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: 10578810
• 负责人: Daniel Abate-Daga
• 金额: $ 46.29万
• 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/10578810
• 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; CAR T cell therapy; CD28 gene; CD3 Antigens; CD8B1 gene; CTLA4 gene; CWR22Rv1; Cancer Patient; Castration; Cell Differentiation process; Cell Line; Cells; Clinical; Clinical Trials; Cytokeratin; Cytokine Receptors; Data; Deterioration; Disease; Drug usage; Effectiveness; Engineering; Flow Cytometry; Frustration; Generations; Genetic; Goals; Growth; Histologic; Homing; Human; Immune; Immunocompetent; Immunotherapy; In Vitro; Infiltration; Luciferases; Malignant Bone Neoplasm; Malignant Neoplasms; Malignant neoplasm of prostate; Maps; Membrane; Metastatic Prostate Cancer; Modeling; Molecular; Monitor; Morbidity - disease rate; Mus; Myeloid Cells; Osteoblasts; Osteoclasts; Osteogenesis; Pain; Patients; Performance; 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; bisphosphonate; bone; cancer cell; carcinogenesis; 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; pharmacologic; 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.

虽然免疫疗法在治疗其他癌症方面取得了长足的进步，但castrate抗性前列腺癌 （CRPC）在很大程度上没有反应症，强调了对新方法的需求。一种这样的方法依赖 关于自体T细胞进行基因修饰以表达嵌合抗原受体（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）在 前列腺癌，我们以前已经开发了一种靶向具有体内效率的PSCA的汽车。到 靶骨转移性CRPC（MCRPC），我们建议操纵特定的T细胞子集，称为γδ 可以通过双膦酸盐（例如唑来膦酸盐（ZOL））将其驱动到骨骼 临床用来限制骨MCRPC男性癌症引起的骨病。重要的是，ZOL处理 诱导肿瘤细胞中磷酸剂的积累，这些肿瘤细胞被γδT细胞检测到。我们的初步 调查结果表明，ZOL可以增强γδT细胞的归巢，在那里它们可以通过 CAR和通过内源性T细胞受体（TCR）识别。在存在或不存在的情况下，γδCAR-T处理 ZOL可以减轻癌症引起的骨骼恶化。此外，我们发现由 骨髓衍生的间充质基质细胞（MSC）可以增加γδCART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 FUCTIONαβ和γδT细胞差异化，需要设计针对γδT细胞优化的汽车。基于这些 初步发现，我们假设γδCAR-T细胞的归巢和细胞毒性活性 通过遗传，药物和 微环境方法。我们将通过测试我们的假设； 1）定义最佳γδCAR-T设计 这将显着增强CRPC细胞毒性。我们将测试选择替代方案的生物学含义 汽车跨膜和共刺激域，特别关注其调节能力 细胞因子受体的表达。我们还将剖析可以控制γδcar-的特定信号通路 T细胞持久性。最后，我们将确定由不同的汽车触发的分子信号通路 共刺激域。 2）确定ZOL是否可以驱动γδCAR-T募集和抗骨MCRPC活性 体内。我们将使用骨MCRPC的Xenograponic和PDX模型来表征生物利用度和治疗 γδCAR-T + ZOL的功效；以及用于绘制磷脂积累部位的免疫能力模型。 3）剖析γδCAR-T对骨MCRPC微环境的相互影响。我们将分析 γδCAR-T + ZOL处理对骨骼结构和生理的影响以及MSC对 γδCAR-T细胞在体内的性能。根据预期的结果，表征特定的结果 骨/肿瘤微环境的特性将揭示新的见解，从而提供强大的理由 用于量身定制的免疫疗法，以消除当前无法治愈的骨MCRPC。