Role of hypoxia in fibroblast reprogramming in pancreatic cancer

缺氧在胰腺癌成纤维细胞重编程中的作用

• 批准号: 10748608
• 负责人: Tenzin Ngodup
• 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: 2023-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10748608
• 关键词: 3-Dimensional; Accounting; Area; Blood Vessels; Cancer Control; Cells; Coculture Techniques; Data; Desmoplastic; Diagnosis; Disease; Extracellular Matrix; Fibroblasts; Goals; Human; Hypoxia; Hypoxia Inducible Factor; Immune; Incidence; Inflammatory; Interleukin-1; Knock-out; Link; M cell; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Mediator; Molecular; Mus; Mutation; Oxygen; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Patients; Play; Production; Proteins; Regulation; Resistance; Role; Sampling; Signal Pathway; Signal Transduction; Stromal Cells; System; Testing; Therapeutic; Tissues; Wit; Work; cancer cell; cancer therapy; cytokine; density; design; genetic signature; hypoxia inducible factor 1; in vivo; insight; mortality; mouse model; neoplastic cell; normoxia; novel therapeutic intervention; pancreatic cancer cells; pancreatic tumorigenesis; response; single-cell RNA sequencing; therapy resistant; tumor; tumor hypoxia; tumor initiation; tumor microenvironment; tumorigenesis

PROJECT SUMMARY Pancreatic ductal adenocarcinoma (PDAC) is a prevalent form of pancreatic cancer, accounting for about 90% of all cases of this disease. PDAC is highly fatal, where mortality closely parallels incidence, primarily due to the inability to diagnose PDAC at early stages and resistance to all existing treatments. Besides genetic changes in tumor cells, the tumor microenvironment plays a key role in tumor initiation, progression, and therapeutic resistance. PDAC is notable for its dense desmoplastic stroma, consisting of extracellular matrix, cancer- associated fibroblasts (CAFs), and immune cells. Another notable feature of the PDAC microenvironment is hypoxia, a condition of insufficient oxygen availability. Although hypoxia leads to adaptive responses in both cancer cells and stromal cells, the effects of hypoxia and hypoxia-inducible factor 1ɑ (HIF1ɑ), a master regulator of hypoxic adaptation, on the PDAC stroma and tumor-stroma interactions are not fully understood. We have recently found that inflammatory CAFs (iCAFs), a CAF subset producing high levels of inflammatory cytokines, are linked to the hypoxia-related gene signature and the HIF1 signaling pathway in human and mouse PDAC. By exposing 3D cocultures of pancreatic tumor cells and fibroblasts to either hypoxia or normoxia, we demonstrated that hypoxia promotes iCAF formation via IL1ɑ secreted from tumor cells. Importantly, the presence of hypoxic fibroblasts further elevated IL1ɑ levels in tumor cells, implicating hypoxia as a modulator of bidirectional interactions between tumor cells and fibroblasts. Based on these data, I hypothesize that hypoxia promotes iCAF formation by modulating bidirectional interactions between tumor cells and fibroblasts in PDAC. In Specific Aim 1, I will determine how hypoxia induces IL1ɑ expression in tumor cells via fibroblasts by using a three-dimensional (3D) coculture system of pancreatic tumor cells and fibroblasts and exposing the coculture to either hypoxia or normoxia. In Specific Aim 2, I will determine whether and how fibroblast HIF1ɑ regulates tumor cell-fibroblast interactions and pancreatic tumorigenesis by using 3D cocultures of tumor cells and fibroblasts deficient of HIF1ɑ, and using mouse models lacking fibroblast expression of HIF1ɑ. My work will identify the molecular mechanisms underlying hypoxic regulation of cancer cell-CAF crosstalk. In addition, my studies will determine how activation of HIF1ɑ within CAFs regulates the tumor stroma and PDAC progression, which will inform targeting fibroblast HIF1ɑ as a potential therapeutic avenue to treat PDAC patients.

项目摘要 胰腺导管腺癌（PDAC）是一种普遍的胰腺癌形式，约占90％ 在所有这种疾病的病例中。 PDAC高度致命，死亡率紧密相似，主要是由于 无法在早期诊断PDAC并对所有现有治疗方法进行抵抗。除了遗传变化 肿瘤细胞，肿瘤微环境在肿瘤起始，进展和治疗中起关键作用 反抗。 PDAC以其致密的脱肿瘤基质而闻名，由细胞外基质，癌症组成 相关的成纤维细胞（CAF）和免疫细胞。 PDAC微环境的另一个值得注意的特征是 缺氧，氧气可用性不足。尽管缺氧导致两者都有自适应反应 癌细胞和基质细胞，缺氧和缺氧诱导因子1（HIF1）的作用，主要调节剂 在PDAC基质和肿瘤中相互作用上的缺氧适应性尚未完全了解。我们有 最近发现，炎症CAF（ICAFS）是一种CAF子集，产生了高水平的炎性细胞因子， 与人和小鼠PDAC中与缺氧相关的基因信号和HIF1信号通路有关。 通过暴露胰腺肿瘤细胞和成纤维细胞的3D共培养物中的缺氧或正常氧 证明缺氧通过肿瘤细胞分泌的IL1促进ICAF形成。重要的是， 缺氧成纤维细胞的存在进一步升高了肿瘤细胞中的IL1水平，隐式缺氧作为调节剂 肿瘤细胞与成纤维细胞之间的双向相互作用。基于这些数据，我假设缺氧 通过调节PDAC中肿瘤细胞与成纤维细胞之间的双向相互作用来促进ICAF的形成。 在特定的目标1中，我将通过使用A使用成纤维细胞来确定缺氧如何在肿瘤细胞中诱导IL1表达 胰腺肿瘤细胞和成纤维细胞的三维（3D）共培养系统，并将共培养暴露于 缺氧或正常氧。在特定目标2中，我将确定是否以及如何调节肿瘤的成纤维细胞HIF1 通过使用肿瘤细胞和成纤维细胞的3D共培养物和成纤维细胞共培养的细胞成纤维细胞相互作用和胰腺肿瘤发生 HIF1的缺陷，并使用鼠标模型缺乏HIF1的成纤维细胞表达。我的工作将确定 癌细胞-CAF串扰低氧调节的分子机制。此外，我的学习将 确定CAF中HIF1的激活如何调节肿瘤基质和PDAC进展，这将是 告知靶向成纤维细胞HIF1的潜在治疗途径以治疗PDAC患者。