Understanding the influence of bone-metastatic prostate cancer and mesenchymal stromal cells on γδ T cells, in the bone microenvironment.

了解骨微环境中骨转移性前列腺癌和间充质基质细胞对 γT 细胞的影响。

• 批准号: 10356098
• 负责人: Daniel Abate-Daga
• 金额: $ 48.91万
• 依托单位: H. LEE MOFFITT CANCER CTR & RES INST
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-09 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10356098
• 关键词: Acute; Adoptive Transfer; Affect; Anatomy; Antigen Targeting; Antigens; Apoptosis; Apoptotic; Attention; Autologous; Binding; Biological Availability; Biological Testing; Bone Diseases; Bone Marrow; Bone Matrix; Bone neoplasms; CAR T cell therapy; CD28 gene; CD3 Antigens; CD8B1 gene; CTLA4 gene; CWR22Rv1; Cell Differentiation process; Cell Line; Cells; Clinical; Clinical Trials; Cytokeratin; Cytokine Receptors; Data; Deterioration; Disease; Drug usage; Effectiveness; Engineering; Flow Cytometry; Generations; Genetic; Goals; Growth; Histologic; Homing; Human; Immune; Immunocompetent; Immunotherapy; In Vitro; Luciferases; Malignant Bone Neoplasm; Malignant Neoplasms; Malignant neoplasm of prostate; Maps; Metastatic Prostate Cancer; Modeling; Molecular; Monitor; Morbidity - disease rate; Mus; Myeloid Cells; Osteoblasts; Osteoclasts; Osteogenesis; Pain; Patients; Performance; Pharmacology; Phenotype; Physiology; Process; Production; Property; Randomized; Reagent; Regulatory T-Lymphocyte; Resolution; Roentgen Rays; Safety; Signal Pathway; Signal Transduction; Site; Skeleton; Solid; Stains; Structure; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Subsets; Techniques; Testing; Therapeutic Effect; Time; Tissues; Toxic effect; Translations; Transmembrane Domain; Treatment Efficacy; Tumor Antigens; Xenograft Model; Zoledronate; advanced prostate cancer; androgen deprivation therapy; anti-cancer; antitumor effect; base; bisphosphonate; bone; cancer cell; castration resistant prostate cancer; chimeric antigen receptor; chimeric antigen receptor T cells; cytotoxic; cytotoxicity; design; early phase clinical trial; enhancing factor; experimental study; genetically modified cells; immune checkpoint blockade; in vitro testing; in vivo; indexing; insight; isopentenyl pyrophosphate; luminescence; men; mesenchymal stromal cell; microSPECT; mortality; mouse model; neoplastic cell; novel; novel strategies; osteogenic; overexpression; patient derived xenograft model; phosphoproteomics; pre-clinical; prevent; programmed cell death ligand 1; prostate cancer cell; prostate stem cell antigen; recruit; response; subcutaneous; success; targeted cancer therapy; tumor; tumor growth; tumor microenvironment; tumor progression; γδ T cells

While immunotherapies have made strides in the treatment of other cancers, castrate resistant prostate cancer (CRPC) remains largely unresponsive, underscoring the need for novel approaches. One such approach relies on the administration of autologous T cells genetically modified to express a chimeric antigen receptor (CAR) that recognize specific tumor-associated antigens. Prostate Stem Cell Antigen (PSCA) is widely expressed in prostate cancer and we have previously developed a CAR targeting PSCA that has potent in vivo efficacy. To target bone metastatic CRPC (mCRPC), we propose to manipulate a specific subset of T cells, called γδ that can be driven to the skeleton via systemic treatment with bisphosphonates such as zoledronate (ZOL) that is clinically used to limit cancer-induced bone disease in men with bone mCRPC. Importantly, ZOL treatment induces accumulation of phosphoantigens in tumor cells, which are detected by γδ T cells. Our preliminary findings show that ZOL can enhance γδ T-cells' homing to bone where they can prevent cancer growth via CAR and via endogenous T-cell receptor (TCR) recognition. γδ CAR-T treatment, in presence or absence of ZOL, can mitigate cancer-induced bone deterioration. Moreover, we found that soluble factors secreted by bone marrow derived mesenchymal stromal cells (MSC) can increase the cytotoxic potential of γδ CART cells. Finally, we found that the choice of CAR structural and costimulatory moieties affects the phenotype and fuction αβ and γδ T cells differentially, requiring the design of CARs optimized for γδ T cells. Based on these preliminary findings we hypothesize that the homing and cytotoxic activity of γδ CAR-T cells for the treatment of bone metastatic CRPC can be greatly enhanced through genetic, pharmacological, and microenvironmental approaches. We will test our hypothesis by; 1) Defining the optimum γδ CAR-T design that will significantly enhance CRPC cytotoxicity. We will test the biological implications of choosing alternative CAR transmembrane and costimulatory domains, with a specific focus on their ability to modulate the expression of cytokine receptors. We will also dissect the specific signaling pathways that can govern γδ CAR- T cell persistence. Finally, we will identify the molecular signaling pathways triggered by CARs with different costimulatory domains. 2) Determining if ZOL can drive γδ CAR-T recruitment and anti-bone mCRPC activity in vivo. We will use xenograft and PDX models of bone mCRPC to characterize the bioavailability and therapeutic efficacy of γδ CAR-T + ZOL; and an immunocompetent model to map sites of phosphoantigen accumulation. 3) Dissecting the reciprocal effects of γδ CAR-T on the bone mCRPC microenvironment. We will analyze the impact of γδ CAR-T + ZOL treatment on the structure and physiology of the bone, and the effects of MSC on the performance of γδ CAR-T cells in vivo. Based on the anticipated results, characterizing the specific properties of the bone/tumor microenvironment will reveal novel insights thereby providing a strong rationale for the translation of immunotherapies tailored to eliminate currently incurable bone mCRPC.

虽然免疫疗法在其他癌症的治疗方面取得了长足进步，但去势抵抗性前列腺癌 （CRPC）在很大程度上仍然没有反应，这凸显了对新方法的需求。其中一种方法依赖于 关于使用经过基因改造表达嵌合抗原受体（CAR）的自体 T 细胞 识别特定的肿瘤相关抗原。前列腺干细胞抗原（PSCA）广泛表达于 前列腺癌，我们之前开发了一种针对 PSCA 的 CAR，具有强大的体内功效。到 针对骨转移性 CRPC (mCRPC)，我们建议操纵 T 细胞的特定子集，称为 γδ， 可以通过双膦酸盐（例如唑来膦酸盐（ZOL））的全身治疗将其驱动至骨骼 临床用于限制患有骨 mCRPC 的男性癌症诱发的骨疾病。重要的是，ZOL治疗 诱导肿瘤细胞中磷酸抗原的积累，由 γδ T 细胞检测到。我们的初步 研究结果表明，ZOL 可以增强 γδ T 细胞向骨骼的归巢，从而防止癌症生长 CAR 通过内源性 T 细胞受体 (TCR) 识别。 γδ CAR-T 治疗，无论是否存在 ZOL，可以减轻癌症引起的骨质恶化。此外，我们发现，可溶性因子 骨髓来源的间充质基质细胞（MSC）可以增加γδ CART细胞的细胞毒性潜力。 最后，我们发现CAR结构和共刺激部分的选择会影响表型和 αβ 和 γδ T 细胞的功能存在差异，需要设计针对 γδ T 细胞优化的 CAR。基于这些 根据初步结果，我们假设 γδ CAR-T 细胞的归巢和细胞毒活性 骨转移性 CRPC 的治疗可以通过遗传、药物和 微环境方法。我们将通过以下方式检验我们的假设： 1) 定义最佳γδ CAR-T设计 这将显着增强CRPC的细胞毒性。我们将测试选择替代方案的生物学意义 CAR 跨膜和共刺激域，特别关注它们调节 细胞因子受体的表达。我们还将剖析可以控制 γδ CAR-的特定信号通路 T细胞持久性。最后，我们将确定不同CARs触发的分子信号通路 共刺激域。 2) 确定 ZOL 是否可以驱动 γδ CAR-T 招募和抗骨 mCRPC 活性 体内。我们将使用骨 mCRPC 的异种移植和 PDX 模型来表征其生物利用度和治疗效果 γδ CAR-T + ZOL 的功效；以及绘制磷酸抗原积累位点的免疫活性模型。 3）剖析γδ CAR-T对骨mCRPC微环境的相互影响。我们将分析 γδ CAR-T + ZOL治疗对骨结构和生理的影响，以及MSC对骨的影响 γδ CAR-T细胞在体内的性能。根据预期结果，描述具体的 骨/肿瘤微环境的特性将揭示新的见解，从而提供强有力的理由 翻译专为消除目前无法治愈的骨 mCRPC 而设计的免疫疗法。