Ethnicity-determined T cell responses and GARP/TGFbeta1 signaling in prostate cancer

前列腺癌中种族决定的 T 细胞反应和 GARP/TGFbeta1 信号传导

• 批准号: 10358338
• 负责人: Anshu Agrawal
• 金额: $ 37.06万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-09 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10358338
• 关键词: Accounting; Affect; African American; Age; American; Anticoagulants; Aspirin; Atrial Fibrillation; Autologous; Autologous Tumor Cell; Benign; Biological Factors; CD8-Positive T-Lymphocytes; Cancer Patient; Cancer Vaccines; Carcinoma; Cells; Clinical Trials; Coculture Techniques; Complex; Cryopreservation; Data; Development; Dissection; Distant; Docking; ERG gene; Enzyme-Linked Immunosorbent Assay; Ethnic Origin; Ethnic group; European; Fibroblasts; Flow Cytometry; Formalin; Gene Fusion; Genetic; Gleason Grade for Prostate Cancer; Goals; Histology; Human; Immune; Immune Evasion; Immune Tolerance; Immune checkpoint inhibitor; Immune response; Immunobiology; Immunohistochemistry; Immunologics; Immunosuppression; Immunotherapy; Incidence; Infiltration; Integrins; Interferons; LRRC32 gene; Lead; Malignant Neoplasms; Malignant neoplasm of prostate; Measures; Mediating; Modeling; Mutation; Organoids; PTEN gene; Paraffin Embedding; Patients; Peripheral Blood Lymphocyte; Persons; Plasma; Play; Prostatic Neoplasms; Provenge; Race; Regulatory T-Lymphocyte; Reporting; Resistance; Resources; Role; Safety; Signal Transduction; Socioeconomic Factors; Stains; Stroke prevention; T cell response; T-Lymphocyte; Testing; Tissue Embedding; Tissues; Transforming Growth Factor beta; Transgenic Organisms; Tumor Escape; Tumor-Derived; Tumor-infiltrating immune cells; Variant; anti-CTLA4; anti-PD-1; cancer health disparity; castration resistant prostate cancer; checkpoint therapy; clinically relevant; cytotoxic CD8 T cells; effector T cell; health disparity; improved outcome; individual patient; men; mortality; mouse model; neoplastic cell; novel strategies; overexpression; prostate cancer progression; racial and ethnic; receptor; response; side effect; tool; tumor; tumor growth; tumor microenvironment; tumor xenograft

Project Summary Prostate cancer (PCa) incidence and mortality rates are the highest in African American (AA) men compared to any other racial/ethnic population. These differences persist even after accounting for socioeconomic factors, suggesting genetics and unknown biological factors may contribute to PCa health disparities. However, common genetic alterations, such as TMPRRSS2-ERG gene fusions and PTEN loss, were found to occur much less frequently in AA PCa than in European American (EA) PCa. Instead, prominent differences in tumor immunobiology between AA vs. EA men were reported in several studies, including a clinical trial with a cancer vaccine, Sipuleucel-T, which AA men had a median nine-month of overall survival advantage over EA men. To mechanistically dissect the immunological and/biological factors that determine tumor cell sensitivity and resistance to immunotherapy of different races, we have developed primary cultures of AA and EA PCa patient-derived tumor organoids, normal organoids, carcinoma associated fibroblasts (CAFs) and benign- associated fibroblasts (BAFs) from many patients and cryopreserved their peripheral blood lymphocytes (PBLs) from the same patient over past years. Our preliminary data show that AA CAFs secrete increased levels of active TGF- in the culture medium than EA CAFs. In addition, we are the first to show that Glycoprotein A repetitions predominant (GARP), the docking receptor for the release of active TGF-β, is over expressed in the adjacent stroma of AA PCa compared to adjacent stroma of EA PCa and AA PCa tissues. Interestingly, the adjacent stroma of AA PCa has increased infiltration of cytotoxic CD8+T cells compared to the adjacent stroma of EA PCa and to the distant stroma of AA PCa, suggesting that immune response is higher in AA stroma but may not be effective due to the increased TGF-β1 and GARP. Our preliminary data of co-culture studies with CAFs and T cells lends further support to the scenario as we observed increased TGF-β1 and reduced IFN-  in these co-cultures. These results suggest that although AA PCa patients may be more responsive to immunotherapies, GARP/TGFβ signaling represents a vulnerable point in AA PCa and may be used as a target for developing more effective immunotherapies. Therefore, we hypothesize that the interaction between tumor and stroma in AA and EA PCa differentially affect the tumor reactivity of T cells and that GARP/TGF-β signaling contribute to the differences in the T cell tumor reactivity among patients. To test the hypothesis, first we will determine whether T cells from AA and EA PCa patients display differences in tumor reactivity in co-cultures with autologous tumor organoids and/or CAFs. Second, our preliminary data have shown that Dabigatran etexilate, an anticoagulant drug for preventing stroke in people with atrial fibrillation, effectively blocks GARP/TGF-1 signaling and that its combination with anti-CTLA4 results in a durable regression of Myc-CaP xenograft tumors. We therefore will determine whether Dabigatran can enhance the anti-PD1/anti-CTLA4’s anti-PCa efficacy in the HiMyc and TRAMP transgenic PCa mouse models. Third, to further enhance the clinical relevance of our study, we will utilize available formalin fixed paraffin embedded tissue blocks from 141 AA and 141 EA matched PCa cases by age and Gleason score and determine the relationship between GARP/TGF-β signaling and various infiltrating immune cells in tumor and stroma of AA and EA PCa. Impact: This proposal capitalizes on the development of unique organoids and cell resources, which allow dissecting factors and mechanisms (i.e. GARP/GARP/TGF-β) signaling leading to immune differences between AA and EA PCa. In addition, a novel strategy for enhancing immune checkpoint therapies in PCa in general and AA PCa in particular may be developed by repurposing Dabigatran etexilate through targeting GARP/TGF-β signaling that is highly activated in the tumor microenvironment of AA PCa, as suggested by our preliminary data. Dabigatran etexilate has shown similar safety profiles or side effects as Aspirin in humans.

项目摘要 前列腺癌(PCA)的发病率和死亡率在非裔美国人(AA)男性中最高 与任何其他种族/民族人口相比。即使在考虑到以下因素之后，这些差异仍然存在 社会经济因素，表明遗传和未知的生物因素可能有助于前列腺癌的健康 差距。然而，常见的遗传改变，如TMPRRSS2-ERG基因融合和PTEN丢失， 在AA-PCa中发生的频率比在欧美(EA)PCa中少得多。相反， 在几项研究中，报告了AA和EA男性在肿瘤免疫生物学方面的显著差异， 包括一项癌症疫苗SiPuleucel-T的临床试验，AA患者的总体寿命中位数为9个月 比艺人更有生存优势。 从机械上剖析决定肿瘤细胞敏感性的免疫和/生物因素 和不同种族对免疫治疗的抵抗力，我们建立了AA和EA PCA的原代培养体系 患者来源的肿瘤器官、正常器官、肿瘤相关成纤维细胞(CAF)和良性- 许多患者的相关成纤维细胞(BAFs)及其外周血淋巴细胞的冷冻保存 (PBL)来自同一患者在过去几年。 我们的初步数据显示，AA CAF在培养中分泌活性的转化生长因子-水平增加。 比EA CAF中等。此外，我们还首次发现糖蛋白A重复序列占主导地位 GARP是释放活性转化生长因子-β的对接受体，在邻近间质中过度表达 AA-PCa与EA-PCa和AA-PCa组织的邻近间质比较。有趣的是，邻近的基质 与相邻间质相比，AA-PCa的细胞毒性CD8+T细胞的浸润增加。 AA-PCa的远处间质，提示AA间质的免疫反应较高，但可能不是。 这是由于转化生长因子-β-1和GARP升高所致。我们与CAF共培养研究的初步数据 而T细胞进一步支持了这一假设，因为我们观察到转化生长因子-β-1增加，干扰素-1减少。 在这些共培养中。 这些结果表明，尽管再生障碍性前列腺癌患者可能对免疫治疗更敏感， GARP/转化生长因子β信号转导通路是AA-PCa的一个薄弱环节，可作为研究的靶点 更有效的免疫疗法。因此，我们假设肿瘤和间质之间的相互作用 在AA和EA中，PCA不同地影响T细胞的肿瘤活性以及GARP/转化生长因子-β信号转导 导致患者之间T细胞肿瘤反应性的差异。 为了验证这一假设，我们首先将确定AA和EA PCA患者的T细胞是否显示 与自体肿瘤器官和/或CAF共培养时肿瘤反应性的差异。第二，我们的 初步数据显示，达比加特兰依昔拉特是一种预防中风的抗凝药 在房颤时，有效阻断GARP/转化生长因子-CTLA1信号转导及其与抗CTLA4的结合 结果：Myc-Cap异种移植瘤持久消退。因此我们将决定是否 达比加兰增强hMyc和TRAMP转基因细胞中抗PD1/抗CTLA4‘S抗PCa的作用 建立PCA小鼠模型。第三，为了进一步加强我们研究的临床相关性，我们将利用现有的 来自141例AA和141例EA的福尔马林固定石蜡包埋组织块与年龄和 Gleason评分，确定GARP/转化生长因子-β信号转导通路与各种浸润性免疫的关系 AA和EA Pca的肿瘤和间质中的细胞。 影响：这项提议利用了独特的有机化合物和细胞资源的开发，这 允许剖析导致免疫差异的因素和机制(即GARP/GARP/转化生长因子-β) 在AA和EA PCA之间。此外，在前列腺癌中加强免疫检查点治疗的新策略 一般的和特别的AA PCA可以通过通过靶向重新调整达比卡特兰的用途来开发 GARP/转化生长因子-β信号在AA-PCa的肿瘤微环境中高度激活，如 我们的初步数据。Dabigatran etexilate已显示出与阿司匹林类似的安全性或副作用 在人类身上。