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: 9294990
• 负责人: Jaehyuk Choi
• 金额: $ 15.24万
• 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/9294990
• 关键词: Achievement; Adult T-Cell Leukemia/Lymphoma; Advisory Committees; Apoptosis; Binding; Binding Proteins; Binding Sites; Bioinformatics; Blood; CD4 Positive T Lymphocytes; CD47 gene; CD8-Positive T-Lymphocytes; Cause of Death; Cell Line; Cell physiology; Cells; ChIP-seq; Clinical; Clone Cells; Complex; Cutaneous; Cutaneous Lymphoma; Dermatologic; Dermatology; Disease; Down-Regulation; Educational Curriculum; Epigenetic Process; Exhibits; Foundations; Funding; Gene Targeting; Genes; Genetic; Genetic Recombination; Genetic Transcription; Germ-Line Mutation; Goals; Hereditary Disease; Homeobox; Homing; Human; Immune; Immune system; Immunobiology; Immunophenotyping; 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; Receptor Activation; Recurrence; Reporting; Research; Resistance; Resources; Role; Sampling; Scientist; Sezary Syndrome; Skin; Small Interfering RNA; Structure; Sum; T-Cell Lymphoma; T-Cell Proliferation; T-Cell Receptor; T-Lymphocyte; Time; Tissues; Training; Transcript; Transforming Growth Factor beta; Tumor Suppressor Proteins; United States National Institutes of Health; Visceral; Zinc Fingers; career; cohort; cytokine; exome sequencing; experience; genome sequencing; genome-wide; immunoregulation; immunosuppressed; in vivo; loss of function mutation; lymph nodes; mouse model; mutant; neoplastic cell; novel; novel therapeutics; professor; promoter; public health relevance; statistics; thymocyte; transcription factor; transcriptome; transcriptome sequencing; translational scientist; whole genome

 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 CTCLs. 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, epigenetic, 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）：美国国立卫生研究院K 08指导职业发展奖为我提供了必要的基础，以实现我的长期职业目标，成为一名独立资助的翻译研究者，临床和科学重点是皮肤T细胞淋巴瘤（CTCL）。CTCL是一种皮肤归巢的CD 4 + T细胞的不可治愈的非霍奇金淋巴瘤。患者最初仅在皮肤中存在淋巴瘤细胞;然而，随着疾病的进展，肿瘤细胞扩散到血液，淋巴结和内脏器官。恶性T细胞克隆产生细胞因子，显著改变免疫系统;相应地，CTCL的常见死亡原因是致命的免疫抑制。作为耶鲁大学的助理教授，我把我的临床工作集中在照顾这种恶性肿瘤的病人上。由于癌症从根本上说是一种遗传性疾病，我将实验室的工作重点放在确定CTCL的遗传基础上。据我们所知，我已经进行了CTCL的第一个外显子组测序。对40个来自晚期白血病患者的CTCL进行测序，我们已经高度清晰地确定了癌症促进突变的景观。特别是，我们发现ZEB 1是CTCL中的关键肿瘤抑制因子，在65%的患者中存在功能丧失突变。ZEB 1编码一个锌指E盒结合同源盒转录因子，有趣的是，该因子在多个基因上具有推定的结合位点，这些基因被认为介导CTCL独特的免疫表型。突出了其在T细胞中的肿瘤抑制功能，84%的ZEB 1生殖系突变小鼠自发发展CD 4+外周T细胞淋巴瘤（PTCL）。在这个提议中，我们的目标是阐明ZEB 1突变在促进CD 4 + T细胞恶性转化中的作用。在目标1中，我们将通过对早期皮肤局限性CTCL中的ZEB 1进行测序和对白血病CTCL中的ZEB 1进行全基因组测序来确定CTCL中ZEB 1突变的发作时间。在目标2中，我们将鉴定ZEB 1在CTCL中的转录靶点，使用1）RNA-Seq以发现在ZEB 1-/- CTCL中改变的转录物，和2）ChIP-Seq以鉴定ZEB 1在ZEB 1充满的CD 4 + T细胞中的结合位点。在目标3中，我们将通过仔细分析ZEB 1-/-小鼠PTCL的表型和转录组来分离ZEB 1突变对体内淋巴瘤发生的贡献。为了获得成功实现所提出的目标所需的专业知识，我已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全面的培训计划。我将遵循一个结构化的课程，包括免疫生物学，统计学和生物信息学的研讨会和课程。此外，我将得到一位高素质的初级导师（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CTCL、表观遗传学和免疫生物学方面的指导和非重叠专业知识的良好记录精心挑选的。总之，这一建议将有望阐明一个关键的机制CTCL发病机制，同时准备我作为一个成功的独立的职业生涯医生科学家。