Probing the biological consequences of a previously undescribed de novo mutation of ZMYND11 in a schizophrenia patient by CRISPR genome editing and induced pluripotent stem cell based in vitro disease-modeling

Csongor Tordai; Edit Hathy; Hella Gyergyák; Katalin Vincze; Máté Baradits; Júlia Koller; Ádám Póti; Bálint Jezsó; László Homolya; Mária Judit Molnár; László Nagy; Dávid Szüts; Ágota Apáti; János M Réthelyi

doi:10.1016/j.schres.2024.01.024

Probing the biological consequences of a previously undescribed de novo mutation of ZMYND11 in a schizophrenia patient by CRISPR genome editing and induced pluripotent stem cell based in vitro disease-modeling

Schizophr Res. 2024 Jan 29:S0920-9964(24)00024-0. doi: 10.1016/j.schres.2024.01.024. Online ahead of print.

Authors

Affiliations

¹ Institute of Molecular Life Sciences, Research Center for Natural Sciences, 1117 Budapest, Magyar tudósok körútja 2, Budapest, Hungary; Molecular Psychiatry Research Group, Semmelweis University, 1083 Budapest, Balassa utca 6, Budapest, Hungary.
² Molecular Psychiatry Research Group, Semmelweis University, 1083 Budapest, Balassa utca 6, Budapest, Hungary.
³ Institute of Molecular Life Sciences, Research Center for Natural Sciences, 1117 Budapest, Magyar tudósok körútja 2, Budapest, Hungary.
⁴ Molecular Psychiatry Research Group, Semmelweis University, 1083 Budapest, Balassa utca 6, Budapest, Hungary; Department of Psychiatry and Psychotherapy, Semmelweis University, 1083 Budapest, Balassa utca 6, Budapest, Hungary.
⁵ Molecular Psychiatry Research Group, Semmelweis University, 1083 Budapest, Balassa utca 6, Budapest, Hungary; Institute of Genomic Medicine and Rare Disorders, Semmelweis University, 1083 Budapest, Balassa utca 6, Budapest, Hungary.
⁶ Institute of Molecular Life Sciences, Research Center for Natural Sciences, 1117 Budapest, Magyar tudósok körútja 2, Budapest, Hungary; Doctoral School of Biology and Institute of Biology, Eötvös Loránd University, 1117 Budapest, Pázmány Péter sétány 1/c, Budapest, Hungary.
⁷ Institute of Genomic Medicine and Rare Disorders, Semmelweis University, 1083 Budapest, Balassa utca 6, Budapest, Hungary.
⁸ Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, 4032 Debrecen, Egyetem tér 1, Debrecen, Hungary.
⁹ Institute of Molecular Life Sciences, Research Center for Natural Sciences, 1117 Budapest, Magyar tudósok körútja 2, Budapest, Hungary. Electronic address: szuts.david@ttk.hu.
¹⁰ Institute of Molecular Life Sciences, Research Center for Natural Sciences, 1117 Budapest, Magyar tudósok körútja 2, Budapest, Hungary. Electronic address: apati.agota@ttk.hu.
¹¹ Molecular Psychiatry Research Group, Semmelweis University, 1083 Budapest, Balassa utca 6, Budapest, Hungary; Department of Psychiatry and Psychotherapy, Semmelweis University, 1083 Budapest, Balassa utca 6, Budapest, Hungary. Electronic address: rethelyi.janos@med.semmelweis-univ.hu.

PMID: 38290943
DOI: 10.1016/j.schres.2024.01.024

Abstract

Background: Schizophrenia (SCZ) is a severe neuropsychiatric disorder of complex, poorly understood etiology, associated with both genetic and environmental factors. De novo mutations (DNMs) represent a new source of genetic variation in SCZ, however, in most cases their biological significance remains unclear. We sought to investigate molecular disease pathways connected to DNMs in SCZ by combining human induced pluripotent stem cell (hiPSC) based disease modeling and CRISPR-based genome editing.

Methods: We selected a SCZ case-parent trio with the case individual carrying a potentially disease causing 1495C > T nonsense DNM in the zinc finger MYND domain-containing protein 11 (ZMYND11), a gene implicated in biological processes relevant for SCZ. In the patient-derived hiPSC line the mutation was corrected using CRISPR, while monoallelic or biallelic frameshift mutations were introduced into a control hiPSC line. Isogenic cell lines were differentiated into hippocampal neuronal progenitor cells (NPCs) and functionally active dentate gyrus granule cells (DGGCs). Immunofluorescence microscopy and RNA sequencing were used to test for morphological and transcriptomic differences at NPC and DGCC stages. Functionality of neurons was investigated using calcium-imaging and multi-electrode array measurements.

Results: Morphology in the mutant hippocampal NPCs and neurons was preserved, however, we detected significant transcriptomic and functional alterations. RNA sequencing showed massive upregulation of neuronal differentiation genes, and downregulation of cell adhesion genes. Decreased reactivity to glutamate was demonstrated by calcium-imaging.

Conclusions: Our findings lend support to the involvement of glutamatergic dysregulation in the pathogenesis of SCZ. This approach represents a powerful model system for precision psychiatry and pharmacological research.

Keywords: CRISPR; Precocious neuronal differentiation; Schizophrenia; ZMYND11; hiPSC.