Dissecting the role of GPR34 in cDC1 migration and function

项目总结/摘要 作为针对细胞内病原体和癌症的免疫的关键介质，1型常规 树突状细胞（cDC 1）交叉呈递外源性细胞相关抗原至CD 8 + T细胞， 这两种细胞类型在次级淋巴组织的T细胞区中的共定位。这种显微解剖学 组织是由CCR 7和XCR 1协调的，CCR 7和XCR 1是cDC 1上的两个G蛋白偶联受体（GPCR）， 引导它们向化学引诱物配体的梯度迁移。最近的报告表明， 肿瘤相关cDC 1在癌症免疫中的作用。然而，介导cDC 1募集的GPCR和 在炎症部位和肿瘤中的定位还不完全清楚。 初步研究表明，GPR 34是一种未充分研究的X连锁GPCR， cDC 1，体外研究表明，这种溶血磷脂酰丝氨酸（lysoPS）反应性受体支持 细胞迁移本文提供的初步数据表明，GPR 34促进cDC 1在细胞中的积累。 炎症小鼠腹腔（PerC）在细胞内的方式。PerC是临床上重要的部位， 结肠癌和卵巢癌转移。在一项初步实验中， 积累和癌症结果，GPR 34缺陷导致皮下（SQ）肿瘤生长增加 模型该提案将检验GPR 34调节cDC 1组织和功能的假设 在炎症和癌症期间。目标1将机械地定义GPR 34如何控制丰度， 运输和定位PerC cDC 1，使用包括体内动力学免疫在内的技术组合 表型分析、体外迁移测定和免疫荧光显微术。目标2将研究 使用异位SQ和PerC肿瘤模型沿着工具研究癌症免疫中的GPR 34和lysoPS 抗原特异性T细胞应答和操纵配体水平。完成这些目标将阐明关键 cDC 1 s上的GPR 34影响炎症和恶性肿瘤反应的机制， 这为开发调节lysoPS-GPR 34化学引诱物系统的治疗剂提供了机会。 这些研究目标将与全面的培训计划一起进行， 申请人作为一个独立的职业生涯作为一个学术物理学家，科学家。培训包括严格的 导师在科学技能从一个高素质的赞助商，博士贾森Cyster;技术教育， 申请人实验室和合作实验室的博士后研究员;研讨会，期刊俱乐部，研讨会， 会议;和临床活动。研究和培训将在哥伦比亚大学进行。 加州，旧金山弗朗西斯科，提供高度创新和协作的免疫学研究环境 以及一所提供临床培训的优秀医学院

PROJECT SUMMARY/ABSTRACT As critical mediators of immunity against intracellular pathogens and cancer, type 1 conventional dendritic cells (cDC1s) cross-present exogenous, cell-associated antigens to CD8+ T cells, facilitated by the colocalization of these two cell types in the T cell zone of secondary lymphoid tissues. This microanatomical organization is orchestrated by CCR7 and XCR1, two G protein-coupled receptors (GPCRs) on cDC1s that guide their migration towards gradients of chemoattractant ligand. Recent reports have established a crucial role for tumor-associated cDC1s in cancer immunity. However, the GPCRs mediating cDC1 recruitment and positioning at sites of inflammation and in tumors are incompletely understood. Preliminary work has revealed that GPR34, an understudied X-linked GPCR, is highly expressed by cDC1s, and in vitro studies have shown that this lysophosphatidylserine (lysoPS)-responsive receptor supports cell migration. Preliminary data presented herein indicate that GPR34 promotes the accumulation of cDC1s in the inflamed mouse peritoneal cavity (PerC) in a cell-intrinsic manner. The PerC is a clinically important site for colon and ovarian cancer metastasis. In a preliminary experiment inspired by the connection between cDC1 accumulation and cancer outcomes, GPR34-deficiency led to increased growth of a subcutaneous (SQ) tumor model. This proposal will test the hypothesis that GPR34 regulates cDC1 organization and function during inflammation and cancer. Aim 1 will mechanistically define how GPR34 controls the abundance, trafficking, and positioning of PerC cDC1s using a combination of techniques including in vivo kinetic immune phenotyping, in vitro migration assays, and immunofluorescence microscopy. Aim 2 will investigate the role of GPR34 and lysoPS in cancer immunity using ectopic SQ and PerC tumor models along with tools to study antigen-specific T cell responses and to manipulate ligand levels. Completion of these Aims will elucidate key mechanisms by which GPR34 on cDC1s influences the response to inflammation and malignancy, providing opportunities for the development of therapeutics that modulate the lysoPS-GPR34 chemoattractant system. These research goals will be conducted in conjunction with a comprehensive training plan to prepare the applicant for an independent career as an academic physician-scientist. Training includes rigorous mentorship in scientific skills from a highly qualified sponsor, Dr. Jason Cyster; technical education from postdoctoral fellows in the applicant’s lab and collaborating labs; seminars, journal clubs, workshops, and conferences; and clinically geared activities. Research and training will take place at the University of California, San Francisco, which offers a highly innovative and collaborative immunology research environment and an exceptional medical school for clinical training.