The Role of ZEB1 Mutations in Cutaneous T Cell Lymphoma

ZEB1 突变在皮肤 T 细胞淋巴瘤中的作用

基本信息

批准号：
9129253
负责人：
Jaehyuk Choi
金额：
$ 13.53万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-12-08 至 2020-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9129253
关键词：
Achievement Adult T-Cell Leukemia/Lymphoma Advisory Committees Apoptosis Binding Binding Sites Bioinformatics Blood Boxing CD4 Positive T Lymphocytes CD47 gene CD8B1 gene Cause of Death Cell Line Cell physiology Cells ChIP-seq Clinical Complex Cutaneous Dermatology Disease Down-Regulation Educational Curriculum Epigenetic Process Exhibits Foundations Funding Gene Expression Profile Gene Targeting Genes Genetic Genetic Recombination Genetic Transcription Germ-Line Mutation Goals Health Hereditary Disease Homeobox Homing Human Immune Immune system Immunobiology Immunosuppression In Vitro Interleukin-13 Interleukin-4 Interleukin-5 K-Series Research Career Programs Laboratories LacZ Genes Lymphoma Lymphomagenesis Lymphopenia Malignant - descriptor Malignant Neoplasms Mediating Mentors Mentorship Mus Mutation Mycosis Fungoides Non-Hodgkin&apos s Lymphoma Non-Malignant Organ Pathogenesis Pathway interactions Patient Care Patients Peripheral Phenotype Physicians Production Protein Binding Qualifying Receptor Activation Recurrence Reporting Research Research Personnel Resistance Resources Role Sampling Scientist Sezary Syndrome Skin Small Interfering RNA Staging Structure Sum T-Cell Lymphoma T-Cell Proliferation T-Cell Receptor T-Lymphocyte Time Tissues Training Transcript Tumor Suppressor Proteins United States National Institutes of Health Visceral Zinc Fingers base career cytokine exome sequencing experience genome sequencing genome-wide immunosuppressed in vivo loss of function mutation lymph nodes mouse model mutant neoplastic cell novel novel therapeutics professor promoter statistics thymocyte transcription factor transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): The NIH K08 mentored career development award provides the necessary foundation for me to fulfill my long- term career goal of becoming an independently funded translational investigator with a clinical and scientific focus on cutaneous T cell lymphoma (CTCL). CTCL is an incurable non-Hodgkin lymphoma of the skin-homing CD4+ T cell. Patients initially present with lymphoma cells exclusively in the skin; however, as disease progresses, the tumor cells spread to the blood, lymph nodes, and visceral organs. The malignant T cell clone produces cytokines that dramatically alter the immune system; correspondingly, a common cause of death from CTCL is fatal immunosuppression. As an Assistant Professor at Yale, I have focused my clinical efforts on caring for patients with this malignancy. Since cancer is fundamentally a genetic disease, I have focused my laboratory efforts on identifying the genetic basis of CTCL. To our knowledge, I have performed the first exome sequencing of CTCL. Sequencing 40 CTCLs from patients with advanced leukemic disease, we have identified the landscape of cancer promoting mutations with high clarity. In particular, we found that ZEB1 is a critical tumor suppressor in CTCL, subject to loss-of-function mutations in 65% of patients. ZEB1 encodes a zinc finger E-box binding homeobox transcription factor that interestingly has putative binding sites at multiple genes thought to mediate CTCL's distinctive immune phenotype. Highlighting its tumor suppressor function in T cells, 84% of mice with germline mutations in ZEB1 spontaneously develop CD4+ peripheral T cell lymphomas (PTCL). In this proposal, our objectives are to elucidate the role of ZEB1 mutations in promoting the malignant transformation of CD4+ T cells. In Aim 1, we will determine the time of onset of ZEB1 mutations in CTCL by sequencing ZEB1 in early-stage skin-limited CTCL and whole genome sequencing ZEB1 in leukemic CTCL. In Aim 2, we will identify the transcriptional targets of ZEB1 in CTCL, using 1) RNA-Seq to find transcripts that are altered in ZEB1-/- CTCL and 2) ChIP-Seq to identify ZEB1's binding sites in ZEB1-replete CD4+ T cells. In Aim 3, we will isolate the contribution of ZEB1 mutations to lymphomagenesis in vivo by carefully analyzing the phenotypes and transcriptomes of ZEB1-/- murine PTCLs. To achieve the expertise necessary for the successful achievement of the proposed aims, I have committed to a comprehensive training plan utilizing the extensive scientific and clinical resources available in Yale's world-renowned Departments of Genetics, Immunobiology, and Dermatology. I will follow a structured curriculum consisting of seminars and coursework in immunobiology, statistics, and bioinformatics. In addition, I will be closely mentored by a highly qualified primary mentor (Dr. Richard Lifton) and an advisory committee, which was carefully chosen for their proven track record for mentorship and non-overlapping expertise in CTCL, epigenetics, and immunobiology. In sum, this proposal will hopefully elucidate a critical mechanism underlying CTCL pathogenesis while preparing me for a successful independent career as a physician scientist.

描述（由申请人提供）：NIH K08 指导的职业发展奖为我实现长期职业目标提供了必要的基础，即成为一名独立资助的转化研究人员，专注于皮肤 T 细胞淋巴瘤 (CTCL) 的临床和科学研究。 CTCL 是一种无法治愈的皮肤归巢 CD4+ T 细胞非霍奇金淋巴瘤。患者最初仅在皮肤中出现淋巴瘤细胞；然而，随着疾病的进展，肿瘤细胞会扩散到血液中。恶性 T 细胞克隆会产生显着改变免疫系统的细胞因子；相应地，CTCL 死亡的常见原因是致命的免疫抑制。由于癌症从根本上来说是一种遗传性疾病，因此我的实验室工作重点是确定 CTCL 的遗传基础，据我们所知，我对患者的 40 例 CTCL 进行了首次外显子组测序。对于晚期白血病，我们已经高度明确地确定了癌症促进突变的情况，特别是，我们发现 ZEB1 是 CTCL 中的关键肿瘤抑制因子，在 65% 的患者中，ZEB1 编码 a 基因，会发生功能丧失突变。有趣的是，锌指 E-box 结合同源盒转录因子在多个基因上具有假定的结合位点，这些基因被认为介导 CTCL 独特的免疫表型，突显其在 T 细胞中的肿瘤抑制功能，84% 的小鼠具有种系突变。 ZEB1 自发形成 CD4+ 外周 T 细胞淋巴瘤 (PTCL) 在本提案中，我们的目标是阐明 ZEB1 突变在促进 CD4+ T 细胞恶性转化中的作用。在目标 1 中，我们将确定 ZEB1 突变的发生时间。在目标 2 中，我们将通过对早期皮肤限制性 CTCL 中的 ZEB1 进行测序和对白血病 CTCL 中的全基因组测序 ZEB1 来识别 CTCL 中的转录靶点。 CTCL 中的 ZEB1，使用 1) RNA-Seq 来查找 ZEB1-/- CTCL 中发生改变的转录本，以及 2) ChIP-Seq 来识别 ZEB1 在充满 ZEB1 的 CD4+ T 细胞中的结合位点。在目标 3 中，我们将分离贡献。通过仔细分析 ZEB1-/- 鼠 PTCL 的表型和转录组，研究 ZEB1 突变对体内淋巴瘤发生的影响，以获得成功实现所需的专业知识。为了实现拟议的目标，我致力于利用耶鲁大学世界著名的遗传学、免疫生物学和皮肤病学系提供的广泛科学和临床资源制定全面的培训计划，我将遵循由免疫生物学、统计学研讨会和课程组成的结构化课程。此外，我将得到一位高素质的主要导师（理查德·利夫顿博士）和一个咨询委员会的密切指导，该委员会是根据他们在指导和不重叠方面的良好记录而精心挑选的。总之，这项提案有望阐明 CTCL 发病机制的关键机制，同时为我作为一名成功的独立医师职业生涯做好准备。