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.

概括激素受体（HR）+乳腺癌（BC）在美国导致大多数与BC相关的死亡，这反映了未满足创新治疗方法的需求。实际上，对标准治疗和转移性的抵抗力传播仍然是主要的挑战，并显示了诸如免疫检查点阻滞剂（ICB）之类的新颖方法到目前为止的有效性有限。该提案的长期目标是机械剖析BCl2的影响，一种抗凋亡蛋白，有利于化学疗法和放射疗法（RT），关于幼稚和辐照的HR+ BC的免疫构型。具体而言，该项目将测试BCl2可能代表治疗的高度创新假设可操作的免疫检查点，因为它具有维护线粒体完整性的能力具体目的：1）确定Bcl2水平对治疗的免疫环境的影响人和小鼠HR+ BC； 2）定义BCL2对HR+ BCS免疫微环境的影响在体外和体内响应RT； 3）阐明Bcl2的值作为提升的目标 RT在HR+ BC的小鼠模型中的免疫刺激作用，包括模仿钥匙的创新模型人HR+ BC的特征。 BCl2是一个特别重要的目标，因为大约80％的HR+ BC病例过表达BCL2和BCL2抑制剂Venetoclax批准用于临床使用。为了实现我们的目标来自HR+ BC女性的诊断活检将由codex评估Bcl2表达，肿瘤浸润通过调节抗癌免疫力的关键免疫电池，并表达免疫抑制蛋白（如MHC） 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在体外，通过遗传（缺失，过表达）和药物（例如，维内托克斯给药）方法如果显微镜和ELISA进行评估抗癌免疫的关键调节剂，则与流式细胞仪结合。将利用类似的遗传和药物方法来改变Bcl2能力并描述 BCl2对在治疗中的免疫TME的影响，未经治疗和受照射的小鼠HR+ BCS建立基于IHC，流式细胞术和单细胞RNA测序的免疫能力宿主。流量研究基于小鼠HR+ BC细胞在生长的小鼠HR+ BC细胞的体内研究以及体内研究免疫缺陷与免疫能力小鼠将采用BCl2对固有的影响放射敏性与免疫依赖性肿瘤控制。最后，涉及RT和的不同组合方案 Venetoclax将进行研究以提高效率（在没有治疗和治疗性环境中）和在HR+ BC的内源性小鼠模型中，抗性的潜在机制模仿了关键特征人HR+ BC。我们的发现将阐明BCL2对HR+ BC的免疫TME的影响并确定临床试验测试RT和HR+女性的临床试验测试和Venetoclax的最佳方法卑诗省是一种毁灭性疾病，在美国每年仍会影响> 200,000，杀死> 25,000名新女性。