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Targeting the BCL2 immune checkpoint to enhance the immunostimulatory effects of radiation in breast cancer

靶向 BCL2 免疫检查点增强放射对乳腺癌的免疫刺激作用

基本信息

批准号：
10734237
负责人：
Lorenzo Galluzzi
金额：
$ 70.31万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2028-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10734237
关键词：
Affect Apoptotic Attention BCL2 gene Biological Assay Biopsy Breast Cancer Cell Breast Cancer Patient CD8-Positive T-Lymphocytes Cancer Etiology Cell Membrane Permeability Cells Cessation of life Chemotherapy and/or radiation Chronic Lymphocytic Leukemia Clinical Clinical Management Clinical Trials Clinical Trials Design Competence Coupled Cytosol Data Diagnosis Diagnostic Disease Disease model Dose Enzyme-Linked Immunosorbent Assay Exhibits FDA approved FOXP3 gene Flow Cytometry Genetic Goals Hormone Receptor Human Immune Immune Targeting Immune response Immunocompetent Immunologic Deficiency Syndromes Immunologics Immunotherapy In Vitro Infiltration Interferon Type I Knock-out Link MHC Class I Genes Methods Microscopy Mitochondria Mitochondrial DNA Modeling Mus Patients Process Proteins Radiation Tolerance Radiation therapy Regimen Regulation Regulatory T-Lymphocyte Resistance Role Signal Transduction Testing Therapeutic Translating Translations Treatment Efficacy Tumor Immunity Woman XCR1 gene anti-cancer cancer cell cancer survival clinically relevant combinatorial ds-DNA human disease immune cell infiltrate immune checkpoint immune checkpoint blockers immunogenicity in vivo inhibitor innovation malignant breast neoplasm mitochondrial membrane mouse model novel strategies overexpression pharmacologic pre-clinical preservation prevent programmed cell death ligand 1 programmed cell death protein 1 radiation effect radiation response research clinical testing resistance mechanism single-cell RNA sequencing standard care therapy resistant tool transcriptomics tumor tumor microenvironment tumor-immune system interactions virtual

项目摘要

SUMMARY Hormone receptor (HR)+ breast cancer (BC) causes the majority of BC-related deaths in the US, reflecting an unmet need for innovative therapeutic approaches. Indeed, resistance to standard treatments and metastatic spread remain major challenges, and novel approaches such as immune checkpoint blockers (ICBs) have shown limited efficacy so far. The long-term objective of this proposal is to mechanistically dissect the impact of BCL2, an antiapoptotic protein that favors the resistance of cancer cells to death imposed by chemotherapy and radiation therapy (RT), on the immunological configuration of treatment naïve and irradiated HR+ BCs. Specifically, this project will test the highly innovative hypothesis that BCL2 may represent a therapeutically actionable immune checkpoint because of its ability to preserve mitochondrial integrity, based on these specific aims: 1) determining the influence of BCL2 levels on the immune microenvironment of treatment-naïve human and mouse HR+ BCs; 2) defining the impact of BCL2 on the immune microenvironment of HR+ BCs responding to RT in vitro and in vivo; and 3) elucidating the value of BCL2 as a target to boost the immunostimulatory effects of RT in mouse models of HR+ BCs, including an innovative model that mimics key features of human HR+ BC. BCL2 is a particularly significant target because ~80% of HR+ BC cases overexpress BCL2, and the BCL2 inhibitor venetoclax, is approved for clinical use. To achieve our goals, diagnostic biopsies from women with HR+ BC will be evaluated by CODEX for BCL2 expression, tumor infiltration by key immune cells that regulate anticancer immunity, and expression of immunosuppressive proteins like MHC Class I and PD-L1. The impact of BCL2 on the immunological response of HR+ BC cells to RT will be interrogated in vitro, by genetic (deletion, overexpression) and pharmacological (e.g., venetoclax administration) methods coupled to flow cytometry, IF microscopy and ELISA for the assessment of key regulators of anticancer immunity. Similar genetic and pharmacological approaches will be harnessed to alter BCL2 competence and delineate the influence of BCL2 on the immunological TME of treatment-naïve and irradiated mouse HR+ BCs established in immunocompetent hosts, based on IHC, flow cytometry and single-cell RNA sequencing. In vitro studies by flow cytometry and clonogenic assays, as well as in vivo studies based on mouse HR+ BC cells growing in immunodeficient vs immunocompetent mice, will be employed to dissect the impact of BCL2 on intrinsic radiosensitivity vs immune-dependent tumor control. Finally, different combinatorial regimens involving RT and venetoclax will be investigated for efficacy (in both treatment-naïve and treatment resistance settings) and potential mechanisms of resistance in an endogenous mouse model of HR+ BCs that mimics key features of human HR+ BC. Our findings will elucidate the impact of BCL2 on the immune TME of HR+ BC and identify the best approach to inform the initiation of clinical trials testing RT plus venetoclax in women with HR+ BC, a devastating disease that still affects >200,000 and kills >25,000 new women every year in the US.

概括在美国，激素受体 (HR)+ 乳腺癌 (BC) 导致了大多数与 BC 相关的死亡，这反映了对创新治疗方法的需求未得到满足。事实上，对标准治疗和转移的耐药性传播仍然是主要挑战，免疫检查点阻断剂 (ICB) 等新方法已表明迄今为止疗效有限。该提案的长期目标是机械地剖析 BCL2 的影响，一种抗凋亡蛋白，有利于癌细胞抵抗化疗造成的死亡，放射治疗（RT），对未接受治疗和接受辐射的 HR+ BC 的免疫结构的影响。具体来说，该项目将测试高度创新的假设，即 BCL2 可能代表一种治疗方法。可操作的免疫检查点，因为它能够保持线粒体完整性，基于这些具体目标：1) 确定 BCL2 水平对未接受治疗的免疫微环境的影响人类和小鼠 HR+ BC； 2) 定义BCL2对HR+ BC免疫微环境的影响体外和体内对 RT 的反应； 3) 阐明 BCL2 作为提升目标的价值 RT 在 HR+ BC 小鼠模型中的免疫刺激作用，包括模仿关键的创新模型人类 HR+ BC 的特征。 BCL2 是一个特别重要的目标，因为约 80% 的 HR+ BC 病例过表达 BCL2 和 BCL2 抑制剂 Venetoclax 已被批准用于临床。为了实现我们的目标， HR+ BC 女性的诊断性活检将由 CODEX 评估 BCL2 表达、肿瘤浸润通过调节抗癌免疫的关键免疫细胞以及 MHC 等免疫抑制蛋白的表达 I 类和 PD-L1。 BCL2 对 HR+ BC 细胞对 RT 的免疫反应的影响将受到质疑体外，通过遗传（缺失、过度表达）和药理学（例如维奈托克给药）方法与流式细胞术、IF 显微镜和 ELISA 结合用于评估抗癌免疫的关键调节因子。类似的遗传和药理学方法将被用来改变 BCL2 能力并描述 BCL2 对未接受治疗和接受辐射的小鼠 HR+ BC 免疫学 TME 的影响基于 IHC、流式细胞术和单细胞 RNA 测序的免疫活性宿主。体外流式研究细胞计数和克隆形成测定，以及基于小鼠 HR+ BC 细胞生长的体内研究免疫缺陷小鼠与免疫功能正常小鼠的比较，将用于剖析 BCL2 对内在的影响放射敏感性与免疫依赖性肿瘤控制。最后，涉及 RT 和将研究维奈托克的疗效（在未接受治疗和治疗耐药的情况下）和 HR+ BC 的内源性小鼠模型中的潜在耐药机制，模拟了人类 HR+ BC。我们的研究结果将阐明 BCL2 对 HR+ BC 免疫 TME 的影响，并确定通知启动针对 HR+ 女性进行 RT 加维奈托克临床试验测试的最佳方法 BC 是一种毁灭性的疾病，在美国每年仍影响超过 200,000 人，并导致超过 25,000 名新女性死亡。