DOCSLIB.ORG

Notch-Mediated Expansion of Cord Blood Progenitors

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Notch-Mediated Expansion of Cord Blood Progenitors Letters to the Editor 1948 patient number places some limitation on interpretation, there is a REFERENCES suggestion that this treatment could still be considered for patients 1 Auner HW, Szydlo R, Rone A, Chaidos A, Giles C, Kanfer E et al. Salvage autologous who may have not previously been referred for auto-HCT but who stem cell transplantation for multiple myeloma relapsing or progressing after up- now have relapsed disease. front autologous transplantation. Leuk Lymphoma 2013; 54: 2200–2204. Our adaptive study design ultimately determined that the 2 Shah N, Ahmed F, Bashir Q, Qureshi S, Dinh Y, Rondon G et al. Durable remission highest doses of lenalidomide (75 and 100 mg) were equivalent to with salvage second autotransplants in patients with multiple myeloma. Cancer 118 – achieve a balance between tolerability and efficacy. Owing to 2012; : 3549 3555. relatively small patient numbers, a longer follow-up time may 3 Lemieux E, Hulin C, Caillot D, Tardy S, Dorvaux V, Michel J et al. Autologous stem cell transplantation: an effective salvage therapy in multiple myeloma. Biol Blood further differentiate the true clinical effect of the dose escalation Marrow Transplant 2013; 19:445–449. and the impact of post-auto-HCT maintenance therapy, which 4 Cook G, Williams C, Brown JM, Cairns DA, Cavenagh J, Snowden JA et al. High- 63% of patients received. Unfortunately (although not surpris- dose chemotherapy plus autologous stem-cell transplantation as consolidation ingly), poor-risk cytogenetics independently predicted for poorer therapy in patients with relapsed multiple myeloma after previous autologous OS, reminding us of the continued need for innovative treatment stem-cell transplantation (NCRI myeloma × relapse (Intensive trial)): a randomised, options for these patients. open-label, phase 3 trial. Lancet Oncol 2014; 15: 874–885. In conclusion, combination of high-dose melphalan with high- 5 Kumar SK, Lacy MQ, Hayman SR, Stewart K, Buadi FK, Allred J et al. Lenalidomide, cyclophosphamide and dexamethasone (CRd) for newly diagnosed multiple dose lenalidomide appears to be a well tolerated and safe 86 – preparative regimen in the setting of salvage auto-HCT. As novel myeloma: results from a phase 2 trial. Am J Hematol 2011; :640 645. 6 van de Donk NW, Wittebol S, Minnema MC, Lokhorst HM. Lenalidomide (Revlimid) agents, maintenance regimens and immunotherapies emerge, it is combined with continuous oral cyclophosphamide (endoxan) and prednisone conceivable that they too may be combined with this auto-HCT (REP) is effective in lenalidomide/dexamethasone-refractory myeloma. Br J platform to offer the relapsed patient yet another chance at a Haematol 2010; 148: 335–337. durable response. 7 Durie BG, Harousseau JL, Miguel JS, Blade J, Barlogie B, Anderson K et al. International uniform response criteria for multiple myeloma. Leukemia 2006; 20:1467–1473. 8 Thall PF, Cook JD. Dose-finding based on efficacy-toxicity trade-offs. Biometrics CONFLICT OF INTEREST 2004; 60:684–693. χ2 Drs N Shah, Bashir, JJ Shah, Hosing and Orlowski receive research funding from 9 Fisher R. On the interpretation of from contingency tables, and the 85 – Celgene. Drs N Shah, JJ Shah and Orlowski have served on Celgene Advisory Boards. calculation of P. J R Stat Soc 1922; :87 94. 10 Freeman GH, Halton JH. Note on exact treatment of contingency, goodness of fit fi 38 – 1 2 2 1 3 1 4 and other problems of signi cance. Biometrika 1951; :141 149. N Shah , PF Thall , PS Fox , Q Bashir , JJ Shah , S Parmar , P Lin , 11 Gelman ACJ, Stern HS, Rubin DB. Bayesian Data Analysis, 2nd edn. Chapman & 1 1 1 1 1 P Kebriaei , Y Nieto , UR Popat , CM Hosing , A Cornelison , Hall/CRC Press: New York, NY, USA, 2004. 1 3 1 1 EJ Shpall , RZ Orlowski , RE Champlin and MH Qazilbash 12 Ibrahim JG, Chen M-H, Sinha D. Bayesian Survival Analysis.Springer:NewYork,NY,2001. 1Department of Stem Cell Transplantation and Cellular Therapy, The 13 Doo NW, Thompson PA, Prince HM, Seymour JF, Ritchie D, Stokes K et al. University of Texas MD Anderson Cancer Center, Houston, TX, USA; Bortezomib with high dose melphalan conditioning for autologous transplant is 2Department of Biostatistics, The University of Texas MD Anderson safe and effective in patients with heavily pretreated and high risk multiple 54 – Cancer Center, Houston, TX, USA; myeloma. Leuk Lymphoma 2013; : 1465 1472. 3Department of Lymphoma and Myeloma, The University of Texas 14 Orlowski RZ. Novel agents for multiple myeloma to overcome resistance in phase III clinical trials. Semin Oncol 2013; 40:634–651. MD Anderson Cancer Center, Houston, TX, USA and 4 15 Kumar SK, Lee JH, Lahuerta JJ, Morgan G, Richardson PG, Crowley J et al. Risk of Department of Hematopatholoy, The University of Texas MD progression and survival in multiple myeloma relapsing after therapy with IMiDs Anderson Cancer Center, Houston, TX, USA and bortezomib: a multicenter international myeloma working group study. E-mail: [email protected] Leukemia 2012; 26:149–157. Supplementary Information accompanies this paper on the Leukemia website (http://www.nature.com/leu) Notch-mediated expansion of cord blood progenitors: maintenance of transcriptional and epigenetic fidelity Leukemia (2015) 29, 1948–1951; doi:10.1038/leu.2015.61 provide more rapid short-term myeloid reconstitution when co- infused with a non-manipulated CB unit(s). However, infusion of ex vivo generated HSPCs for long-term hematopoietic reconstitu- Recent advances have been made towards successful generation tion remains a goal in the context of stem cell transplantation and of human hematopoietic stem and progenitor cells (HSPCs) gene therapy, thus the development of methods to assess the derived from embryonic, induced pluripotent or hematopoietic safety of these cultured products are essential. stem cells. However, whether these stem cell populations, which Our group and others have demonstrated successful expansion are generated after ex vivo manipulation, are suitable for clinical of CB HSPCs for both pre-clinical and clinical applications. application depends on the faithful preservation of their Standardized methods for assessing transcriptional quality control transcriptional, epigenetic and functional properties with respect of pluripotent cell lines have been developed; however, there has to their primary cell counterparts. been no standard approach to assessing long-term safety of To overcome the significant delay in neutrophil recovery cultured progenitor cells infused in humans.1 Here we investi- following cord blood (CB) transplantation (CBT), we developed gated the transcriptional and epigenetic states of CB HSPC methods for the ex vivo expansion of CD34+ CB-derived HSPC by generated after ex vivo expansion on Delta1. We found that the culture with the Notch-ligand Delta1. When infused into patients transcriptomes and methylomes of the expanded HSPC overall undergoing a myeloablative CBT, these cells have been shown to recapitulate those of their primary cell counterparts. Accepted article preview online 6 March 2015; advance online publication, 27 March 2015 Leukemia (2015) 1939 – 1958 © 2015 Macmillan Publishers Limited Letters to the Editor 1949 CD34+ CB HSPCs were cultured under conditions identical to cells. To determine whether BEX2 downregulation was the result those in our ongoing clinical trials.2 We validated in vitro and of altered expression of transcription factors known to regulate in vivo growth characteristics of cells used for transcriptional and BEX2, we looked at the expression levels of SOX2, β-catenin, p65 methylation studies to ensure their similarity with our previously and ERBB2 and found no difference between primary and published results. Mean CD34+ fold expansion was 191 ± 62-fold cultured cells. as compared with 184 ± 35 previously. In vivo repopulation in We compared genes upregulated in the cultured cells with an sublethally irradiated (275 rad) NOD-SCID IL-2Rγ-null mice (NSG) established list of proto-oncogenes (www.uniprot.org/uniprot, approved for use by the Fred Hutchinson Cancer Research Center search terms ‘proto-oncogene’, ‘human’) as upregulation of these Institutional Animal Care and Use Committee was at least as genes might be expected to result in unintended cell proliferation. robust as our previously published results. Among the list of 232 proto-oncogenes, 11 were upregulated in For transcriptional analysis, CD34+ CB HSPCs were isolated from cultured cells (Figure 1c). Although many of these genes have six individual CB units within 24 h of collection. Freshly isolated been implicated in hematologic malignancies, they also have CD34+ CB HSPCs were then frozen for subsequent RNA extraction, important roles in normal hematopoiesis, and thus may be whereas remaining cells were placed in culture for 14 days as upregulated in this setting. Because the CB HSPCs were cultured in previously described.2 Upon cell harvest at day 14 of culture, the presence of Delta1, we also determined whether there was CD34+ cells were selected for RNA extraction and analysis on the persistent altered expression of Notch downstream target genes Illumina HT12v3 platform (Illumina Inc, San Diego, CA, USA). All as some of these have known roles in cell cycle entry and data were analyzed using the Bioconductor framework.3 Lumi proliferation. We found no difference in the expression of known package4 was used for reading and normalizing the raw data Notch targets SKP2, HES1, HEY1, CCND1 or BCL2. As shown in output by GenomeStudio software (Illumina Inc.). Limma package3 Figure 1c, MYC expression was upregulated following culture on was used for identifying differentially expressed genes between Delta1 ligand; however, Notch is one of many regulators of MYC freshly isolated and cultured CD34+ CB HSPCs. Significant genes expression. were defined as those with false discovery rate (FDR) o1% and Whereas transcriptional activity represents cellular function, we 4twofold change between non-manipulated and ex vivo expected that DNA methylation states would offer a more detailed expanded cells. picture of the fidelity of the expanded populations with respect to Of 24 912 genes analyzed, 668 genes were upregulated in the cell identity and thus safety for clinical use.
Load more

Recommended publications
  • Genes in a Refined Smith-Magenis Syndrome Critical Deletion Interval on Chromosome 17P11.2 and the Syntenic Region of the Mouse
    Downloaded from genome.cshlp.org on September 25, 2021 - Published by Cold Spring Harbor Laboratory Press Article Genes in a Refined Smith-Magenis Syndrome Critical Deletion Interval on Chromosome 17p11.2 and the Syntenic Region of the Mouse Weimin Bi,1,6 Jiong Yan,1,6 Paweł Stankiewicz,1 Sung-Sup Park,1,7 Katherina Walz,1 Cornelius F. Boerkoel,1 Lorraine Potocki,1,3 Lisa G. Shaffer,1 Koen Devriendt,4 Małgorzata J.M. Nowaczyk,5 Ken Inoue,1 and James R. Lupski1,2,3,8 Departments of 1Molecular & Human Genetics, 2Pediatrics, Baylor College of Medicine, 3Texas Children’s Hospital, Houston, Texas 77030, USA; 4Centre for Human Genetics, University Hospital Gasthuisberg, Catholic University of Leuven, B-3000 Leuven, Belgium; 5Department of Pathology and Molecular Medicine, McMaster University, Hamilton, Ontario L8S 4J9, Canada Smith-Magenis syndrome (SMS) is a multiple congenital anomaly/mental retardation syndrome associated with behavioral abnormalities and sleep disturbance. Most patients have the same ∼4 Mb interstitial genomic deletion within chromosome 17p11.2. To investigate the molecular bases of the SMS phenotype, we constructed BAC/PAC contigs covering the SMS common deletion interval and its syntenic region on mouse chromosome 11. Comparative genome analysis reveals the absence of all three ∼200-kb SMS-REP low-copy repeats in the mouse and indicates that the evolution of SMS-REPs was accompanied by transposition of adjacent genes. Physical and genetic map comparisons in humans reveal reduced recombination in both sexes. Moreover, by examining the deleted regions in SMS patients with unusual-sized deletions, we refined the minimal Smith-Magenis critical region (SMCR) to an ∼1.1-Mb genomic interval that is syntenic to an ∼1.0-Mb region in the mouse.
    [Show full text]
  • Metastatic Adrenocortical Carcinoma Displays Higher Mutation Rate and Tumor Heterogeneity Than Primary Tumors
    ARTICLE DOI: 10.1038/s41467-018-06366-z OPEN Metastatic adrenocortical carcinoma displays higher mutation rate and tumor heterogeneity than primary tumors Sudheer Kumar Gara1, Justin Lack2, Lisa Zhang1, Emerson Harris1, Margaret Cam2 & Electron Kebebew1,3 Adrenocortical cancer (ACC) is a rare cancer with poor prognosis and high mortality due to metastatic disease. All reported genetic alterations have been in primary ACC, and it is 1234567890():,; unknown if there is molecular heterogeneity in ACC. Here, we report the genetic changes associated with metastatic ACC compared to primary ACCs and tumor heterogeneity. We performed whole-exome sequencing of 33 metastatic tumors. The overall mutation rate (per megabase) in metastatic tumors was 2.8-fold higher than primary ACC tumor samples. We found tumor heterogeneity among different metastatic sites in ACC and discovered recurrent mutations in several novel genes. We observed 37–57% overlap in genes that are mutated among different metastatic sites within the same patient. We also identified new therapeutic targets in recurrent and metastatic ACC not previously described in primary ACCs. 1 Endocrine Oncology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. 2 Center for Cancer Research, Collaborative Bioinformatics Resource, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. 3 Department of Surgery and Stanford Cancer Institute, Stanford University, Stanford, CA 94305, USA. Correspondence and requests for materials should be addressed to E.K. (email: [email protected]) NATURE COMMUNICATIONS | (2018) 9:4172 | DOI: 10.1038/s41467-018-06366-z | www.nature.com/naturecommunications 1 ARTICLE NATURE COMMUNICATIONS | DOI: 10.1038/s41467-018-06366-z drenocortical carcinoma (ACC) is a rare malignancy with types including primary ACC from the TCGA to understand our A0.7–2 cases per million per year1,2.
    [Show full text]
  • Genomic Organization of the Approximately 1.5 Mb Smith
    European Journal of Human Genetics (2001) 9, 892 ± 902 ã 2001 Nature Publishing Group All rights reserved 1018-4813/01 $15.00 www.nature.com/ejhg ARTICLE Genomic organisation of the ~1.5 Mb Smith-Magenis syndrome critical interval: Transcription map, genomic contig, and candidate gene analysis Rebecca E Lucas1, Christopher N Vlangos1, Parimal Das4, Pragna I Patel4 and Sarah H Elsea*,1,2,3 1Genetics Graduate Program, Michigan State University, East Lansing, Michigan, MI 48824, USA; 2Department of Zoology, Michigan State University, East Lansing, Michigan, MI 48824, USA; 3Department of Pediatrics and Human Development, Michigan State University, East Lansing, Michigan, MI 48824, USA; 4Department of Neurology, Baylor College of Medicine, Houston, Texas, TX 77030, USA Smith-Magenis syndrome (SMS) is a multiple congenital anomalies/mental retardation syndrome associated with an interstitial deletion of chromosome 17 involving band p11.2. SMS is hypothesised to be a contiguous gene syndrome in which the phenotype arises from the haploinsufficiency of multiple, functionally-unrelated genes in close physical proximity, although the true molecular basis of SMS is not yet known. In this study, we have generated the first overlapping and contiguous transcription map of the SMS critical interval, linking the proximal 17p11.2 region near the SMS-REPM and the distal region near D17S740 in a minimum tiling path of 16 BACs and two PACs. Additional clones provide greater coverage throughout the critical region. Not including the repetitive sequences that flank the critical interval, the map is comprised of 13 known genes, 14 ESTs, and six genomic markers, and is a synthesis of Southern hybridisation and polymerase chain reaction data from gene and marker localisation to BACs and PACs and database sequence analysis from the human genome project high-throughput draft sequence.
    [Show full text]
  • Delineation of Key Regulatory Elements Identifies Points Of
    DELINEATION OF KEY REGULATORY ELEMENTS IDENTIFIES POINTS OF VULNERABILITY IN THE MITOGEN-ACTIVATED SIGNALING NETWORK SUPPLEMENTARY MATERIALS List of contents Supplementary Figures with legends 1. Figure S1: Distribution of primary siRNA screen data, and standardization of assay procedure. 2. Figure S2: Scatter plot of screen data. 3. Figure S3: Functional relevance of the identified targets and Calculation of residence time from PDT and cell cycle distribution. 4. Figure S4: FACS profiles for ABL1 and AKT1. Table for data in Figure 5B. 5. Figure S5: Venn diagram showing the results of the comparative analysis of other screen results 6. Figure S6: Dose response profiles for the AKT1 + ABL1 inhibitor combination for CH1, list of the 14 cell lines and their description, effect of ABL1+AKT1 inhibitor combination on increase in apoptotic cells and G1 arrest in 14 cell lines, effects of CHEK1 inhibitor on combination C1,C2 on 4 cell lines. Supplementary Tables 1. Table S1: siRNA screen results for targeted kinases and phosphatases. 2. Table S2: Gene expression status of the validated hits. 3. Table S3: Role played by identified RNAi hits in regulation of cell cycle, the effect on PDTs along with phase-specific RTs. 4. Table S4: List of molecules classified as cell cycle targets. 5. Table S5: High confidence network used for graph theory analysis. 6. Table S6: Occurrences of nodes in shortest path networks. 7. Table S7: Network file used as SNAVI background. 8. Table S8: Classification of nodes present in modules according to specificity. Legends for tables Supplementary Experimental Procedures References Figure S1 A 450 400 G1 S 350 G2 300 250 200 150 100 50 Distribution of molecules Distribution 0 -6-4-20246 Z-score 350 200 400 G1 S 300 G2 150 300 250 200 100 200 150 100 50 100 Distribution of molecules 50 0 0 0 -4 -2 0 2 4 -4-20246 -4-20246 Z-score B PLK1 GAPDH PLCg BTK PLCg CDC2A PLCg CHEK1 PLCg MET Distribution profiles of complete primary screen and western blots showing knockdown efficiency.
    [Show full text]
  • The Proteomic Landscape of Resting and Activated CD4+ T Cells Reveal Insights Into Cell Differentiation and Function
    International Journal of Molecular Sciences Article The Proteomic Landscape of Resting and Activated CD4+ T Cells Reveal Insights into Cell Differentiation and Function Yashwanth Subbannayya 1 , Markus Haug 1, Sneha M. Pinto 1, Varshasnata Mohanty 2, Hany Zakaria Meås 1, Trude Helen Flo 1, T.S. Keshava Prasad 2 and Richard K. Kandasamy 1,* 1 Centre of Molecular Inflammation Research (CEMIR), Department of Clinical and Molecular Medicine (IKOM), Norwegian University of Science and Technology, 7491 Trondheim, Norway; [email protected] (Y.S.); [email protected] (M.H.); [email protected] (S.M.P.); [email protected] (H.Z.M.); trude.fl[email protected] (T.H.F.) 2 Center for Systems Biology and Molecular Medicine, Yenepoya (Deemed to be University), Mangalore 575018, India; [email protected] (V.M.); [email protected] (T.S.K.P.) * Correspondence: [email protected] Abstract: CD4+ T cells (T helper cells) are cytokine-producing adaptive immune cells that activate or regulate the responses of various immune cells. The activation and functional status of CD4+ T cells is important for adequate responses to pathogen infections but has also been associated with auto-immune disorders and survival in several cancers. In the current study, we carried out a label-free high-resolution FTMS-based proteomic profiling of resting and T cell receptor-activated (72 h) primary human CD4+ T cells from peripheral blood of healthy donors as well as SUP-T1 cells. We identified 5237 proteins, of which significant alterations in the levels of 1119 proteins were observed between resting and activated CD4+ T cells.
    [Show full text]
  • Downloaded from Ftp://Ftp.Uniprot.Org/ on July 3, 2019) Using Maxquant (V1.6.10.43) Search Algorithm
    bioRxiv preprint doi: https://doi.org/10.1101/2020.11.17.385096; this version posted November 17, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-ND 4.0 International license. The proteomic landscape of resting and activated CD4+ T cells reveal insights into cell differentiation and function Yashwanth Subbannayya1, Markus Haug1, Sneha M. Pinto1, Varshasnata Mohanty2, Hany Zakaria Meås1, Trude Helen Flo1, T.S. Keshava Prasad2 and Richard K. Kandasamy1,* 1Centre of Molecular Inflammation Research (CEMIR), and Department of Clinical and Molecular Medicine (IKOM), Norwegian University of Science and Technology, N-7491 Trondheim, Norway 2Center for Systems Biology and Molecular Medicine, Yenepoya (Deemed to be University), Mangalore, India *Correspondence to: Professor Richard Kumaran Kandasamy Norwegian University of Science and Technology (NTNU) Centre of Molecular Inflammation Research (CEMIR) PO Box 8905 MTFS Trondheim 7491 Norway E-mail: [email protected] (Kandasamy R K) Tel.: +47-7282-4511 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.11.17.385096; this version posted November 17, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-ND 4.0 International license. Abstract CD4+ T cells (T helper cells) are cytokine-producing adaptive immune cells that activate or regulate the responses of various immune cells.
    [Show full text]
  • The Human Gene Connectome As a Map of Short Cuts for Morbid Allele Discovery
    The human gene connectome as a map of short cuts for morbid allele discovery Yuval Itana,1, Shen-Ying Zhanga,b, Guillaume Vogta,b, Avinash Abhyankara, Melina Hermana, Patrick Nitschkec, Dror Friedd, Lluis Quintana-Murcie, Laurent Abela,b, and Jean-Laurent Casanovaa,b,f aSt. Giles Laboratory of Human Genetics of Infectious Diseases, Rockefeller Branch, The Rockefeller University, New York, NY 10065; bLaboratory of Human Genetics of Infectious Diseases, Necker Branch, Paris Descartes University, Institut National de la Santé et de la Recherche Médicale U980, Necker Medical School, 75015 Paris, France; cPlateforme Bioinformatique, Université Paris Descartes, 75116 Paris, France; dDepartment of Computer Science, Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel; eUnit of Human Evolutionary Genetics, Centre National de la Recherche Scientifique, Unité de Recherche Associée 3012, Institut Pasteur, F-75015 Paris, France; and fPediatric Immunology-Hematology Unit, Necker Hospital for Sick Children, 75015 Paris, France Edited* by Bruce Beutler, University of Texas Southwestern Medical Center, Dallas, TX, and approved February 15, 2013 (received for review October 19, 2012) High-throughput genomic data reveal thousands of gene variants to detect a single mutated gene, with the other polymorphic genes per patient, and it is often difficult to determine which of these being of less interest. This goes some way to explaining why, variants underlies disease in a given individual. However, at the despite the abundance of NGS data, the discovery of disease- population level, there may be some degree of phenotypic homo- causing alleles from such data remains somewhat limited. geneity, with alterations of specific physiological pathways under- We developed the human gene connectome (HGC) to over- come this problem.
    [Show full text]
  • Human Social Genomics in the Multi-Ethnic Study of Atherosclerosis
    Getting “Under the Skin”: Human Social Genomics in the Multi-Ethnic Study of Atherosclerosis by Kristen Monét Brown A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy (Epidemiological Science) in the University of Michigan 2017 Doctoral Committee: Professor Ana V. Diez-Roux, Co-Chair, Drexel University Professor Sharon R. Kardia, Co-Chair Professor Bhramar Mukherjee Assistant Professor Belinda Needham Assistant Professor Jennifer A. Smith © Kristen Monét Brown, 2017 [email protected] ORCID iD: 0000-0002-9955-0568 Dedication I dedicate this dissertation to my grandmother, Gertrude Delores Hampton. Nanny, no one wanted to see me become “Dr. Brown” more than you. I know that you are standing over the bannister of heaven smiling and beaming with pride. I love you more than my words could ever fully express. ii Acknowledgements First, I give honor to God, who is the head of my life. Truly, without Him, none of this would be possible. Countless times throughout this doctoral journey I have relied my favorite scripture, “And we know that all things work together for good, to them that love God, to them who are called according to His purpose (Romans 8:28).” Secondly, I acknowledge my parents, James and Marilyn Brown. From an early age, you two instilled in me the value of education and have been my biggest cheerleaders throughout my entire life. I thank you for your unconditional love, encouragement, sacrifices, and support. I would not be here today without you. I truly thank God that out of the all of the people in the world that He could have chosen to be my parents, that He chose the two of you.
    [Show full text]
  • Views for Entrez
    BASIC RESEARCH www.jasn.org Phosphoproteomic Analysis Reveals Regulatory Mechanisms at the Kidney Filtration Barrier †‡ †| Markus M. Rinschen,* Xiongwu Wu,§ Tim König, Trairak Pisitkun,¶ Henning Hagmann,* † † † Caroline Pahmeyer,* Tobias Lamkemeyer, Priyanka Kohli,* Nicole Schnell, †‡ †† ‡‡ Bernhard Schermer,* Stuart Dryer,** Bernard R. Brooks,§ Pedro Beltrao, †‡ Marcus Krueger,§§ Paul T. Brinkkoetter,* and Thomas Benzing* *Department of Internal Medicine II, Center for Molecular Medicine, †Cologne Excellence Cluster on Cellular Stress | Responses in Aging-Associated Diseases, ‡Systems Biology of Ageing Cologne, Institute for Genetics, University of Cologne, Cologne, Germany; §Laboratory of Computational Biology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland; ¶Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand; **Department of Biology and Biochemistry, University of Houston, Houston, Texas; ††Division of Nephrology, Baylor College of Medicine, Houston, Texas; ‡‡European Molecular Biology Laboratory–European Bioinformatics Institute, Hinxton, Cambridge, United Kingdom; and §§Max Planck Institute for Heart and Lung Research, Bad Nauheim, Germany ABSTRACT Diseases of the kidney filtration barrier are a leading cause of ESRD. Most disorders affect the podocytes, polarized cells with a limited capacity for self-renewal that require tightly controlled signaling to maintain their integrity, viability, and function. Here, we provide an atlas of in vivo phosphorylated, glomerulus- expressed
    [Show full text]
  • WO 2016/040794 Al 17 March 2016 (17.03.2016) P O P C T
    (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date WO 2016/040794 Al 17 March 2016 (17.03.2016) P O P C T (51) International Patent Classification: AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, C12N 1/19 (2006.01) C12Q 1/02 (2006.01) BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, C12N 15/81 (2006.01) C07K 14/47 (2006.01) DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KN, KP, KR, (21) International Application Number: KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, PCT/US20 15/049674 MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, (22) International Filing Date: PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, 11 September 2015 ( 11.09.201 5) SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (25) Filing Language: English (84) Designated States (unless otherwise indicated, for every (26) Publication Language: English kind of regional protection available): ARIPO (BW, GH, (30) Priority Data: GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, 62/050,045 12 September 2014 (12.09.2014) US TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, (71) Applicant: WHITEHEAD INSTITUTE FOR BIOMED¬ DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, ICAL RESEARCH [US/US]; Nine Cambridge Center, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, Cambridge, Massachusetts 02142-1479 (US).
    [Show full text]
  • Datasheet: AHP653 Product Details
    Datasheet: AHP653 Description: GOAT ANTI HUMAN PRAM1 Specificity: PRAM1 Format: Purified Product Type: Polyclonal Antibody Isotype: Polyclonal IgG Quantity: 0.1 mg Product Details Applications This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit www.bio-rad-antibodies.com/protocols. Yes No Not Determined Suggested Dilution Flow Cytometry Immunohistology - Frozen Immunohistology - Paraffin ELISA Immunoprecipitation Western Blotting 1ug/ml - 3ug/ml Where this antibody has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. It is recommended that the user titrates the antibody for use in their own system using appropriate negative/positive controls. Target Species Human Product Form Purified IgG - liquid Antiserum Preparation Antisera to human PRAM1 were raised by repeated immunisations of goats with highly purified antigen. Purified IgG was prepared from whole serum by affinity chromatography. Buffer Solution TRIS buffered saline Preservative 0.02% Sodium Azide Stabilisers 0.5% Bovine Serum Albumin Approx. Protein IgG concentration 0.5 mg/ml Concentrations Immunogen Peptide sequence DFCDPLENQPLPLGR corresponding to the C-terminus of PRAM1 (NP_115528). External Database Links UniProt: Q96QH2 Related reagents Entrez Gene: Page 1 of 2 84106 PRAM1 Related reagents Specificity Goat anti Human PRAM1 antibody recognises PML-RARA target gene encoding protein, which is expressed and regulated during normal human myelopoiesis. PRAM-1 is a ~97 kDa adaptor protein, which is expressed and regulated during normal human myelopoiesis.
    [Show full text]
  • DRG2 Antibody Cat
    DRG2 Antibody Cat. No.: 60-339 DRG2 Antibody Specifications HOST SPECIES: Rabbit SPECIES REACTIVITY: Mouse HOMOLOGY: Predicted species reactivity based on immunogen sequence: Bovine This DRG2 antibody is generated from rabbits immunized with a KLH conjugated synthetic IMMUNOGEN: peptide between 154-180 amino acids from the Central region of human DRG2. TESTED APPLICATIONS: WB APPLICATIONS: For WB starting dilution is: 1:1000 PREDICTED MOLECULAR 41 kDa WEIGHT: Properties This antibody is purified through a protein A column, followed by peptide affinity PURIFICATION: purification. CLONALITY: Polyclonal ISOTYPE: Rabbit Ig CONJUGATE: Unconjugated September 27, 2021 1 https://www.prosci-inc.com/drg2-antibody-60-339.html PHYSICAL STATE: Liquid BUFFER: Supplied in PBS with 0.09% (W/V) sodium azide. CONCENTRATION: batch dependent Store at 4˚C for three months and -20˚C, stable for up to one year. As with all antibodies STORAGE CONDITIONS: care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures. Additional Info OFFICIAL SYMBOL: DRG2 ALTERNATE NAMES: Developmentally-regulated GTP-binding protein 2, DRG-2, DRG2 ACCESSION NO.: P55039 PROTEIN GI NO.: 1706518 GENE ID: 1819 USER NOTE: Optimal dilutions for each application to be determined by the researcher. Background and References The DRG2 gene encodes the developmentally regulated GTP-binding protein 2, a name derived from the fact that it shares significant similarity to known GTP-binding proteins. DRG2 was identified because it is expressed in normal fibroblasts but not in SV40- BACKGROUND: transformed fibroblasts. Read-through transcripts containing this gene and a downstream gene have been identified, but they are not thought to encode a fusion protein.
    [Show full text]