SBIR-STTR Award

This study aims to prove the concept and feasibility that TendonCure, extracellular vesicles that contain signaling molecules (i.e. exosomes), secreted by mesenchymal stem cells (i.e. tendon stem/progenitor cells [TSPCs] or adipose-derived stem cells [ADSCs]) from individual patients (donors) cultured on a novel scaffold exert therapeutic efficacy on tendinopathy when injected into the diseased tendon of the same donor. Tendinopathy is a common chronic tendon disorder that affects 30-50% of individuals over 60 years old. It is characterized by pain, swelling, loss of function, and impaired performance. There is currently no cure for tendinopathy. Spontaneous repair or treatment typically leads to scar formation, resulting in a weakened tissue with reduced function and mechanical properties that may ultimately rupture with further use. Our previous and preliminary studies show that MSCs grown on the TendonCure scaffold are modulated towards tenocyte differentiation with enhanced tenogenesis-related gene expression. Furthermore, TSPCs cultured with exogenous TendonCure-T or TendonCure-A, TendonCure derived from TSPCs or ADSCs, respectively, exhibited tenogenesis-related gene expression profile changes, with increased expression of tenogenic markers and decreased expression of gene markers for the adipo- and chondrogenic lineages. Importantly, local injection of TendonCure showed therapeutic efficacy in an overuse-induced supraspinatus tendinopathy rat model in vivo. While the results are promising, the suggested efficacy of TendonCure was based on statistically significant results in TendonCure from MSCs of a young to middle-aged group. However, whether TendonCure derived from MSCs of aged individuals exerts such therapeutic effects is unknown. We therefore hypothesize that TendonCure from MSCs of aged individuals exerts therapeutic efficacy on tendinopathy. TendonCure-T and TendonCure-A will be harvested from TSPCs and ADSCs, respectively, which are isolated from aged patients at 65-80 y.o. and cultured on TendonCure scaffolds. Control exosomes will be derived from TSPCs and ADSCs from the same patients as the TendonCure groups and grown in regular 2D culture. Adult nude rats subjected to decline treadmill at two weeks (with expected mild tendinopathy) will be injected weekly in the supraspinatus tendons with TendonCure or control exosomes, or placebo (phosphate buffered saline), or kept for normal cage activity (control). Four weeks after injection, animals will be evaluated for efficacy on pain behaviors, and supraspinatus tendons will be dissected for efficacy evaluation with assays for histology and mechanical properties testing. Upon successful completion of Phase I and II studies, we will carry out clinical trials focusing on common sites of tendinopathy. TendonCure will be marketed as a biologic for treating tendinopathy and other tendon disorders. 1

Public Health Relevance Statement:
Project Narrative Tendinopathy is a common chronic tendon disorder, especially in the aged population, characterized by pain and impaired performance, and currently has no cure or effective treatments. Our preliminary studies show exosomes, extracellular vesicles secreted by mesenchymal stem cells (MSCs) from young donors, when grown on a novel scaffold, exert therapeutic efficacy in healing diseased tendon tissue by carrying specific signaling molecules as active ingredients. The proposed study will determine whether exosomes from MSCs of aged donors grown on this novel scaffold also have therapeutic efficacy in treating tendinopathy.

Project Terms:
achilles tendon; Adipose tissue; Adult; Affect; age group; aged; aging population; Alpha Cell; Animal Model; Animals; Anti-inflammatory; Anti-Inflammatory Agents; Apoptotic; Autologous; base; Behavior; Behavior assessment; Biological Assay; Biological Products; Businesses; cell age; Cells; Characteristics; Chronic; Cicatrix; Clinical; Clinical Trials; Collagen; collagenase; commercialization; Communication; Control Groups; Cultured Cells; Data; design; Disease; Dose; effective therapy; efficacy evaluation; Evaluation; Exhibits; exosome; extracellular vesicles; Frequencies; Gene Expression; Gene Expression Profile; Harvest; healing; Histologic; Histology; Immunohistochemistry; Impairment; improved; in vivo Model; Individual; individual patient; Injectable; Injection of therapeutic agent; Intervention; Investigation; Legal patent; Locomotion; loss of function; mechanical properties; Medical center; Mesenchymal Stem Cells; MicroRNAs; middle age; Modeling; Morphology; nanoscale; New York; novel; Nude Rats; Operative Surgical Procedures; Oryctolagus cuniculus; Outcome; Pain; pain behavior; patellar tendon; Patients; Performance; Phase; Phenotype; Phosphate Buffer; Placebo Control; Placebos; Play; protein biomarkers; Proteins; Protocols documentation; Quality Control; Rattus; repaired; research and development; Role; Rotator Cuff; Rupture; Saline; scaffold; scleraxis; Shapes; Signaling Molecule; Silk; Site; stem; Stem cells; Stress; supraspinatus muscle; Surface; Swelling; Tendinopathy; Tendon Injuries; Tendon structure; Testing; Therapeutic; Therapeutic Effect; Tissues; tool; translational medicine; Travel; treadmill; Treatment Efficacy; two-dimensional; Vesicle

Tendinopathy is a tendon disorder characterized by tendon deterioration that often leads to tendon rupture, and is associated with pain, swelling and impaired performance. There is currently no cure for tendinopathy. Therefore, there is an urgent need for effective treatments for tendinopathy. Exosomes are specialized membranous nano-sized extracellular vesicles derived from endocytic compartments that are released by many cell types. Our initial discovery was that exosomes secreted from tendon derived stem/progenitor cells (TSPCs) cultured on a novel scaffold, when injected into a tendinopathic tendon, mitigated pathology and pain in a rat tendinopathy model. We therefore designated the exosomes derived from MSCs cultured on this novel scaffold as “TenoGen,” and established a project to develop TenoGen as an FDA-approved biologic for tendinopathy treatment. In our Phase I study, we found that the exosomes derived from human adipose-derived stem cells (ADSCs) exert an efficacy on tendinopathy that was comparable to that derived from TSPCs, and revealed that TenoGen produced by TSPCs or ADSCs from aged donors exerts a therapeutic effect on tendinopathy. Furthermore, we examined and found no general signs of toxicity in the rats treated with TenoGen. These exciting results in the Phase I study encouraged us to further test our hypothesis that TenoGen exerts a therapeutic effect in mitigating tendinopathy pathology and relieving tendinopathy-related pain and symptoms with no or minimal adverse effects. The Phase II study will focus on providing further critical evidence towards developing TenoGen as an FDA-approved biologic for the autologous or allogeneic treatment of tendinopathy. We will first determine the efficacy and safety of TenoGen derived from human ADSCs in a tendinopathy model in nude rats (Aim 1). Specifically, we will first determine the optimal dose and dosing schedule of TenoGen and determine the efficacy of TenoGen on pathology of tendinopathy. By using the selected optimal dose and optimal dosing schedule. We will further determine efficacies of TenoGen on improving the mechanical properties of the diseased tendon, and on relieving pain and behaviors related to tendinopathy. Furthermore, in alignment with the regulatory requirements for TenoGen as a novel biologic, the safety of human TenoGen will be evaluated in this immunodeficient animal model. In Aim 2, we will determine the efficacy and safety of TenoGen for autologous and allogeneic treatment in a tendinopathy model in rabbits, which allows for the evaluation of TenoGen on tendinopathy in a mid-sized animal model that closely mimics the future treatment protocol in humans. Successful completion of these studies will provide critical preclinical evidence to support the efficacy and safety of TenoGen as a biologic for the allogeneic treatment of tendinopathy. The data will be instrumental for a FDA IND application, and the future R&D towards initiating clinical trials in humans. 1

Public Health Relevance Statement:
Project Narrative Tendinopathy is a common chronic tendon disorder, characterized by pain and impaired performance, and currently has no cure or effective treatments. Our Phase I studies showed that exosomes, nanosized extracellular vesicles secreted by mesenchymal stem cells, when grown on the TenoGen scaffold, a novel 3- dimensional scaffold, exert therapeutic efficacy in healing diseased tendon tissue in rats. The proposed study will test the efficacy and safety of the intended clinical product in an immunodeficient rodent model, and of an analogous product in a rabbit model via autologous and allogeneic treatment.

Project Terms:
3-Dimensional; Address; Adipose tissue; Adverse effects; aged; Allogenic; Animal Model; Animals; Attention; Autologous; base; Biological; Biological Response Modifier Therapy; Businesses; cell age; cell type; Chronic; Cicatrix; Clinical; Clinical Research; Clinical Trials; collagenase; commercial application; commercialization; Cultured Cells; Data; Deterioration; Disease; Dose; effective therapy; efficacy testing; Evaluation; exosome; extracellular vesicles; FDA approved; Fiber; Future; Harvest; healing; Histopathology; Human; immunogenic; Impairment; improved; Inflammatory Infiltrate; Investigation; Investments; Legal patent; mechanical properties; Mesenchymal Stem Cells; Modeling; Morbidity - disease rate; nanosized; New York; novel; Nude Rats; Operative Surgical Procedures; Oryctolagus cuniculus; Outcome; Pain; pain behavior; pain relief; pain symptom; Pathology; Performance; phase 1 study; phase 2 study; pre-clinical; Rattus; repaired; research and development; Rodent Model; Rupture; Safety; scaffold; Schedule; Silk; Site; stem; stem cells; Swelling; Tendinopathy; tendon rupture; Tendon structure; Testing; Therapeutic; Therapeutic Effect; Tissues; Toxic effect; translational medicine; Treatment Efficacy; Treatment Protocols; United States