Characterization and Early Assays Development in FOXG1 Deficient Neurons

Boston Children'S Hospital, Boston MA

Investigators

Abstract

PROJECT SUMMARY FOXG1 syndrome is a rare neurological disorder that often causes severe intellectual disability, microcephaly and neurological symptoms including hypotonia, seizures, and stereotypic movements. FOXG1 syndrome is caused by heterozygous mutation or deletion of the forkhead box 1 protein (FOXG1), a transcription factor that controls expression of key cortical development proteins. FOXG1 is specifically important for telencephalon neural progenitor cell (NPC) proliferation and differentiation to GABAergic interneurons. We have prioritized FOXG1 syndrome based on a framework we developed to evaluate monogenetic neurodevelopmental disorders for therapeutic development based on generic, preclinical validation, clinical and ethical considerations. We have also created three pluripotent stem cell (PSC) lines from FOXG1 syndrome patients and used CRISPR/Cas9 gene editing to create isogenic control lines. We hypothesize that a drug-like small molecule that increases FOXG1 expression in patients during early postnatal development would improve symptoms in these individuals. In the proposed research, we will use FOXG1 PSC lines to create an endogenous locus gene reporter line by using CRISPR/Cas9 to insert a HaloTag reporter protein at the C-terminus of the FOXG1 gene. A method will be developed whereby these modified FOXG1 PSCs with be differentiated to cortical neural progenitor cells (NPCs) and treated with HaloTag ligand tethered dyes to measure accurately measure FOXG1 expression. The primary goal of this project will be to demonstrate that this FOXG1 reporter system is suitable for screening small molecules for their ability to increase FOXG1 expression.