Role of TRPV4 and YAP/TAZ in Tendon Fibrosis and Engineered Tendon Development

TRPV4 和 YAP/TAZ 在肌腱纤维化和工程肌腱发育中的作用

• 批准号: 10546427
• 负责人: Thomas Lee Jenkins
• 金额: $ 4.77万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10546427
• 关键词: Affect; Agonist; Animal Model; Architecture; Attenuated; Biocompatible Materials; Bioreactors; Bone Development; Cadaver; Calcium Signaling; Cations; Cicatrix; Collagen; Control Groups; Data; Data Set; Deposition; Development Plans; Disease; Drug Design; Engineering; Environment; Exhibits; Extracellular Matrix; Faculty; Fibroblasts; Fibrosis; Future; Gene Expression; Goals; Harvest; Health; Human; Image; In Vitro; Injury; Institution; Knock-out; Liver; Liver Fibrosis; Lung; Manuscripts; Mechanical Stimulation; Mechanics; Mentors; Molecular Biology; Motion; Mus; Myofibroblast; Natural regeneration; Nuclear; Operative Surgical Procedures; Outcome; Physiological; Play; Polarization Microscopy; Postoperative Period; Process; Production; Proteins; Proteomics; Publications; Pulmonary Fibrosis; Quality of life; Research; Research Design; Research Personnel; Rodent Model; Role; Rotator Cuff; Signal Transduction; Sirius Red F3B; Stains; Structure; Tendon Injuries; Tendon structure; Testing; Tissue Engineering; Tissues; Training; Transcription Coactivator; Transcriptional Activation; Transcriptional Coactivator with PDZ-Binding Motif; Verteporfin; Work; antagonist; biceps brachii muscle; caprolactone; career; career development; clinical translation; coronary fibrosis; design; healing; improved; in vivo; infraspinatous muscle; keratinocyte; kidney fibrosis; mechanical properties; mechanotransduction; member; overexpression; pharmacologic; preclinical study; prevent; protein expression; receptor; repaired; response; rotator cuff tear; scaffold; skills; small hairpin RNA; supraspinatus muscle; symposium; tendon development; transcriptome sequencing

Project Summary/Abstract More than 300,000 rotator cuff surgeries occur annually in the US. However, treatment is often complicated by disorganized collagen matrix formed via fibrosis and results in high re-tear rates. Tendon tissue engineering seeks to solve the problem using biomaterials to promote neo-tendon formation to augment repair or regenerate tendon. However, while current biomaterials provide the opportunity to improve tendon healing, they frequently still exhibit fibrosis in preclinical studies. Therefore, a critical need exists to understand the mechanisms of aligned collagen formation when designing biomaterials for tendon tissue engineering. Matrix architecture and transient receptor potential cation channel subfamily V member 4 (TRPV4) regulate aligned collagen formation during tenogenesis in vitro, but the mechanism remains to be determined. Recently, TRPV4 stimulation was found to induce nuclear localization and activation of transcriptional co-activators Yes-associated protein (YAP) and transcriptional co-activator with PDZ-binding motif (TAZ) in keratinocytes and myofibroblasts during fibrosis. YAP expression is upregulated during tendon development, a process characterized by aligned collagen formation, and in response to physiological mechanical stimulation, suggesting it could play an important role in tendon. Both TRPV4 and YAP/TAZ are dysregulated in lung, liver, kidney, and cardiac fibrosis. However, the role of TRPV4 and YAP/TAZ, and their interactions, have not been investigated in tendon fibrosis or during engineered tendon development. The overall hypothesis is that TRPV4 regulates collagen alignment via YAP/TAZ during tenogenesis on biomaterials and stimulates neo-tendon formation to a similar extent as tensile loading. In contrast, dysregulation of this relationship between YAP/TAZ and TRPV4 will promote fibrotic matrix deposition. Aim 1 will compare the role of YAP/TAZ and TRPV4 in healthy and fibrotic rotator cuff tendon. Aim 2 will investigate the interaction between TRPV4 and YAP/TAZ in aligned collagen formation during tenogenesis on electrospun biomaterials in ‘healthy’ and ‘fibrotic’ conditions. Aim 3 will evaluate how TRPV4 stimulation affects neo-tendon formation on biomaterials. This information will directly inform planned preclinical studies of rotator cuff repair initially in rodent models, before, if successful, moving to large animal models and clinical translation. Completing the proposed studies will provide me with the opportunities and training to support my career goals and develop the skills I need to achieve my long-term goal of an independent faculty career at a research-intensive institution.

项目概要/摘要 美国每年进行超过 300,000 例肩袖手术，但治疗往往很复杂。 通过纤维化形成混乱的胶原基质，导致肌腱组织工程的高再撕裂率。 寻求使用生物材料促进新肌腱形成以增强修复或再生来解决问题 然而，虽然目前的生物材料提供了改善肌腱愈合的机会，但它们经常发生。 在临床前研究中仍然表现出纤维化，因此迫切需要了解其机制。 在设计用于肌腱组织工程的生物材料和基质时对齐胶原蛋白的形成。 瞬时受体电位阳离子通道亚家族 V 成员 4 (TRPV4) 调节排列胶原形成 最近，TRPV4 刺激在体外进行，但其机制仍有待确定。 发现可诱导转录辅激活因子 Yes 相关蛋白 (YAP) 的核定位和激活 以及纤维化过程中角化细胞和肌成纤维细胞中具有 PDZ 结合基序 (TAZ) 的转录共激活因子。 YAP 表达在肌腱发育过程中上调，这一过程的特点是胶原蛋白对齐 形成，并响应生理机械刺激，表明它可以在 TRPV4 和 YAP/TAZ 在肺、肝、肾和心脏纤维化中均失调。 TRPV4 和 YAP/TAZ 的作用及其相互作用尚未在肌腱纤维化或肌腱纤维化过程中进行研究。 总体假设是 TRPV4 通过调节胶原排列。 YAP/TAZ 在生物材料的肌腱形成过程中刺激新肌腱形成，其程度与拉伸相似 相反，YAP/TAZ 和 TRPV4 之间这种关系的失调会促进基质纤维化。 目标 1 将比较 YAP/TAZ 和 TRPV4 在健康和纤维化肩袖肌腱中的作用。 将研究肌腱形成过程中 TRPV4 和 YAP/TAZ 之间在排列胶原蛋白形成中的相互作用 目标 3 将评估 TRPV4 刺激的效果。 该信息将直接影响生物材料上的新肌腱形成。 肩袖修复最初在啮齿动物模型中进行，如果成功，然后转移到大型动物模型和临床 完成拟议的研究将为我提供支持我的机会和培训。 职业目标并培养我实现独立教师职业的长期目标所需的技能 研究密集型机构。

项目成果

Tendon-derived matrix crosslinking techniques for electrospun multi-layered scaffolds.

• DOI: 10.1002/jbm.a.37588
• 发表时间: 2023-07
• 期刊: Journal of biomedical materials research. Part A
• 作者: Thomas L Jenkins;Paula A Sarmiento Huertas;Kentaro Umemori;F. Guilak;D. Little