Interaction of fibroblasts with cell corpses increases collagen synthesis during lung repair

成纤维细胞与细胞尸体的相互作用增加了肺修复过程中的胶原蛋白合成

• 批准号: 10736792
• 负责人: Alexandra Leigh McCubbrey
• 金额: $ 61.93万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10736792
• 关键词: Address; Alveolar; Apoptotic; Basement membrane; Binding; Biological Process; Bleomycin; Blood capillaries; Cell Death; Cell Proliferation; Cells; Coculture Techniques; Collagen; Cues; Data; Deposition; Development; Enzymes; Extracellular Matrix; Fibroblasts; Gene Deletion; Health; Human; In Vitro; Inflammation; Injury; Isotopes; Lipids; Liposomes; Lung; Macrophage; Maintenance; Molecular Conformation; Mus; Ornithine Decarboxylase; Phagocytes; Phosphatidylserines; Polyamines; Production; Proliferating; Protein Biosynthesis; Proteins; Proteomics; Pulmonary Inflammation; Research; Ribosomes; Role; Signal Transduction; Small Interfering RNA; Spermidine; Tamoxifen; Testing; Translations; Work; deoxyhypusine synthase; eIF-5A; healing; hypusine; in vivo; inhibitor; lung health; lung injury; lung repair; metabolomics; novel; novel strategies; novel therapeutics; phosphatidylserine receptor; polyproline; prevent; receptor; reconstitution; repair function; repaired; response; restoration; scaffold; seal; tissue injury; tissue repair; transcriptome sequencing; wound closure

PROJECT SUMMARY Repair after injury is a fundamental biologic process that is critical for maintaining lung health. Fibroblasts must perform key pro-repair functions after injury including proliferation and synthesis of new extracellular matrix, particularly collagens, that scaffold wound closure. Dead cells (collectively termed cell corpses) are produced during lung injury and inflammation and have been shown to promote the transition from inflammation to repair. Prior research has focused on understanding macrophage interactions with apoptotic corpses. However, non- professional phagocytes including fibroblasts can also interact with cell corpses and the consequence of these interactions in the lung has not been studied. This proposal seeks to address unknowns regarding fibroblast- corpse interactions and their role in healthy lung repair. We have found that recognition of cell corpses by fibroblasts causes fibroblasts to increase collagen protein synthesis. Our preliminary data support a mechanism where the phosphatidylserine receptor Axl binds cells corpses, activates polyamine metabolite synthesis, increasing intracellular spermidine levels, which fuel hypusination. Hypusination occurs on a single protein: the ribosomal subunit eiF5a. When hypusinated, eiF5a is able to stabilize translation of poly-proline-repeat proteins including collagen. Hypusination is also important for cell proliferation. Blocking polyamine synthesis prevented hypusination of eiF5a in response to corpse recognition. Blocking Axl or hypusine-eiF5a prevented the increase in collagen in response to corpse recognition. We also show that, in vivo, hypusine-eiF5a is increased in fibroblasts after lung injury and that collagen deposition and alveolar-capillary repair are reduced when hypusination is blocked with the inhibitor GC7 or through targeted deletion of the hypusination enzyme deoxyhypusine synthase in fibroblasts. This led us to hypothesize that interaction of fibroblasts with cell corpses is a central cue that activates fibroblast proliferation and collagen synthesis following lung injury, facilitating repair. We will use three aims to test if 1) corpse recognition via Axl triggers a translationally-regulated increase in collagen protein, 2) corpse recognition drives polyamine synthesis that increases spermidine that increases hypusine-eiF5a and stabilizes collagen translation, and 3) to fully activate pro-repair programming fibroblasts require the ability to sense corpses via Axl, synthesize polyamines via ornithine decarboxylase, and hypusinate eiF5a via deoxyhypusine synthase in order to proliferate and synthesize new extracellular matrix after lung injury.

项目摘要 受伤后修复是一种基本的生物学过程，对于维持肺部健康至关重要。成纤维细胞必须 损伤后执行关键的促进功能，包括增殖和合成新的细胞外基质， 特别是胶原蛋白，脚手架伤口闭合。产生死细胞（集体称为细胞尸体） 在肺损伤和炎症期间，已被证明可以促进从炎症到修复的过渡。 先前的研究集中在理解与凋亡尸体的巨噬细胞相互作用。但是，非 包括成纤维细胞在内的专业吞噬细胞也可以与细胞尸体相互作用，而这些吞噬细胞的结果 尚未研究肺部的相互作用。该建议旨在解决有关成纤维细胞的未知数 尸体相互作用及其在健康肺修复中的作用。我们发现，通过 成纤维细胞导致成纤维细胞增加胶原蛋白合成。我们的初步数据支持一种机制 其中磷脂酰丝氨酸受体AXL结合细胞尸体，激活多胺代谢产物合成， 增加细胞内的精子素水平，从而燃料降低。无偶然发生在单个蛋白质上： 核糖体亚基EIF5A。当量不足时，EIF5A能够稳定多生酸酯蛋白的翻译 包括胶原蛋白。非胰腺对细胞增殖也很重要。阻止多胺合成阻止 EIF5A响应尸体识别的不为性。阻止Axl或dellusine-EIF5A阻止了增加 在胶原蛋白的响应中。我们还表明，在体内，hybusine-eif5a在 肺损伤后的成纤维细胞以及胶原蛋白沉积和肺泡毛细血管修复时会减少 通过抑制剂GC7或通过靶向删除非偶然性酶阻断非uSANDIND酶。 成纤维细胞中的脱氧类合酶。这导致我们假设成纤维细胞与细胞尸体的相互作用 是一种中央提示，可激活肺损伤后成纤维细胞增殖和胶原蛋白合成 维修。我们将使用三个目标来测试1）通过AXL识别尸体是否触发翻译的增加 在胶原蛋白中，2）尸体识别驱动多胺合成，增加了精子的增加 无蛋白-EIF5A并稳定胶原蛋白翻译，3）完全激活培养基编程成纤维细胞 需要能够通过AXL吸引尸体，通过鸟氨酸脱羧酶合成多胺和降低的能力 EIF5A通过脱氧蛋白酶合酶通过肺损伤后增殖和合成新的细胞外基质。