A T cell STAT3/BATF-axis regulates intestinal γδ T cell homeostasis and disease

项目总结 我的长期职业目标是作为一名大学的独立调查员进行假设驱动的研究 跨越炎症性肠病(IBD)的基础和临床免疫学。为了实现这一目标，我 组建了一个由马修·奥尔森博士(赞助人)、蒂莫西·拉特利夫博士(共同赞助人)、 慈文博士(合作者)、马吉德·卡兹米安博士(合作者)、匡世欢博士和韦恩博士 坎贝尔为体外细胞培养技术、在体小鼠肠道模型提供技术培训 疾病，灵知生物训练，以及科学写作和科学写作的其他专业发展技能 沟通。我在这项提案中概述的初步工作表明，患有常规T细胞的小鼠 两个重要的T细胞转录因子STAT3和BATF的细胞特异性缺失导致侵袭性 自发性结肠炎，以微生物区系失调和γδT细胞数量增加为特征 肠道与在人类病人身上观察到的相似。鉴于目前可供选择的治疗方法 IBD在长期治疗中并不完全有效，我的数据表明通过破坏STAT3/BATF轴 靶向失调的肠道微生物区系或γδT细胞可能代表一种新的治疗策略。为了进一步 了解肠道微生物区系失调和非常规γδT细胞之间的这种复杂关系 对于IBD样结肠炎，这项建议将解决以下问题：(1)阐明微生物区系在调节 肠道疾病时肠道γδT细胞的动态平衡。16S rRNA测序将用于定义 每种小鼠基因中的微生物群落。为了确定这个微生物区系是否需要并且足以 诱导γδT细胞反应和疾病我们将用抗生素治疗动物并进行粪便微生物区系 分别转化为无菌动物。(2)探讨γδT细胞在IBD发病机制中的作用。从治疗上讲， 我将用阻断γδT细胞受体激活的单抗治疗小鼠，并监测其对 肠道的炎症反应与临床疾病的发展。为了确定γδT细胞 可以传播疾病，我会从Stat3fl/flBatffl/flCD4Cre+小鼠中分离γδT细胞并将它们转移到结肠炎中 易感(RAG-/-)小鼠。这一提议的结果将进一步阐明γδT细胞在 Stat3fl/flBatffl/flCD4Cre+小鼠，可作为研究γδT细胞在IBD样结肠炎中的有用模型。考虑到 目前对IBD患者的治疗只有部分有效，这项研究的结果数据和 在IBD样结肠炎中建立这种独特的γδT细胞小鼠模型将有可能确定新的治疗方法 目标是对全球680万IBD患者产生积极影响。

PROJECT SUMMARY My long-term career goal is to do hypothesis-driven research as an independent investigator at a university spanning basic and clinical immunology in inflammatory bowel diseases (IBD). In order to achieve this goal, I have put together an advisory team composed of Dr. Matthew Olson (sponsor), Dr. Timothy Ratliff (co-sponsor), Dr. Tzu-wen Cross (collaborator), Dr. Majid Kazemian (collaborator), Dr. Shihuan Kuang, and Dr. Wayne Campbell to provide technical training for in vitro cell culture techniques, in vivo mouse models of intestinal disease, and gnotobiotic training, as well as other professional development skills in scientific writing and communication. My preliminary work outlined in this proposal has demonstrated that mice with conventional T cell-specific deletions in two important T cell transcription factors, STAT3 and BATF, developed an aggressive spontaneous colitis that was marked by a dysregulated microbiota and elevated numbers of γδ T cells in the intestines similar to what is observed in human patients. Given that the current treatment options available for IBD are not fully effective in long term treatment, my data indicate that disrupting this STAT3/BATF-axis through targeting the dysregulated gut microbiota or γδ T cells may represent a novel therapeutic strategy. To further understand this complex relationship between dysregulated gut microbiota and non-conventional γδ T cells in IBD-like colitis, this proposal will address the following: (1) Elucidate the role of the microbiota in regulating intestinal γδ T cell homeostasis during intestinal disease. 16S rRNA sequencing will be used to define shifts in the microbial community in each mouse genotype. To determine if this microbiota is required and sufficient to induce γδ T cell responses and disease we will treat animals with antibiotics and perform fecal microbiota transfers into germ-free animals, respectively. (2) Determine the role of γδ T cells in driving IBD. Therapeutically, I will treat mice with monoclonal antibodies that block γδ T cell receptor activation and monitor its impact on inflammatory responses in the intestines and clinical disease development. In order to determine if γδ T cells can confer disease, I will isolate γδ T cells from Stat3fl/flBatffl/flCd4Cre+ mice and transfer them into colitis susceptible (Rag-/-) mice. The results of this proposal will further elucidate the role of γδ T cells in Stat3fl/flBatffl/flCd4Cre+ mice, which may serve as useful model to study γδ T cells in IBD-like colitis. Given that current therapies for IBD patients are only partially effective, the resulting data from this study and the establishment of this unique mouse model of γδ T cells in IBD-like colitis will potentially identify novel therapeutic targets to make a positive impact on the 6.8 million people living with IBD worldwide.