Development of a High-content drug repurposing screen using CDKL5 patient derived iPSC neurons

Heterozygous or homozygous de novo loss-of-function mutations in the Xlinked gene CDKL5 are associated with a rare neurodevelopmental disorder characterized by early neonatal/infantile onset of epilepsy, cortical visual impairment, severe developmental delay and intellectual disability. Boston Children’s Hospital (BCH) is home to one of three CDKL5 Centers of Excellence sponsored by the International Foundations for CDKL5 Research (IFCR) dedicated to providing comprehensive multidisciplinary care to patients suffering from CDKL5 related neurodevelopmental disorder. The Human Neuron Core at BCH has collected CDKL5 patient samples, made iPSC derived neurons and developed several medium throughput cellular assays for neuronal phenotypes with potential relevance to CDKL5 pathology. Validated assays developed in the Human Neuron Core include measures of neuronal morphology, neuronal hyper excitability and mitochondrial biology via collaboration with a network of faculty in the BCH Kirby Neurobiology Center. We hypothesize that patient-derived neurons bearing mutations in the CDKL5 gene will exhibit changes in gene expression and cell autonomous deficits related to CDKL5 function. Drugs that reverse these cell biological deficits may provide therapeutic benefit for patients expressing defective alleles of CDKL5. We have the following aims;

  • Create state-of-the-art isogenic pairs of patient iPSC lines engineered to express a fluorescent reporter to indicate the expression of a mutant allele in CDKL5 in neurons.
  • Differentiate and phenotype cortical neurons from isogenic pairs of iPSC lines in collaboration withan interdisciplinary team of Boston Children's Hospital (BCH) subject matter experts (H. Olson, C.Woolf, M. Sahin, T. Schwartz) in order to optimize cell type and endpoint measures for a robust image-based High-content drug screen (HCS) in the Human Neuron Core at BCH.
  • We will collaborate closely with M. Johnson (Imperial College, London) who will develop network based predictions for potential repurposed drug treatments based on transcriptional analysis of differentiated neurons from isogenic iPSC lines.