DPYSL2/CRMP2 isoform B knockout in human iPSC-derived glutamatergic neurons confirms its role in mTOR signaling and neurodevelopmental disorders
The DPYSL2/CRMP2 gene encodes a microtubule-stabilizing protein crucial for neurogenesis and is associated with numerous psychiatric and neurodegenerative disorders including schizophrenia, bipolar disorder, and Alzheimer’s disease. DPYSL2 generates multiple RNA and protein isoforms, but few studies...
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| Published in: | Molecular psychiatry Vol. 28; no. 10; pp. 4353 - 4362 |
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| Main Authors: | , , , |
| Format: | Journal Article |
| Language: | English |
| Published: |
London
Nature Publishing Group UK
01-10-2023
Nature Publishing Group |
| Subjects: | |
| Online Access: | Get full text |
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| Summary: | The
DPYSL2/CRMP2
gene encodes a microtubule-stabilizing protein crucial for neurogenesis and is associated with numerous psychiatric and neurodegenerative disorders including schizophrenia, bipolar disorder, and Alzheimer’s disease.
DPYSL2
generates multiple RNA and protein isoforms, but few studies have differentiated between them. We previously reported an association of a functional variant in the
DPYSL2-B
isoform with schizophrenia (SCZ) and demonstrated in HEK293 cells that this variant reduced the length of cellular projections and created transcriptomic changes that captured schizophrenia etiology by disrupting mTOR signaling-mediated regulation. In the present study, we follow up on these results by creating, to our knowledge, the first models of endogenous
DPYSL2-B
knockout in human induced pluripotent stem cells (iPSCs) and neurons. CRISPR/Cas9-faciliated knockout of
DPYSL2-B
in iPSCs followed by
Ngn2
-induced differentiation to glutamatergic neurons showed a reduction in
DPYSL2-B
/CRMP2-B RNA and protein with no observable impact on
DPYSL2-A
/CRMP2-A. The average length of dendrites in knockout neurons was reduced up to 58% compared to controls. Transcriptome analysis revealed disruptions in pathways highly relevant to psychiatric disease including mTOR signaling, cytoskeletal dynamics, immune function, calcium signaling, and cholesterol biosynthesis. We also observed a significant enrichment of the differentially expressed genes in SCZ-associated loci from genome-wide association studies (GWAS). Our findings expand our previous results to neuronal cells, clarify the functions of the human
DPYSL2-B
isoform and confirm its involvement in molecular pathologies shared between many psychiatric diseases. |
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| Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 AUTHOR CONTRIBUTIONS KLF performed all the laboratory work, performed bioinformatics analysis, and contributed significantly to writing and editing the manuscript. XP performed RNA sequencing data analysis and differential expression analysis and proofreading of the manuscript. CKY provided support for all laboratory aspects of the project (reagent orders and preparation, support with tissue cultures) and proofreading of the manuscript. DA conceptualized and designed the project, supervised and interpreted experiments, contributed to writing and editing the manuscript and secured the necessary funding. |
| ISSN: | 1359-4184 1476-5578 1476-5578 |
| DOI: | 10.1038/s41380-023-02186-w |