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Preclinical evaluation of CRISPR-based therapies for Noonan syndrome caused by deep-intronic LZTR1 variants

Preclinical evaluation of CRISPR-based therapies for Noonan syndrome caused by deep-intronic LZTR1 variants

Authors :
Carolin Knauer
Henrike Haltern
Eric Schoger
Sebastian Kügler
Lennart Roos
Laura C. Zelarayán
Gerd Hasenfuss
Wolfram-Hubertus Zimmermann
Bernd Wollnik
Lukas Cyganek
Source :
Molecular Therapy: Nucleic Acids, Vol 35, Iss 1, Pp 102123- (2024)
Publication Year :
2024
Publisher :
Elsevier, 2024.

Abstract

Gene variants in LZTR1 are implicated to cause Noonan syndrome associated with a severe and early-onset hypertrophic cardiomyopathy. Mechanistically, LZTR1 deficiency results in accumulation of RAS GTPases and, as a consequence, in RAS-MAPK signaling hyperactivity, thereby causing the Noonan syndrome-associated phenotype. Despite its epidemiological relevance, pharmacological as well as invasive therapies remain limited. Here, personalized CRISPR-Cas9 gene therapies might offer a novel alternative for a curative treatment in this patient cohort. In this study, by utilizing a patient-specific screening platform based on iPSC-derived cardiomyocytes from two Noonan syndrome patients, we evaluated different clinically translatable therapeutic approaches using small Cas9 orthologs targeting a deep-intronic LZTR1 variant to cure the disease-associated molecular pathology. Despite high editing efficiencies in cardiomyocyte cultures transduced with lentivirus or all-in-one adeno-associated viruses, we observed crucial differences in editing outcomes in proliferative iPSCs vs. non-proliferative cardiomyocytes. While editing in iPSCs rescued the phenotype, the same editing approaches did not robustly restore LZTR1 function in cardiomyocytes, indicating critical differences in the activity of DNA double-strand break repair mechanisms between proliferative and non-proliferative cell types and highlighting the importance of cell type-specific screens for testing CRISPR-Cas9 gene therapies.

Details

Language :
English
ISSN :
21622531
Volume :
35
Issue :
1
Database :
Directory of Open Access Journals
Journal :
Molecular Therapy: Nucleic Acids
Publication Type :
Academic Journal
Accession number :
edsdoj.8ebba1ffab3c41c4a0a2fa619386fcc5
Document Type :
article
Full Text :
https://doi.org/10.1016/j.omtn.2024.102123