Modeling Immune Cell Recruitment and its Impact on Triple Negative Breast Cancer Recurrence in the Irradiated Microenvironment

模拟免疫细胞募集及其对辐射微环境中三阴性乳腺癌复发的影响

• 批准号: 10321214
• 负责人: Benjamin Christian Hacker
• 金额: $ 3.09万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10321214
• 关键词: 4T1; American; Biological Assay; Breast; Breast Cancer Cell; Breast Cancer Model; Breast Cancer Patient; Breast Cancer therapy; CD8-Positive T-Lymphocytes; Cancer Etiology; Cell Communication; Cells; Cessation of life; Chemotaxis; Chemotherapy and/or radiation; Coculture Techniques; Diagnosis; Face; Fatty acid glycerol esters; Flow Cytometry; Goals; Human; Immune; Immunocompetent; Immunocompromised Host; Immunological Models; Infiltration; Laboratories; Lead; Literature; MDA MB 231; Malignant Neoplasms; Mediating; Modeling; Monitor; Monoclonal Antibodies; Mus; Neoplasm Circulating Cells; Normal tissue morphology; Operative Surgical Procedures; Organoids; Patients; Phenotype; Play; Primary Neoplasm; Process; Prognosis; Radiation; Radiation induced damage; Radiation therapy; Recurrence; Research; Role; Site; Study models; Testing; Tissues; Training; Tumor Cell Invasion; Tumor Cell Migration; Tumor-infiltrating immune cells; Woman; Work; blocking factor; cancer recurrence; cell behavior; cytokine; effective therapy; experience; high risk; immunosuppressive macrophages; improved; in vitro Model; in vivo; interest; macrophage; malignant breast neoplasm; mammary; mammary epithelium; migration; mouse model; neoplastic cell; orthotopic breast cancer; pre-clinical; preclinical study; radiation response; recruit; small molecule inhibitor; triple-negative invasive breast carcinoma; tumor; tumor growth; tumor progression; wound healing

PROJECT SUMMARY Approximately half of triple negative breast cancer (TNBC) patients receive breast conserving therapy, which includes irradiating the primary tumor site. Although this therapy improves prognosis overall, patients continue to experience locoregional recurrences at high rates. The mechanisms controlling recurrence following therapy are poorly understood. Previous research suggests that normal tissue radiation damage may be correlated to recurrence. Pre-clinical studies have shown that tumor cells migrate into irradiated mammary tissue and that macrophages play a critical role in the tumor cell recruitment process. While blockade of macrophage infiltration was shown to eliminate tumor cell migration to irradiated sites, the impact of macrophage phenotype has not been investigated. Macrophages with a wound healing phenotype (M2) are known to promote tumor growth and progression, but whether M2 macrophages influence tumor cell recruitment and proliferation in irradiated tissues is unknown. The central hypothesis of this proposal is that radiation damage in the tissue microenvironment promotes a pre-metastatic niche through immunosuppressive cell infiltration. I will test this hypothesis using pre- clinical orthotopic breast cancer models as well as organoids to study the normal tissue response to radiation and its influence on recurrence. This project is guided by two specific aims: 1) To characterize macrophage phenotypes associated with RT- induced recurrence and 2) to identify targetable secreted factors that contribute to tumor cell recruitment in irradiated tissues. In Aim 1, I will rigorously examine the immune cell infiltrate into irradiated mammary tissue using flow cytometry. To confirm the importance of M2 macrophages in recurrence, I will develop an M2 macrophage-depleted mouse model by blocking polarization with small molecule inhibitors or monoclonal antibodies. Finally, I will determine the impact of M2 macrophage depletion on tumor cell recruitment to irradiated tissues. In Aim 2, I will use an organoid model developed by our laboratory that recapitulates pre-clinical observations. I will analyze secreted pro-tumor factors by co-culturing organoids with macrophages, tumor cells, and CD8+ T cells and characterizing secreted cytokines in conditioned media. I will then evaluate how these factors contribute to tumor cell recruitment in vivo. This study will establish the conditions that lead to recurrence following radiotherapy, including composition of macrophage infiltrate and mechanisms of tumor regrowth, which will have significant implications for TNBC patients vulnerable to recurrence.

项目概要 大约一半的三阴性乳腺癌（TNBC）患者接受保乳治疗，这 包括照射原发肿瘤部位。尽管这种疗法总体上改善了预后，但患者仍继续 局部复发率很高。治疗后控制复发的机制 人们了解甚少。先前的研究表明，正常组织的辐射损伤可能与 复发。临床前研究表明，肿瘤细胞会迁移到受辐射的乳腺组织中，并且 巨噬细胞在肿瘤细胞招募过程中发挥着关键作用。同时阻断巨噬细胞浸润 已被证明可以消除肿瘤细胞向照射部位的迁移，但巨噬细胞表型的影响尚未消除 被调查。已知具有伤口愈合表型 (M2) 的巨噬细胞可促进肿瘤生长并 进展，但 M2 巨噬细胞是否影响受辐射组织中肿瘤细胞的募集和增殖 未知。该提案的中心假设是组织微环境中的辐射损伤 通过免疫抑制细胞浸润促进转移前生态位。我将使用预检验这个假设 临床原位乳腺癌模型以及类器官，用于研究正常组织对辐射的反应 及其对复发的影响。 该项目有两个具体目标：1）表征与 RT-相关的巨噬细胞表型 诱导复发，2) 识别有助于肿瘤细胞募集的靶向分泌因子 受辐射的组织。在目标 1 中，我将严格检查免疫细胞浸润到受辐射的乳腺组织中 使用流式细胞术。为了确认 M2 巨噬细胞在复发中的重要性，我将开发一个 M2 通过用小分子抑制剂或单克隆抗体阻断极化来建立巨噬细胞耗尽的小鼠模型 抗体。最后，我将确定 M2 巨噬细胞耗竭对肿瘤细胞募集到辐射中的影响 组织。在目标 2 中，我将使用我们实验室开发的类器官模型，该模型概括了临床前的情况 观察。我将通过将类器官与巨噬细胞、肿瘤细胞、 和 CD8+ T 细胞并表征条件培养基中分泌的细胞因子。然后我将评估这些 因素有助于体内肿瘤细胞的募集。这项研究将确定导致复发的条件 放疗后，包括巨噬细胞浸润的组成和肿瘤再生的机制， 将对易复发的 TNBC 患者产生重大影响。