An in vitro human cell assay to investigate the effects of psychiatric risk gene CACNA1C dosage on neuronal network activity

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There is a strong need in the pharmaceutical industry for disease relevant neuronal cell-based assays of high construct validity for accelerating medicine development in mental health. It is well established that clinical genetic evidence for disease risk significantly de-risks early stage drug discovery, delivering significant benefits to the development pipeline. Here, we bring the world-leading mental health genomics of the MRC Centre for Neuropsychiatric Genetics and Genomics together with our expertise in human neurocellular biology to create a cell-based tool that places patient genetics in close apposition with the drug discovery process. Many current preclinical drug screens are based on rodent animal and cell models, or non-neuronal cultured human cells, and this limits their predictive capacity to truly model human neuropsychiatric conditions. However, a convergence of new technologies for neuronal development of human stem cells, genome editing via CRISP and new cell phenotying techniques promises the creation of a new generation of disease-relevant cell-based assays. This project focuses on the development of a human cell assay platform based on CACNA1C, a risk gene associated with psychiatric disorders, including autism, schizophrenia and bipolar disorder. We will use adaptations of CRISPR technology (CRISPRi and CRISPRa) to control levels of gene CACNA1C expression in neurons differentiated from human stem cells and model the range of altered expression seen in psychiatric disorders. These cells will be tested for altered neuronal network activity using microelectrode array (MEA) technology. We have a fully functional MEA protocol established for neuronal networks derived from human induced pluripotent stem cells (hiPSC), and previously have shown that pharmacological blockade of L-type voltage-gated calcium channels, encoded by the CACNA1 gene family, alters the pattern of neuronal firing on MEAs in a dose dependent manner. This project will build onthese results to establish specificity of CACNA1C gene for this altered pattern of network behaviour and investigate the effect of gene dosage on neuronal activity, in turn creating an assay for further pharmacological testing and drug discovery. The student will develop skills in stem cell culture, in vitro neuronal differentiation; genome editing and cell analysis. They will also develop the quantitative biological and computational skills needed to analyse MEA outputs and parameterise network activity. Our AstraZeneca partner will bring knowledge of commercial assay development, and help translate the experimental findings into an industrial standard prototype assay. By working closely with our industrial partner, including exchange trips, we will design the laboratory-based experiments with the aim of testing the company's in-house compounds and potential drugs for beneficial pharmacological activity.