Generation of induced pluripotent stem cell lines from nonaffected parents and monozygotic triplets affected with autism spectrum disorder and epilepsy

Characterization of a loss-of-function NSF attachment protein beta mutation in monozygotic triplets affected with epilepsy and autism using cortical neurons from proband-derived and CRISPR-corrected induced pluripotent stem cell lines

“…Peripheral blood mononuclear cells extracted from the probands and both parents were transduced with non-integrating Sendai virus expressing OCT4, SOX2, c-MYC and KLF4 transcription factors to generate induced pluripotent stem cell (iPSC) lines for each subject ( Ali et al, 2022 ). Selected iPSC lines had typical human embryonic stem cell (hESC)-like morphology, expressed defining markers of pluripotency, and differentiated into all three germ layers in vitro ( Ali et al, 2022 ). Targeted sequencing of genomic DNA confirmed the expected genotypes of VPS13B and NAPB in all iPSC lines ( Supplementary Figures S1A,B ).…”

“…In the present study, we used induced pluripotent stem cells (iPSCs) derived from monozygotic triplets having homozygous splice donor mutation in NAPB (c.354+2T>G) and missense variant in VPS13B c.8516G>A 9 (p.Arg2839Gln) with varying degree of neurodevelopmental disorder (NDD) and their heterozygous parents ( Ali et al, 2022 ; AbdelAleem et al, 2023 ). Parents derived control iPSCs (CtrlF and CtrlM) and probands derived iPSCs (NDD_01, NDD_04 and NDD_05) were maintained under a feeder free condition in mTeSR1 medium (Stemcell Technologies, Vancouver) on Matrigel (1:80, BD Biosciences) coated plate cultured in a 37°C incubator with humidified atmosphere and 5% CO 2 .…”

“…To investigate the molecular and biological consequence of NAPB LoF in human neurons, we generated induced pluripotent stem cell (iPSC) lines from all three probands and their parents ( Ali et al, 2022 ). Isogenic iPSC lines were also generated by CRISPR gene editing to correct the NAPB mutation in proband-derived iPSC.…”

NAPB and developmental and epileptic encephalopathy: Description of the electroclinical profile associated with a novel pathogenic variant

“…3 In conclusion, the description of a fourth family that associates DEE and NAPB variants strongly supports that NAPB is a novel DEE-related gene. An extended study of the βSnap-KO mice and in vitro analyses of induced pluripotent stem-derived neurons 18 would help to better understand the pathophysiological mechanism involved in NAPB-related DEE. Because multigene panels are still often used as first-tier testing, 19,20 we suggest including NAPB but strongly recommend using exome and genome sequencing to identify and describe more NAPB patients and better understand the associated phenotypes.…”