Controlling Adipocyte-Myofibroblast Interactions to Improve Healing

控制脂肪细胞-肌成纤维细胞相互作用以改善愈合

• 批准号: 10582858
• 负责人: James Allen Ankrum
• 金额: $ 33.3万
• 依托单位: UNIVERSITY OF IOWA
• 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 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10582858
• 关键词: 3-Dimensional; Adipocytes; Adipose tissue; Appearance; Attention; Autologous; Back; Behavior; Biological Assay; Blood Vessels; Cell Lineage; Cells; Cicatrix; Clinical; Communication; Complement; Complex; Cost of Illness; Coupled; Data; Dermal; Development; Engineering; Extracellular Matrix; Extracellular Matrix Proteins; Family suidae; Fatty acid glycerol esters; Fibroblasts; Fibrosis; Fractionation; Genetic Transcription; Genomics; Goals; Human; In Vitro; Injury; Mechanics; Myofibroblast; Operative Surgical Procedures; Phenotype; Prevention; Process; Production; Proteomics; Quality of life; Reconstructive Surgical Procedures; Role; Site; Source; Surgeon; System; Techniques; Testing; Therapeutic Intervention; Tissues; Transplantation; Work; Wound healing therapy; Wound models; adipokines; adipose derived stem cell; cell type; design; effective therapy; fat grafting; healing; improved; in vitro Model; innovation; interest; knowledge base; mechanical properties; novel; novel therapeutic intervention; prevent; response; stem cells; subcutaneous; targeted treatment; wound bed; wound healing

PROJECT SUMMARY/ABSTRACT Fibrosis and scarring from injury and disease costs $20 billion annually to treat in the US, and even when treated remains a persistent problem that significantly diminishes quality of life. The use of autologous adipose micrografts used in reconstructive surgeries appears beneficial for reducing scar. Furthermore, intradermal adipocytes have also recently gained attention for their role in the early stages of normal wound healing. The objective of this proposal is to determine how human adipocyte lineage cells impact fibroblast-myofibroblast conversion and extracellular matrix (ECM) remodeling in a healing wound. Our long-term goal is to engineer more effective and improved clinical therapies for wound healing. To achieve this objective, we propose three aims that are designed to unravel the communication between adipose lineage cells and fibroblasts. In Aim 1, we will determine how secreted factors from adipocyte linage cells regulate fibroblast-myofibroblast conversion, phenotype, and ECM remodeling. We hypothesize that adipocyte lineage cells differentially regulate inhibition, induction, and reversal of fibroblast to myofibroblast conversion, which in turn impacts the composition of the ECM produced. We also hypothesize that adipokine-induced myofibroblasts (compared to TGF-β1-induced myofibroblasts) produce distinct ECM that results in less scaring. We will test these hypotheses and identify how one-way communication from adipocyte lineage cells regulates fibroblast-myofibroblast activity using a myofibroblast conversion assay coupled with fractionation, proteomics, computational genomics, and inhibition/add-back assays. In Aim 2 we will determine how adipocyte lineage cells coordinate with fibroblasts to remodel ECM. We hypothesize that reciprocal communication between adipocyte lineage cells and fibroblasts- myofibroblasts impacts wound healing by altering cell activity and the composition of the ECM proteins produced by both fibroblasts and adipocyte lineage cells. We will use our 3D adipose spheroids and our 3D in vitro models to interrogate the interactions between the relevant cell types and the composition and mechanics of the ECM produced. In Aim 3, we will complement the in vitro work of Aims 1 and 2 by determining how transplantation of adipocyte lineage cells alter fibroblast behavior and ECM remodeling in a porcine wound model. We hypothesize that wound bed fibroblasts will differentially respond to adipose-based treatments depending on the cellular composition delivered. We will test this hypothesis by assessing the effect of transplanted adipose stem cells, preadipocytes, or adipocytes on fibroblast/myofibroblast behavior and wound healing. Successful completion of these aims will substantially improve our understanding of adipose-fibroblast interactions that will be critical for the development of more effective and improved clinical therapies for wound healing.

项目摘要/摘要 纤维化和因受伤和疾病的疤痕每年花费200亿美元用于治疗，即使在治疗时 仍然是一个持久的问题，可大大降低生活质量。自体脂肪的使用 重建手术中使用的微移植物似乎有益于减少疤痕。此外，皮内 脂肪细胞最近也因其在正常伤口愈合的早期阶段而引起了人们的关注。 该建议的目的是确定人类脂肪细胞谱系细胞如何影响成纤维细胞肌纤维细胞 转换和细胞外基质（ECM）在愈合伤口中重塑。我们的长期目标是设计 更有效，改善了伤口愈合的临床疗法。为了实现这一目标，我们提出了三个 旨在阐明脂肪谱系细胞和成纤维细胞之间的通信的目的。在AIM 1中， 我们将确定来自脂肪细胞弦细胞的分泌因子如何调节成纤维细胞果纤维细胞转换， 表型和ECM重塑。我们假设脂肪细胞谱系细胞对抑制作用不同， 诱导和成纤维细胞向肌纤维细胞转换的逆转，这反过来影响 ECM产生。我们还假设脂肪因子诱导的肌纤维细胞（与TGF-β1诱导的 肌纤维细胞）产生独特的ECM，从而减少恐惧。我们将检验这些假设，并确定 来自脂肪细胞谱系细胞的单向通信调节成纤维细胞元素纤维细胞活性 肌成纤维细胞转换测定法以及分馏，蛋白质组学，计算基因组学和 抑制/添加测定。在AIM 2中，我们将确定脂肪细胞谱系细胞如何与成纤维细胞协调 重塑ECM。我们假设脂肪细胞谱系细胞与成纤维细胞之间的相互交流 - 肌纤维细胞通过改变细胞活性和产生的ECM蛋白的组成来影响伤口愈合 通过成纤维细胞和脂肪细胞谱系细胞。我们将使用3D脂肪球体和3D体外模型 询问相关细胞类型与ECM的组成和力学之间的相互作用 生产。在AIM 3中，我们将通过确定如何移植来完成AIM 1和2的体外工作 脂肪细胞谱系细胞改变猪伤口模型中的成纤维细胞行为和ECM重塑。我们假设 那个伤口床的成纤维细胞会根据细胞对基于脂肪的治疗的反应不同 构图交付。我们将通过评估移植脂肪干细胞的影响来检验这一假设， 成纤维细胞/肌纤维细胞行为和伤口愈合的前脂肪细胞或脂肪细胞。成功完成 这些目标将大大提高我们对脂肪纤维细胞相互作用的理解，这对 开发更有效和改进的伤口愈合临床疗法。