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

• 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
• 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' 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

 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(由申请者提供)：NIH K08导师职业发展奖为我实现长期职业目标提供了必要的基础，我希望成为一名独立资助的翻译研究员，专注于皮肤T细胞淋巴瘤(CTCL)的临床和科学研究。CTCL是一种不可治愈的皮肤归巢CD4+T细胞非霍奇金淋巴瘤。患者最初的淋巴瘤细胞仅存在于皮肤；然而，随着疾病的发展，肿瘤细胞扩散到血液、淋巴结和内脏器官。恶性T细胞克隆产生的细胞因子可以极大地改变免疫系统；相应地，CTCL的一个常见死亡原因是致命性免疫抑制。作为耶鲁大学的一名助理教授，我一直致力于临床护理这种恶性肿瘤的患者。由于癌症从根本上说是一种遗传性疾病，我的实验室工作重点是确定CTCL的遗传基础。据我们所知，我已经完成了CTCL的第一个外显子组测序。对来自晚期白血病患者的40个CTCL进行了测序，我们已经以高清晰度确定了癌症促进突变的图景。特别是，我们发现ZEB1是CTCL中的一个关键的肿瘤抑制因子，在65%的患者中会发生功能丧失突变。ZEB1编码一种锌指E盒结合同源盒转录因子，有趣的是，该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了CTCL独特的免疫表型。突显其在T细胞中的肿瘤抑制功能的是，ZEB1胚系突变的小鼠中有84%会自发发展为CD4+外周T细胞淋巴瘤(PTCL)。在这项建议中，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。在目的1中，我们将通过对早期皮肤受限CTCL的ZEB1测序和白血病CTCL的全基因组测序来确定CTCL中ZEB1突变的发生时间。在目标2中，我们将确定ZEB1在CTCL中的转录靶点，使用1)RNA-SEQ来寻找ZEB1-/-CTCL中发生改变的转录本，以及2)CHIP-SEQ来确定ZEB1‘S结合位点在ZEB1-完全的CD_4+T细胞中。在目标3中，我们将通过仔细分析ZEB1-/-小鼠PTCL的表型和转录本来分离ZEB1突变在体内淋巴肿瘤发生中的作用。为了获得成功实现拟议目标所需的专业知识，我承诺利用耶鲁大学世界知名的遗传学、免疫生物学和皮肤病学系的广泛科学和临床资源，制定一项全面的培训计划。我将遵循一个由免疫生物学、统计学和生物信息学的研讨会和课程组成的有组织的课程。此外，我将得到一位非常合格的初级导师(Richard Lifton博士)和一个咨询委员会的密切指导，该委员会是精心挑选的，因为他们在CTCL、表观遗传学和免疫生物学方面的导师和不重叠的专业知识方面的记录得到了证明。总而言之，这项建议有望阐明CTCL发病的关键机制，同时为我作为一名内科科学家成功的独立职业生涯做好准备。