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 ?1 receptor (?1R) 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. Public Health Relevance Statement Tendon and ligament injuries are an acute healthcare burden in the United States, costing >$30 billion annually. The goal of this research is to develop a regenerative product for tendon repair.
Project Terms: Arthritis ; arthritic ; Biological Assay ; Assay ; Bioassay ; Biologic Assays ; Western Blotting ; Western Immunoblotting ; protein blotting ; Cells ; Cell Body ; Cicatrix ; Scars ; Connective Tissue ; Disease ; Disorder ; Pharmaceutical Preparations ; Drugs ; Medication ; Pharmaceutic Preparations ; drug/agent ; Ethers ; Fibrin ; Genes ; Goals ; Grant ; Heart ; Heart Rate ; Cardiac Chronotropism ; Human ; Modern Man ; In Vitro ; nerve supply ; innervation ; Investments ; arthropathies ; Joint Diseases ; arthropathic ; arthropathy ; joint disorder ; Laboratories ; Libraries ; Maleates ; Musculoskeletal System ; locomotor system ; Neurons ; Nerve Cells ; Nerve Unit ; Neural Cell ; Neurocyte ; neuronal ; Legal patent ; Patents ; Drug Kinetics ; Pharmacokinetics ; Pharmacology ; Plasma ; Blood Plasma ; Plasma Serum ; Reticuloendothelial System, Serum, Plasma ; Play ; Privatization ; Rattus ; Common Rat Strains ; Rat ; Rats Mammals ; Muscarinic Acetylcholine Receptor ; Muscarinic Receptors ; Nicotinic Receptors ; Nicotinic Acetylcholine Receptors ; Natural regeneration ; Regeneration ; regenerate ; Research ; research and development ; Development and Research ; R & D ; R&D ; Risk ; Role ; social role ; Safety ; Signal Transduction ; Cell Communication and Signaling ; Cell Signaling ; Intracellular Communication and Signaling ; Signal Transduction Systems ; Signaling ; biological signal transduction ; stem cells ; Progenitor Cells ; Technology ; Temporomandibular Joint ; Jaw Joint ; Mandibular joint ; TMJ ; Tendon Injuries ; Tendon structure ; Tendons ; Tensile Strength ; Testing ; Tissues ; Body Tissues ; United States ; Universities ; Vimentin ; oxotremorine M ; Tenascin ; Cytotacin ; Cytotactin ; Hexabrachion ; Tenascin-C ; Measures ; Drug Delivery Systems ; Drug Delivery ; Businesses ; Healthcare ; health care ; Injectable ; base ; dosage ; improved ; Area ; Acute ; Encapsulated ; repaired ; repair ; Phase ; Histologic ; Histologically ; Research Peer Review ; Fostering ; Licensing ; Technology Transfer ; Funding ; Agonist ; scleraxis ; Therapeutic ; damage to ligament ; ligament damage ; ligamentous injury ; ligament injury ; Inferior ; Operative Procedures ; Surgical ; Surgical Interventions ; Surgical Procedure ; surgery ; Operative Surgical Procedures ; innovative technologies ; Lytotoxicity ; cytotoxicity ; physical property ; Receptor Protein ; receptor ; Hydrogels ; Toxicities ; Toxic effect ; Therapeutic Index ; novel ; Agreement ; Negotiating ; Negotiation ; Mediation ; Topical Drug Administration ; administer topically ; apply topically ; deliver topically ; topical administration ; topical delivery ; topical drug application ; topical treatment ; topically administered ; topically applied ; topically delivered ; topically treated ; treat topically ; Topical application ; Cardiac Toxicity ; Cardiotoxic ; Cardiotoxicity ; Short interfering RNA ; siRNA ; Small Interfering RNA ; Collagen Fibril ; small molecule ; CD146 ; Chicken Homolog of Gicerin ; MCAM ; MUC18 ; Melanoma Adhesion Molecule ; Melanoma-Associated Glycoprotein MUC18 ; MCAM gene ; Beta-TG ; CTAP III ; CTAP3 ; CTAPIII ; CXC Chemokine Ligand 7 ; CXCL7 ; Connective Tissue-Activating Peptide III ; LA-PF4 ; NAP-2 ; NAP2 ; Neutrophil-Activating Peptide 2 ; PBP gene ; PBP protein ; PPBP ; Platelet Basic Protein ; Pro-Platelet Basic Protein ; SCYB7 ; Small Inducible Cytokine Subfamily B, Member 7 ; TC1 ; TC2 ; TGB1 ; Thrombocidin 1 ; Thrombocidin 2 ; Thromboglobulin, Beta-1 ; beta-Thromboglobulin ; β-TG ; β-Thromboglobulin ; PPBP gene ; ERK 1 ; ERK1 ; ERK1 Kinase ; Extracellular Signal-Regulated Kinase 1 ; MAP Kinase 3 ; MAPK3 ; MAPK3 Mitogen-Activated Protein Kinase ; Mitogen-Activated Protein Kinase 3 ; Mitogen-Activated Protein Kinase 3 Gene ; P44ERK1 ; PSTkinase p44mpk ; p44 MAPK ; MAPK3 gene ; FADK ; FAK ; FAK1 ; PTK2 ; pp125FAK ; PTK2 gene ; Dose ; DNA Alteration ; DNA mutation ; Genetic mutation ; Sequence Alteration ; genomic alteration ; DNA Sequence Alteration ; Data ; NIDCR ; NIDR ; National Institute of Dental Research ; National Institute of Dental and Craniofacial Research ; in vivo ; Small Business Technology Transfer Research ; STTR ; Process ; Development ; developmental ; tissue regeneration ; regenerate new tissue ; regenerate tissue ; regenerating damaged tissue ; regenerating tissue ; tissue renewal ; tissue specific regeneration ; Pathway interactions ; pathway ; cost ; healing ; knock-down ; knockdown ; stem ; regenerative therapy ; regeneration based therapy ; regeneration therapy ; regenerative therapeutics ; FDA approved ; regenerative ; in vitro testing ; patellar tendon ; safety assessment ; Regenerative pathway ; regeneration pathway ; Regenerative engineering ;
