Role of Cilp1 in Post-Natal Heart Response to Injury

Cilp1 在产后心脏损伤反应中的作用

• 批准号: 10580830
• 负责人: Zhi-Ping Liu
• 金额: $ 54.47万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10580830
• 关键词: Adult; Affect; Binding; Biological Assay; Blood; Blood Vessels; Bone Development; CD36 gene; Cardiac; Cartilage; Cell Communication; Cell Proliferation; Cell Proliferation Regulation; Cicatrix; Clinical; Collagen; Conditioned Culture Media; Cues; DNA cassette; Deposition; Development; Disease; Electric Conductivity; Embryonic Development; Enterobacteria phage P1 Cre recombinase; Extracellular Matrix; Fibroblasts; Fibrosis; Future; Gene Expression; Gene Expression Regulation; Genetic Transcription; Growth Factor; Heart; Heart Diseases; Heart failure; Infarction; Inflammation; Inflammatory; Injury; Integrins; Interleukin-6; Knock-out; Knockout Mice; Left Ventricular Hypertrophy; Liver; Liver Fibrosis; Macrophage; Measures; Molecular; Molecular Profiling; Mus; Mutagenesis; Myocardial Infarction; Myocardium; Myofibroblast; N-terminal; NFKB Signaling Pathway; Necrosis; Outcome; Pathologic; Pathway interactions; Pharmaceutical Preparations; Phenotype; Physiological; Process; Production; Proliferating; Property; Protein Secretion; Proteins; Proteomics; Public Health; Reagent; Receptor Cell; Recombinants; Regenerative capacity; Research; Risk Factors; Role; Shapes; Signal Pathway; Structural Protein; Tamoxifen; Testing; Therapeutic; Tissues; Transgenic Mice; angiogenesis; autocrine; cartilage development; cell behavior; cell type; coronary fibrosis; cytokine; design; differential expression; effective therapy; experimental study; genomic locus; heart cell; heart function; heart preservation; inducible Cre; insight; ischemic injury; organ growth; overexpression; paracrine; periostin; postnatal; prevent; promoter; receptor; receptor-mediated signaling; response to injury; single nucleus RNA-sequencing; success; tendon development; therapeutic target

Matricellular proteins are constituents of the extracellular matrix (ECM). They are normally expressed highly during embryonic development but absent/low in adult tissues unless activated by cues for tissue remodeling. Matricellular proteins shape ECM properties through interactions with structural proteins, growth factors, and cell receptors during organ development and differentiation. In an attempt to identify molecular signatures unique to irreversible cardiac fibrosis, we performed proteomics of fibrotic heart and liver and found that cartilage intermediate layer protein 1 (Cilp1) is differentially upregulated in the infarcted heart. Cilp1 is normally associated with bone and cartilage development. Its function and mechanism of action in adult heart diseases are unknown. We generated Cilp1 knockout (KO) mice from commercial Cilp1fl/fl mice and transgenic (Tg) mice with Cilp1 overexpressed in myofibroblasts. While deletion of Cilp1 reduced adverse cardiac remodeling upon myocardial infarction (MI), overexpression of Cilp1 worsened it. Cilp1 is expressed predominantly in cardiac fibroblasts. We hypothesize that fibroblast Cilp1 promotes inflammation and myofibroblast proliferation upon MI injury. We now generated fibroblast conditional fbKO mice (PostnMCM;Cilp1fl/fl and Tcf21MCM;Cilp1fl/fl that contain a tamoxifen inducible Cre-recombinase expression cassette within Periostin (Postn) and Tcf21 genetic locus, respectively). Aim 1. To determine the cell-type specific function of Cilp1 in post-MI cardiac remodeling. We will delete Cilp1 in cardiac fibroblasts before and post-MI day (d) 1 & d4 to investigate its effect on cardiac remodeling, including cardiac function, inflammation, myofibroblast proliferation/differentiation, and collagen remodeling. We will also perform proteomics of myofibroblasts isolated from these mouse hearts. The role of fibroblast Cilp1 in regulation of gene transcription in various heart cell types will be investigated with single-nuclei RNA-seq of infarcted WT and Cilp1 fbKO hearts at post-MI d3. Aim 2. To establish the molecular function of Cilp1 and its mechanism of action. Preliminary studies showed that Cilp1 protein in culture medium promotes myofibroblast proliferation via the mTORC1 pathway and binds scavenge receptor CD36. Cilp1 may interact with cell receptor/growth factor, promoting cell proliferation and inflammatory gene expression via receptor-mediated signaling pathways. To test this hypothesis, we will identify the minimal functional domain(s) of Cilp1 via mutagenesis and potential Cilp1- binding partners using both screen- and candidate-based assays and will establish how Cilp1 may act as a paracrine factor to regulate the cellular phenotypes of various heart cell types via receptor-mediated signaling pathways. We will also measure blood level of Cilp1 in mice before and after an anti-fibrogenic therapy upon MI injury. Matricellular proteins are clinically tractable owing to their accessibility to systemically delivered therapeutic reagents. Our preliminary studies established a pathological role of Cilp1 in post-MI remodeling. This proposal will establish how Cilp1 instructs development and differentiation of heart cells and gene expression to promote adverse remodeling, thus providing mechanistic insight on how to target this protein.

细胞蛋白是细胞外基质（ECM）的组成部分。它们通常高度表达