 1993; 13:680-686 21.6005 at 2.7193272 TNC Tenascin ECM Dev Dyn. 2000 Remodelling June: 218(2): 235 59. 211958 at 2.71.18793 IGFBP5 insulin-like Collagen Arthritis Rheum. growthfactor Deposition 2006 September; binding 54(9): 3001-10 protein 5 214702 at 2.70994.62 fibronectin 1 Structural Exp Cell Res. protein 990 November: 91(1): 8-13 206026 s at 2.519742 TNFAIP6 O Induces BMC Immunol. necrosis MMPS 2009 Mar. 19: actor, alpha induced protein 6 204790 at 2.1506758 up SMAD7 SMAD Negative Biol Chem. amily regulator of 2005 Mar. 4; member 7 collagen 280(9): 8079-85. synthesis Epub 2004 Dec. 3 213093 at up PRKCA protein kinase Stimulates Free Radic Biol C, alpha Collagenase Med. 1999 October: 27(7-8): 729-37 202724 sat 2.0982,356 up FOXO1 orkhead box induces Am J Physiol Cell O1 Collagen Physiol 292: Synthesis C850-C856, 2007 201150 s at 2.0579739 up TIMP3 TIMP inhibits Invest metallopepti MMPS Dermatol. 1998 daseinhibitor 3 April:110(4): 416-21 203184 at 2.0282688 up fibrillin 2 Elastin Invest Fibre Dermatol. 1996 Formation May: 106(5): 1090-5 207 720 at 32.353355 down LOR loricrin Component Biol Chem. of Cornefied 992 Sep. 5; envelope 267(25): 18060-6. 1552319 a. at 4.8667984 down KLK8 kallikrein Break ournal of related down of nvestigative peptidase 8 cellular Dermatology adhesion (2005) 125, proteins 182-1189 206033 s at 4.676169 down DSC3 desmocollin 3 Cellular FASEB.J. 1996 Adhesion une; 10(8): 871-81. protein 2098.73 s at 4.6149898 down PKP3 plakophilin 3 Cellular FASEB.J. 1996 Adhesion une; 10(8): 871-81. protein US 2012/0034613 A1 Feb. 9, 2012 67

TABLE 3-continued

Probe Fold Gene Set ID change Direction Symbol Gene Title Function Reference 209792 s at 4.4899607 down KLK10 kallikrein related Cellular J Invest peptidase Adhesion Dermatol 10 protein 2003 121: 542-549 214370 at 2.9802194 up S100A8 S100 calcium Increases Arch Dermatol binding with Res. 2009 protein A8 photoaging August: 301 (7): (calgranulim destroys 523-9. Epub 2009 A or cystatin proteins May 23. A) 205207 at 3.1970735 up IL6 Interleukin 6 Stimulates ournal of MMPS nvestigative Dermatology (2004) 123, O12-1019 204990s at 4.5270753 down ITGB4 integrin, beta 4 Required Cell Biol. 1996 for cell to ul. 2; 134(2): cell 559-572 adhesion 203535 at 2.590224 down S100A9 S100 calcium Mediates Biochem Biophys binding fibronectin Res Commun. protein A9 adhesion 2007 Mar. 2: (calgranulim 354(1): 84-89 B) 1561042 at 2.1507823 up ITGB1 integrin, beta 1 Marker for Br J Dermatol. skin 2003 April; structural 48(4): 770-8 damage caused by UV 211981 at 2.121121 up COL4A1 Collagen Skin Clin Invest. Type 4 Alpha 1 Structural 989 March; protein 83 (3): 791-795

0134. As a gene associated with a skin aging attribute can level for gene f is not consistent with the data from the be a part of a specific biochemical pathway involved in the literature. According to the (hypothetical) literature, down physical appearance of skin aging that improves the skin regulation of gene fprovides a less youthful skin appearance. attribute in a youthful direction or one that can be a part of a Therefore, geneb is kept in the group, while gene fis dropped specific biochemical pathway that increases the appearance from the group at this time. of skin aging (i.e., changes skin appearance in a non-youthful 0.136 For the skin pigmentation attribute 128 of the sim direction), the intersection processing requires additional plified example, genes band c have a fold change greater than logic to include in the gene assembly for a particular skin 2, and the up regulation of genes band c from the (hypotheti attribute only those genes that are regulated in a direction cal) gene expression level are consistent with the data from reflective of youthful skin appearance. Thus, the genes of the literature indicating that up regulation of both genes band Table 3 (see also, FIG. 7 at 756) are processed by a module c provide more youthful skin structure. Therefore, both genes identifying which genes were reported in the microarray a b and c are kept in the group. regulated in a direction (up or down regulated) that is the I0137 In the skin hydration attribute 130 of the simplified desired, more youthful direction. This criterion removes from example, genes c and g have a fold change greater than 2, but the preliminary gene assembly of Table 3 any gene for which the up regulation shown by the (hypothetical) gene expres the microarray data shows that it up-regulates a pathway that sion level is not consistent with the data from the literature. decreases youthful appearance or that down-regulates a path According to the (hypothetical) literature, down regulation of way that increases youthful appearance. This directional genes c and g provide a more youthful skin appearance. Gene logic is step 134 of FIG. 1G. e has a fold change greater than 2, and the up regulation of that 0135 Returning to the simplified, hypothetical example, gene from the (hypothetical) gene expression level is consis FIGS. 1E and 1F, rows 215a-d show for each skin attribute the tent with the data from the (hypothetical) literature indicating results or the answer to the question, whether or not the gene that up regulation of that gene provides more youthful skin stays in the group. In the skin structure attribute example 126, structure. Therefore, genes c and g are dropped from the geneb has a fold change greater than 2, and the up regulation group at this time, but gene e is kept in the group. of that gene from the (hypothetical) gene expression level is (0.138. In the cell turnover attribute 132 of the simplified consistent with the data from the literature that indicates that example, gene e has a fold change greater than 2, and the up up regulation of that gene provides more youthful skin struc regulation of that gene from the (hypothetical) gene expres ture. However, gene fhas a fold change greater than 2, but the sion level is consistent with the data from the (hypothetical) down regulation shown by the (hypothetical) gene expression literature indicating that up regulation of that gene provides US 2012/0034613 A1 Feb. 9, 2012 68 more youthful skin structure. Genefhas a fold change greater expression levels in a direction reflective of youthful skin than 2, and the down regulation of that gene from the (hypo appearance for the “skin structure' attribute. The up/down thetical) gene expression level is consistent with the data from gene regulation shown by Affymetrix testing is thus for some the literature that indicates that down regulation of that gene genes consistent with Scientific literature as to regulation of provides more youthful skin structure. Therefore, both genes the gene in a more “youthful direction. Table 4 is a subgroup e and fare kept in the group. of Table 3 and is a second subset of genes, further defined by 0139 Table 4 shows the result when the processing logic applying the method steps sketched in FIGS. 1A-1G of the of the simplified example is applied to the data from actual simplified example (through step 134) for the skin attribute microarray testing of tissue exposed to Salicin and when the “skin structure'. skin attribute is “skin structure', which is the focus of the data 0.141 Genes in Table 3 not chosen for Table 4 based on the set in Table 3. The intersection processing module using the logic requiring the alignment of the Affymetrix testing-de data of Table 3 identifies those genes that not only have an rived data with the literature's position on regulation of the association with skin structure but also have been found in the gene in a “youthful direction may nonetheless be considered test data to be up-regulating a pathway that provides more for additional research on “skin structure' but that must be youthful skin structure or genes down-regulating a pathway based on secondary research factors. Table 4 shows only the that provides less youthful skin structure. genes that have the required alignment of “youthful' direc 0140 Table 4 (see also, FIG. 7 at 758) shows for the tion for “skin structure' in both the literature and the Affyme Affymetrix testing-derived data a group of genes exhibiting trix testing-derived data.

TABLE 4

Probe Fold Gene Set ID change Direction Symbol Gene Title Function Reference 239272 at 4.074,428 down MMP28 matrix Destroys Matrix Biol. 2009 metallopepti- Collagen March; 28(2):74-83. dase 28 Epub 2009 Jan. 20 208188 at 3.736902 down KRT9 keratin 9 structural Mech Ageing protein Dew. 2008 October: 129(10): 563-71. Epub 2008 un. 3. 201820 at 3.736902 down KRTS keratin 5 Structural Mech Ageing protein Dew. 2008 October: 129(10): 563-71. Epub 2008 un. 3. 202643 s at 6.915339 up TNFAIP3 tumor Negative FEBS Lett. 2003 necrosis Feedback Feb. 11:536(1-3): factor, alpha- of NFKA 35-40 induced (aging protein 3 marker) 215078 at 5.712224 up SOD2 Superoxide Protects ournal of dismutase 2, from UV nvestigative mitochondrial damage Dermatology (1994) 102,476 480 21591.0 s at 3.663163 up FNDC3A fibronectin Collagen FEBS Lett. 1994 type III Assembly Apr. 18; 343(1): domain 47-SO containing 3A 21.6005 at 2.719327 TNC Tenascin ECM Dev Dyn. 2000 Remodelling June: 218(2): 235 59. 211958 at 2.711879 up IGFBP5 insulin-like Collagen Arthritis Rheum. growth factor Deposition 2006 September; binding 54(9): 3001-10 protein 5 214702 at 2.709946 up FN1 fibronectin 1 Structural Exp Cell Res. protein 1990 November; 191 (1): 8-13 202724 S at 2.098236 up FOXO1 forkhead box induces Am J Physiol Cell O1 Collagen Physiol 292: Synthesis C850-C856, 2007 201150 s at 2.057974 up TIMP3 TIMP inhibits J Invest metallopepti- MMP's Dermatol. 1998 dase inhibitor 3 April; 110(4): 416 21 203184 at 2.028269 up FBN2 fibrillin 2 Elastin J Invest Dermatol. Fibre 1996 May; Formation 106(5): 1090-5 US 2012/0034613 A1 Feb. 9, 2012 69

TABLE 4-continued

Probe Fold Gene Set ID change Direction Symbol Gene Title Function Reference 1561042 at 2.150782 up ITGB1 integrin, beta 1 Marker for Br J Dermatol. skin 2003 April; structural 148(4): 770-8 damage caused by UV 211981 at 2.121121 up COL4A1 Collagen Skin J. Clin Invest. Type 4 Alpha 1 Structural 1989 March; protein 83 (3): 791-795

Youthful direction based on published literature. sensitive than Northern blot, and can be used to quantify 0142. As can be seen, with a data set like Table 3 derived mRNAs that are expressed at low levels. (Applied Biosys for other skin attributes, a table like Table 4 can be derived for tems, Inc., The Basics: What is a Nuclease Protection skin attributes other than “skin structure'. Assay?(C2010, last accessed May 18, 2010, from http://www. ambion.com/techlib/basics/npa/index.html). Confirmation 0148 Real Time Polymerase Chain Reaction is a labora Determination of RNA Quantification for Second tory technique used for DNA quantification, which measures Subset of Genes the accumulation of DNA product after each round of PCR amplification. This laboratory technique is also known as 0143 A gene assembly developed to the status of Table 4 quantitative real time polymerase chain reaction (RTO-PCR/ may be further confirmed and refined by a different method Q-PCR/qPCR) or kinetic polymerase chain reaction, which is ology with a different gene analysis tool, in particular, by used to amplify and simultaneously quantify a targeted DNA performing further skin model testing that takes advantage of molecule. The technique enables both detection and quanti the narrowing of focus to a list of genes as in Table 4. fication (as absolute number of copies or relative amount 0144. Different methodologies include determining RNA when normalized to DNA input or additional normalizing types and levels by RNA quantification metrics including, for genes) of one or more specific sequences in a DNA sample. example, Northern blot technique, Ribonuclease Protection 014.9 The amplified DNA is detected as the reaction Assay (RPA) and Real Time Polymerase Chain Reaction progresses in real time, as compared to standard PCR, where (RT-PCR). the product of the reaction is detected at its end. Two common 0145 Northern blot is a well-known process for detecting methods for detection of products in real-time PCR are: (1) and quantifying mRNA levels. The northern blotting tech non-specific fluorescent dyes that intercalate with any nique is often used for comparison of gene expression pat double-stranded DNA, and (2) sequence-specific DNA terns for different tissue types. In terms of skin genomics, it is probes consisting of oligonucleotides that are labeled with a less used than the modern techniques but can be used as fluorescent reporter which permits detection only after confirmation step in understanding gene expression in the hybridization of the probe with its complementary DNA tar skin. get. 0146 Northern blots start with the extraction and isolation of mRNA from the sample. RNA samples are then separated (O150 Reverse Transcription PCR (RT-PCT) is used for by gel electrophoresis. Once separated, the RNA is then trans amplifying DNA from RNA. Reverse transcriptase reverse ferred to a positively charged membrane, most often made of transcribes RNA into cDNA, which is then amplified by PCR. nylon. Once transferred to the membrane, RNA is then immo RT-PCR allows for a high sensitivity detection technique, bilized to the membrane through covalent linkage with the where low copy number or less abundant RNA molecules can use of UV light or heat. Hybridization probes (fragment of be detected. RT-PCR is widely used in expression profiling, to DNA or RNA used to detect the presence of specific determine the expression of a gene or to identify the sequence sequences) to be used for the experiments are labeled and of an RNA transcript, including transcription start and termi placed on the membrane for hybridization. The membrane is nation sites. then washed to ensure probe binding is strong as well to avoid 0151 Real-time PCR may be combined with reverse tran background signals. The signals are then detected by X-ray scription to quantify messenger RNA and Non-coding RNA film and can be quantified by densitometry. (Alberts, B., et al. in cells or tissues. Real-time reverse-transcription PCR is also Molecular Biology of the Cell, 5th ed. pp. 538-539, New York: known as qRT-PCR, RRT-PCR, or RT-rt PCR. Taylor & Francis Group (2008)). 0152. In some embodiments, Real Time Reverse Tran 0147 Ribonuclease protection assay is a sensitive tech scriptase Polymerase Chain Reaction (RT-rt-PCR) experi nique for detection, quantification and characterization of ments on the second Subset of genes are performed to confirm RNA. Isolated RNA is hybridized to a single stranded cDNA activity of the skin anti-aging agent acting on the gene. of the gene of interest. After annealing, the sample is subject 0153. In some embodiments, determining the levels of to enzymatic digestion to remove all single Stranded nucleic expression for the second Subset of genes is done by using an acids, leaving only double-stranded RNA. The double RNA quantification metric. Selecting a further set of genes Stranded nucleic acid fragments are then separated on high from a previous set of genes in a second sample of human skin resolution polyacylamide gels. Quantification is carried out tissue is based on measured levels of expression, which meet similar to that of Northern Blot. The assay is much more a criterion of biological relevance. US 2012/0034613 A1 Feb. 9, 2012 70

0154 As seen in FIG. 1G of the simplified example, to Universal MasterMix without UNG and loaded into the wells perform confirmation of a skin attribute Subset, or a prelimi of the TLDA cards. The cards are run using an Applied Bio nary functional youth gene assembly the next step is to expose systems 7900HT instrument according to the manufacturer's a second sample of human or human equivalent skin tissue to cycling parameters. the agent 136. Preferably this is done with the same skin model as used with the microarray technology for measuring 0160. As with the microassay, the analysis is done with a global gene expression. The qualities of the agent (concen skin model exposed to the agent and a reference that is not tration, solvent, etc.) should normally be the same. exposed to the agent. The skin model not exposed to the agent 0155. In some embodiments, the agent tested is salicinata may be used for calibration. The skin model may be human concentration of 0.5% salicin, available from Symrise Cor equivalent skin tissue. The target genes get normalized to a poration (Teterborro, N.J.). The salicin is dissolved in water. stable endogenous control (genes that are invariants in all cell Affymetrix microarray testing provides results for thousands types such as f-actin). This normalization is to account for of genes, whereas RT-rt-PCR testing provides results for a variations that may occur during sample loading. The unex smaller gene group. For RT-rt-PCR testing, about 90 genes posed information gathered is used for comparison against are tested at a time for this particular experimental design the tested Sample. (other designs may test as many as 390 at a time on the (0161 The formula for AC (delta cycle threshold) is C. equipment identified below). The experimenter may choose (target) minus C (endogenous control gene). (AC, C (tar this number based on cost. get)-C (endogenous control gene)). The test system, data 0156 To start a process of confirmation using a second processing system 710 stores the data then uses process appli gene analysis tool that works with Smaller arrays and a dif cation modules 720 to take the C values for both the exposed ferent, perhaps more sensitive measurement of regulation by and unexposed and get a AAC value, which is reported for the agent, expose a second sample of human skin tissue to the that specific gene in a log ratio Scale. Once this data is col agent and select a set of candidate genes for confirmation. For lected and stored, such as in database 730 (see FIG. 7), it is example, the set of candidate genes may be the genes of a analyzed by conducting bioinformatics statistical analysis on preliminary functional youth gene assembly of a particular data 140. In one embodiment, data analysis is carried out skin attribute, supplemented with a few other genes that are of according to the RQanalysis method using RQManager and interest based on secondary research. More that one candidate StatMiner (v3.1) software programs. group may of course be explored by confirmation. For 0162. At least part of the comparing of the data is per example, a candidate group may be built around the prelimi formed by a computer system, such as the data processing nary functional youth gene assembly of each of the skin system 710 (see FIG. 7) for performing data access and stor attributes discussed above: skin structure, skin pigmentation, age and various computations specified by Software (process skin hydration and cell turnover. application modules 720) corresponding to the functions 0157. The subsets of genes related to a particular skin occurring at various described steps of this method. Addi attribute are conveniently tested on one test card. Referring tional process application modules 722 perform a parametric again to the process Schematically shown for a simplified t-test with a Benjamini and Hochberg false discovery rate hypothetical gene set in FIGS. 1A-1G, the next step is to correction is performed to identify genes with a statistically determine quantification of RNA and directions of regulation significant p value equal to or less than 0.05. The up or down in the second exposed sample for each of the skin attribute regulation of the gene is also identifiable from the RT-rt-PCR Subdivisions of the second Subset of genes using a determi analysis. Results of the change in threshold cycle (AC) val nation method different than for the first exposed sample 138, ues between the exposed and unexposed human skin tissues for example, using an RT-rt-PCR method, for each attribute. are calculated by a software program. The selected genes will For the simplified example set of genes of FIG. 1G, we may have a cycle threshold of biological relevance. For these assume all genes from all four attributes are tested on one experiments, the selected genes have a cycle threshold of less chip. than about <35, which is a value of biological relevance. In 0158 To implement this step, RT-rt-PCR is conducted for every cycle of PCR (C, value) the amount of DNA is approxi specific genes known to be involved in skin aging. In one mately duplicated, thus, the C is in the logarithmic scale and embodiment Custom TaqMan R. Low Density Arrays (TL inversely proportional to the quantity of DNA/RNA. There DAs) were configured using Applied Biosystems validated fore, high C values represent low expression while highly gene expression assays. The validated gene expression assays expressed genes have low C values. Comparing the normal contain a TaqMan(R) fluorescent probe and primers for each ized expression of the two conditions it is possible to calculate target gene. Genes for the TLDA cards are selected based on the fold change of the expression of the gene, AAC value. either, published literature describing the genes functional The fold change is the expression ratio: if the fold change is role in skin cell biology and aging and/or the previous positive it means that the gene is up-regulated; if the fold Affymetrix testing results. (See Tables 1-4). Five endogenous change is negative it means it is down-regulated. This is control genes may be included on each card. Thus, when a represented in a log scale. particular gene assembly is tested with RT-rt-PCR, the data 0163 The Corcycle threshold is defined as the number of resulting may cover more than just the set of genes as in Table cycles required for the fluorescent signal to cross the thresh 3 old. C levels are inversely proportional to the amount of 0159 cDNA is synthesized from an aliquot of total RNA target DNA in the same. Standard real time reactions undergo using the High Capacity cloNA reverse transcription kit from 40 cycles of amplification. C-20 indicate strong positive Applied Biosystems (Foster City, Calif.) according to the reactions and an abundance of the targeted DNA. C. values of manufacturer's Suggested protocols. (High-Capacity cDNA 30-37 are positive reactions indicative of moderate amounts Reverse Transcription Kits for 200 and 1000 Reactions Pro of target DNA. C. values of 38-40 are weak reactions indica tocol (October, 2006)). cDNA was mixed with TaqMan(R) tive of minimal amounts of target DNA. The C values are an US 2012/0034613 A1 Feb. 9, 2012 criterion of biological relevance. The experimenter optionally identified with a strong anti-aging effect or is otherwise chooses a criterion based on biological relevance for gene known to be of possible interest for this skin attribute. It also expression in aging skin. may be useful to add to a test card a gene that showed a 0164. Table 5 (see also, FIG. 7 at 762) shows a testing derived example of quantitative measurements of gene Sufficient fold change in microarray data but has not yet been expression and the direction of regulation for genes associ identified with a strong anti-aging effect. Optionally, the ated with the skin attribute “skin structure', derived from experimenter could add to a test card a gene that has a fold RT-rt-PCR analysis. As can be seen, the number of genes in change greater than the selected fold change criterion (e.g., Table 5 exceeds that in Table 4. This reflects that, in some 2), but the regulation direction shown by the first gene expres cases, it may be useful to add to a test card a gene that did not sion level testing is not consistent with the data from the show a Sufficient fold change in the microarray data but is literature. This can provide a useful second look at a gene.

TABLE 5

Gene Gene Symbol Function Direction p-value Log1ORG Reference COL4A6 Structural no data The Journal of Cell protein in Biology, Vol 130, skin 1219-1229, Copyright (C) 1995 SDC1 Cell- down 0.002811879 -0.396098529 Arch Dermatol Res. extracellular 2008 August: 300(7): matrix 393-395 interactions KRT1 Epidermal down 0.001131397 -1.111598451 Gene Expr Patterns. structural 2005 Aug. 5(6): 801-8 protein MMP28 Destroys no data Matrix Biol. 2009 Collagen Mar: 28(2):74-83. Epub 2009 Jan 20 KRT6B Epidermal no data Acta Derm Venereol structural 2004: 84:18-22 protein KRT1S Epidermal no data J Invest Dermatol. structural 1999 Mar: 112(3): 362-9 protein COL17A1 Structural no data http://ghr.nlm.nih.gov/gene=col17a1 protein in the skin FILIP1L Cytoskeleton no data Nature Genetics 33, remodelling 487-491 (2003) Published online: March 2003: doi:10.1038/ng1119 DSP Cellular down O.O1726O177 -0.222288576 FASEB.J. 1996 Adhesion Jun; 10(8): 871-81. protein KRT9 Structural no data Mech Ageing Dev. protein 2008 Oct: 129(10): 563-71. Epub 2008 Jun 3. KRTS Structural down 0.001982.117 -0.444955798 Mech Ageing Dev. protein 2008 Oct: 129(10): 563-71. Epub 2008 Jun 3. CAT Protects down 0.000701114 -0.476760771 Journal of rom UV nvestigative damage Dermatology (2006) 26, 182-190 TNFAIP3 Negative no data FEBS Lett. 2003 Feb Feedback of 1:536(1-3): 135-40 NFKA (aging marker) SOD2 Protects up 0.027437424 0.302541956 Journal of rom UV nvestigative damage Dermatology (1994) O2,476-480 FNDC3A Collagen no data FEBS Lett. 1994 Apr Assembly 8:343(1): 47-50 FGF2 Inhibits no data Arteriosclerosis, matrix Thrombosis, and collagen Vascular Biology. synthesis 993; 13:680-686 US 2012/00346 13 A1 Feb. 9, 2012 72

TABLE 5-continued

Gene Gene Symbol Function Direction p-value Log1ORG Reference TNC ECM O.OO1112139 0.142852906 Dev Dyn. 2000 Remodelling un: 218(2): 235-59. IGFBP5 Collagen Arthritis Rheum. 2006 Deposition Sep; 54(9): 3001-10 FN1 Structural O.O27437424 O.241.122656 Exp Cell Res. 1990 protein Nov. 191 (1): 8-13 TNFAIP6 Induces BMCImmunol. 2009 MMPS Mar 19; 10: 15 SMAD7 Negative O.OO1982.117 O.2325298.68 Biol Chem. 2005 regulator of Mar 4: 280(9): 8079-85. collagen Epub 2004 Dec 3 synthesis PRKCA Stimulates O.274898O33 Free Radic BioMed. Collagenase 999 Oct; 27(7-8): 729-37 FOXO1 Induces O.O3684.8433 O.238284.357 Am J Physiol Cell Collagen Physiol 292: C850-C856, Synthesis 2007 TIMP3 Inhibits no data Invest Dermatol. MMPS 998 Apr: 110(4): 416-21 FBN2 Elastin Fibre no data Invest Dermatol. Formation 996 May: 106(5): 1090-5 LOR Component down O.OOO701 114 -192078.4953 Biol Chem. 1992 of Cornefied Sep 5: 267 (25): 18060-6. envelope KLK8 Breakdown no data ournal of of cellular nvestigative adhesion Dermatology (2005) proteins 25, 1182-1189 DSC3 Cellular down O.OOS260102 -O.293.838.188 FASEB.J. 1996 Adhesion un; 10(8): 871-81. protein PKP3 Cellular no data FASEB.J. 1996 Adhesion un; 10(8): 871-81. protein KLK10 Cellular no data Invest Dermatol Adhesion 2003 121 : 542-549 protein S100A8 C3S(S down O.OO1131397 -0.63072926 Arch Dermatol Res. with 2009 Aug. 301(7): 523-9. photoaging Epub 2009 May 23. destroys proteins IL6 Stimulates up O.OO1131397 Journal of MMPS Investigative Dermatology (2004) 123, 1012-1019 ITGB4 Required for down O.OO1131397 -O.828.309922 J Cell Biol. 1996 July cell to cell 2: 134(2): 559-572 adhesion S100A9 Mediates down O.OO1131397 -0.6S 1297398 Biochem Biophys Res fibronectin Commun. 2007 March adhesion 2: 354(1): 84-89 ITGB1 Marker for up O.O1401738 Br J Dermatol. 2003 skin Apr. 148(4): 770-8 structural damage caused by UV COLAA1 Skin up O.2O8638.283 J. Clin Invest. 1989 Structural March; 83(3): 791-795 protein Taqman TLDA Analysis Relative Quantition - T-test with Benjamini and Hochberg correction HPRT as Endogenous Control Gene Untreated Control as Calibrator p value <0.05 Salicin 0.5% ws. Untreated Control US 2012/0034613 A1 Feb. 9, 2012 73

Selecting Third Subset of Genes row 216-218 to confirm/reject the results shown in the selec tion of genes in the table at 126 (corresponding to Table 4 Functional Youth Gene Assembly derived from Affymetrix microarray testing data) to arrive at the following results for the simplified hypothetical: (0165. Results from the RT-rt-PCR experiments may or 0171 Functional youth gene assembly for skin structure may not confirm that the candidate genes Subject to confir 144: Gene b is confirmed because it meets the AC criterion mation testing are regulated in the direction reflective of and matches the direction of expression associated with more youthful appearing skin based on the published scientific youthful skin structure per the literature data set on skin literature. Structure. 0166 The RT-rt-PCR data thus provide an additional basis 0172 Functional youth gene assembly for skin pigmenta for refining a gene functional youth gene assembly that is tion 146: Gene b is confirmed because it meets the AC derived from steps 102-134 of FIGS. 1A-1G. If a gene as criterion and matches the direction of expression associated tested for a gene assembly is not confirmed as having the with more youthful skin pigmentation per the literature data same direction that led to its selection from steps 102-134, it set on skin pigmentation. Gene c is not confirmed because it may now be removed as a member of the gene assembly. does not meet the AC criterion. Because the RT-rt-PCR data provide a new reading on the 0173 Functional youth gene assembly for skin hydration level of expression, a further expression level threshold can 148: Gene e is not confirmed because it does not meet the AC applied as a criterion for membership in an assembly. Genes criterion. Geneg is added because it meets the AC criterion are selected from those with expression levels determined and matches the direction of expression associated with more based on a relative quantification analysis. youthful skin hydration per the literature data set on skin 0167 Referring now to the simplified example shown in hydration. This is contrary to the microarray data. FIG. 1G, the next step is to select from the previously subdi 0.174 Functional youth gene assembly for cell turnover vided groups from the second Subset of genes (the functional 150: Gene e is not confirmed because it does not meet the AC youth gene assemblies), a third Subset of genes also Subdi criterion; however, if the criterion had been set at 38, it would vided by skin attribute regulated in a direction reflective of have met that level. Gene fis confirmed because it meets the youthful skin appearance, with an appropriately selected bio AC criterion and matches the direction of expression asso logical relevance level, e.g., a cycle criterion level (for ciated with more youthful skin cell turnover per the literature example, the criterion could be selected at 35 cycles or less). data set on skin cell turnover. (For PCR data, the lower the cycle number, the stronger the 0.175 Genes not chosen for the third subset of genes (gene gene expression.) h) may be considered for additional research based on sec 0.168. In FIG. 1G is a table showing a set of hypothetical ondary research factors 152. results for the simplified sample set of genes a-h; in particular, 0176 Turning to the testing-derived data example (actual it shows in row 216 AC values for genes b, c, e, f, g and h. (In Affymetrix data for salicin exposed tissue and data from the simplified example, we have assumed that genes a and d RT-rt-PCR testing), Table 6 (see also, FIG. 7 at 770) illustrates have no values from RT-rt-PCR, as those genes were previ one embodiment of a functional youth gene assembly ously filtered out of the second subset.) selected for the skin attribute “skin structure.’ Table 6 thus 0169. As noted, some genes dropped at earlier stages of represents a comparative analysis of the results of Table 5 and the process outlined in FIGS. 1G-1K may be reconsidered by Table 4, by confirmation of direction of regulation criterion adding them back into the RT-rt-PCR testing. In the simplified and the AC criterion applied to the hypothetical data. Thus, a example, gene h shows a Sufficient fold change in the hypo gene appears in Table 6, only if (a) it appears in Table 4 and the thetical microarray data but has not yet been identified with a data of Table 5 confirm that it have a strong-enough level of strong anti-aging effect. In the simplified example, geneg has expression based on the AC criterion and that the data of a fold change greater than 2, but the regulation direction Table 5 do not show a regulation direction that contradicts the shown by the first gene expression level testing was not con more youthful direction that was the basis for inclusion in sistent with the data from the literature. Assuming geneg and Table 4, or (b) although not in Table 4, it was added to the geneh were reconsidered during the RT-rt-PCR testing, this candidate list for secondary reasons and the result of the results in hypothetical data in row 216-218. RT-rt-PCR testing provides strong evidence that it should be (0170 Referring to FIGS. 1G-1K describing the simplified added, including a regulation direction that is consistent the example, the confirmation of direction of regulation criterion more youthful direction and a strong AC value, exceeding and the AC criterion are applied to the hypothetical data in the AC criterion.

TABLE 6

Gene Gene Symbol Description Direction p-value Log1ORG Reference

FOXO1 forkhead up 0.036848.433 0.238284.357 Am J Physiol Cell box O1 Physiol 292: C850-C856, 2007 ITGB1 integrin, beta 1 up 0.01401738 0.093562957 Br Dermatol. 2003 Apr. 148(4): 770-8 fibronectin 1 up 0.027437424 0.241 122656 Exp Cell Res. 1990 Nov. 191 (1): 8-13 US 2012/0034613 A1 Feb. 9, 2012 74

TABLE 6-continued

Gene Gene Symbol Description Direction p-value Log1ORG Reference SOD2 Superoxide up 0.027437424 0.302541956Journal of dismutase 2, Investigative mitochondrial Dermatology (1994) 102,476-480 COLAA1 Collagen Type up 0.016310135 0.208.638283J Clin Inwest. 1989 4 Alpha 1 March; 83(3): 791-795 S100A8 S100 calcium down 0.001131397 -0.63072926 Arch Dermatol binding Res. 2009 protein A8 Aug. 301 (7): 523-9. (calgranulim Epub 2009 May A or cystatin 23. A)

0177. The RT-rt-PCR methodology permits not only a sec- yet reported in the literature. These may be genes that are not ond reading on the activity of genes that have met the criteria currently associated with any biochemical pathway associ for a gene assembly in steps 102-134, it provides an oppor ated with skin, but may be in the future, as there are advances tunity to test a gene that has not met these criteria, but might in technology and further research studies. These genes may meet certain secondary research factors that Suggest it may be optionally be added to an appropriate functional youth gene of interest for a particular gene assembly. Secondary research assembly. factors may suggest further testing of genes that have a high 0181 Genes with non-skin related anti-aging mechanisms fold change without any literature support for their relevance of action may be subjected to further testing to determine the in skin tissue, genes associated with anti-aging mechanisms gene's effect, if any, on skin aging. For example, Scientific of action but not thought of as skin-related, or genes that are literature Suggests that the B-klotho gene appears to be strongly supported by literature as having an effect on skin involved in the aging process. See, U.S. Pat. No. 7,537.903. aging, but not achieving a significant fold change cutoff in 0182 Genes that are supported by literature as having an testing as described in steps 102-134 of the simplified effect on skin aging, but not achieving the biologically rel example. Genes with significant fold change values that are evant fold change cutoff in micro-array testing may be Sub not identified in the literature as having a favorable impact on jected to another round of micro-array testing with different a skin attribute may be considered for additional research. In concentrations of the agent or with different anti-aging agent the simplified sample set of genes, geneh not chosen for the (s). third subset of genes is considered for additional research 0183 In some embodiments, the functional youth gene based on secondary research factors 152. See FIGS. 1G-1K. assemblies, the groups of genes identified for a skin attribute For example, genes such as Klotho (KL) which have pub in genome-wide microarray tests, are optionally refined lished anti-aging benefits in mice may be of interest. Genes based on the results from the RT-rt-PCR experiments. If the Such as these may provide insights on identifying and discov literature discloses that a gene with 'up' regulation results in ering new novel pathways in the skin aging process. better skin structure, and the RT-rt-PCR data shows "down” 0.178 A favorable impact on a skin attribute is a biologi regulation for this gene, the gene may be set aside for possible cally relevant change that establishes a state of an attribute further research at a later date. Alternately, if the literature more similar to the non-aged state of the attribute. For discloses a gene with 'up' regulation results in better skin example, a favorable impact is recognized when an agent that structure, and the RT-rt-PCR data shows 'up' regulation for is applied to the skin results in a more youthful appearance. this gene, then the gene may be added to the functional youth The present system and method may be used to extend more gene assembly. efficiently the search for agents that cause a favorable impact. 0184. After application of secondary research factors, For example if the potential of a possible useful agent needs more genes are optionally added into one or more functional basic exploration, it can be run through the entire method of youth gene assembly 154. FIGS. 1A-1K to see how the resulting functional youth gene 0185. A method that utilizes the results of the groups of assembly compares to that of other agents. Using more than genes, the functional youth gene assemblies, may be used to one agent to execute the method decreases possible agent bias guide further research on aging of the skin. resulting from using a singular agent to determine a func tional youth gene assembly. Data Processing System Implementation 0179 For greater efficiency, once researchers have confi dence in one or more functional youth gene assemblies, test Data Sets ing of an agent my be done by omitting full genome microar 0186. As discussed above, FIG. 7 shows a schematic dia ray studies and using only more limited Studies for the genes gram of a system for carrying out the method disclosed, included in one of more of the functional youth gene assem including data set developed and used as the method proceeds blies. and the test equipment used to develop various sets of data 0180. One outcome of using the entire method described from the tissue samples of the skin models. This system can to screen agents that triggera relevant change in gene expres now be further explained with reference to FIG. 8, that shows sion is to identify genes for further study, even if they are not the data sets used in the system and method and how they are US 2012/0034613 A1 Feb. 9, 2012

transformed into the functional youth gene assemblies. The may be written in any suitable language, such as C++, or other system 700 broadly comprises a data processing system 710 languages suitable for the analysis and steps discussed above with a CPU and memory, in which there is an operating and shown in FIGS. 1A-1K. system 712, and the test equipment that develops data, includ 0191 FIG. 8 shows in simplified form the progression of ing a full genome microarray device 780 and a PCR testing data sets as the system proceeds to execute the method. In device 790. The test equipment is supplied, and the materials particular, FIG. 8 traces the test results data sets 802, 804, to be tested, prepared per the Supplier's instructions, include 806, 810,812 and shows the effect of the literature-based data the samples of agent exposed skin model 782 and the non sets and user selected parameters. FIG. 8 shows how the data exposed skin model 784. sets Stemming from the full genome microarray data 802. 804, 806, 808 and data set 810, stemming from the PCR 0187. The data processing system 710 includes a database instrument, are modified to obtain a confirmed skin attribute 730 that receives and stores the data used in the process subset 812, that is a function youth gene assembly by the data described above. The process applications modules 720 processing steps outlined in the simplified hypothetical execute, including statistics modules 722 and applications example of FIGS. 1A-1K. FIG. 8 also references Tables 1-6 using user selected process parameters 724 to perform the that are based on actual whole genome microarray and PCR flowchart (see FIGS. 1A-1K) processes. The process appli testing. cations modules 720 access the database 730 using suitable database management protocols 732. Screening Method 0188 The database stores the various data sets involved 0.192 While a primary use of the present methodology is including the full genome data sets 750a, 750b developed at to develop the functional youth gene assemblies that provide the full genome microarray device 780, the calculated ratio a focus for further gene-level research on skin attributes, the data 750c and the fold criterion result data set 752, developed methodology may also be used to Screen agents for effective by application of the fold change criterion. The database 730 ness to reduce skin aging. An agent may be chosen for testing also stores the pathway criterion data set 754 that identifies to assess the efficacy of the particular agent and to explore the the association between a gene and one or more biological genetic pathway focus of its action. Known anti-aging agents pathways and the intersection dataset 755 resulting from the have shown significantly different levels of gene expression intersection of the fold criterion result data set 752 and the in genes associated with a plurality of biochemical pathways pathway criterion data set 754. The database 730 further of the skin. A screening method of this type could signifi stores the skin attribute focus data sets 756 that defines the cantly lessen the number of costly and lengthy in vivo testing association between a particular skin attribute that is a under procedures done on many anti-aging product candidates. For study and genes that are associated with that attribute in the example, many consumer studies on facial anti-aging prod literature. After the intersection analysis of a particular skin ucts run for at least 12 weeks. Provided a reliable functional attribute focus data set 756 with the skin attribute/regulation youth gene assembly is identified, testing the effects of an direction data and the fold criterion result data 752 (which agent on the biochemical pathways associated with particular includes determining alignment of the more youthful regula genes provides a focused way to develop data on the action of tion direction for the particular biochemical pathways), the the an anti-aging candidate on a much shorter time frame and developed skin attribute subset 758, representing a prelimi provides quantitative data for comparison to other agents. nary functional youth gene assembly for a particular skin 0193 A screening approach may be used to assess the attribute is stored in database 730. likelihood of another agent working well in an anti-aging skin 0189 The data processing system's database 730 also care product. A new agent triggering levels of gene expres receives and holds data relevant to the PCR testing and results sion to a functional youth gene assembly similar to or Superior of the confirmation analysis for the preliminary functional to a known skin anti-aging agent may be considered for youth gene assembly. This includes storing the PCR candi further study, while a new agent that does not trigger similar date data 760, i.e., the listing of the genes based on the levels of gene expression in those genes in a functional youth preliminary functional youth gene assembly as Supplemented gene assembly may not be considered for further research with genes of secondary interest that will be subject to PCR investment. testing under the cycle level criterion or other parameters 0194 The screening method may also be used for improv used in the analysis of the PCR testing data. After the PCR ing the effective properties of existing anti-aging skin care tests have been run, the database 730 receives the PCR cycle products, selecting new anti-aging ingredients for products, data 762 including the associated up/down regulation direc and selecting blends of anti-aging ingredients for products. tion observed from testing. From the PCR cycle data set 762, From an understanding of which genes and which biochemi the processing applications 724 derives the Final Attribute cal pathways have skin anti-aging effects, the properties of an Data 770, 772 for one or more skin attributes. agent as a promoter of a biochemical pathway associated with 0190. As noted, stored in memory are the process appli more youthful appearance or an inhibitor of a biochemical cation modules 720. These are software generally in two pathway associated with less youthful appearance may be categories. A first category is the conventional statistical improved. Using the screening method on many possible analysis programs 722. Such as GeneSpring GX Software agent candidates instead of time-consuming clinical testing (version 10) or other commercial software to perform a para on fewer agent candidates is both a time-efficient and cost metric t-test with a Benjamini and Hochberg false discovery effective way of performing research and development. The rate correction. The StatMiner (Version 3) software may be method helps to provide consumers with anti-aging products used for analysis of the PCR data. The second category is the based on the most recent scientific research. flowchart process applications that implement the analysis 0.195 Other agents and agent blends including, for and steps discussed above and shown in FIGS. 1A-1K. The example, arNOX inhibitory agents derived from process application modules 720 that are custom-developed extracts may be tested. The plant for extract is optionally US 2012/0034613 A1 Feb. 9, 2012 76 selected from broccoli, shitake, coleus, rosemary, lotus, arti Surement of skin parameters confirm changes in particular choke, Sea rose tangerine, Oenothera biennis, astaxanthin, skin aging attributes as regulated by a particular functional red orange, Schisandra chinensis, Lonicera, Fagopyrum, car youth gene assembly. rot, Narcissus tazetta or olive. The arNOX inhibitory agents 0199 After a group of genes are selected as a functional optionally include salicylates, for example, Salicin, salicylic youth gene assembly, in vitro studies including the assay acid, salicyl hydroxamate, derivatives or combinations methods discussed above, are used to screen agent candidates thereof. and limit the amount of in vivo studies used in product devel 0196. While one embodiment of the present methodology opment. is to develop the functional youth gene assemblies that pro 0200. A skin care product, ageLOC(R) Future Serum, vide a focus for further gene-level research on skin attributes, including 0.5% salicin is the finished formulation used for when the methodology is used to screen agents for effective evaluation in clinical testing. The ageLOC(R) Future Serum is ness to reduce skin aging, it can assist in the formulation of a commercially available from Nu Skin Enterprises, Inc. composition to reduce skin aging. Once an agent has been (Provo, Utah). identified in testing to have efficacy as a promoter of a bio 0201 Twenty-nine out of thirty subjects completed the chemical pathway associated with more youthful appearance clinical study. Table 7 summarized the demographics of the or an inhibitor of a biochemical pathway associated with less study participants. youthful appearance for at least one skin attribute, that agent can be a candidate for an active ingredient in a composition to TABLE 7 reduce skin aging. Provided a reliable functional youth gene assembly has been identified and efficacy of an agent on the Study Participant's demographic summary. biochemical pathways associated with particular genes in that Demographic All Subjects assembly has been found, the composition can be targeted Summary (n = 30) specifically to improvement of the skin attribute associated Age (Years) Mean Age + Standard 56.06 7.76 with that functional youth gene assembly. A composition can Deviation be formulated that addresses multiple skin attributes, once Minimum Age 40.44 effective agents for the multiple skin attributes are found by Maximum Age 70.28 the process and system disclosed herein. A composition can Ethnicity Caucasian SO% Asian SO% also include a pharmaceutically acceptable carrier. A phar Fitzpatrick I S.10% maceutical acceptable carrier refers to a carrier medium that SkinType II 42.40% does not interfere with the effectiveness of the biological III 47.50% activity of the active ingredient, is chemically inert, and is not Demographic All Subjects toxic to the patient to whom it is administered. The type of the Summary (n = 30) carrier may include powders, emollients, lotions, creams, liquids and the like. 0202 The Fitzpatrick skin classification is based on the 0.197 Thus, the understanding of which genes and which skin's unprotected response to the first30 to 45 minutes of sun biochemical pathways have skin anti-aging effects and the exposure after a winter season without Sun exposure: properties of an agent as a promoter of a biochemical pathway 0203 I Always burns easily; never tans; II—Always associated with more youthful appearance oran inhibitor of a burns easily; tans minimally; III—Burns moderately, tans biochemical pathway associated with less youthful appear gradually; IV—Burns minimally, always tans well; ance is improved by the methods discussed herein, this under V rarely burns; tans profusely: VI Never burns; deeply standing can be translated into compositions that are directed pigmented. to one or more skin attributes associated with a functional 0204 Clinical Procedures youth gene assembly. It is expected that agents showing an 0205 At baseline (Visit 1), each prospective subject com anti-aging efficacy will be derived from broccoli, shitake, pleted an Eligibility and Health Questionnaire, and read and coleus, rosemary, lotus, artichoke, Sea rose tangerine, signed a Confidentially Agreement, a Photography Release Oenothera biennis, astaxanthin, red orange, Schisandra chin Form and an Informed Consent Agreement. Each Subject was ensis, Lonicera, Fagopyrum, carrot, Narcissus tazetta or explained the type of study, the detailed procedures and mate olive. They also may be derived from arNOX inhibitory rials to be tested, along with any known adverse reactions that agents that include Salicylates, for example, salicin, salicylic may result from participation. Subjects arrived at the clinic acid, salicyl hydroxamate, derivatives or combinations having refrained from applying any moisturizer to the face at thereof. These agents and their derivatives may then be least 3 to 5 days prior to visit and having cleansed the face to deployed in skin anti-aging formulations with a Sound basis in remove all makeup at least 30 minutes prior to visit. Subjects research at the genetic level. were consequently not allowed to use any other topical cos meceuticals, topical retinoid, or moisturizers during the 12 In Vivo Testing week duration of the study. Evaluation of Composition Including Anti-Aging 0206. Subjects used the study product on their face twice Agent daily for 12 weeks. Ordinal grading on a 9-point scale (0-none, 1-3-mild, 4-6-moderate, 7-9-severe) of facial fine 0198 For further confirmation of the effects of an agent lines, mottled pigmentation, uneven skin tone, tactile rough that is viewed as regulating in a youthful direction the path ness, global firmness appearance, jaw-line contour, radiance ways of a functional youth gene assembly for a skin attribute, and overall appearance was performed by investigator at consumer clinical studies may be conducted with a skin care baseline, week 1, week 4 week 8 and week 12. product including the anti-aging agent tested with in vitro 0207 Digital high-resolution photography was performed methods. Clinical studies with trained observation and mea on the front, right and left sides of the face. Each image was US 2012/0034613 A1 Feb. 9, 2012 77 taken while the subject's eyes were open. Each subject's of wrinkles evaluated against baseline at week 1, week 4, baseline photograph was compared to each post-baseline week 8 and week 12 time points. photograph to ensure consistent placement and lighting. 0217 FIG. 3 depicts the result of clinical investigator Color standards were imaged prior to each study visit. grading showing change of each investigated parameter (ex 0208 Corneometry measurements were taken on each cept wrinkles) against baseline at week 1, week 4 week 8 and subject’s left ocular bone (in line with pupil) to measure the week 12 time points. moisture content of the stratum corneum. 0218. The clinical investigator's facial assessments indi 0209 Ultrasound measurements were taken on the left cated a statistically significant improvement in facial fine side of each subject's face to measure density of the facial lines, tactile roughness, pore size, radiance and overall skin in the crow's feet area. Measurements were taken with appearance at week 1 time point (Ps0.05). All of the benefits the probe oriented perpendicular to the body axis while the continued into weeks 4, 8 and 12. Statistically significant Subjects were resting Supine on a padded patient table. improvement in mottled pigmentation, global firmness, Sub 0210. A single cutometer measurement was taken on the categories of wrinkles and jaw-line contour was recorded at right side of the subject's face, in line with the corner of the week 4 time point (Ps0.05) and continued through to week eye and the edge of the nostril, to measure the extensibility of 12. the skin. 0219 Corneometer measurements indicated statistical 0211 All clinical and corneometer measurements and improvement in hydration at week 1 time point (Ps0.05). evaluations were taken at baseline, week 1 (visit 2), week 4 This improvement continued through week 8. Hydration of (visit 3), week 8 (visit 4) and week 12 (visit 5). Ultrasound the stratum corneum was decreased significantly (Ps0.05) at and cutometer measurements and evaluations were taken at the week 12 time point. baseline, week 4 (visit 3), week 8 (visit 4) and week 12 (visit 0220 FIG. 4 depicts the result of corneometer grading 5). Completed patient's diaries were reviewed for compliance showing moisture content of the Stratum corneum, at week 1, at each visit. week 4 week 8 and week 12 time points. 0212 Biostatistics 0221 Cutometer measurements indicated statistical 0213 Mean clinical grading and instrumentation scores at improvement in extensibility of the skin at week 12 time point each visit were statistically compared to baseline scores using (Ps0.05). paired t-test. Changes from baseline were considered signifi 0222 FIG. 5 depicts the results of cutometer readings cant at the ps0.05 level. Mean percent change from baseline showing extensibility measurements of the skin, evaluated and incidence of positive responders were calculated for all against baseline at week 4 week 8 and week 12 time points. attributes. Comparisons, based on the average from baseline, 0223 Ultrasound measurements indicated statistical were made between the test materials using analysis of Vari improvement in density of the skin at week 4 time point ance (ANOVA). (Ps0.05). This improvement continued through week 12. 0214) Results 0224 FIG. 6 depicts the result of density evaluation from 0215. Twenty-nine of thirty subjects successfully com ultrasound, against baseline at week 4 week 8 and week 12 pleted the study with one subject unable to complete due to time points. personal reasons. Compliance assessments indicated that 0225. No tolerability issues related to erythema, edema Subjects were following test formulation use instructions. and Scaling were observed by the investigator. Three Subjects 0216 FIG. 2 depicts the result of clinical investigator reported a slight stinging at week 8 (Ps0.05) and slight grading showing a breakdown of the different Sub-categories itching at week 12 (Ps0.05).

TABLE 8 Mean values of Clinical Grading and Instrumentation

Base line Week 1 Week 4 Week 8 Week 12

Mean Mean % Change Mean % Change Mean % Change Mean % Change Crows Feet Wrinkles 3.29 3.21 (-0.5%) 3.00 (-8.9%) 2.71 0 (-17.8%) 2.48 (-24.6%) Under-Eye Wrinkles 3.19 3.07 (-1.7%) 2.79 0 (-12.4%) 2.550 (-20.0%) 2.240 (-29.7%) Area Cheek Wrinkles 2.41 2.29 (-1.5%) 2.26 (-6.4%) 2.12 0 (–12.1%) 2.020 (-16.4%) Face Fine Lines 4.29 4.14 (-3.7%) 3.76 0 (-12.4%) 3.47 0 (-19.2%) 3.100 (-27.7%) (Overall) Mottled 5.05 5.05 (-0.7%) 4.740 (-6.1%) 4.57 (-9.5%) 4.33 (-14.3%) Pigmentation Uneven Skin 5.05 5.05 (-0.7%) 4.74 0. (-6.1%) 4.57 (-9.5%) 4.340 (-13.9%) Tone Tactile 3.48 2.41 (-30.7%) 2.020 (-42.0%) 1.43 0 (-58.9%) 0.980 (-71.7%) Roughness Smoothness Global S.O.3 S.04 (0.3%) 4.860 (-3.4%) 4.640 (-7.8%) 4.48 0 (-10.9%) Firmness Appearance Jawline S.16 5.14 . (0.0%) S.10 (-1.0%) 4.95 U. (-4.0%) 4.74 U. (-8.0%) Contour US 2012/0034613 A1 Feb. 9, 2012 78

TABLE 8-continued Mean values of Clinical Grading and Instrumentation Base line Week 1 Week 4 Week 8 Week 12 Mean Mean % Change Mean % Change Mean % Change Mean % Change Pore Size 4.41 4.30 (-2.4%) 4.050 (-8.2%) 3.869 (-12.5%) 3.71 0 (-16.0%) Radiance 5.81 5.20 (-11.0%) 4.90 C (-15.7%) 4.72 (-18.6%) 4.59 0 (-21.0%) Overall 547 5.21 (-4.2%) 4.91 0 (-10.0%) 4.670 (-14.5%) 4.500 (-17.6%) Appearance Left Ocular Corneometer 56.49 60.541 (7.3%) 62.491 (10.6%) 63.91 ff (13.1%) 52.23 (-7.5%) Bone Cutometer Extensibility 1.15 NA NA 1.17 (1.8%) 1.17 (1.3%) 1.261. (9.6%) Ultrasound Density 18.55 NA NA 21.38 ft (15.2%) 23.52 ft. (26.7%) 25.21 ft. (35.8%)

Indicates a statistically significant (ps 0.05) increase compared to Baseline Indicates a statistically significant (ps 0.05) decrease compared to Baseline

0226 All references disclosed herein, whether patent or 2. The method of claim 1, wherein the biochemical path non-patent, are hereby incorporated by reference as if each way associated with the physical appearance of skin aging was included at its citation, in its entirety. comprises at least one of skin structural protein synthesis, 0227. Although the present disclosure has been described skin structural degradation and maintenance, extracellular with a certain degree of particularity, it is understood the matrix assembly, cellular differentiation, skin barrier compo disclosure has been made by way of example, and changes in nent synthesis, skin barrier integrity, water regulation, or detail or structure may be made without departing from the regulation of melanin production and control. spirit of the disclosure as defined in the appended claims. 3. The method of claim 1, wherein the skin attribute for the Aspects of the disclosure include: at least one skin attribute Subset of genes is skin structure, skin 1. A method of testing to identify genes associated with one pigmentation, skin hydration or cell turnover. or more physical attributes of skin aging comprising: 4. The method of claim 1, wherein the first, selected bio exposing a first sample of human skin tissue to an agent; logical relevance level is about a two fold difference between determining a first set of expression levels of a plurality of the exposed and unexposed samples. genes in the first sample of human skin; 5. The method of claim 1, wherein the human skin tissue comparing the first set of expression levels to a second set comprises skin cells comprising at least one of keratinocytes, of expression levels, the second set of expression levels fibroblasts, adipocytes, melanocytes or combinations thereof. corresponding to expression levels of human skin tissue 6. The method of claim 1, wherein the first set of expression not exposed to the agent, to identify a first Subset of levels of a plurality of genes comprises expression levels for genes having a fold change difference in expression essentially the full human genome. level between the exposed and unexposed samples that 7. The method of claim 1, wherein the method for deter meets a first, selected biological relevance level; mining expression levels that is different than that used for the Selecting from the first Subset of genes a second Subset of first sample of human tissue is a method using an RNA genes, each gene being associated with a biochemical quantification metric. pathway associated with physical appearance of skin 8. The method of claim 1, wherein the step of selecting aging: from the second Subset of genes, at least one skin attribute Selecting from the second Subset of genes, at least one skin Subset of genes, each gene in the skin attribute Subset being attribute Subset of genes, each gene in the skin attribute associated with a biochemical pathway relating to the skin Subset being associated with a biochemical pathway attribute that is shown in the comparing step to have been relating to the skin attribute that is shown in the compar regulated in a more youthful direction for that biochemical ing step to have been regulated in a more youthful direc pathway and skin attribute comprising performing this step tion for that biochemical pathway and skin attribute: for a plurality of skin attribute Subsets of genes; and exposing a second sample of human skin tissue to the the step selecting a third Subset of genes from the at least agent, one skin attribute Subset of genes whose expression lev determining the levels of expression for the at least one skin els in the second sample of human skin tissue meet a attribute Subset of genes in the second sample of human second, selected biological relevance level and whose skin tissue using a method for determining expression direction of regulation conforms to the more youthful levels that is different than that used for the first sample direction used in selecting the at least one skin attribute of human skin tissue; and Subset of genes comprising performing this step for a Selecting a third Subset of genes from the at least one skin plurality of skin attribute Subsets of genes. attribute subset of genes whose expression levels in the 9. The method of claim 8, wherein the plurality of skin second sample of human skin tissue meet a second, attribute subsets of genes are two or more skin attribute subset selected biological relevance level and whose direction of genes selected from the group consisting of skin structure, of regulation conforms to the more youthful direction skin pigmentation, skin hydration and cell turnover. used in selecting the at least one skin attribute Subset of 10. The method of claim 1 further comprising determining genes. the levels of expression for additional genes associated with a US 2012/0034613 A1 Feb. 9, 2012 79 biochemical pathway associated with skin aging in the sec 15. The system of claim 11, wherein the first set of expres ond sample of human skin tissue using a method for deter Sion, levels of a plurality of genes comprises expression levels mining expression levels that is different than that used for the for essentially the full human genome. first sample of human tissue; and 16. The system of claim 11, wherein the second instrument Selecting for the third Subset of genes those genes from the for determining expression levels that is different than that used for the first sample of human tissue is an instrument additional genes associated with a biochemical pathway using an RNA quantification metric. associated with skin aging whose expression levels in 17. A method of assessing the efficacy of a skin anti-aging the second sample of human skin tissue meet a second, agent comprising: selected biological relevance level and whose direction exposing a first sample of human skin tissue to an agent; ofregulation conforms to the more youthful direction of determining a first set of expression levels of a plurality of regulation of the associated biochemical pathway. genes in the first sample of human skin; 11. A computer based system of testing to identify genes comparing the first set of expression levels to a second set associated with one or more physical attributes of skin aging of expression levels, the second set of expression levels comprising: corresponding to expression levels of human skin tissue a first instrument for exposing a first sample of human skin not exposed to the agent, to identify a first Subset of tissue to an agent and determining a first set of expres genes having a fold change difference in expression sion levels of a plurality of genes in the first sample of level between the exposed and unexposed samples that human skin; meets a first, selected biological relevance level; a computer module for comparing the first set of expression selecting from the first Subset of genes a second Subset of levels to a second set of expression levels, the second set genes, each gene being associated with a biochemical of expression levels corresponding to expression levels pathway associated with physical appearance of skin of human skin tissue not exposed to the agent to identify aging: a first Subset of genes having a fold change difference in selecting from the second Subset of genes, at least one skin expression level between the exposed and unexposed attribute Subset of genes, each gene in the skin attribute samples that meet a first, selected biological relevance Subset being associated with a biochemical pathway level; relating to the skin attribute that is shown in the compar a computer module for accessing a stored data set identi ing step to have been regulated in a more youthful direc fying genes, each gene being associated with a bio tion for that biochemical pathway and skin attribute; chemical pathway associated with physical appearance exposing a second sample of human skin tissue to the of skin aging and for selecting from the first Subset a agent, second Subset comprising those genes also in the second determining the levels of expression for the at least one skin Subset; attribute Subset of genes in the second sample of human a computer module for selecting from the second Subset of skin tissue using a method for determining expression genes, at least one skin attribute Subset of genes, each levels that is different than that used for the first sample gene in the skin attribute Subset being associated with a of human skin tissue; biochemical pathway relating to the skin attribute that is selecting a third Subset of genes from the at least one skin shown in the comparing step to have been regulated in a attribute subset of genes whose expression levels in the more youthful direction for that biochemical pathway second sample of human skin tissue meet a second, selected biological relevance level and whose direction and skin attribute; of regulation conforms to the more youthful direction a second instrument for exposing a second sample of used in selecting the at least one skin attribute Subset of human skin tissue to the agent and for determining the genes; and levels of expression for the at least one skin attribute comparing the third Subset of genes to a previously deter Subset of genes in the second sample of human skin mined third Subset of genes for a second agent, thereby tissue using a method for determining expression levels showing the efficacy of the skin anti-aging agent. that is different than that used for the first sample of 18. The method of claim 17, wherein the skin attribute for human tissue; and the at least one skin attribute Subset of genes is skin structure, a computer module for selecting a third Subset of genes skin pigmentation, skin hydration or cell turnover. from the at least one skin attribute subset of genes whose 19. The method of claim 17, wherein the method for deter expression levels in the second sample of human skin mining expression levels that is different than that used for the tissue meet a second, selected biological relevance level first sample of human tissue is a method using an RNA and whose direction of regulation conforms to the more quantification metric. youthful direction used in selecting the at least one skin 20. A method of formulating a composition with a plurality attribute Subset of genes. of skin anti-aging agents, comprising: 12. The system of claim 11, wherein the skin attribute for assessing the efficacy of each of two or more skin anti the at least one skin attribute Subset of genes is skin structure, aging agents by a method comprising: skin pigmentation, skin hydration or cell turnover. exposing a first sample of human skin tissue to an agent; 13. The system of claim 11, wherein the first, selected determining a first set of expression levels of a plurality of biological relevance level is about a two fold difference genes in the first sample of human skin; between the exposed and unexposed samples. comparing the first set of expression levels to a second set 14. The system of claim 11, wherein the human skin tissue of expression levels, the second set of expression levels comprises skin cells comprising at least one of keratinocytes, corresponding to expression levels of human skin tissue fibroblasts, adipocytes, melanocytes or combinations thereof. not exposed to the agent, to identify a first Subset of US 2012/0034613 A1 Feb. 9, 2012

genes having a fold change difference in expression of regulation conforms to the more youthful direction level between the exposed and unexposed samples that used in selecting the at least one skin attribute Subset of meets a first, selected biological relevance level; genes; and Selecting from the first Subset of genes a second Subset of comparing the third Subset of genes to a previously deter genes, each gene being associated with a biochemical mined third Subset of genes for a second agent, thereby pathway associated with physical appearance of skin showing the efficacy of the skin anti-aging agent; aging: selecting from the two or more skin anti-aging agents Selecting from the second Subset of genes, at least one skin assessed two or more agents found to have efficacy for at attribute Subset of genes, each gene in the skin attribute least one skin attribute; and Subset being associated with a biochemical pathway formulating a composition with Such two or more agents relating to the skin attribute that is shown in the compar found to have efficacy as active ingredients and a phar ing step to have been regulated in a more youthful direc maceutically acceptable carrier. tion for that biochemical pathway and skin attribute: 21. The method of claim 20, wherein the skin attribute for exposing a second sample of human skin tissue to the the at least one skin attribute Subset of genes is skin structure, agent, skin pigmentation, skin hydration or cell turnover and the determining the levels of expression for the at least one skin composition is directed to regulation of the genes in the at attribute Subset of genes in the second sample of human least one skin attribute Subset of genes in a more youthful skin tissue using a method for determining expression direction. levels that is different than that used for the first sample 22. The method of claim 20, wherein the method for deter of human skin tissue; mining expression levels that is different than that used for the Selecting a third Subset of genes from the at least one skin first sample of human tissue is a method using an RNA attribute subset of genes whose expression levels in the quantification metric. second sample of human skin tissue meet a second, selected biological relevance level and whose direction