Engineering T cells to overcome inhibitory receptor signals that limit the efficacy of adoptive cell therapy against ovarian cancer

改造 T 细胞以克服抑制性受体信号，这些信号限制了过继性细胞疗法对卵巢癌的疗效

• 批准号: 10526155
• 负责人: Kristin Gail Anderson
• 金额: $ 19.22万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-05 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10526155
• 关键词: Address; Adoptive Cell Transfers; Adoptive Transfer; Affinity; Antibodies; Antigen Targeting; Antigens; Autoantigens; Autologous; Blocking Antibodies; CD8-Positive T-Lymphocytes; Cancer cell line; Cell Culture Techniques; Cell Line; Cell physiology; Cells; Clinical; Cytotoxic T-Lymphocytes; Data; Diagnosis; Engineering; Flow Cytometry; Functional disorder; Gene Modified; Genes; Goals; Human; Immune; Immune response; Immunocompetent; Immunohistochemistry; Immunosuppression; Immunotherapy; In Situ; Infiltration; Knowledge; Ligands; Ligation; Malignant Neoplasms; Malignant neoplasm of ovary; Mediating; Methods; Modeling; Molecular Profiling; Mus; Myeloid Cells; Outcome; Pathway interactions; Patient-Focused Outcomes; Phenotype; Pre-Clinical Model; Production; Proteins; Publishing; Receptor Signaling; Research; Resistance; Sampling; Signal Pathway; Signal Transduction; Slice; Solid Neoplasm; System; T cell response; T cell therapy; T-Cell Receptor; T-Lymphocyte; Testing; Therapeutic; Tissues; Toxic effect; Tumor Debulking; Tumor Escape; Tumor-infiltrating immune cells; United States; Woman; Work; anti-PD-1; anti-cancer; cancer cell; cancer infiltrating T cells; cancer type; chemotherapy; chimeric antigen receptor; clinical development; clinically relevant; cytokine; cytotoxic; effective therapy; engineered T cells; immune checkpoint; immune checkpoint blockade; improved; in vivo; insight; knock-down; mesothelin; mouse model; novel; novel strategies; ovarian neoplasm; overexpression; pre-clinical; programmed cell death protein 1; receptor; receptor expression; response; side effect; systemic toxicity; transcriptomics; tumor; tumor growth; tumor microenvironment

PROJECT SUMMARY Over 20,000 women are diagnosed with ovarian cancer in the United States annually, and over half will die within five years. Outcomes have changed little in the last 20 years, highlighting the need for more effective therapies. One promising new strategy employs immune T cells engineered to target proteins uniquely overexpressed in tumors; such T cell immunotherapies have the potential to control tumor growth without toxicity to healthy tissues. My ongoing work targets the antigen mesothelin (Msln), which contributes to malignancy and invasive progression in ovarian cancer but has limited expression in healthy cells. I showed that T cells engineered to express a human or mouse Msln-specific high-affinity T cell receptor (TCRMsln) can kill human ovarian cancer cell lines or the murine ID8 cell line, respectively. In a disseminated ID8 tumor model, adoptively transferred TCRMsln T cells preferentially accumulated within established tumors, delayed ovarian tumor growth and significantly prolonged mouse survival. However, data also revealed that the ovarian tumor microenvironment (TME) limits engineered T cell persistence and anti-cancer efficacy. Tumor-specific T cells express inhibitory receptors upon encountering antigen, reducing antitumor cytokine production. I detected the ligands for the PD-1, Tim-3 and Lag-3 inhibitory receptors (immune checkpoints) expressed in human and ID8 ovarian tumors. Moreover, tumor-infiltrating TCRMsln T cells expressed PD-1, Tim- 3 and Lag-3, which correlated with reduced cytokine production. I hypothesized that immune checkpoint blockade could overcome inhibitory receptor ligation-driven suppression of engineered T cells. Therefore, I treated tumor-bearing mice with TCRMsln T cells plus anti-PD-1, anti-Tim-3 and/or anti-Lag-3 checkpoint- blocking antibodies, targeting up to three inhibitory receptors simultaneously. Triple checkpoint blockade dramatically increased anti-tumor cytokine production by intratumoral TCRMsln T cells, but treatment also produced greater off-tumor toxicities. I now propose to use preclinical mouse models (Aim 1) and a novel human slice culture system (Aim 2) to interrogate the transcriptomic and functional changes that occur in engineered T cells and the ovarian TME after checkpoint blockade. I plan to use T cell engineering to knock down endogenous inhibitory receptor expression in tumor-specific T cells and determine if anti-tumor function is improved without the immune- related toxicities observed with systemic combination checkpoint blockade. Many solid tumors overexpress Msln and share the same T cell inhibitory pathways. Therefore, the findings from these new studies will likely inform the development of clinically-relevant T cell engineering strategies that are more resistant to immune suppression within the solid TME of many malignancies.

项目摘要 每年在美国被诊断出20,000多名妇女被诊断出患有卵巢癌，一半以上会死亡 五年之内。在过去的20年中，结果发生了很少的变化，强调了需要更有效的需求 疗法。一种有前途的新策略采用工程设计的免疫T细胞，以唯一靶向蛋白质 过表达肿瘤；这种T细胞免疫疗法有可能控制肿瘤生长而没有 对健康组织的毒性。我正在进行的工作针对的是抗原间皮素（MSLN），这有助于 卵巢癌的恶性肿瘤和侵入性进展，但在健康细胞中的表达有限。我展示了 T细胞设计为表达人或小鼠MSLN特异性高亲和力T细胞受体（TCRMSLN）可以 分别杀死人类卵巢癌细胞系或鼠ID8细胞系。在传播的ID8肿瘤中 模型，采用转移的TCRMSLN T细胞优先积聚在已建立的肿瘤中，延迟 卵巢肿瘤的生长并显着延长小鼠的生存率。但是，数据还显示卵巢 肿瘤微环境（TME）限制了设计的T细胞持久性和抗癌功效。 肿瘤特异性T细胞在遇到抗原时表达抑制性受体，减少抗肿瘤细胞因子 生产。我检测到PD-1，TIM-3和LAG-3抑制受体的配体（免疫检查点） 用人和ID8卵巢肿瘤表达。此外，肿瘤浸润的TCRMSLN T细胞表达PD-1，Tim- 3和LAG-3，与细胞因子产生减少有关。我假设免疫检查点 封锁可以克服抑制性受体连接驱动的工程T细胞的抑制。因此，我 用TCRMSLN T细胞治疗了肿瘤的小鼠，加上抗PD-1，抗TIM-3和/或抗LAG-3检查点 - 阻断抗体，同时靶向三个抑制受体。三重检查点封锁 肿瘤内TCRMSLN T细胞产生抗肿瘤细胞因子的抗肿瘤细胞因子，但也治疗 产生更大的肿瘤毒性。 我现在建议使用临床前小鼠模型（AIM 1）和一种新型的人类切片培养系统（AIM 2） 询问工程T细胞和卵巢TME中发生的转录组和功能变化 检查点封锁后。我计划使用T细胞工程来击倒内源性抑制受体 在肿瘤特异性T细胞中的表达，并确定抗肿瘤功能是否得到改善，而没有免疫 使用全身组合检查点阻滞观察到的相关毒性。许多实体瘤过表达 MSLN并共享相同的T细胞抑制途径。因此，这些新研究的发现可能会 告知与免疫力更具耐药性的临床临床T细胞工程策略的发展 在许多恶性肿瘤的固体TME中抑制。