Identification of Novel Epigenetic Regulators of lymphocyte Development

项目总结/摘要 T细胞在免疫中起着核心作用，具有重要的治疗意义。T细胞研究 发展提供了对各种疾病的病理生理学的直接见解， 促进过继性T细胞免疫疗法。尽管已经做了很大的努力来确定 这些研究主要集中在分泌蛋白、细胞因子和细胞因子上， 表面受体和已知直接驱动发育过程的转录因子。 最近，我们发现，操纵淋巴潜能的表观遗传调节因子， 从人诱导多能干细胞（iPSC）产生成熟的功能性T细胞， 证明了表观遗传调节剂在T细胞发育和功能中的重要作用。到 迄今为止，T细胞发育的表观遗传调控仍然难以捉摸。在本申请中，我们提出 为了验证iPSC提供了一个平台来识别调节T细胞的新机制的假设， 发展，以及对T细胞分化的表观遗传调控的新见解，可用于 促进产生用于过继性T细胞免疫疗法的更稳健的iPSC-T细胞。在此之前， 我们已经建立了一个无基质的分化系统，忠实地再现了T细胞 文化的发展。在目标1中，我们将在iPSC的不同阶段进行小分子筛选， T细胞分化，发现新的表观遗传调节因子，可以影响淋巴规范和T细胞分化。 细胞成熟我们已经通过使用一个库进行试点屏幕提供了一个概念验证， 调节表观遗传修饰酶活性的小分子。识别的屏幕 几种表观遗传调节剂，如组蛋白甲基转移酶G9 a，促进T细胞命运 承诺.在目标2中，我们将把我们的研究转移到机械地研究表观遗传如何影响我们的研究。 调节通过调节染色质结构来控制淋巴细胞的命运决定。的影响 表观遗传调节剂，包括G9 a，对淋巴细胞生成的影响将在iPSC衍生的血液和骨髓中进行探测。 细胞和斑马鱼模型。最后，Aim 3将专注于使用小分子介导的表观遗传 在一些实施方案中，iPSC-T细胞可以通过调节来促进具有增强功能的成熟iPSC-T细胞的产生。成功 拟议的研究不仅将提高我们对潜在的分子机制的理解， 它不仅可以促进免疫细胞的形成，而且还为基于干细胞的免疫疗法开辟了新的途径。的 职业发展奖将使我能够发展必要的新技能，以实现所提出的目标 培养我成为一名独立调查员

Project Summary/Abstract T cells play a central role in immunity and are of great therapeutic importance. Studies on T cell development provide direct insights into the pathophysiology of a wide variety of diseases and can facilitate adoptive T cell immunotherapies. Though a large effort has been made to identify pathways that regulate T cell differentiation, these studies have mostly focused on secreted proteins, cell surface receptors, and transcription factors known to directly drive developmental processes. Recently, we discovered that manipulation of an epigenetic regulator of lymphoid potential promotes the generation of mature, functional T cells from human induced pluripotent stem cells (iPSCs), demonstrating an important role for epigenetic modulators in T cell development and function. To date, the epigenetic regulation of T cell development remains elusive. In this application, we propose to test the hypothesis that iPSCs provide a platform to identify novel mechanisms regulating T cell development, and new insights into epigenetic regulations of T cell differentiation can be used to facilitate the generation of more robust iPSC-T cells for adoptive T cell immunotherapy. Previously, we have established a stroma-free differentiation system that faithfully recapitulates T cell development in culture. In Aim 1, we will perform small molecule screens at different stages of iPSC- T cell differentiation to discover new epigenetic regulators that can affect lymphoid specification and T cell maturation. We have provided a proof-of-concept by conducting a pilot screen using a library of small molecules that modulate the activity of epigenetic modification enzymes. The screen identified several epigenetic modulators, such as histone methyltransferase G9a, that promote T cell fate commitment. In Aim 2, we will transition our study to mechanistically examine how epigenetic regulations govern lymphocyte fate decisions via modulating chromatin structure. The impact of epigenetic modulators, including G9a, on lymphopoiesis will be probed in both iPSC-derived blood cells and zebrafish models. Lastly, Aim 3 will focus on using small molecule-mediate epigenetic modulation to facilitate the production of mature iPSC-T cells with enhanced function. Success in the proposed study will not only improve our understanding of the molecular mechanisms underlying the formation of immune cells but also open new avenues for stem cell-based immunotherapies. The career development award will allow me to develop new skills necessary to fulfill the proposed goals and foster my development into an independent investigator.