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

• 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
• 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

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.

摘要 激素受体(HR)+乳腺癌(BC)是美国大多数BC相关死亡的原因，反映了 对创新治疗方法的需求尚未得到满足。事实上，对标准治疗和转移性疾病的耐药性 传播仍然是主要的挑战，免疫检查点阻滞剂(ICBS)等新方法已经表明 到目前为止，疗效有限。这项提案的长期目标是机械地剖析BCL2的影响， 一种抗凋亡蛋白，它有助于癌细胞抵抗化疗和药物诱导的死亡 放射治疗(RT)，对治疗单纯和照射后的HR+BCS的免疫学形态的影响。 具体地说，这个项目将测试高度创新的假设，即bcl2可能代表着一种治疗 可操作的免疫检查点，因为它有能力保持线粒体的完整性，基于这些 具体目标：1)测定bcl2水平对治疗免疫微环境的影响--幼稚 2)确定bcl2对人和小鼠HR+BCS免疫微环境的影响 在体外和体内对RT的反应；3)阐明bcl2作为促进肿瘤生长的靶点的价值。 RT对HR+BCS小鼠模型的免疫刺激作用，包括一种模仿KEY的创新模型 人类HR+BC的特点。BCL2是一个特别重要的目标，因为大约80%的HR+BC病例 过度表达bcl2和bcl2抑制剂ventoclax已被批准用于临床。为了实现我们的目标， 对HR+BC患者的诊断活检将通过CODEX评估bcl2的表达、肿瘤的侵袭 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级和PD-L1级。Bcl2对HR+BC细胞对RT免疫应答的影响将被问及 在体外，通过遗传(缺失、过表达)和药理(例如，万乃馨给药)方法 再结合流式细胞仪、IF显微镜和ELISA法对抗癌免疫的关键调节因子进行评估。 类似的遗传和药理学方法将被用来改变bcl2的能力并描绘出 BCL2对未处理和照射的小鼠HR+BCS免疫TME的影响 免疫活性宿主，基于免疫组化、流式细胞术和单细胞RNA测序。用Flow进行体外研究 细胞学和克隆分析，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活性小鼠，将被用来解剖bcl2对内源性的影响。 放射敏感性与免疫依赖性肿瘤控制。最后，不同的联合方案涉及RT和 将研究万乃馨的疗效(在治疗幼稚和治疗抵抗的情况下)和 内源性HR+BCS小鼠模型中潜在的耐药机制 人HR+BC。我们的发现将阐明bcl2对HR+BC免疫TME的影响并确定 启动RT加万乃馨对HR+妇女进行临床试验的最佳途径 不列颠哥伦比亚省是一种毁灭性的疾病，在美国每年仍有20万人感染并导致2.5万名新女性死亡。