Age related ubiquitin defect on T-cell response to tumor

年龄相关的泛素缺陷影响 T 细胞对肿瘤的反应

• 批准号: 7060416
• 负责人: HUANG-GE ZHANG
• 金额: $ 22.45万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7060416
• 关键词: Age; related; ubiquitin; defect; cell

DESCRIPTION (provided by applicant): The overall goal of this revised proposal is to determine the biologic effects of a novel breast tumor antigen containing F-box and WD40 domains, with an emphasis on its roles in age- and T cell-dependent susceptibility to tumor growth. We have identified a novel human breast tumor antigen (HBTA1) that strongly inhibits proliferation of T cells from aged, but not young, mice. Immunization of 2-mo-old BXD 12 mice with HBTA1 protects against the tumor challenge at 16-mo-of-age. Sucrose gradient purification of a mouse homologue HBTA1 (mHBTA1) followed by electromicroscope examination of the purified samples led to the realization that mHBTA1 is packed in the exosomes of the TS/A tumors. The TS/A exosomal mHBTA1 strongly inhibits the proliferation of T cells isolated from aged, but not young, BXD12 mice. The ability of T cells to proliferate in the presence of TS/A exosomes is correlated with the ubiquitination status of exosomal mHBTA1. Our aims are (1) To determine if ubiquitination of mHBTA1 is correlated with susceptibility to tumor growth and the tumor-specific cytotoxic T-cell response. Aged-mice will be immunized with ubiquitinated exosomal mHBTA1 to determine its effects on enhancement of protection against the tumor growth. In parallel, we will determine if the production of endogenous ubiquitinated mHBTA1 in TS/A exosomes is correlated with the tumor susceptibility. (2) To use siRNA technology to determine if knockout of exosomal mHBTA1 of tumor cells is sufficient to reverse the inhibition of T cell proliferation mediated by TS/A exosomes or whether other exosomal proteins are required. (3) To determine if the age-dependent T cell factors we recently identified play a role in ubiquitination of exosomal mHBTA1, and thus inactivation of mHBTA1-mediated inhibition of T-cell activation; we will further determine the pathway by which TS/A exosomal mHBTA1 interacts with T-cell factors. (4) To determine if an age-related T-cell factor(s) regulates the ubiquitination of exosomal HBTA1 of human breast tumors, and, further to determine its possible association with the age-related incidence of breast cancers. The data generated should elucidate the cellular and molecular basis for the role of tumor exosomes and exosomal mHBTA1 in the development of age- and T cell-dependent breast cancer.

描述（由申请人提供）：本修订提案的总体目标是确定含有F-box和WD 40结构域的新型乳腺肿瘤抗原的生物学效应，重点是其在肿瘤生长的年龄和T细胞依赖性易感性中的作用。我们已经确定了一种新的人乳腺肿瘤抗原（HBTA 1），它强烈抑制老年小鼠T细胞的增殖，而不是年轻小鼠。用HBTA 1免疫2月龄BXD 12小鼠可在16月龄时保护其免受肿瘤攻击。小鼠同源物HBTA 1（mHBTA 1）的蔗糖梯度纯化，然后对纯化样品进行电镜检查，结果发现mHBTA 1被包装在TS/A肿瘤的外泌体中。TS/A外泌体mHBTA 1强烈抑制从老年而非年轻BXD 12小鼠分离的T细胞的增殖。T细胞在TS/A外泌体存在下增殖的能力与外泌体mHBTA 1的泛素化状态相关。我们的目的是（1）确定mHBTA 1的泛素化是否与肿瘤生长的易感性和肿瘤特异性细胞毒性T细胞反应相关。将用泛素化的外泌体mHBTA 1免疫BALB/c小鼠，以确定其对增强针对肿瘤生长的保护的作用。同时，我们将确定TS/A外泌体中内源性泛素化mHBTA 1的产生是否与肿瘤易感性相关。(2)利用siRNA技术确定敲除肿瘤细胞外泌体mHBTA 1是否足以逆转TS/A外泌体介导的T细胞增殖抑制，或者是否需要其他外泌体蛋白。(3)为了确定我们最近发现的年龄依赖性T细胞因子是否在外泌体mHBTA 1的泛素化中发挥作用，从而使mHBTA 1介导的T细胞活化抑制失活;我们将进一步确定TS/A外泌体mHBTA 1与T细胞因子相互作用的途径。(4)确定年龄相关的T细胞因子是否调节人乳腺肿瘤外泌体HBTA 1的泛素化，并进一步确定其与年龄相关的乳腺癌发病率的可能关联。产生的数据应该阐明肿瘤外泌体和外泌体mHBTA 1在年龄和T细胞依赖性乳腺癌发展中作用的细胞和分子基础。