Parkinson's Disease (PD) affects more than 1 million adults in the U.S, and cell replacement therapy(CRT) using human embryonic stem cell (hESC) derived midbrain dopaminergic (DA) neurons has showngreat promise in animal studies in alleviating symptoms of PD. However, current manufacturing strategies forcell therapies are constrained by the industry standard format - monolayer, 2-dimensional (2D), cell culture -that additionally suffer from the lot-to-lot variability of animal-derived materials such as Matrigel as well asharsh enzymatic cell retrieval methods. Axent Biosciences Inc. is developing a proprietary 3-dimensional (3D)hydrogel based culture method to offer a critical extra dimension for cells to expand during production andthereby significantly increase the cell quantity produced per culture volume. Furthermore, our proprietaryhydrogel technology is fully synthetic and avoids animal derived products, and its temperature responsivephase change enables gentle, high viability cell harvesting by cooling. We have shown that our 3D cultureapproach enables highly scalable expansion of hESCs, followed by 3D differentiation to DA neurons. Notably,we have achieved up to 25 times the quantity of potent DA neurons produced per culture volume with our 3Dculture format compared to standard 2D formats. Moving forward, our overall objective is to scale up our 3Dcell manufacturing process to produce high-purity, functional hESC-derived DA neurons and validate their invitro and in vivo functionality to develop our Chemistry, Manufacturing, and Controls (CMC) towards filing anInvestigational New Drug (IND) application. In Aim 1, we will scale up production of hESC-derived DA neuronsto a pilot bioreactor capable of suppling sufficient numbers of cells for a Phase I clinical trial. In Aim 2, we willdevelop a purification strategy to remove residual contaminating cell types. In Aim 3, we will validate in vivofunctionality of DA neurons produced in 3D bioreactor process and will benchmark against DA neuronsgenerated in a conventional 2D production format. Successful completion of these aims will yield the basis fora cGMP-compliant manufacturing platform capable of producing higher quantities of functional DA neurons perculture volume compared to current 2D culture formats and thereby overcome a significant bottleneck for thedevelopment of PD cell therapies. Furthermore, our manufacturing process is generalizable and can be tunedto manufacture numerous cell types from hESCs. Finally, the purification process will minimize the risk ofcontaminating cell types in the final formulation, thereby increasing the safety profile of our candidate therapyin preparation for an FDA INTERACT meeting and subsequent filing of an IND application. To build toward thisgoal further in a Phase II SBIR proposal, we will include IND-enabling preclinical studies - particularly a 9-month animal study to validate safety - to progress translation of our DA neuron therapy for PD to the clinic.
Public Health Relevance Statement: PROJECT NARRATIVE
Parkinson's disease (PD) affects more than 1 million adults in the U.S, and cell replacement therapy (CRT)
using human dopaminergic (DA) neurons has fortunately shown great promise in alleviating symptoms of PD in
animal studies and human trials. However, manufacturing sufficient numbers of cells for clinical development
and commercialization is a major challenge. To overcome this severe bottleneck, Axent Biosciences is
developing an innovative 3D manufacturing strategy for producing consistent and high-quality hESC-derived
DA neurons that will make cell therapies accessible to the large patient population in need.
Project Terms: Adult ; 21+ years old ; Adult Human ; adulthood ; Affect ; Animals ; Antibodies ; Biological Sciences ; Biologic Sciences ; Bioscience ; Life Sciences ; Brain ; Brain Nervous System ; Encephalon ; Cell Count ; Cell Number ; Cell Culture Techniques ; cell culture ; Cell Survival ; Cell Viability ; Cells ; Cell Body ; Chemistry ; Corpus striatum structure ; Corpus Striatum ; Striate Body ; Striatum ; striatal ; Disease ; Disorder ; Animal Disease Models ; Dopamine ; Hydroxytyramine ; Investigational Drugs ; Investigational New Drugs ; Excipients ; Goals ; Cyclic GMP ; Guanosine Cyclic Monophosphate ; cGMP ; Human ; Modern Man ; In Vitro ; Industrialization ; Midbrain structure ; Mesencephalon ; Mid-brain ; Midbrain ; Methods ; Neuroglia ; Glia ; Glial Cells ; Kolliker's reticulum ; Neuroglial Cells ; Non-neuronal cell ; Nonneuronal cell ; nerve cement ; Parkinson Disease ; Paralysis Agitans ; Parkinson ; Parkinson's disease ; Parkinsons disease ; Primary Parkinsonism ; Patients ; Production ; Rattus ; Common Rat Strains ; Rat ; Rats Mammals ; Risk ; Rotation ; Running ; Safety ; Technology ; Temperature ; Time ; Tissues ; Body Tissues ; Translations ; matrigel ; Neurites ; Oxidopamine ; 6-OHDA ; 6-hydroxydopamine ; Injectable ; Investigational New Drug Application ; base ; density ; Area ; Clinical ; Residual state ; Residual ; Phase ; Histologic ; Histologically ; Undifferentiated ; CD140a Antigens ; PDGF alpha Receptor ; PDGF receptor α ; PDGF-R-alpha ; PDGFR-α ; PDGFRα ; Platelet-Derived Growth Factor Receptor Alpha Polypeptide ; platelet-derived growth factor receptor α ; Platelet-Derived Growth Factor alpha Receptor ; cell mediated therapies ; cell-based therapeutic ; cell-based therapy ; cellular therapy ; Cell Therapy ; Bioreactors ; Dimensions ; Contralateral ; Clinic ; Protocol ; Protocols documentation ; cell type ; System ; 3-D ; 3D ; three dimensional ; 3-Dimensional ; fetal ; Best Practice Analysis ; Benchmarking ; meetings ; phase change ; Hydrogels ; sorting ; Sorting - Cell Movement ; Touch ; tactile sensation ; Touch sensation ; motor deficit ; DA Neuron ; Dopamine neuron ; dopaminergic neuron ; Modeling ; Pluripotent Stem Cells ; flasks ; stem cell based therapy ; stem cell mediated therapy ; stem cell therapeutics ; stem cell treatment ; stem cell-based treatment ; stem cell therapy ; monolayer ; Early-Stage Clinical Trials ; Phase 1 Clinical Trials ; phase I protocol ; Phase I Clinical Trials ; cell dimension ; hESC ; human ES cell ; human ESC ; human embryonic stem cell ; Dose ; Dopaminergic Cell ; Harvest ; Motor ; Retrieval ; in vivo ; Small Business Innovation Research Grant ; SBIR ; Small Business Innovation Research ; Preparation ; Process ; Development ; developmental ; preclinical study ; pre-clinical study ; design ; designing ; manufacturing process ; scale up ; Population ; innovation ; innovate ; innovative ; combinatorial ; Implant ; two-dimensional ; 2-dimensional ; implantation ; dopaminergic differentiation ; commercialization ; patient population ; risk minimization ; Industry Standard ; Formulation ; reduce symptoms ; alleviate symptom ; ameliorating symptom ; decrease symptom ; fewer symptoms ; relieves symptoms ; symptom alleviation ; symptom reduction ; symptom relief ; experimental study ; experiment ; experimental research ; clinical development ; cell replacement therapy ;
