Human ESC-based modeling of H3.3G34R-mutant high grade glioma

H3.3G34R-mutant gliomas are lethal tumors of the cerebral hemispheres, with unknown mechanisms of topographic specificity and tumorigenicity. We developed a human embryonic stem cell (hESC)-based model of H3.3G34R-mutant glioma that recapitulates the anatomical and genetic characteristics of the patient tumors. Overall design: A series of engineered hESC lines, which harbor p53 and/or ATRX mutations and express wild type or G34R-mutant histone H3.3 variant under HES5 promoter, were differentiated into ventral forebrain neural progenitor cells (NPCs) as well as ventral hindbrain NPCs and maintained in neural differentiation condition as 3D spheroids. At day 46 of differentiation, HES5-positive NPCs were isolated and expression profiles were obtained by RNA-sequencing.