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.

抽象的 在美国，肌腱和韧带损伤是一项严重的医疗负担，造成的损失超过 300 亿美元 肌腱损伤经常导致物理性能较差的疤痕样组织 - 但并非如此。 迄今为止，再生疗法已经存在，我们已经识别并表征了血管周围（CD146+）。 肌腱干/祖细胞 (TSC) 通过 FAK 和 ERK1/2 在肌腱愈合中发挥重要作用 发信号。 在我们的初步研究中，我们从 FAK 和 ERK1/2 激动剂库中筛选了小分子，并 鉴定出 Oxotremorine M (Oxo-M) 和 PPBPmaleate (4-PPBP) 可刺激 TSC 再生 Oxo-M 和 4-PPBP 最初是为治疗神经元疾病而开发的，但从未开发过。 在体外肌肉骨骼系统中进行了测试，Oxo-M 和 4-PPBP 的组合可产生显着的效果。 参与肌腱修复的肌腱相关基因的表达增加。 高达 10 倍工作剂量时没有细胞毒性，Western blot 和 siRNA 敲低 (KD) 证实 FAK 和 ERK1/2 信号传导在体内直接调节 Oxo-M 和 4-PPBP 诱导的 TSC 肌腱分化。 将 Oxo-M 和 4-PPBP 局部递送到全横断的大鼠髌腱 (PT) 上的效果显着改善 肌腱愈合，组织学上观察到密集重组的胶原纤维，功能上观察到 显着增强的拉伸强度是由快速但短暂的增加引导的。 此外，Oxo-M 和 4-PPBP 还特别强调了正在进行肌腱分化的内源性 TSC 的数量。 通过毒蕈碱乙酰胆碱受体 (AChR) 和 σ1 受体 (σ1R) 靶向 CD146+ TSC 这些发现证明了一种新颖且对其他类型肌腱细胞的影响。 Oxo-M 和 4-PPBP 的组合通过特异性靶向作用在肌腱愈合中具有良好的活性 内源性 TSC。 该 STTR 赠款的总体目标是开发一种可靠且有效的基于小分子的 通过短暂激活内源性再生途径来治疗肌腱损伤 STTR 第一阶段资助的总体目标是优化肌腱撕裂。 Oxo-M 和 4-PPBP 这 2 种化合物的组合并确定它们组合的成药性。 第一阶段 STTR 的目标是获得可靠的概念验证和初步安全数据，以建立 创新技术的技术优点、可行性和商业潜力。