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.

总结 在美国，激素受体（HR）+乳腺癌（BC）导致大多数BC相关死亡，反映了 对创新治疗方法的需求未得到满足。事实上，对标准治疗的抗性和转移性 传播仍然是主要挑战，诸如免疫检查点阻断剂（ICB）等新方法已经显示 到目前为止，疗效有限。该提案的长期目标是机械地剖析BCL 2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），对未经治疗和经辐照的HR+ BC的免疫学构型。 具体来说，该项目将测试高度创新的假设，即BCL 2可能代表治疗上的 可操作的免疫检查点，因为它能够保持线粒体的完整性，基于这些 具体目的：1）确定BCL 2水平对未经治疗的免疫微环境的影响， 人和小鼠HR+ BC; 2）确定BCL 2对HR+ BC的免疫微环境的影响 在体外和体内响应RT;和3）阐明BCL 2作为靶点的价值，以增强免疫应答。 RT在HR+ BC小鼠模型中的免疫刺激作用，包括一种模拟关键免疫刺激的创新模型。 人类HR+ BC的特征。BCL 2是一个特别重要的靶点，因为约80%的HR+ BC病例 过表达BCL 2和BCL 2抑制剂venetoclax被批准用于临床。为了实现我们的目标， 来自HR+ BC女性的诊断性活检将通过CODEX评估BCL 2表达、肿瘤浸润 调节抗癌免疫的关键免疫细胞，以及免疫抑制蛋白如MHC的表达， I类和PD-L1。将研究BCL 2对HR+ BC细胞对RT的免疫应答的影响 在体外，通过遗传（缺失，过表达）和药理学（例如，venetoclax给药）方法 结合流式细胞术、IF显微术和ELISA，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利用来改变BCL 2的能力，并描绘出BCL 2的表达。 BCL 2对在小鼠中建立的未经处理和经辐射的小鼠HR+ BC的免疫TME的影响 免疫活性宿主，基于IHC，流式细胞术和单细胞RNA测序。按流量进行的体外研究 细胞计数和克隆形成测定，以及基于在小鼠中生长的HR+ BC细胞的体内研究。 免疫缺陷小鼠与免疫活性小鼠，将用于剖析BCL 2对内源性免疫缺陷的影响。 放射敏感性与免疫依赖性肿瘤对照。最后，不同的组合方案涉及RT和 将研究维奈托克的疗效（在初治和耐药情况下）， 在HR+ BC的内源性小鼠模型中， 人类HR+ BC。我们的研究结果将阐明BCL 2对HR+ BC的免疫TME的影响，并确定 为在HR+女性中启动RT+维奈托克临床试验提供信息的最佳方法 BC是一种毁灭性的疾病，在美国每年仍有超过20万人受到影响，并有超过25,000名新妇女死亡。