Small Molecules Promote Tendon Regeneration by Targeting Endogenous Stem Cells

小分子通过靶向内源干细胞促进肌腱再生

• 批准号: 10258102
• 负责人: Mo Chen
• 金额: $ 25.21万
• 依托单位: WNT SCIENTIFIC, LLC
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-05 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10258102
• 关键词: Acute; Agonist; Agreement; Area; Arthritis; Biological Assay; Businesses; Cardiotoxicity; Cells; Cicatrix; Collagen Fibril; Connective Tissue; DNA Sequence Alteration; Data; Development; Disease; Dose; Drug Delivery Systems; Drug Kinetics; Encapsulated; Ethers; FDA approved; Fibrin; Fostering; Funding; Genes; Goals; Grant; Healthcare; Heart; Heart Rate; Histologic; Human; Hydrogels; In Vitro; Inferior; Injectable; Investments; Laboratories; Legal patent; Libraries; Licensing; MAPK3 gene; MCAM gene; Maleates; Measures; Mediation; Muscarinic Acetylcholine Receptor; Musculoskeletal System; National Institute of Dental and Craniofacial Research; Natural regeneration; Neurons; Nicotinic Receptors; Operative Surgical Procedures; PPBP gene; PTK2 gene; Pathway interactions; Pharmaceutical Preparations; Pharmacology; Phase; Plasma; Play; Privatization; Process; Rattus; Regenerative engineering; Regenerative pathway; Research; Research Peer Review; Risk; Role; Safety; Signal Transduction; Small Business Technology Transfer Research; Small Interfering RNA; Technology; Technology Transfer; Temporomandibular Joint; Tenascin; Tendon Injuries; Tendon structure; Tensile Strength; Testing; Therapeutic; Therapeutic Index; Tissues; Topical application; Toxic effect; United States; Universities; Vimentin; Western Blotting; arthropathies; base; cost; cytotoxicity; dosage; healing; improved; in vitro testing; in vivo; innovative technologies; knock-down; ligament injury; nerve supply; novel; oxotremorine M; patellar tendon; physical property; receptor; regenerative; regenerative therapy; repaired; research and development; safety assessment; scleraxis; small molecule; stem; stem cells; tissue regeneration

Abstract Tendon and ligament injuries represent an acute healthcare burden in the United States, costing >$30 billion annually. Tendon injuries frequently result in scar-like tissue with inferior physical properties - however no regenerative therapy exists to date. Recently, we have identified and characterized perivascular (CD146+) tendon stem/progenitor cells (TSCs) that play an essential role in tendon healing via FAK and ERK1/2 signaling. In our preliminary study, we screened small molecules from a library of FAK and ERK1/2 agonists and identified Oxotremorine M (Oxo-M) and PPBP maleate (4-PPBP) that stimulated TSCs toward regenerative tendon healing. Oxo-M and 4-PPBP were originally developed for treating neuronal diseases but have never been tested in the musculoskeletal system. In vitro, a combination of Oxo-M and 4-PPBP induced significant increases in the expression of tendon-related genes involved in tendon repair. Oxo-M and 4-PPBP showed no cytotoxicity up to 10X working doses. Western blot and siRNA knockdown (KD) confirmed that FAK and ERK1/2 signaling regulate Oxo -M & 4-PPBP-induced tenogenic differentiation of TSCs. In vivo, direct topical delivery of Oxo-M and 4-PPBP onto full-transected rat patellar tendons (PT) significantly improved tendon healing, as observed histologically as densely reorganized collagen fibrils, and functionally as significantly enhanced tensile strength. This process was guided by a rapid but transient increase in the number endogenous TSCs undergoing tenogenic differentiation. In addition, Oxo-M and 4-PPBP specifically targeted CD146+ TSCs through muscarinic acetylcholine receptors (AChRs) and σ1 receptor (σ1R) pathways, with minimal effect on other types of tendon cells. These findings demonstrate a novel and promising activity of the combination of Oxo-M and 4-PPBP in tendon healing by specifically targeting endogenous TSCs. The overall objectives of this STTR grant are to develop a reliable and effective small molecule-based regenerative therapy for tendon injuries by transiently activating regenerative pathways of endogenous TSCs and repair tendon tears. The overarching goal of the STTR phase I grant is to optimize the combination of Oxo-M and 4-PPBP, the 2 compounds and determine their drugability in combination. The 2 aims of the phase I STTR are to obtain robust proof-of-concept and preliminary safety data to establish technical merit, feasibility, and commercial potential of the innovative technology.

摘要