Hypoxic regulation of macrophage migration and function via fibroblast reprogramming in pancreatic cancer

通过胰腺癌成纤维细胞重编程对巨噬细胞迁移和功能的缺氧调节

• 批准号: 10677376
• 负责人: Sean Matthew Hannifin
• 金额: $ 4.08万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 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 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10677376
• 关键词: Area; Blood Vessels; CCL2 gene; Cancer Etiology; Cells; Cessation of life; Coculture Techniques; Communication; Data; Development; Disease; Disease Progression; Environment; Exclusion; Fibrinogen; Fibroblasts; Gene Expression; Goals; HIF1A gene; Human; Hypoxia; Hypoxia Inducible Factor; Immune; Immune System Diseases; Immunosuppression; Immunotherapeutic agent; Immunotherapy; Infiltration; Inflammatory; Macrophage; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Modeling; Mus; Myelogenous; Outcome; Oxygen; Pancreas; Pancreatic Ductal Adenocarcinoma; Patients; Phenotype; Play; Population; Regulation; Resistance; Role; Signal Transduction; Stress; Stromal Cells; Survival Rate; T-Lymphocyte; Testing; United States; cell type; chemokine; cytokine; density; effective therapy; experience; immunosuppressive macrophages; insight; migration; monocyte; neoplastic cell; neutralizing antibody; normoxia; pancreatic cancer cells; pancreatic neoplasm; pancreatic tumorigenesis; recruit; response; therapy resistant; transcription factor; tumor; tumor hypoxia; tumor progression; tumor-immune system interactions

PROJECT SUMMARY Pancreatic ductal adenocarcinoma (PDAC) is the third leading cause of cancer-related deaths in the United States despite representing only 2% of all cancer cases. PDAC is characterized by an extensive fibroinflammatory stroma and a hypoxic microenvironment, which contributes to disease progression and therapeutic resistance. Macrophages and cancer-associated fibroblasts (CAFs) are the predominant cell types within the PDAC stroma. Macrophages are a major immunosuppressive population in PDAC. These cells are highly plastic and, as a result, their environment plays an important role in regulating their function. Macrophages adapt to hypoxia, a condition of low oxygen availability, primarily through the stabilization of oxygen-liable transcription factors called hypoxia-inducible factors (HIFs). Although PDAC is extremely hypoxic and both the tumor cells and stromal cells experience hypoxia, the effects of hypoxia and HIFs on macrophages and their communication with other stromal cells in PDAC remain largely unknown. Our group has recently found that hypoxia promotes the acquisition of inflammatory cancer-associated fibroblasts (inflammatory CAFs), a recently defined PDAC fibroblast subtype that produces high levels of inflammatory cytokines and chemokines. By injecting a hypoxia probe into mice bearing PDAC, we have observed that inflammatory CAFs and macrophages predominantly reside in hypoxic tumor regions compared with normoxic regions whereas T cells are largely excluded from hypoxic tumor areas. Based on these preliminary data, I hypothesize that inflammatory fibroblasts induced by hypoxia promote macrophage infiltration into hypoxic tumor regions and facilitate an immunosuppressive macrophage phenotype. This hypothesis will be investigated with the following two Aims: (1) to determine the role of hypoxia-induced fibroblast-secreted factors in regulating macrophage migration and function in PDAC and (2) to define the role of macrophage HIFs in regulating macrophage migration and function in PDAC. To complete Aim 1, I will culture macrophages with conditioned media derived from mono- and co- cultures of pancreatic tumor cells and fibroblasts under either hypoxia or normoxia, and then assess the expression of immunosuppressive macrophage markers as well as their migration. To complete Aim 2, I will culture HIF1α-deficient and HIF2α-deficient macrophages under hypoxia with fresh media or conditioned media from co-cultures of pancreatic tumor cells and fibroblasts under hypoxia. Then I will assess the expression of immunosuppressive markers in these macrophages and evaluate their migration. Finally, I will define the role of macrophage HIFs in mediating pancreatic tumorigenesis using orthotopic tumors from mice lacking myeloid expression of HIF1ɑ or HIF2ɑ. My proposal will provide new insights into how hypoxia promotes an immunosuppressive PDAC microenvironment by modulating the macrophage-fibroblast crosstalk, and ultimately inform strategies to overcome resistance to immunotherapy.

项目概要 胰腺导管腺癌 (PDAC) 是美国癌症相关死亡的第三大原因 尽管各州仅占所有癌症病例的 2%。 PDAC 的特点是广泛 纤维炎症基质和缺氧微环境，这有助于疾病进展和 治疗抵抗。巨噬细胞和癌症相关成纤维细胞 (CAF) 是主要的细胞类型 在 PDAC 基质内。巨噬细胞是 PDAC 中的主要免疫抑制群体。这些细胞是 高度可塑性，因此，它们的环境在调节它们的功能方面发挥着重要作用。巨噬细胞 适应缺氧，即氧气供应量低的情况，主要是通过稳定氧气不稳定的状态来适应 转录因子称为缺氧诱导因子（HIF）。尽管 PDAC 极度缺氧并且 肿瘤细胞和基质细胞经历缺氧，缺氧和HIF对巨噬细胞及其作用的影响 PDAC 中与其他基质细胞的通讯仍然很大程度上未知。我们小组最近发现 缺氧促进炎症性癌症相关成纤维细胞（炎症性 CAF）的获得，这是最近发现的一种 确定的 PDAC 成纤维细胞亚型产生高水平的炎症细胞因子和趋化因子。经过 通过向携带 PDAC 的小鼠注射缺氧探针，我们观察到炎症性 CAF 和巨噬细胞 与含氧量正常的区域相比，T 细胞主要存在于缺氧的肿瘤区域，而 T 细胞则主要分布在缺氧的肿瘤区域。 排除缺氧肿瘤区域。根据这些初步数据，我假设炎症成纤维细胞 缺氧诱导促进巨噬细胞浸润缺氧肿瘤区域并促进 免疫抑制巨噬细胞表型。该假设将通过以下两个目标进行研究： (1)确定缺氧诱导的成纤维细胞分泌因子在调节巨噬细胞迁移和迁移中的作用 (2) 定义巨噬细胞 HIF 在调节巨噬细胞迁移和功能中的作用 在 PDAC 中。为了完成目标 1，我将使用来自单和共-的条件培养基培养巨噬细胞 在缺氧或常氧条件下培养胰腺肿瘤细胞和成纤维细胞，然后评估 免疫抑制巨噬细胞标记物的表达及其迁移。为了完成目标2，我会 用新鲜培养基或条件培养基在缺氧条件下培养 HIF1α 缺陷型和 HIF2α 缺陷型巨噬细胞 来自缺氧条件下胰腺肿瘤细胞和成纤维细胞的共培养。然后我将评估的表达 这些巨噬细胞中的免疫抑制标记并评估它们的迁移。最后，我将定义角色 巨噬细胞 HIF 在利用缺乏骨髓小鼠的原位肿瘤介导胰腺肿瘤发生中的作用 HIF1ɑ 或 HIF2ɑ 的表达。我的建议将为缺氧如何促进 通过调节巨噬细胞-成纤维细胞串扰来抑制免疫抑制 PDAC 微环境，最终 提供克服免疫治疗耐药性的策略。