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.

项目总结/摘要 在美国，每年治疗损伤和疾病引起的纤维化和疤痕花费200亿美元，即使在治疗时， 仍然是一个持续存在的问题，大大降低了生活质量。使用自体脂肪 用于重建手术的微型移植物似乎有利于减少疤痕。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中的作用而受到关注。的 本提案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-肌成纤维细胞 转化和细胞外基质（ECM）重塑。我们的长期目标是 用于伤口愈合的更有效和改进的临床疗法。为了实现这一目标，我们提出三个 旨在解开脂肪谱系细胞和成纤维细胞之间的通讯。在目标1中， 我们将确定来自脂肪细胞谱系细胞的分泌因子如何调节成纤维细胞-成肌纤维细胞转化， 表型和ECM重塑。我们假设脂肪细胞谱系细胞差异调节抑制， 诱导和逆转成纤维细胞向肌成纤维细胞的转化，这反过来又影响了细胞的组成。 ECM生产。我们还假设脂肪因子诱导的肌成纤维细胞（与TGF-β1诱导的相比） 肌成纤维细胞）产生不同的ECM，导致较少的瘢痕形成。我们将测试这些假设，并确定如何 来自脂肪细胞谱系细胞的单向通讯利用一种 肌成纤维细胞转化试验结合分级分离、蛋白质组学、计算基因组学和 抑制/加回测定。在目标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， 改造ECM。我们假设脂肪细胞系细胞和成纤维细胞之间的相互交流- 肌成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 由成纤维细胞和脂肪细胞谱系细胞组成。我们将使用我们的3D脂肪球体和3D体外模型 询问相关细胞类型与ECM的组成和机制之间的相互作用 制作。在目标3中，我们将通过确定如何移植 脂肪细胞谱系细胞改变猪创伤模型中的成纤维细胞行为和ECM重塑。我们假设 伤口床成纤维细胞对基于脂肪的治疗的反应会有所不同， 组成交付。我们将通过评估移植脂肪干细胞的效果来验证这一假设， 前脂肪细胞或脂肪细胞对成纤维细胞/肌成纤维细胞行为和伤口愈合的影响。成功完成 这些目标将大大提高我们对脂肪成纤维细胞相互作用的理解， 开发更有效和改进的伤口愈合临床疗法。