Primorigen will use SBIR SHIFT funds to develop an array system that can identify chemically-defined surfaces to optimize induced pluripotent stem cell (hiPSC) differentiation and maturation. The platform initially will be validated for its capacty to help increase functional maturity of stem cell-derived hepatocytes, particularly cytochrome p450 (CYP) activity, and subsequently for large scale production of hiPSC-derived hepatocytes. Despite the widely acknowledged promise of human pluripotent stem cells to provide new therapies for numerous devastating degenerative diseases and new screening tools for more rapid, cost effective drug development, there are no reports of cells with sufficient functional maturity for these applications. Primorigen's technology will address this need by developing a novel peptide array for assessing the impact of insoluble cell signals on maturation of stem-cell derived lineages. Phase I studies will validate array stability and capacity to identify conditions that improve function and CYP activity of primary and hiPSC-derived hepatocytes. Phase II will apply this approach to other lineages including hPSC-derived cardiomyocytes, and adapt the optimal conditions for high-capacity production of hepatocytes. Planned products include: 1) a user- friendly array kit to identify optimal surface treatments, and ultimately, 2) functionally mature cells produced in large volumes to support drug screening applications.
Public Health Relevance Statement: Public Health Relevance: Functional hepatocytes have enormous value in the screening of new drugs for adsorption, distribution, metabolism, excretion (ADME), and toxicity. The limited availability of cadaveric primary hepatocytes has fostered the use of inadequate alternatives such as animal cell sources, which do not accurately model human responses, as well as immortalized cell lines like Hep-G2 that are deficient in full functionality, particularly cytochrome P450 (CYP) activity. Up to 60 percent of drug failures are due to poor ADME/Tox profiles which could be avoided if more accurate models were available. Stem cell derived hepatocytes represent a renewable, readily available source of hepatocytes for drug screening or therapeutics. Primorigen's hepatocyte differentiation protocol provides 95% pure populations of hepatocyte-like cells that have many primary cell functions such as albumin and urea secretion, lipid profiles, and engraftment, but lack optimal CYP activity. To address these deficiencies, Primorigen will take the novel approach of defining an optimal adhesion substrate to explore and exploit the impact of existing cell-extracellular matrix interactions on stem cell differentiation and function. The result will be a defined screening tool that enables customers to optimize differentiation protocols for reliable and successful production of functional hepatocytes, and that provides the basis for developing similar screening tools for other lineages.
NIH Spending Category: Stem Cell Research
Project Terms: Address; Adhesions; Adsorption; Albumins; alpha-Fetoproteins; Animals; base; Behavior Control; Biomedical Engineering; Cardiac Myocytes; Cell Adhesion; cell behavior; Cell Differentiation process; Cell Line; Cell physiology; Cells; Clinical; cost effective; Culture Media; Cytochrome P450; cytokine; Degenerative Disorder; density; Dependence; Derivation procedure; drug development; Drug Metabolic Detoxication; drug use screening; Engraftment; Environment; Enzymes; Evaluation; Excretory function; Exhibits; Extracellular Matrix; Failure (biologic function); Fostering; Funding; Government; Growth Factor; Hepatocyte; Human; immortalized cell; improved; improved functioning; In Vitro; induced pluripotent stem cell; infrared spectroscopy; Label; large scale production; Legal patent; Ligands; Lipids; Metabolism; Modeling; monolayer; novel; novel strategies; Peptide Library; Peptides; Persons; Pharmaceutical Preparations; Phase; phase 1 study; Phenotype; physical property; Pluripotent Stem Cells; Population; Preclinical Drug Evaluation; Process; Production; Proteins; Protocols documentation; public health relevance; Reporting; Research; response; screening; Signal Transduction; Small Business Innovation Research Grant; Source; stem cell differentiation; Stem cells; Substrate Interaction; Surface; System; Technology; Testing; Therapeutic; Time; tool; Toxic effect; Universities; Urea; user-friendly; Wisconsin; Xenobiotics
