Regulation of signaling in tumor infiltrating CD8+ T cells by Protocadherin-18

Protocadherin-18 对肿瘤浸润 CD8 T 细胞信号传导的调节

基本信息

批准号：
8132458
负责人：
Edwin J. Vazquez-Cintron
金额：
$ 0.03万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-01 至 2011-09-06
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8132458
关键词：
Affect Antibodies Binding CD8B1 gene Cancer Patient Cells Characteristics Coculture Techniques Complex Data Defect Detergents Development Dissociation Down-Regulation Epitopes Functional disorder Genes Genetic Transcription Goals Immune response Immunity Immunoblot Analysis Immunoblotting Immunotherapeutic agent In Situ In Vitro Lytic Malignant Neoplasms Mediating Memory Modeling Molecular Weight Mus Myelogenous Nonlytic Peptides Phase Phenotype Phosphopeptides Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Protein Dephosphorylation Protein Tyrosine Kinase RNA Interference Recovery Recruitment Activity Regulation Role Signal Transduction Site Suppressor-Effector T-Lymphocytes T cell response T-Lymphocyte Testing Tumor Escape Tumor-Infiltrating Lymphocytes Tyrosine Phosphorylation base cancer therapy design in vitro Assay neoplastic cell novel prevent programs protein aminoacid sequence protocadherin 18 release of sequestered calcium ion into cytoplasm restoration src-Family Kinases synthetic peptide tool tumor tumor growth

项目摘要

DESCRIPTION (provided by applicant): Systemic T cell immunity is intact in both cancer patients and in murine models but Tumor Infiltrating Lymphocytes (TIL) are defective in cytolytic function. Available data describing TIL dysfunction suggests that tumor-induced suppression of TIL effector phase function is a common characteristic of cancer which may contribute to tumor escape from antitumor immune response. The Frey lab has determined that TCR-mediated signaling is defective in both nonlytic CD8+ TIL in situ and when freshly-isolated and assayed in vitro: upon conjugation with cognate tumor cells in vitro TIL do not activate tyrosine kinases or flux calcium. Since signaling is required for T cell effector phase function, defective TIL signal transduction is undoubtedly the basis for the nonlytic phenotype. The signaling and lytic defects are transient: when TIL are purified and briefly cultured in vitro signaling is restored coincident with recovery of lytic function. The signaling defect is proximal in the TCR-mediated signaling cascade: contact with tumor cells in vitro causes rapid dephosphorylation of the activation motif in p56lck (Y394) therein preventing propagation of the activating signal. In nonlytic TIL, but not in lytic TIL, the inhibitory phosphatase Shp-1 is activated and co-localizes with p56lck at the TIL:target cell contact site. TIL signaling and lytic defects are completely restored upon purification and brief culture in vitro including dissociation of Shp-1 from the TCR complex and reduction of Shp-1 phosphatase activity. Recovery of defective TIL signaling in vitro is blocked by co-culture with tumor cells implying a contact-dependent, fast-acting 'switch'. Also, if TIL are first 'recovered' prior to co-culture with tumor cells (by brief culture in the absence of tumor cells), the TIL proximal TCR signaling defect is re-introduced. Upon culture of TIL with non-cognate tumor cells or tumor associated myeloid derived suppressor cells, TIL signaling and lytic functions are not affected. These observations show that tumor cells deliver an inhibitory signal to TIL resulting in recruitment of activated Shp-1 to the TCR complex which in turn inactivates proximal TCR-mediated signal transduction and therein effector phase function. Identification of the mechanism by which Shp-1 is recruited to the TIL TCR is a major objective of the proposed project. My long-term goal is to define the mechanism by which tumors escape elimination by the immune response which will have implications for the rational design of effective immunotherapeutic tools for treatment of cancer, as well as contribute to our understanding of how antitumor T cell response is regulated during tumor growth.

描述（由申请人提供）：癌症患者和鼠模型中的全身性T细胞免疫都是完整的，但是肿瘤浸润的淋巴细胞（TIL）在细胞溶解功能方面有缺陷。描述TIL功能障碍的可用数据表明，肿瘤诱导的TIL效应相位功能的抑制是癌症的常见特征，这可能导致肿瘤从抗肿瘤免疫反应中逃脱。 Frey Lab已确定TCR介导的信号传导在原位和新鲜分离并在体外进行测定时在非赖以晶状的CD8+ TIL中均有缺陷：与同源肿瘤细胞结合后，体外TIL会激活酪氨酸激酶或通量钙。由于T细胞效应器相函数所需的信号传导，因此TIL信号转导有缺陷无疑是非宽度表型的基础。信号传导和裂解缺陷是短暂的：纯化纯化并在体外信号中短暂培养时，与恢复裂解功能的恢复相吻合。信号缺陷在TCR介导的信号传导级联反应近端：在体外与肿瘤细胞接触，导致p56LCK（Y394）激活基序的快速去磷酸化，从而阻止了激活信号的传播。在非散型TIL中，但在裂解TIL中，抑制性磷酸酶SHP-1被激活，并在TIL：靶细胞接触位点与p56LCK共定位。 TIL信号传导和裂解缺陷在体外纯化和短暂培养后完全恢复，包括SHP-1与TCR复合物的解离以及SHP-1磷酸酶活性的还原。与肿瘤细胞共培养的恢复在体外的恢复，这意味着接触依赖性，快速作用的“开关”。同样，如果在与肿瘤细胞共培养之前首先“回收”（在没有肿瘤细胞的情况下通过短暂培养），则重新引入TIL近端TCR信号缺陷。在与非共物肿瘤细胞或肿瘤相关的髓样抑制细胞的TIL培养后，TIL信号传导和裂解功能受到影响。这些观察结果表明，肿瘤细胞发出了抑制信号，直到导致活化的SHP-1募集到TCR复合物中，而TCR络合物又会使TCR近端TCR介导的信号转导和其中效应子相位函数失活。识别将SHP-1募集到TIL TCR的机制是拟议项目的主要目标。我的长期目标是定义肿瘤通过免疫反应消除消除的机制，这将对有效的免疫治疗工具的理性设计产生影响，并有助于我们理解在肿瘤生长过程中如何调节抗肿瘤T细胞反应。