Tissue Homeostatic Control by CSF-dependent Macrophage-Stromal Cell Interactions

脑脊液依赖性巨噬细胞-基质细胞相互作用的组织稳态控制

• 批准号: 8718854
• 负责人: JEREMY B JACOX
• 金额: $ 2.73万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8718854
• 关键词: Ablation; Address; Adult; Affect; Atherosclerosis; Attenuated; Binding; Binding Sites; Biochemical; Biological Assay; Biotinylation; Bone Marrow; Breast Carcinoma; CSF1 gene; Carcinogens; Cell Communication; Cell Death; Cell physiology; Cells; Coculture Techniques; Conditioned Culture Media; Data; Dependence; Development; Diagnostic Neoplasm Staging; Diethylnitrosamine; Differentiation and Growth; Disease; EGF gene; Embryo; Endopeptidase K; Epithelial Cells; Evolution; Feedback; Fibroblasts; Flow Cytometry; Fractionation; Genetic; Genetic Models for Cancer; Genetic Transcription; Glycoside Hydrolases; Growth; Growth Factor; Homeostasis; Hormones; Human Development; Hypoxia; Immune system; Immunity; Immunofluorescence Immunologic; Immunohistochemistry; Immunoprecipitation; In Vitro; Inflammatory; Knock-out; Life; Liquid substance; Liver; Logic; Lupus; Macrophage Colony-Stimulating Factor; Maintenance; Malignant Epithelial Cell; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Membrane; Mesenchymal; Messenger RNA; Methods; Modeling; Molecular; Mus; NMR Spectroscopy; Necrosis; Neoplasms; Normal tissue morphology; Nuclear Extract; Phenotype; Physiological; Primary carcinoma of the liver cells; Production; Recruitment Activity; Reporting; Resistance development; Role; Serum; Signal Transduction; Starvation; Stimulus; Streptavidin; Stromal Cells; Surface; System; Testing; Tissues; Transplantation; Tumor Cell Line; Tumor Pathology; Tumor stage; Western Blotting; biophysical techniques; cancer therapy; cytokine; fast protein liquid chromatography; genetic analysis; in vivo; in vivo Model; macrophage; neoplastic; neoplastic cell; nuclease; overexpression; paracrine; promoter; public health relevance; repaired; research study; response; small hairpin RNA; tumor; tumor growth; tumor microenvironment; tumor progression; tumorigenesis; vector

DESCRIPTION (provided by applicant): All tissues contain stromal and parenchymal cells and tissue-resident macrophages, which are critical for tissue homeostasis, repair and immunity. Macrophage colony stimulating factor is a critical cytokine for macrophage growth, differentiation, and development, and its deficiency leads to substantial macrophage deficits in the CSF1op/op mouse. While stromal cell-secreted CSF1 is known to be upregulated in response to various cytokines and hormones, very little is understood about how it may be regulated locally by interactions with macrophages, the primary MCSF consumer. Less is understood about how tumor cells can manipulate MCSF synthesis, except for a putative EGF-CSF1 paracrine positive feedback loop between macrophages and invasive breast carcinoma cells. Such a feedback loop cannot explain the evident stability of tissue-specific ratios of macrophages to stromal cells in the tissue microenvironment, nor how adaptive physiologic demands for macrophage functions or macrophage-produced growth factors affect the local MCSF production set point. We hypothesize that macrophages produce a signal reporting their MCSF requirements to stromal cells, and that stromal cells respond to this signal by inducing CSF1 transcription, inhibiting macrophage signal production once through feedback inhibition, and postulate that a reciprocal negative feedback loop exists reporting stromal cell demand for macrophage functions or growth factors. Consistent with our hypothesis, our preliminary data in a mouse embryonic fibroblast - macrophage co-culture system suggests that stromal cell CSF1 expression may be induced significantly by as yet unknown intercellular signals, and conditioned media transfer experiments suggest that this CSF1-inducing signal may be mediated by a soluble factor. Furthermore, little is known of how a functional mismatch between MCSF production by tumors and tumor- associated macrophage (TAM) MCSF requirements leads to tumor pathology. Although some tumors are known to overexpress CSF1 successfully to recruit TAMs, we hypothesize that tumors fail to adequately regulate local MCSF production to TAM demands, leading to increased necrosis, hypoxia, pro-inflammatory TAM and tumor cell death. In this project, we combine in vitro macrophage and stromal cell co-culture systems, transcriptional analysis, with biophysical and biochemical methods for molecular signal isolation. Additionally, we develop in vivo models of CSF1-attenuated tumor cell lines and carcinogen-induced hepatocellular carcinoma on a liver-specific CSF1 conditional knockout model to test these hypotheses, to together better elucidate the CSF1-dependent signaling mechanisms between macrophages and stromal cells in normal tissue homeostasis and tumorigenesis.

描述（由申请人提供）：所有组织都含有基质细胞和实质细胞以及组织驻留巨噬细胞，它们对于组织稳态、修复和免疫至关重要。巨噬细胞集落刺激因子是巨噬细胞生长、分化和发育的关键细胞因子，其缺乏会导致 CSF1op/op 小鼠巨噬细胞严重缺陷。虽然已知基质细胞分泌的 CSF1 会因各种细胞因子和激素的反应而上调，但人们对它如何通过与巨噬细胞（MCSF 的主要消费者）相互作用进行局部调节却知之甚少。除了巨噬细胞和侵袭性乳腺癌细胞之间推定的 EGF-CSF1 旁分泌正反馈环之外，人们对肿瘤细胞如何操纵 MCSF 合成了解甚少。这种反馈回路无法解释组织微环境中巨噬细胞与基质细胞的组织特异性比率的明显稳定性，也无法解释对巨噬细胞功能或巨噬细胞产生的生长因子的适应性生理需求如何影响局部 MCSF 产生设定点。我们假设巨噬细胞产生向基质细胞报告其 MCSF 需求的信号，并且基质细胞通过诱导 CSF1 转录、通过反馈抑制抑制巨噬细胞信号产生来响应该信号，并假设存在相互负反馈循环，报告基质细胞对巨噬细胞功能或生长因子的需求。与我们的假设一致，我们在小鼠胚胎成纤维细胞-巨噬细胞共培养系统中的初步数据表明，基质细胞 CSF1 表达可能是由迄今未知的细胞间信号显着诱导的，并且条件培养基转移实验表明，这种 CSF1 诱导信号可能是由可溶性因子介导的。此外，人们对肿瘤产生的 MCSF 与肿瘤相关巨噬细胞 (TAM) MCSF 需求之间的功能不匹配如何导致肿瘤病理学知之甚少。尽管已知某些肿瘤会过度表达 CSF1 以成功招募 TAM，但我们假设肿瘤无法根据 TAM 需求充分调节局部 MCSF 的产生，从而导致坏死、缺氧、促炎 TAM 和肿瘤细胞死亡的增加。在这个项目中，我们将体外巨噬细胞和基质细胞共培养系统、转录分析以及用于分子信号分离的生物物理和生化方法结合起来。此外，我们在肝脏特异性CSF1条件敲除模型上开发了CSF1减毒肿瘤细胞系和致癌物诱导的肝细胞癌的体内模型来测试这些假设，以更好地阐明正常组织稳态和肿瘤发生中巨噬细胞和基质细胞之间的CSF1依赖性信号传导机制。