Neuron-Based High Throughput/High Content Screening (HTS/HCS) Platforms for Parkinson's Disease, Amyotrophic Lateral Sclerosis, Fronto-Temporal Dementia and Huntington's Disease. INVENTORS

SUMMARY Cell-based screening is a mainstay of the drug discovery process. However, there are major limitations accompanying the use of conventional cell-based screens to identify potential new therapeutic agents for neurological disorders. The routine use of cell lines in testing often presents problems because of the myriad cellular, molecular, and physiologic differences that exist between immortalized cell lines and the neurons that they are meant to model. Importantly, the identification of genetic mutations associated with specific familial forms of many neurological diseases and disorders, has helped dissect the molecular mechanisms underlying the pathologies observed and facilitated the creation of more robust cell and animal models. We have developed faithful and reliable ex-vivo cell-based models of Parkinson’s disease (PD), amyotrophic lateral sclerosis (ALS), fronto-temporal dementia (FTD), Huntington’s disease (HD) and Alzheimer’s disease (AD) using primary neurons from rodent models. These neuron-based models together with our automated robotic microscope system enable sophisticated experimental designs to monitor multiple changes over time with greater precision and throughput than existing technologies. Recently, we began using human neurons differentiated from induced pluripotent stem cells (iPSCs) to create additional cell-based models of disease. Reprogrammed patient-derived somatic cells provide iPSCs for differentiation into various types of primary neurons. Subclasses of primary neurons reproduce the unique cell type context within which a specific mutation drives cell pathology. When fluorescent reporter proteins are introduced into live cells to probe the activity of biological pathways of interest, large data sets can be automatically analyzed at great speed with the same rigorous statistical tools used for clinical studies. The creation of the desired iPSCs is facilitated by the recent set up of an open-access, mutation-defined fibroblast resource for neurological disease research. The National Institute for Neurological Disorders and Stroke (NINDS) Repository, at the Coriell Institute for Medical Research, maintains the deposited cell lines. This is a significant expanding resource that will advance the use of patient cells as disease models; mutation-defined cell lines are available upon request from a wide range of neurological disorders. Together with our automated robotic microscope system’s longitudinal imaging and analysis capabilities for multiple functions (including survival) in single cells, these models form powerful and flexible HTS/HCS platforms for detecting small molecule modifiers of various neurological disease or disorder phenotypes.