SBIR-STTR Award

Peripheral nerve injury (PNI) is a common consequence of trauma or surgery, and the extent of functional recovery is often unsatisfactory. Moreover, a particular challenge is the repair of “major” PNI, defined as injuries where a large segment of a nerve is lost (i.e., >5 cm) or more proximal injuries (e.g., brachial plexus or upper arm) that necessitate extremely long distances for axonal regeneration to distal end targets (e.g., hand). Patients who experience major PNI face the high probability of a significant residual functional deficit, even following state-of-the-art surgical reconstruction. To address this gap in care, Axonova Medical has developed a revolutionary approach using tissue engineered nerve grafts (TENGs). TENGs are living three-dimensional nerve constructs that consist of longitudinally aligned axonal tracts spanning discrete neuronal populations. In rat and pig models of PNI, we have shown that allogeneic TENGs can circumvent the key limitations of current reconstructive strategies, notably the ability to (1) accelerate and direct axon regeneration across the zone of injury, and (2) sustain the regenerative environment of distal nerve structures and receptiveness of target muscles to enable host axons to reinnervate long-distance targets. Notably, in an exciting recent study, we found that TENGs serve as a “living scaffold” to promote rapid nerve regeneration and functional restoration in an extremely challenging major nerve gap (5 cm) model in pigs. The scope of the current proposal is to translate our preclinical protocols to create TENGs using a starting biomass suitable for eventual clinical testing in humans. Through a partnership with Revivicor, Axonova will pursue a xenogeneic product development strategy to build TENGs using so-called “humanized” pig neurons. Specifically, neurons used to generate TENGs will be sourced from Revivicor's custom and proprietary GalSafe® pigs that are genetically engineered to be non-immunogenic when transplanted into humans. These GalSafe® TENGs will then be tested in an established xenogeneic PNI repair model using athymic “nude” rats, and the regenerative efficacy will be compared to benchmarks established using allogeneic TENGs. Axonova's ultimate product will be a combination product (GalSafe® TENGs encased in a biodegradable wrap) that will be regulated under a Biologics License Application. Based on the company's preclinical data, TENGs not only represent a superior option to current reconstructive strategies, but a transformative, off-the-shelf solution for repair of currently untreatable major PNI. Successful execution of the milestones in this SBIR proposal will accelerate Axonova's path to clinical testing in humans. GalSafe® TENGs represent an innovative product and potentially transformative strategy for major PNI repair that can greatly improve the quality of life for hundreds of thousands of patients in the U.S alone.

Public Health Relevance Statement:
NARRATIVE Peripheral nerve injury (PNI) is a common consequence of trauma or surgery, and the extent of functional recovery is often unsatisfactory. Axonova Medical has developed a revolutionary approach to repair major PNI lesions using tissue engineered nerve grafts that serve as a “living scaffold” to promote rapid nerve regeneration and functional restoration. The scope of the current proposal is to translate our preclinical protocols to create a biological product using a source material suitable for eventual clinical testing in humans. Axonova's tissue engineered nerve grafts represent an innovative product and potentially transformative strategy for major PNI repair that can greatly improve the quality of life for hundreds of thousands of patients in the U.S alone.

NIH Spending Category:
Bioengineering; Biotechnology; Injury (total) Accidents/Adverse Effects; Neurodegenerative; Neurosciences; Peripheral Neuropathy; Regenerative Medicine; Transplantation

Project Terms:
Acute; Address; afferent nerve; Allogenic; Antigens; Autologous Transplantation; Axon; axon regeneration; base; Benchmarking; Biological Products; Biomass; Brachial plexus structure; Caring; Cells; Chemistry; clinical translation; Clinical Trials; complement system; Custom; Cyclic GMP; Data; density; Dimensions; Distal; Distant; Environment; Evaluation; experience; experimental study; Facial Injuries; Family suidae; Farming environment; fetal; Fetus; functional outcomes; functional restoration; Galactose; Generations; Genetic Engineering; Gold; Hand; Harvest; Health; Homologous Transplantation; Human; immunogenic; Immunosuppression; improved; In Vitro; in vivo; in vivo evaluation; Infiltration; Injury; injury and repair; innovation; Left; Length; Lesion; Licensing; manufacturing process; Medical; meetings; Modality; Modeling; Motor; Muscle; Natural regeneration; Nerve; nerve gap; Nerve Regeneration; Neurons; Nude Rats; Operative Surgical Procedures; Outcome; Patients; Peripheral nerve injury; Phase; Population; pre-clinical; Probability; Process; product development; Production; programs; Protocols documentation; Quality of life; Rattus; reconstruction; Recovery of Function; regenerative; reinnervation; repaired; research clinical testing; Residual state; Risk; Rodent; scaffold; Schwann Cells; sensor; Shoulder; Small Business Innovation Research Grant; Source; Structure; Target Populations; Testing; Tissue Engineering; Tissues; Translating; Transplantation; Trauma; Upper arm; Xenograft procedure

Tissue engineered medical products have tremendous promise to address the currently irreparable effects of neurotrauma and/or neurodegenerative diseases. Axonova Medical’s lead product, the tissue engineered nerve graft (TENG), is a bioactive “living scaffold” proven to promote rapid nerve regeneration and functional restoration when implanted to repair major peripheral nerve injury (PNI). Implementing living neuronal cells as the starting biomass, Axonova routinely biomanufactures TENGs comprised of living, long, aligned axon tracts through the proprietary process of axon “stretch-growth”. To date, TENGs have been shown to consistently promote host axon regeneration and improve functional recovery compared to current clinical practices in both small and large animal models of PNI, including the repair of 5 cm segmental defects. In order to move to a manufacturing- compliant biomass, Axonova has partnered with Revivicor to generate GalSafe® TENGs using their genetically engineered porcine neurons to mitigate a host immune response upon eventual transplantation in humans. Through the Phase I efforts, GalSafe® TENGs were successfully stretch-grown to create products at lengths of 1 cm, 3 cm, and 5 cm with a health and axon density that met or exceeded established quality benchmarks. Moreover, 1 cm TENGs were transplanted into a rodent PNI model to demonstrate regenerative efficacy, revealing that GalSafe® TENGs survived as well as actively drove host axonal regeneration via the proprietary mechanism-of-action of TENGs, referred to as axon facilitated axon regeneration (AFAR). Along with choosing a safe and readily available starting biomass, other product development challenges include biopreservation and quality assurance criteria to allow increased shelf life, transportation, and maintained efficacy of the product. Accordingly, the current Phase II SBIR program addresses a key facet of clinical grade biomanufacturing, specifically the validation of an effective storage protocol and release criteria ensuring product health, sterility, and potency. TENGs will be biofabricated from a cGMP-compliant cell source (GalSafe® porcine neurons), cold- stored using a cGMP-compliant preservation protocol, and then tested in small and large animal models of PNI to demonstrate regenerative efficacy. Specifically, Axonova will systematically complete the objectives of this program across 3 Specific Aims: (1) determine the efficacy of fresh GalSafe® TENGs in an established porcine PNI model; (2) effectively biopreserve GalSafe® TENGs to increase shelf-life and validate non-invasive release criteria; and (3) determine the potency and efficacy of biopreserved GalSafe® TENGs in vivo in rodent and porcine PNI models. This program addresses a major challenge in the translation of next-generation tissue engineered medical products by validating manufacturing, storage and release criteria for a living biological product to enable eventual testing in humans. The ability to store and distribute GalSafe® TENGs as an “off-the- shelf” product will vastly increase commercial potential by reducing costs and expanding patient access, thereby improving recovery and quality of life for patients suffering from the debilitating effects of major PNI.

Public Health Relevance Statement:
NARRATIVE Axonova Medical’s lead product, the tissue engineered nerve graft (TENG), is a bioactive “living scaffold” proven to promote rapid nerve regeneration and functional restoration in challenging large animal models of peripheral nerve injury (PNI). This Phase II SBIR addresses a key facet of clinical grade biomanufacturing – specifically the validation of an effective storage protocol and release criteria ensuring product health, sterility, and potency – where TENGs will be biofabricated from a cGMP-compliant cell source, cold-stored using a cGMP-compliant preservation protocol, and then tested in a large animal model of PNI to demonstrate regenerative efficacy. This program addresses a major challenge in the translation of next-generation tissue engineered medical products by validating storage and release criteria for a living biological product – necessary for eventual testing in humans – and enabling the ability to store and distribute TENGs as an “off- the-shelf” product that will increase commercial potential by reducing costs and expanding patient access.

Project Terms:
3-Dimensional; Address; afferent nerve; Animal Model; Antigens; Autologous Transplantation; Axon; axon regeneration; base; Benchmarking; biofabrication; Biological; Biological Products; Biomanufacturing; Biomass; Brachial plexus structure; Caring; Cells; Clinical; clinical practice; clinical translation; Clinical Trials; complement system; cost; Cryopreservation; Cyclic GMP; Data; Defect; density; Distal; Distant; efficacy study; efficacy testing; Ensure; Face; Family suidae; functional restoration; Galactose; Generations; Genetic Engineering; Gold; Graft Survival; Growth; Hand; Harvest; Health; Human; Image; Immune response; immunocytochemistry; Implant; improved; In Vitro; in vivo; Injury; injury and repair; Lactate Dehydrogenase; Lead; Length; Lesion; Licensing; Life; Measures; Medical; meetings; Metabolic; Methods; Modeling; Motor; Nerve; nerve autograft; Nerve Regeneration; Nervous System Trauma; Neurodegenerative Disorders; Neurons; next generation; Operative Surgical Procedures; Outcome; Patients; Peripheral nerve injury; Phase; Population; pre-clinical; preservation; Probability; Process; product development; programs; Protocols documentation; quality assurance; Quality of life; reconstruction; Recovery; Recovery of Function; regenerative; reinnervation; repaired; research clinical testing; Rodent; safety study; scaffold; Small Business Innovation Research Grant; Source; Stains; Sterility; Stretching; success; Target Populations; Techniques; Testing; Tissue Engineering; Translations; Transplantation; Transportation; Trauma; Upper arm; Validation