A current major challenge in pharmaceutical industry is to develop treatments for brain diseases and mental disorders such as depression, anxiety, schizophrenia, epilepsy, addiction and many more. Over the last decade, the drug discovery process has evolved dramatically following the continued advancement of technologies. In addition to increasing the efficiency of high-throughput screening (HTS) by assay miniaturization and multiplexing, various initiatives have been taken to improve the science of predicting toxicity and improving extrapolation to humans. With recent advances in genomics and proteomics, a number of novel technologies have made the utilization of live cells with overexpressed drug targets in assays an important aspect in the drug discovery process. Nevertheless, an appropriate cellular platform for HTS of neural targets has not been available. This in part can be attributed to difficulties in transfecting and maintaining neurons in culture. Recently, a breakthrough was made in human embryonic stem cell research that allows in vitro differentiation of neurons from a renewable source of progenitor cells. However, limitations of the current differentiation protocol and poor neuroprogenitor cell transfection techniques have restricted the use of these cells in assays. Originus, Inc. can contribute to the development of sophisticated neuronal cell-based assay platforms that address these issues by using a proprietary solid phase cell transfection technology termed Surface Transfection and Expression Protocol (STEP) exclusively licensed from the University of Michigan. We have successfully developed multiple G- protein-coupled receptor (GPCR) assays in neuronal and non-neuronal cell lines using STEP. The proposed new STEP transfection and differentiation platform will drastically accelerate human stem cell neural differentiation process while simultaneously introducing the target of interest and necessary components for a cell-based assay. Our goal is to develop not only more reliable and robust assays that increase the screening efficiency in early preclinical phase, but also more physiologically relevant assays that may boost clinical success rates. During Phase I of this application we will initially examine the biological responses of the neural progenitor cells in assays, and thoroughly characterize the progenitor cell neuronal differentiation induced by transient over-expression of selected basic helix-loop-helix (bHLH) transcription factors. Further, the potential implication of such a platform for cell-based assays will be demonstrated by examining the response of metabotropic glutamate receptors in differentiated human neurons. During the Phase II, we plan to apply this technology to other human pluripotent cell lines, expand the spectrum of bHLH factors tested to fine tune the differentiation process, and to further develop assays for different drug targets. This novel platform will make feasible HTS of drug targets expressed in human neurons. In addition, it may also be used for toxicology, neural differentiation and degeneration studies.
Public Health Relevance: The proposed studies aim to greatly improve the effectiveness of the drug discovery process for brain diseases and mental disorders by taking advantage of research advances in human stem cells and genetic control of neuronal development. Specifically, the goal of this application is to develop physiologically relevant drug screening platforms for human neurons derived from embryonic stem cells.
Public Health Relevance Statement: The proposed studies aim to greatly improve the effectiveness of the drug discovery process for brain diseases and mental disorders by taking advantage of research advances in human stem cells and genetic control of neuronal development. Specifically, the goal of this proposal is to develop physiologically relevant drug screening platforms for human neurons derived from embryonic stem cells.
NIH Spending Category: Bioengineering; Biotechnology; Brain Disorders; Genetics; Mental Health; Neurosciences; Regenerative Medicine; Stem Cell Research; Stem Cell Research - Embryonic - Human
Project Terms: ACSL1; ACSL1 Gene; ASCL1 gene; ASH1; Address; Adherent Culture; Adhesion Molecule; Anaplastic; Anxiety; Assay; BHLH Protein; Basic HLH Protein; Basic Helix-Loop-Helix Protein; Basic Helix-Loop-Helix Transcription Factors; Bioassay; Biologic Assays; Biological; Biological Assay; Brain Diseases; Brain Disorders; Cell Adhesion Molecules; Cell Line; Cell Lines, Strains; Cell surface; CellLine; Cells; Clinical; Complex; Development; Drug Delivery; Drug Delivery Systems; Drug Evaluation, Preclinical; Drug Industry; Drug Screening; Drug Targeting; Drug Targetings; ES cell; Effectiveness; Elements; Encephalon Diseases; Environment; Epilepsy; Epileptic Seizures; Epileptics; Evaluation Studies, Drug, Pre-Clinical; Evaluation Studies, Drug, Preclinical; G Protein-Complex Receptor; G-Protein-Coupled Receptors; G-Proteins; GFAC; GTP-Binding Proteins; GTP-Regulatory Proteins; Galactosidase; Gene Expression; Gene Expression Monitoring; Gene Expression Pattern Analysis; Gene Expression Profiling; Genetic; Genomics; Glia; Glial Cells; Glutamates; Goals; Growth Agents; Growth Factor; Growth Factors, Proteins; Growth Substances; Guanine Nucleotide Coupling Protein; Guanine Nucleotide Regulatory Proteins; HASH1; HTH DNA Binding Domain; HTH Motifs; Helix-Turn-Helix Motifs; High Throughput Assay; Human; Human, General; Immunologic, Luciferase; In Vitro; Industry; Industry, Pharmaceutic; Intracranial CNS Disorders; Intracranial Central Nervous System Disorders; Ion Channel; Ionic Channels; Kolliker's reticulum; L-Glutamate; Laboratories; Licensing; Life; Luc Gene; Luciferase Gene; Luciferases; MASH1; Man (Taxonomy); Man, Modern; Measures; Membrane Channels; Mental disorders; Mental health disorders; Metabotropic Glutamate Receptors; Methods and Techniques; Methods, Other; Michigan; Microarray Analysis; Microarray-Based Analysis; Miniaturisations; Miniaturization; Miniaturizations; Monolayer culture; Mother Cells; Nerve Cells; Nerve Transmitter Substances; Nerve Unit; Nervous; Neural Cell; Neural Stem Cell; Neurocyte; Neuroglia; Neuroglial Cells; Neuronal Differentiation; Neurons; Neurosciences; Neurotransmitters; Non-neuronal cell; Pathway interactions; Pharmaceutical Industry; Phase; Physiologic; Physiological; Population; Preclinical Drug Evaluation; Process; Production; Profilings, Gene Expression; Progenitor Cells; Proteins; Proteomics; Protocol; Protocols documentation; Psychiatric Disease; Psychiatric Disorder; Regulation; Reporter; Reproducibility; Research; SBIR; SBIRS (R43/44); Schizophrenia; Schizophrenic Disorders; Science; Screening procedure; Seizure Disorder; Signal Pathway; Small Business Innovation Research; Small Business Innovation Research Grant; Solid; Source; Stem Cell Research; Stem cells; Surface; Techniques; Technology; Testing; Time; Toxic effect; Toxicities; Toxicology; Transcript Expression Analyses; Transcript Expression Analysis; Transcription Activation; Transcriptional Activation; Transfection; Undifferentiated; Universities; Unspecified Mental Disorder; Up-Regulation; Work; addiction; base; calcium flux; calcium mobilization; cell adhesion protein; cholinergic; cultured cell line; dementia praecox; depression; design; designing; drug candidate; drug discovery; embryonic stem cell; epilepsia; epileptiform; epileptogenic; gene product; genetic profiling; hESC; helix loop helix; helix turn helix; high throughput screening; human ES cell; human ESC; human embryonic stem cell; human stem cells; immunocytochemistry; improved; interest; mental illness; microarray technology; nerve cement; nerve stem cell; neural; neural progenitor cells; neuron development; neuronal; neuronal progenitor; neuronal progenitor cells; new technology; novel; overexpression; pathway; pre-clinical; preclinical; prototype; psychological disorder; public health relevance; relating to nervous system; release of sequestered calcium ion into cytoplasm; response; schizophrenic; screening; screenings; small molecule libraries; stem cell differentiation; stem cell of embryonic origin; success; technology development; tool; transcription factor
