Autism-associated protein POGZ controls ESCs and ESC neural induction by association with esBAF

Abstract

Background The POGZ gene has been found frequently mutated in neurodevelopmental disorders (NDDs), particularly autism spectrum disorder (ASD) and intellectual disability (ID). However, little is known about its roles in embryonic stem cells (ESCs), neural development and diseases. Methods We generated Pogz−/− ESCs and directed ESC differentiation toward a neural fate. We performed biochemistry, ChIP-seq, ATAC-seq, and bioinformatics analyses to understand the role of POGZ. Results We show that POGZ is required for the maintenance of ESC identity and the up-regulation of neural genes during ESC differentiation toward a neural fate. Genome-wide binding analysis shows that POGZ is primarily localized to gene promoter and enhancer regions. POGZ functions as both a transcriptional activator and repressor, and its loss leads to deregulation of differentiation genes, including neural genes. POGZ physically associates with the SWI-SNF (esBAF) chromatin remodeler complex, and together they modulate enhancer activities via epigenetic modifications such as chromatin remodeling and histone modification. During ESC neural induction, POGZ-mediated recruitment of esBAF/BRG1 and H3K27ac are important for proper expression of neural progenitor genes. Limitations The genotype and allele relevant to human neurodevelopmental disorders is heterozygous loss of function. This work is designed to study the effects of loss of POGZ function on ESCs and during ESC neural induction. Also, this work lacks of in vivo validation using animal models. Conclusions The data suggest that POGZ is both a transcription factor and a genome regulator, and its loss leads to defects in neural induction and neurogenesis.

Deficiency of CHAMP1, a gene related to intellectual disability, causes impaired neuronal development and a mild behavioural phenotype

“… 49 , 50 Both studies, as well as other studies suggest that POGZ plays a role in transcriptional regulation. 49 , 50 , 61 , 63 The possibility that CHAMP1 also functions in transcriptional regulation is worth considering, which can explain the downregulation of neurotransmitter transport and NDD-related genes in embryonic CHAMP1 −/− brains.…”

POGZ suppresses 2C transcriptional program and retrotransposable elements

“…We have shown that POGZ is required for ESC cell survival and genome stability (Sun et al, 2022). However, the exact mechanisms remain unclear.…”

“…ChIP experiments were performed according to the Agilent Mammalian ChIP-on-chip manual as described (Sun et al, 2022). Briefly, 1× 10 For ChIP-PCR, the ChIPed DNA were dissolved in 100 μl distilled water.…”

“…We have recently shown that POGZ maintains ESCs as a chromatin regulator and a transcription factor (Sun et al, 2022). However, the detailed mechanisms remain unclear.…”

“…CUT&TagThe Cleavage Under Targets and Tagmentation (CUT&Tag) experiments were performed using FLAG antibodies (Flag, Sigma, F1804/F3165) in 3× Flag-POGZ restoring Pogz-/-mESCs by the Yingzi and Frasergen Companies (Wuhan, China) (Kaya-Okur et al, 2019). The detailed methods have been recently described(Sun et al, 2022).Quantification and statistical analysisData are presented as mean values ±SD unless otherwise stated. Data were analyzed using Student's t-test analysis.…”