The Generation of Human iPSC Lines from Three Individuals with Dravet Syndrome and Characterization of Neural Differentiation Markers in iPSC-Derived Ventral Forebrain Organoid Model

Cells. 2023 Jan 16;12(2):339. doi: 10.3390/cells12020339.

Abstract

Dravet syndrome (DRVT) is a rare form of neurodevelopmental disorder with a high risk of sudden unexpected death in epilepsy (SUDEP), caused mainly (>80% cases) by mutations in the SCN1A gene, coding the Nav1.1 protein (alfa-subunit of voltage-sensitive sodium channel). Mutations in SCN1A are linked to heterogenous epileptic phenotypes of various types, severity, and patient prognosis. Here we generated iPSC lines from fibroblasts obtained from three individuals affected with DRVT carrying distinct mutations in the SCN1A gene (nonsense mutation p.Ser1516*, missense mutation p.Arg1596His, and splicing mutation c.2589+2dupT). The iPSC lines, generated with the non-integrative approach, retained the distinct SCN1A gene mutation of the donor fibroblasts and were characterized by confirming the expression of the pluripotency markers, the three-germ layer differentiation potential, the absence of exogenous vector expression, and a normal karyotype. The generated iPSC lines were used to establish ventral forebrain organoids, the most affected type of neurons in the pathology of DRVT. The DRVT organoid model will provide an additional resource for deciphering the pathology behind Nav1.1 haploinsufficiency and drug screening to remediate the functional deficits associated with the disease.

Keywords: Dravet syndrome; Nav1.1 haploinsufficiency; Panayiotopoulos syndrome; SCN1A-related disorders; organoids; stem cell reprogramming.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Epilepsies, Myoclonic* / genetics
  • Humans
  • Induced Pluripotent Stem Cells* / metabolism
  • NAV1.1 Voltage-Gated Sodium Channel / genetics
  • NAV1.1 Voltage-Gated Sodium Channel / metabolism
  • Neurons / metabolism
  • Prosencephalon / metabolism

Substances

  • NAV1.1 Voltage-Gated Sodium Channel

Grants and funding

This research was funded by Polish National Science Centre, grants 2019/03/X/NZ3/02139, 2015/17/B/NZ4/02669, and Statutory Funds of MMRI PAS, Warsaw, Poland.