DOCSLIB.ORG

Open Full Page

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Open Full Page Published OnlineFirst March 14, 2013; DOI: 10.1158/1078-0432.CCR-12-1000 Clinical Cancer Predictive Biomarkers and Personalized Medicine Research Molecular Profiling of Aromatase Inhibitor–Treated Postmenopausal Breast Tumors Identifies Immune-Related Correlates of Resistance Anita K. Dunbier1,2,5, Zara Ghazoui1,2, Helen Anderson1,2, Janine Salter1, Ashutosh Nerurkar1, Peter Osin1, Roger A'hern3, William R. Miller4, Ian E. Smith1, and Mitch Dowsett1,2 Abstract Purpose: Estrogen withdrawal by treatment with aromatase inhibitors is the most effective form of endocrine therapy for postmenopausal estrogen receptor–positive (ERþ) breast cancer. However, response to therapy varies markedly and understanding of the precise molecular effects of aromatase inhibitors and causes of resistance is limited. We aimed to identify in clinical breast cancer those genes and pathways most associated with resistance to aromatase inhibitors by examining the global transcriptional effects of AI treatment. Experimental Design: Baseline and 2-week posttreatment biopsies were obtained from 112 postmen- opausal women with ERþ breast cancer receiving neoadjuvant anastrozole. Gene expression data were obtained from 81 baseline and 2-week paired samples. Pathway analysis identified (i) the most prevalent changes in expression and (ii) the pretreatment genes/pathways most related to poor antiproliferative response. Results: A total of 1,327 genes were differentially expressed after 2-week treatment (false discovery rate < 0.01). Proliferation-associated genes and classical estrogen-dependent genes were strongly downregulated whereas collagens and chemokines were upregulated. Pretreatment expression of an inflammatory signature correlated with antiproliferative response to anastrozole and this observation was validated in an inde- pendent study. Higher expression of immune-related genes such as SLAMF8 and TNF as well as lymphocytic infiltration were associated with poorer response (P < 0.001) and validated in an independent cohort. Conclusions: The molecular response to aromatase inhibitor treatment varies greatly between patients consistent with the variable clinical benefit from aromatase inhibitor treatment. Higher baseline expression of an inflammatory signature is associated with poor antiproliferative response and should be assessed further as a novel biomarker and potential target for aromatase inhibitor-treated patients. Clin Cancer Res; 19(10); 2775–86. Ó2013 AACR. Introduction inhibitors is the most effective form of endocrine therapy, Approximately 80% of human breast carcinomas present but response to them varies markedly (1–3). Mechanisms of as estrogen receptor a–positive (ERþ). In postmenopausal resistance appear to be multiple but are poorly character- In vitro women, estrogen withdrawal by treatment with aromatase ized. studies of acquired resistance to estrogen deprivation have identified several putative mechanisms which largely involve growth factor–related signal trans- Authors' Affiliations: 1Royal Marsden Hospital; 2Breakthrough Breast duction pathways (4–7) but there is limited clinical evi- 3 Cancer Research Centre, Institute of Cancer Research; Cancer Research dence to support these. It is also notable that these models UK Clinical Trials and Statistics Unit, Section of Clinical Trials, Institute of Cancer Research, London; 4Breast Research Group, University of Edin- do not involve an assessment of the contribution of human burgh, Edinburgh, United Kingdom; and 5Department of Biochemistry, stromal elements. University of Otago, Dunedin, New Zealand The presurgical/neoadjuvant setting provides an excep- Note: Supplementary data for this article are available at Clinical Cancer tionally valuable scenario for linking biology to clinical Research Online (http://clincancerres.aacrjournals.org/). response and to study mechanisms of resistance; within this A.K. Dunbier, Z. Ghazoui, and H. Anderson contributed equally to this work. setting, the proliferation marker Ki67 is a validated phar- Corresponding Author: Anita K. Dunbier, Department of Biochemistry, macodynamic indicator of response to endocrine therapy University of Otago, PO Box 56, Dunedin 9016, New Zealand. Phone: (8–10). Treatment-dependent changes in sequential mea- 643479-9258; Fax: 643479-7738; E-mail: [email protected] surements of Ki67 in neoadjuvant trials of endocrine ther- doi: 10.1158/1078-0432.CCR-12-1000 apy in postmenopausal women (IMPACT, p024, and Ó2013 American Association for Cancer Research. Z1031; refs. 9, 11, 12) have all revealed differences or lack www.aacrjournals.org 2775 Downloaded from clincancerres.aacrjournals.org on September 27, 2021. © 2013 American Association for Cancer Research. Published OnlineFirst March 14, 2013; DOI: 10.1158/1078-0432.CCR-12-1000 Dunbier et al. Materials and Methods Translational Relevance Patient samples Most postmenopausal women with estrogen receptor Fourteen-gauge core-cut tumor biopsies were obtained (ER)–positive breast cancer receive an aromatase inhib- from 112 postmenopausal women with stage I to IIIB ERþ itor at some stage during their treatment. However, early breast cancer before and after 2-week anastrozole responsiveness to aromatase inhibitors varies greatly. monotherapy in a neoadjuvant trial (17, 18). Patient demo- Previous studies of resistance mechanisms have been graphics are shown in Supplementary Table S1. Tissue was conducted in cell lines and have highlighted the impor- stored in RNAlater at À20C. Two 4-mm sections from the tance of signal transduction pathways, but these studies core were stained with hematoxylin and eosin (H&E) and omit the stromal influences that are increasingly recog- examined by a pathologist (A. Nerurkar and P. Osin) to nized to have a major influence on tumor biology. In confirm the presence of cancerous tissue, assess histopa- contrast, we report genome-wide expression profiling of thology, and the presence or absence of lymphocytic infil- 81 ER-positive breast carcinomas including stromal tis- tration. Tumors were deemed to be positive for lymphocytic sues before and during treatment with an aromatase infiltration if intraepithelial mononuclear cells could be inhibitor in the neoadjuvant setting. We identify an seen within tumor cell nests or in direct contact with tumor inflammatory signature as the strongest correlate of poor cells. Biopsies in which lymphocytic infiltrate could be seen antiproliferative response and confirm this finding in an without direct contact to tumor cells were not considered to independent set of tumors as well as using gross lym- have lymphocytic infiltration. Samples were assessed blind- phocytic infiltration as an alternative measure of inflam- ly by both pathologists and the concordance rate was 81%. matory activity. This work provides a potential new Discordant samples were then reassessed by A. Nerurkar avenue for drug development and for identifying who was blinded to the result of the initial assessment and patients with a reduced likelihood of response to aro- the more frequent of the 3 assessments was used in the matase inhibition. analysis. Total RNA was extracted using RNeasy (Qiagen). RNA quality was checked using an Agilent Bioanalyser: samples with RNA integrity values of less than 5 were excluded from further analysis. ER (H-score) and Ki67 (% cells positive) values by centralized immunohistochem- of differences that were parallel to recurrence-free survival istry were already available (17). (RFS) differences in the equivalent adjuvant trials (ATAC, An additional 71 stored sections from paraffin-embed- BIG 1-98, and MA27; refs. 1, 13, 14), respectively. In ded, formalin-fixed core-cut biopsies were obtained from a addition, Ki67 of patients after 2 weeks’ treatment predicts subset of patients from the IMPACT trial who received RFS more closely than pretreatment values (8). neoadjuvant anastrozole or tamoxifen (15). Sections were Nearly all patients on an aromatase inhibitor show H&E-stained, and the presence or absence of detectable reduction in Ki67 expression, suggesting that some benefit lymphocytic infiltration was assessed by a pathologist (A. is derived, although this may be modest (9). This contin- Nerurkar). Ki67 data were obtained from previous analysis uous marker of response is well-suited to assessment of of these patients (8, 9). mechanisms of resistance that are also likely to have a nonbinary effect. This may explain why changes in Ki67 have proven to be better predictors of benefit from endo- Gene expression analysis and data preprocessing crine therapy than clinical response during neoadjuvant RNA amplification, labeling, and hybridization on endocrine therapy (15, 16). HumanWG-6 v2 Expression BeadChips (Illumina) were Gene expression profiling of tumor biopsies before and conducted according to the manufacturer’s instructions at during treatment has the potential to enable the identifica- a single Illumina BeadStation facility. Tumor RNA of suf- tion of the most important genes/pathways involved in the ficient quality and quantity was available to generate expres- response to estrogen deprivation therapy and the pretreat- sion data from 104 pretreatment biopsies and 85 two-week ment determinants of response and resistance. Availability biopsies (Supplementary Fig. S1). Data were extracted using of comprehensive expression datasets, as provided here, will BeadStudio software and normalized with variance-stabi- allow the evaluation of candidate genes from experimental lizing transformation (VST) and Robust Spline Normaliza- research
Load more

Recommended publications
  • Pulmonary Fibrosis Associated with TINF2 Gene Mutation: Is Somatic Reversion Required?
    Pulmonary fibrosis associated with TINF2 gene mutation: is somatic reversion required? To the Editor: We read with great interest the case reported by FUKUHARA et al. [1] of a 43-year-old female patient with dyskeratosis congenita, pulmonary fibrosis and heterozygous mutation in TINF2 (telomerase repeat binding factor 1-interacting nuclear factor 2). TIN2, the TINF2 gene product, TERT (telomere reverse transcriptase) and TERC (telomerase RNA component) participate in the regulation of telomere elongation, in which mutations have been previously found to be associated with familial pulmonary fibrosis in adults [2]. Indeed mutations of SFTPC, coding for surfactant protein C, were initially described in children before being described in adults as old as 72 years of age who presented with familial pulmonary fibrosis [3]. However, we were surprised that a TINF2 mutation could be evidenced in an adult of that age. As highlighted by FUKUHARA et al. [1], patients with the TINF2 mutation present with severe haematological symptoms before 10 years of age [4]. As mentioned by FUKUHARA et al. [1], the identified mutation is probably not hypomorphic because it is a frame-shift deletion located in the mutational ‘‘hot spot’’ described previously. Furthermore, the patient presented with very short telomeres. The TINF2 mutation was probably inherited from her father because he had abnormal skin pigmentation and aplastic anaemia [1]. Re-analysis of the gene mutation sequencing could provide new hypotheses for this late disease onset. Indeed, the electrophoregram depicted in figure 1b in the study by FUKUHARA et al. [1] probably comes from a PCR product sub-cloned into an expression vector [5], and does not ensure that the deletion is at the heterozygous status usually seen in our patients (fig.
    [Show full text]
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • Genetics of Familial Non-Medullary Thyroid Carcinoma (FNMTC)
    cancers Review Genetics of Familial Non-Medullary Thyroid Carcinoma (FNMTC) Chiara Diquigiovanni * and Elena Bonora Unit of Medical Genetics, Department of Medical and Surgical Sciences, University of Bologna, 40138 Bologna, Italy; [email protected] * Correspondence: [email protected]; Tel.: +39-051-208-8418 Simple Summary: Non-medullary thyroid carcinoma (NMTC) originates from thyroid follicular epithelial cells and is considered familial when occurs in two or more first-degree relatives of the patient, in the absence of predisposing environmental factors. Familial NMTC (FNMTC) cases show a high genetic heterogeneity, thus impairing the identification of pivotal molecular changes. In the past years, linkage-based approaches identified several susceptibility loci and variants associated with NMTC risk, however only few genes have been identified. The advent of next-generation sequencing technologies has improved the discovery of new predisposing genes. In this review we report the most significant genes where variants predispose to FNMTC, with the perspective that the integration of these new molecular findings in the clinical data of patients might allow an early detection and tailored therapy of the disease, optimizing patient management. Abstract: Non-medullary thyroid carcinoma (NMTC) is the most frequent endocrine tumor and originates from the follicular epithelial cells of the thyroid. Familial NMTC (FNMTC) has been defined in pedigrees where two or more first-degree relatives of the patient present the disease in absence of other predisposing environmental factors. Compared to sporadic cases, FNMTCs are often multifocal, recurring more frequently and showing an early age at onset with a worse outcome. FNMTC cases Citation: Diquigiovanni, C.; Bonora, E.
    [Show full text]
  • A Nonsense (C.3978G>A) Abnormal Spindle-Like, Microcephaly Associated (ASPM) Gene Mutation Is a Major Cause of Primary Microc
    African Journal of Biotechnology Vol. 10(34), pp. 6396-6400, 11 July, 2011 Available online at http://www.academicjournals.org/AJB DOI: 10.5897/AJB10.2571 ISSN 1684-5315 © 2011 Academic Journals Full Length Research Paper A nonsense (c.3978G>A) abnormal spindle-like, microcephaly associated (ASPM) gene mutation is a major cause of primary microcephaly in Pashtoon ethnic group of Pakistan Shamim Saleha 1, Muhammad Ajmal 2, Muhammad Jamil 1, Muhammad Nasir 2 and Abdul Hameed 2* 1Department of Biotechnology and Genetic Engineering, Kohat University of Science and Technology, Kohat 26000, Khyber Paktoonkhwa, Pakistan. 2Institute of Biomedical and Genetic Engineering, G.P.O. box 2891, 24-Mauve Area, G-9/1, Islamabad, Pakistan. Accepted 29 April, 2011 Primary microcephaly (MCPH) is an autosomal-recessive congenital disorder characterized by smaller- than-normal brain size and mental retardation. MCPH is genetically heterogeneous with six known loci: MCPH1 to MCPH7. The abnormal spindle-like, microcephaly associated (ASPM) gene at MCPH5 locus, which accounts for 37 to 54% of MCPH, appears to be the most common cause of microcephaly. More than 50% of the MCPH families genetically analyzed in Pakistan were mapped to MCPH5 locus including both families in this study. On mutation screening of ASPM gene by PCR amplification and direct DNA sequencing, a common c.3978G>A transition was identified in exon 17 of ASPM gene to be responsible for diseased phenotype in both families. This change results to the substitution of an amino acid residue at position 1326 from tryptophan to a stop codon (p.Trp1326Stop). The same mutation was also identified in several other families of Pakistani origin.
    [Show full text]
  • Molecular Genetics of Microcephaly Primary Hereditary: an Overview
    brain sciences Review Molecular Genetics of Microcephaly Primary Hereditary: An Overview Nikistratos Siskos † , Electra Stylianopoulou †, Georgios Skavdis and Maria E. Grigoriou * Department of Molecular Biology & Genetics, Democritus University of Thrace, 68100 Alexandroupolis, Greece; [email protected] (N.S.); [email protected] (E.S.); [email protected] (G.S.) * Correspondence: [email protected] † Equal contribution. Abstract: MicroCephaly Primary Hereditary (MCPH) is a rare congenital neurodevelopmental disorder characterized by a significant reduction of the occipitofrontal head circumference and mild to moderate mental disability. Patients have small brains, though with overall normal architecture; therefore, studying MCPH can reveal not only the pathological mechanisms leading to this condition, but also the mechanisms operating during normal development. MCPH is genetically heterogeneous, with 27 genes listed so far in the Online Mendelian Inheritance in Man (OMIM) database. In this review, we discuss the role of MCPH proteins and delineate the molecular mechanisms and common pathways in which they participate. Keywords: microcephaly; MCPH; MCPH1–MCPH27; molecular genetics; cell cycle 1. Introduction Citation: Siskos, N.; Stylianopoulou, Microcephaly, from the Greek word µικρoκεϕαλi´α (mikrokephalia), meaning small E.; Skavdis, G.; Grigoriou, M.E. head, is a term used to describe a cranium with reduction of the occipitofrontal head circum- Molecular Genetics of Microcephaly ference equal, or more that teo standard deviations
    [Show full text]
  • The Genetics and Clinical Manifestations of Telomere Biology Disorders Sharon A
    REVIEW The genetics and clinical manifestations of telomere biology disorders Sharon A. Savage, MD1, and Alison A. Bertuch, MD, PhD2 3 Abstract: Telomere biology disorders are a complex set of illnesses meric sequence is lost with each round of DNA replication. defined by the presence of very short telomeres. Individuals with classic Consequently, telomeres shorten with aging. In peripheral dyskeratosis congenita have the most severe phenotype, characterized blood leukocytes, the cells most extensively studied, the rate 4 by the triad of nail dystrophy, abnormal skin pigmentation, and oral of attrition is greatest during the first year of life. Thereafter, leukoplakia. More significantly, these individuals are at very high risk telomeres shorten more gradually. When the extent of telo- of bone marrow failure, cancer, and pulmonary fibrosis. A mutation in meric DNA loss exceeds a critical threshold, a robust anti- one of six different telomere biology genes can be identified in 50–60% proliferative signal is triggered, leading to cellular senes- of these individuals. DKC1, TERC, TERT, NOP10, and NHP2 encode cence or apoptosis. Thus, telomere attrition is thought to 1 components of telomerase or a telomerase-associated factor and TINF2, contribute to aging phenotypes. 5 a telomeric protein. Progressively shorter telomeres are inherited from With the 1985 discovery of telomerase, the enzyme that ex- generation to generation in autosomal dominant dyskeratosis congenita, tends telomeric nucleotide repeats, there has been rapid progress resulting in disease anticipation. Up to 10% of individuals with apparently both in our understanding of basic telomere biology and the con- acquired aplastic anemia or idiopathic pulmonary fibrosis also have short nection of telomere biology to human disease.
    [Show full text]
  • Structural and Functional Consequences of a Disease Mutation in the Telomere Protein TPP1
    Structural and functional consequences of a disease mutation in the telomere protein TPP1 Kamlesh Bishta,1, Eric M. Smitha,b,1, Valerie M. Tesmera, and Jayakrishnan Nandakumara,b,2 aDepartment of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI 48109; and bProgram in Chemical Biology, University of Michigan, Ann Arbor, MI 48109 Edited by Joachim Lingner, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland, and accepted by Editorial Board Member Dinshaw J. Patel September 29, 2016 (received for review April 8, 2016) Telomerase replicates chromosome ends to facilitate continued cell The discovery of the TEL patch of TPP1 prompted the pre- division. Mutations that compromise telomerase function result in diction that this region could be a hotspot for mutations that stem cell failure diseases, such as dyskeratosis congenita (DC). One cause telomerase-deficiency diseases, such as DC. We recently such mutation (K170Δ), residing in the telomerase-recruitment factor reported a case of a severe variant of DC, Hoyeraal–Hreidarsson TPP1, provides an excellent opportunity to structurally, biochemi- syndrome (23), in which the proband was heterozygous for a cally, and genetically dissect the mechanism of such diseases. We deletion of a single amino acid of the TPP1 protein, namely lysine ACD show through site-directed mutagenesis and X-ray crystallography 170 (K170) (24). Our study placed the gene coding for TPP1 DKC1 TERC TERT that this TPP1 disease mutation deforms the conformation of two protein on a list with 10 other genes ( , , , RTEL1 TINF2 CTC1 NOP10 NHP2 WRAP53 PARN critical amino acids of the TEL [TPP1’s glutamate (E) and leucine-rich , , , , , ,and ) that are found mutated in DC and other telomere-related dis- (L)] patch, the surface of TPP1 that binds telomerase.
    [Show full text]
  • The Molecular Landscape of ASPM Mutations in Primary Microcephaly
    Original article J Med Genet: first published as 10.1136/jmg.2008.062380 on 21 November 2008. Downloaded from The molecular landscape of ASPM mutations in primary microcephaly A K Nicholas,1 E A Swanson,2 J J Cox,1 G Karbani,3 S Malik,3 K Springell,4 D Hampshire,4 M Ahmed,3 J Bond,4 D Di Benedetto,5 M Fichera,5 C Romano,6 W B Dobyns,2 C G Woods1 c Additional tables are ABSTRACT also a diagnosable cause of mental retardation, and published online only at http:// Background: Autosomal recessive primary microcephaly one with a substantial recurrence risk of one in jmg.bmj.com/content/vol46/ (MCPH) is a model disease to study human neurogenesis. issue4 four in subsequent children. In affected individuals the brain grows at a reduced rate 1 The current diagnostic criteria for MCPH are: Department of Medical during fetal life resulting in a small but structurally normal congenital microcephaly more than 23 SD below Genetics, Cambridge Institute for Medical Research, University brain and mental retardation. The condition is genetically age and sex means; mental retardation but no of Cambridge, Cambridge, UK; heterogeneous with mutations in ASPM being most other neurological finding, such as spasticity, 2 University of Chicago, commonly reported. seizures, or progressive cognitive decline; normal Department of Human Genetics, Methods and results: We have examined this further by height and weight, appearance, and results on Chicago, Illinois, USA; studying three cohorts of microcephalic children to extend 6 3 Department of Clinical chromosome analysis and brain scan. Despite this, Genetics, St James’s University both the phenotype and the mutation spectrum.
    [Show full text]
  • (ASPM) Gene in an Iranian Patient with Primary Microcephaly: a Case Report
    Iran J Public Health, Vol. 48, No.11, Nov 2019, pp.2074-2078 Case Report A Novel Frameshift Mutation in Abnormal Spindle-Like Microcephaly (ASPM) Gene in an Iranian Patient with Primary Microcephaly: A Case Report Afsaneh BAZGIR, Mehdi AGHA GHOLIZADEH, Faezeh SARVAR, *Zahra PAKZAD Department of Medical Genetics, Fardis Central Lab, Alborz, Iran *Corresponding Author: Email: [email protected] (Received 10 Mar 2019; accepted 11 Jul 2019) Abstract Autosomal recessive primary microcephaly (MCPH) is a rare genetic disorder, leading to the defect of neurogenic brain development. Individuals with MCPH reveal reduced head circumference and intellectual disability. Several MCPH loci have been identified from several populations. Genetic heterogeneity of this disorder represents mo- lecular testing challenge. An 8 yr old female, born from consanguineous parents, was attended to Fardis Central Lab, Alborz, Iran. Based on the reduced circumference and intellectual disability, MCPH was diagnosed. Whole exome sequencing of the patient identified a novel homozygous frameshift mutation (c.2738dupT, p.Cys914fs) in exon 9 Abnormal Spindle-like Microcephaly )ASPM( gene. By Sanger sequencing, segregation analysis showed that both parents were heterozygous carriers for this variant. The novel frameshift mutation likely truncates the protein, resulting in loss of normal function ASPM in homozygous mutation carriers. The study might add a new pathogenic variant in mutations of the ASPM gene as a causative variant in patients with MCPH and might be helpful in genetic counseling of consanguineous families. Keywords: Autosomal recessive primary microcephaly; ASPM; Whole exome sequencing Introduction Autosomal recessive primary microcephaly ants that resulting in frameshift and protein- (MCPH) is a genetically heterogeneous condition truncating.
    [Show full text]
  • A High-Throughput Approach to Uncover Novel Roles of APOBEC2, a Functional Orphan of the AID/APOBEC Family
    Rockefeller University Digital Commons @ RU Student Theses and Dissertations 2018 A High-Throughput Approach to Uncover Novel Roles of APOBEC2, a Functional Orphan of the AID/APOBEC Family Linda Molla Follow this and additional works at: https://digitalcommons.rockefeller.edu/ student_theses_and_dissertations Part of the Life Sciences Commons A HIGH-THROUGHPUT APPROACH TO UNCOVER NOVEL ROLES OF APOBEC2, A FUNCTIONAL ORPHAN OF THE AID/APOBEC FAMILY A Thesis Presented to the Faculty of The Rockefeller University in Partial Fulfillment of the Requirements for the degree of Doctor of Philosophy by Linda Molla June 2018 © Copyright by Linda Molla 2018 A HIGH-THROUGHPUT APPROACH TO UNCOVER NOVEL ROLES OF APOBEC2, A FUNCTIONAL ORPHAN OF THE AID/APOBEC FAMILY Linda Molla, Ph.D. The Rockefeller University 2018 APOBEC2 is a member of the AID/APOBEC cytidine deaminase family of proteins. Unlike most of AID/APOBEC, however, APOBEC2’s function remains elusive. Previous research has implicated APOBEC2 in diverse organisms and cellular processes such as muscle biology (in Mus musculus), regeneration (in Danio rerio), and development (in Xenopus laevis). APOBEC2 has also been implicated in cancer. However the enzymatic activity, substrate or physiological target(s) of APOBEC2 are unknown. For this thesis, I have combined Next Generation Sequencing (NGS) techniques with state-of-the-art molecular biology to determine the physiological targets of APOBEC2. Using a cell culture muscle differentiation system, and RNA sequencing (RNA-Seq) by polyA capture, I demonstrated that unlike the AID/APOBEC family member APOBEC1, APOBEC2 is not an RNA editor. Using the same system combined with enhanced Reduced Representation Bisulfite Sequencing (eRRBS) analyses I showed that, unlike the AID/APOBEC family member AID, APOBEC2 does not act as a 5-methyl-C deaminase.
    [Show full text]
  • Gene List of the Targeted NGS MCD and CCA Gene Panel AKT3,ALX1
    Gene List of the targeted NGS MCD and CCA gene panel AKT3,ALX1,ALX3,ALX4,AMPD2,ARFGEF2,ARID1B,ARX,ASPM,ATR,ATRX,B3GALTL,BRPF1,c12orf57,C6orf70,CASK,CCND2,CDK5RAP2,CDON,C ENPJ,CEP170,CHMP1A,COL4A1,CREBBP,CYP11A1,DCHS1,DCLK1,DCX,DHCR24,DHCR7,DIS3L2,DISC1,DISP1,DLL1,DMRTA2,DYNC1H1,DYRK1 A,EARS2,EFNB1,EMX1,EOMES,EP300,ERBB4,ERMARD,EXOSC3,FAM36A,FGF8,FGFR1,FGFR2,FLNA,FOXC1,FOXG1,FOXH1,FZD10,GLI2,GLI3,GP R56,GPSM2,HCCS,HESX1,HNRNPU,IGBP1,IGFBP1,ISPD,ITPA,KAL1,KAT6B,KATNB1,KIAA1279,KIF14,KIF1A,KIF1B,KIF21A,KIF2A,KIF5C,KIF7,L1 CAM,LAMB1,LAMC3,LRP2,MCPH1,MED12,MID1,NDE1,NFIB,NPC1,NR2F1,NSD1,NTRK1,NTRK3,OCEL1,OPA1,OTX2,PAFAH1B1,PAX6,PEX1,PHF1 0,PIK3R2,POLR3A,POLR3B,POMT1,POMT2,PTCH1,PTPRS,PYCR1,RAB3GAP1,RARS2,RELN,RFX3,ROBO1,ROBO3,RPS6KA3,RTTN,SATB2,SEPSEC S,SHH,SIX3,SLC12A6,SOX2,SPOCK1,SRPX2,TBCD,TBCE,TCF4,TDGF1,TEAD1,THBS2,TMEM5,TSC1,TSC2,TSEN15,TSEN2,TSEN34,TSEN54,TUBA1 A,TUBA8,TUBB,TUBB2A,TUBB2B,TUBB3,TUBB4A,TUBG1,VAX1,VRK1,WDR47,WDR62,ZBTB18,ZEB2,ZIC2. Gene List of the targeted NGS epilepsy gene panel AARS, ADGRV1, ADRA2B, ADSL, ALDH4A1, ALDH7A1, ALG13, ALPL, ARHGEF15, ARHGEF9, ARX, ASAH1, ATP1A2, ATP1A3, BRD2, CACNA1A, CACNA1H, CACNA2D2, CACNB4, CBL, CDKL5, CERS1, CHD2, CHRNA2, CHRNA4, CHRNB2, CLCN2, CLCN4, CLN8, CLTC, CNKSR2, CNTNAP2, CPA6, CPLX1, CSNK1G1, CSNK2B, CTNND2, DEPDC5, DHDDS, DNM1, DOCK7, DYNC1H1, EEF1A2, EFHC1, EIF2S3, EMC1, EPM2A, FASN, FLNA, FOXG1, GABBR2, GABRA1, GABRA2, GABRA3, GABRB2, GABRB3, GABRD, GABRG2, GAL, GNAO1, GOSR2, GRIA1, GRIN1, GRIN2A, GRIN2B, HCN1, HCN4, HDAC4, HNRNPU, IDH3A, IQSEC2, JRK, KCNA1, KCNA2, KCNB1,
    [Show full text]
  • Supplementary Information
    Osa et al Supplementary Information Clinical implications of monitoring nivolumab immunokinetics in previously treated non– small cell lung cancer patients Akio Osa, Takeshi Uenami, Shohei Koyama, Kosuke Fujimoto, Daisuke Okuzaki, Takayuki Takimoto, Haruhiko Hirata, Yukihiro Yano, Soichiro Yokota, Yuhei Kinehara, Yujiro Naito, Tomoyuki Otsuka, Masaki Kanazu, Muneyoshi Kuroyama, Masanari Hamaguchi, Taro Koba, Yu Futami, Mikako Ishijima, Yasuhiko Suga, Yuki Akazawa, Hirotomo Machiyama, Kota Iwahori, Hyota Takamatsu, Izumi Nagatomo, Yoshito Takeda, Hiroshi Kida, Esra A. Akbay, Peter S. Hammerman, Kwok-kin Wong, Glenn Dranoff, Masahide Mori, Takashi Kijima, Atsushi Kumanogoh Supplemental Figures 1 – 8 1 Osa et al Supplemental Figure 1. The frequency of nivolumab-bound T cells was maintained in patients who continued treatment. Nivolumab binding in CD8 and CD4 T cells was analyzed at two follow-up points, as indicated, in fresh peripheral blood from three representative cases from protocol 1 that continued treatment. 2 Osa et al Supplemental Figure 2. Long-term follow-up of nivolumab binding to T cells from fresh whole blood. Nivolumab binding was followed up in fresh peripheral blood from an additional case, Pt.7. 3 Osa et al Supplemental Figure 3. Long-term duration of nivolumab binding is due to sustained circulation of residual nivolumab in plasma. (A) PBMCs acquired from Pt.8 and 9 at pretreatment (pre PBMCs) and after a single dose (post 1 PBMCs) were cultured in regular medium without nivolumab (top and middle). Pre PBMCs were also incubated with 10 µg/ml nivolumab in vitro before the cultures were started (bottom). Nivolumab binding status was monitored at the indicated time points.
    [Show full text]