Regulation and role of fibroblast-derived interleukin-33 in the pancreatic tumor microenvironment

成纤维细胞来源的白细胞介素33在胰腺肿瘤微环境中的调节和作用

• 批准号: 10604294
• 负责人: Katelyn Lois Donahue
• 金额: $ 4.77万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10604294
• 关键词: Affect; Binding; Biological Assay; CD8-Positive T-Lymphocytes; CRISPR/Cas technology; Cancer Biology; Cancer Etiology; Cells; Cessation of life; Chromatin; Communication; Complement; Complex; Coupled; Cytometry; Data; Disease; Disease Progression; Fibroblasts; Focal Adhesion Kinase 1; Genetic Transcription; Genetically Engineered Mouse; Goals; Growth; Growth Factor; Human; Immunosuppression; Immunotherapy; Implant; In Vitro; Individual; Infiltration; Interleukin-2; Interleukins; Investigation; KRAS oncogenesis; Lesion; Ligands; Literature; Macrophage; Maintenance; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediator; Migration Assay; Modeling; Mus; Nature; Organ; Pancreas; Pancreatic Ductal Adenocarcinoma; Paracrine Communication; Patient-Focused Outcomes; Patients; Phenotype; Population; Regulation; Role; STAT3 gene; Sampling; Signal Pathway; Signal Transduction; Signaling Molecule; Stromal Cells; Subgroup; Survival Rate; T-Cell Activation; Techniques; Testing; Therapeutic; Time; Tissues; Tumor Immunity; Tumor Promotion; Tumor Volume; Tumor-Derived; Tumor-associated macrophages; Tumor-infiltrating immune cells; United States; Up-Regulation; Work; antitumor effect; candidate identification; cytokine; experimental study; extracellular; immune checkpoint blockade; immunosuppressive macrophages; implantation; improved; in vitro Assay; in vivo; mast cell; mouse model; neoplastic cell; novel therapeutics; pancreatic cancer cells; pancreatic ductal adenocarcinoma model; pancreatic neoplasm; premalignant; receptor; recruit; response; single-cell RNA sequencing; trend; tumor; tumor growth; tumor microenvironment

PROJECT SUMMARY/ABSTRACT Pancreatic ductal adenocarcinoma (PDA) is a highly deadly disease with a five-year survival rate of only 10%. The bulk of the PDA tumor volume is composed of a reactive fibroinflammatory tumor microenvironment that assists in the growth and maintenance of the tumor. The highly immunosuppressive nature of the pancreatic tumor microenvironment is also responsible, in part, for the continued inefficacy of immunotherapy treatments in PDA patients. Despite numerous studies that have previously examined different facets of PDA immunosuppression, there is still much that is unknown regarding the complex network of intercellular crosstalk that regulates this phenomenon. Our previous work has demonstrated that infiltrating macrophages are essential mediators of immunosuppression in PDA, and that their polarization is affected by the expression of oncogenic Kras in tumor cells. Other groups have characterized the immunosuppressive impact of certain fibroblast subgroups within the pancreatic tumor microenvironment. Preliminary experiments described in this proposal demonstrate a connection between fibroblasts, macrophages, and tumor cells through fibroblast expression of the cytokine interleukin-33 (IL33). The role of IL33 in cancer biology is controversial, with its overall effect on the tumor thought to be supportive or suppressive depending on the cancer context. Here, results show that IL33 abundance correlates with disease progression and that fibroblast expression of IL33 is dependent on oncogenic Kras-driven signals from tumor cells. Additionally, we have completed a pilot experiment whereby we have orthotopically implanted pancreatic tumor cells into a syngeneic genetically engineered mouse model lacking Il33 in fibroblasts (Il33f/f, Pdgfra-CreERT2/+). Tumors from Il33-deficient mice had slower tumor growth and a trending decrease in macrophage infiltration, suggesting that fibroblast-derived Il33 is both tumor promoting and may play a role in the recruitment of immunosuppressive macrophages. Therefore, the overall objective of this study is to elucidate the cellular interactions that contribute to the establishment and maintenance of immunosuppression in PDA, and the central hypothesis is that fibroblast derived IL33 is a key regulator of immunosuppression in the pancreatic tumor microenvironment through its modulation of macrophage infiltration. This hypothesis will be investigated through the following two Aims: (Aim 1) define the regulation and role of IL33 in cancer-associated fibroblasts, and (Aim 2) assess the impact of IL33 on the recruitment of tumor-associated macrophages. To complete this investigation, a combination of in vitro assays using tumor cell, fibroblast, and macrophage cultures will be performed. We also will continue to utilize the Il33f/f, Pdgfra-CreERT2/+ genetically engineered mouse model to assess Il33-dependent changes in the immunosuppressive nature of pancreatic tumors. The data accumulated by this study will further the field’s understanding of immunosuppression regulation in PDA, and consequentially bring effective immunotherapy strategies one step closer to becoming a viable therapeutic option.

项目摘要/摘要 胰腺导管腺癌(PDA)是一种高度致命的疾病，五年生存率仅为10%。 PDA肿瘤体积的大部分由反应性纤维炎性肿瘤微环境组成， 协助肿瘤的生长和维持。胰腺的高度免疫抑制特性 肿瘤微环境也是免疫治疗持续无效的部分原因。 在PDA患者中。尽管之前已经有大量研究对PDA的不同方面进行了研究 免疫抑制，关于细胞间的复杂网络仍有许多未知之处。 控制这一现象的串扰。我们之前的工作已经证明，渗透的巨噬细胞 是PDA中免疫抑制的重要介质，它们的极化受到表达的影响 致癌基因Kras在肿瘤细胞中的表达。其他小组已经确定了某些特定的免疫抑制作用 胰腺肿瘤微环境中的成纤维细胞亚群。本文件中描述的初步实验 建议证明成纤维细胞、巨噬细胞和肿瘤细胞之间通过成纤维细胞联系 细胞因子白介素33(IL33)的表达。IL33在癌症生物学中的作用是有争议的，其 对肿瘤的总体影响被认为是支持的或抑制的，这取决于癌症的背景。这里, 结果表明，IL33的丰度与疾病的进展有关，成纤维细胞IL33的表达 依赖于来自肿瘤细胞的致癌Kras驱动信号。此外，我们已经完成了一项试点工作 我们将胰腺肿瘤细胞原位移植到同基因的 成纤维细胞缺失IL33的工程化小鼠模型(IL33f/f，PDGFRA-CreERT2/+)。Il33基因缺陷小鼠的肿瘤 肿瘤生长较慢，巨噬细胞浸润呈减少趋势，提示成纤维细胞来源 IL33既具有促肿瘤作用，又可能在免疫抑制巨噬细胞的募集中发挥作用。 因此，这项研究的总体目标是阐明细胞相互作用对 PDA免疫抑制的建立和维持，中心假说是成纤维细胞 派生的IL33是胰腺肿瘤微环境中免疫抑制的关键调节因子，通过其 巨噬细胞浸润的调节。这一假设将通过以下两个目标进行调查： (目标1)确定IL33在癌症相关成纤维细胞中的调节和作用，并(目标2)评估其影响 IL33对肿瘤相关巨噬细胞的募集作用。为了完成这项调查， 将使用肿瘤细胞、成纤维细胞和巨噬细胞培养进行体外分析。我们也将继续 利用Il33f/f，PDGFRA-CreERT2/+基因工程小鼠模型评估IL33依赖于 胰腺肿瘤的免疫抑制性质。这项研究积累的数据将进一步推动该领域的 了解PDA的免疫抑制调节，从而带来有效的免疫治疗 策略离成为可行的治疗选择又近了一步。