SBIR-STTR Award

Human neural cells manufactured using patient-derived induced pluripotent stem cells (iPSCs) hold great promise for modeling neurodevelopmental disorders, discovering new precision therapies, and screening for potential risks from environmental toxins. However, biomanufacturing of neural cells and tissues is severely limited by low efficiency, poor reproducibility, and extended timeframes. Recent scientific discoveries, coupled with the substantial need for iPSC-derived neural cells in pharma applications, frame a significant commercial opportunity. Stem Pharm will address this opportunity by commercializing synthetic hydrogel technologies for biomanufacturing of iPSC-derived neural cells. Specifically, we will develop synthetic hydrogel substrates for iPSC-derived neural cell expansion, differentiation and high throughput screening workflows. The goal of this Phase I project is to develop chemically defined and robust synthetic hydrogels for efficient differentiation of iPSC-derived NPCs into cortical neurons and subsequent maturation to post-mitotic, functionally mature neurons. Specific Aim 1 will use synthetic hydrogel screening technologies to identify formulations that promote efficient differentiation of NPCs into post-mitotic cortical neurons. Specific Aim 2 will use synthetic hydrogel screening technologies to identify formulations that promote viability, retention and accelerated maturation of post-mitotic cortical neurons towards functional maturity. Specific Aim 3 will use a well-characterized neurodevelopmental disease line of Rett Syndrome to demonstrate the applicability and advantages of the identified hydrogel formulations for culturing challenging human disease-derived cells. This work is significant, as there is a critical need for better tools to optimize yields and reduce variability in the differentiation of iPSCs to defined neural subpopulations, support their long-term culture, and reduce the time needed to reach functional maturity. Successful completion of the proposed milestones will result in more effective biomanufacturing of neural cells and cell-based assays for precision medicine. This work is innovative, as our synthetic hydrogel screening technologies will identify hydrogel formulations for optimal differentiation and maturation of neural populations. These unique formulations can be discovered using the proprietary hydrogel screening approach developed and used by Stem Pharm, but once identified they can be used broadly across standard pharmaceutical workflows. Subsequent Phase II activities will demonstrate the performance of the unique hydrogel formulations in high throughput screening for toxicity testing and drug discovery.

Project Terms:
Address; Animals; base; Biochemical; Biological Assay; Biomanufacturing; Calcium Oscillations; Cell Differentiation process; Cell Survival; Cells; Chemicals; cost; Coupled; Disease; Disease model; disease phenotype; drug discovery; Environmental Exposure; Environmental Risk Factor; Etiology; Formulation; Glial Fibrillary Acidic Protein; Goals; Growth; Growth Factor; high throughput screening; Human; human disease; Hydrogels; improved; Individual; induced pluripotent stem cell; innovation; Mechanics; Mitotic; Modeling; nerve stem cell; Neurites; Neurodevelopmental Disorder; Neurons; novel; Patients; Performance; Pharmacologic Substance; Phase; Phenotype; Population; precision medicine; Precision therapeutics; Protocols documentation; public health relevance; relating to nervous system; Reproducibility; Rett Syndrome; screening; self assembly; stem; Technology; Time; Tissues; tool; Toxic Environmental Substances; Toxicity Tests; Toxin; treatment strategy; Tubulin; Work;

Human neural cells manufactured using patient-derived induced pluripotent stem cells (iPSCs) hold great promise for modeling neurodevelopmental disorders, discovering new precision therapies, and screening for potential risks from environmental toxins1-4. There have been significant advances in the last decade in protocols and commercial media systems developed for differentiation into specific neural cell types5-8. However, there remain significant technical challenges to overcome in their generation, manufacturing and assay workflows. iPSCs are typically differentiated on animal-derived substrates that introduce intrinsic variability and lack control over mechanical stiffness and biochemical composition. This often results in low yields and high variability, which may be more pronounced when generating cellular models of diseases. There is a critical need to develop commercial tools that promote differentiation of iPSCs into mature neural cells in a controlled, efficient, and reproducible fashion and that eliminate animal derived products. The resulting cells, associated cell-based assays and cellular therapeutics will have a transformative impact on neural disease modeling, drug and therapeutic discovery and toxin screening. Our Phase I study identified chemically defined and robust synthetic hydrogels for efficient differentiation of iPSC-derived neural progenitor cells (NPCs) into cortical neurons and subsequent maturation to post-mitotic, functionally mature neurons. The highly innovative aspects of this work are that the substrates are employed as thin hydrogel coatings using our proprietary surface-localized polymerization methods which provides several technical and commercialization advantages. In order to bring these novel substrates to market we propose the following specific aims for our Phase II proposal: Specific Aim 1 will further validate the work that demonstrated our optimized synthetic thin hydrogel coatings support neural differentiation and maturation. Including further functional characterization of cells cultured on the substrates by employing microelectrode array analysis and differential transcriptional analysis to compare cells cultured on the substrate. We will characterize of the physical and mechanical properties of the optimized thin hydrogels and develop methods for coating plates using automated systems. Specific Aim 2 will apply the substrates in a Proof-of-Concept demonstration utilizing the substrates to assess cortical neurons from Major Depressive Disorder patient-derived samples compared with controls. Specific Aim 3 will expand the technology platform by optimizing coating techniques on microcarriers suitable for bioreactor scaling, which is a critical step to demonstrate these substrates are applicable to biomanufacturing applications. This work is significant, as there is a critical need for better tools to optimize yields and reduce variability in the differentiation of iPSCs to defined neural subtypes, support their long-term culture, reduce the time needed to reach functional maturity and eliminate animal-derived products in the workflow.

Public Health Relevance Statement:
Public Health Relevance The use of induced pluripotent stem (iPSC) technologies for disease modeling of neurodevelopmental disorders holds great promise for developing treatment strategies, understanding disease etiologies and screening for potential risks from environmental exposures. However, there are current limitations in the differentiation and maturation protocols that need to be improved including growing and differentiating the cells on animal-derived, hard substrates. We aim to develop novel, soft synthetic hydrogels that will provide a defined substrate which is more amenable to the culture and differentiation of neural cells.

Project Terms:
Animals; base; Biochemical; Biological Assay; Biomanufacturing; Bioreactors; Cell Adhesion; Cell Differentiation process; Cell model; Cells; cellular imaging; Chemicals; commercialization; comparative; cost; Coupled; Cultured Cells; Data; design; Development; Disease; Disease model; Elasticity; Environment; Environmental Exposure; Environmental Risk Factor; Etiology; experimental study; Feasibility Studies; Generations; Genetic Transcription; Goals; Growth Factor; Human; Hydration status; Hydrogels; improved; induced pluripotent stem cell; innovation; Luciferases; Major Depressive Disorder; Measurement; mechanical properties; Mechanics; Methodology; Methods; Microelectrodes; Mitotic; Modeling; Morphology; nano; nerve stem cell; Neurodevelopmental Disorder; Neurons; novel; Output; Patients; Pharmaceutical Preparations; Phase; phase 1 study; Phenotype; physical property; polymerization; Polystyrenes; precision medicine; Precision therapeutics; Protocols documentation; public health relevance; Quality Control; relating to nervous system; Reporter; Reproducibility; response; Sampling; screening; self assembly; stem; Surface; System; Techniques; Technology; Therapeutic; Thick; Thinness; Time; time use; tissue culture; Tissues; tool; Toxin; treatment strategy; Validation; Work