Dissecting the role of GPR34 in cDC1 migration and function

剖析 GPR34 在 cDC1 迁移和功能中的作用

• 批准号: 10748747
• 负责人: Hanson Peter Tam
• 金额: $ 3.98万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10748747
• 关键词: Antigen Presentation; Antigens; Area; Biological Assay; Biological Models; Biology; Bone Marrow; CD8-Positive T-Lymphocytes; California; Cell Communication; Cell physiology; Cells; Cellular Assay; Chemotactic Factors; Chimera organism; Clinical; Collaborations; Colon Adenocarcinoma; Colon Carcinoma; Colorectal Cancer; Confocal Microscopy; Cross Presentation; Data; Dendritic Cells; Educational workshop; Engineering; Environment; Enzymes; Epithelial ovarian cancer; Flow Cytometry; G-Protein-Coupled Receptors; Generations; Goals; Greater sac of peritoneum; Growth; Immune; Immune response; Immunity; Immunofluorescence Immunologic; Immunofluorescence Microscopy; Immunology; Implant; In Vitro; Infection; Inflammation; Journals; Kinetics; Ligands; Link; Liquid substance; Lymphoid Tissue; MC38; Malignant Neoplasms; Malignant neoplasm of ovary; Measures; Mediating; Mediator; Mentorship; Microanatomy; Migration Assay; Modeling; Mus; Natural Immunity; Neoplasm Metastasis; Organ; Outcome; Ovalbumin; Peritoneal; Peritonitis; Phenotype; Physicians; Physiology; Positioning Attribute; Postdoctoral Fellow; Process; Proliferating; Qualifying; Reporting; Research; Role; San Francisco; Scientist; Shapes; Signal Transduction; Site; Source; Supporting Cell; System; T cell response; T-Lymphocyte; Techniques; Testing; Therapeutic; Thioglycolates; Time; Training; Tumor Cell Line; Tumor Immunity; Universities; Visualization; Vocational Education; Weight; Work; XCR1 gene; adaptive immunity; antigen-specific T cells; cancer cell; career; cell motility; cell type; clinical training; colon cancer metastasis; druggable target; experimental study; in vivo; innovation; lysophosphatidylserine; medical schools; migration; overexpression; pathogen; receptor; recruit; response; skills; subcutaneous; symposium; therapeutic development; tool; trafficking; tumor; tumor growth

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.

项目摘要/摘要 作为对抗细胞内病原体和癌症免疫的关键介质，1型常规 树突状细胞(CDC1s)向CD8+T细胞递送外源性的细胞相关抗原，这是由 这两种细胞类型共存于次级淋巴组织的T细胞区。这种显微解剖学 组织由CCR7和XCR1协调，这是CDC1s上的两个G蛋白偶联受体(GPCRs)， 引导它们向化学诱导剂配体的梯度迁移。最近的报告已经建立了一个至关重要的 肿瘤相关CDC1s在癌症免疫中的作用。然而，GPCRs调解cDC1的招募和 炎症部位和肿瘤部位的定位尚不完全清楚。 初步工作表明，GPR34是一种未被研究的X连锁GPCR，它在 CDC1s，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。初步数据表明，GPR34促进了cDC1s在体内的积累 炎症的小鼠腹膜腔(Perc)以细胞固有的方式。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，灵感来自于cDc1与 GPR34缺乏导致皮下(SQ)肿瘤生长增加 模特。这一提议将检验GPR34调控cDC1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GPR34如何控制丰度， 利用包括体内运动免疫在内的多种技术组合运输和定位perc cDc1 表型鉴定、体外迁移试验和免疫荧光显微镜。目标2将调查 利用异位SQ和PERC肿瘤模型及研究工具研究GPR34和LlysoPS在肿瘤免疫中的作用 抗原特异性T细胞反应和操纵配体水平。完成这些目标将阐明关键 CDC1s上GPR34影响炎症和恶性肿瘤反应的机制 开发调节LlysoPS-GPR34化学诱导剂系统的治疗药物的机会。 这些研究目标将与全面的培训计划一起进行，以准备 作为一名学术内科医生-科学家的独立职业的申请者。培训包括严格的 来自高资质赞助商Jason Cyster博士的科学技能指导；来自 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、研讨会和 会议；与临床相关的活动。研究和培训将在华盛顿大学进行 加利福尼亚州，旧金山，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特殊医学院。