Aug 2026· Stem Cell Research· Vol 95, pp.
104086
· 0 citations· 13 references
Medicine
TL;DR
Two isogenic induced pluripotent stem cell lines carrying homozygous LMNA variants are generated by prime editing of a healthy donor iPSC line, providing a valuable platform for functional characterization and potential clinical reclassification of LMNA VUS.
Abstract
Variants of uncertain significance (VUS) in the LMNA gene represent a major challenge in clinical genetics, as insufficient functional evidence limits their interpretation and clinical decision-making in laminopathies, including dilated cardiomyopathy (DCM). Here, we generated two isogenic induced pluripotent stem cell (iPSC) lines carrying homozygous LMNA variants, c.293A > G (p.Glu98Gly) and c.439G > A (p.Ala147Thr) by prime editing of a healthy donor iPSC line. Both variants are located within Coil 1B domain of lamin A. The edited iPSC lines retain normal morphology, pluripotency, genomic integrity, and trilineage differentiation capacity, providing a valuable platform for functional characterization and potential clinical reclassification of LMNA VUS.
The GGGGCC hexanucleotide repeat expansion (HRE) within the C9orf72 gene constitutes the leading genetic driver of amyotrophic lateral sclerosis (ALS). This fatal neurodegenerative disorder is characterized by the systematic loss of both the upper and lower motor neurons across both the central and peripheral nervous systems. This work describes the successful reprogramming of two human induced pluripotent stem cell (iPSC) lines originating from two independent ALS patients, both of whom carry a C9orf72 HRE mutation. Validation of the two established iPSC lines confirmed the expression of pluripotency markers, normal karyotypes, and successful trilineage differentiation. Consequently, these lines provide a robust in vitro platform to model ALS and study C9orf72-mediated disease mechanisms.
Dide Wu, A. Kojic, Jay P. Ross et al.· Stem Cell Research· 0 citations
Patient-derived induced pluripotent stem cells (hiPSC) are a valuable approach to model cardiovascular diseases. We nucleofected non-integrating episomal vectors in skin fibroblasts of four family members. Two of them carried the single nucleotide variant (SNV) SCN5A_c.287 T > C, leading to NaV1.5_p.L96P, and two were non-carrier family members. The resulting hiPSC cell lines differentiate into cells of the 3 germ layers, display normal karyotypes and express markers of the undifferentiated hPSC state. Thus, they are a reliable source to study the effect of the identified mutation in a physiologically relevant environment.
E. Selga, R. Martínez-Moreno, Albert Rigat Pujolàs et al.· Stem Cell Research· 0 citations
Fabry disease (FD) is a monogenic, X-linked lysosomal storage disorder originating from mutations in the GLA gene, which encodes alpha-galactosidase A. Impaired enzyme activity leads to accumulation of the substrate globotriaosylceramide (Gb3) and a multisystemic phenotype. Here, we generated two human induced pluripotent stem cell (hiPSC) lines from a female FD patient carrying a heterozygous c.644A > G missense mutation. The hiPSCs displayed normal karyotype, typical morphology, trilineage differentiation capacity and expressed markers of undifferentiated hPSC state. Consequently, MHHi043-A and MHHi043-B provide a valuable resource for studying FD mechanisms and developing therapeutic strategies.
Nick Heise, Carla Borisch, Christopher Jahn et al.· Stem Cell Research· 0 citations
Marfan syndrome is a connective tissue disorder affecting the cardiovascular, skeletal, and ocular systems. Here, we generated and characterized induced pluripotent stem cell (iPSC) lines derived from two Marfan syndrome patients with mutations in the FBN1 gene (c.3333C > A and c.8854_8562delinsTATCAC). Both lines exhibited typical iPSC morphology, normal karyotype, undifferentiated states, and trilineage differentiation capacity. These iPSCs serve to enable investigation into the mechanisms underlying Marfan syndrome for therapeutic discovery.
Byron W H Mui, M. Chorsi, Christopher D. Yan et al.· Stem Cell Research· 0 citations
Background LMNA codes a widely expressed nuclear cytoskeletal protein (lamin A/C) with multiple important functions. Pathogenic LMNA genetic variation may lead to autosomal dominant cardiomyopathy, though the severity and rate of progression can vary with the specific nucleotide change and location. Prior studies showed that induced pluripotent stem cells (iPSC)-derived cardiomyocytes (iCMs) with LMNA R541C exhibited reduced LMNA protein abundance, increased sarcomere disorganization, and abnormal electrophysiology. Methods We investigated the LMNA-R541C variant that exhibits a highly penetrant and severe clinical cardiomyopathy phenotype using transcriptomic analysis of iCMs. Patient-derived iPSCs with CRISPR-corrected (clustered regularly interspersed short palindromic repeats) isogenic control cells and CRISPR knock-in LMNA-R541C heterozygous iPSCs were generated for isogenic controlled experiments. Results In differential gene expression analyses we observed that LMNAR541C/WT iPSC-derived cardiomyocytes had consistent perturbations in 123 genes across CRISPR-corrected and knock-in experiments compared to controls. Pathway analysis identified that the G2M checkpoint and oxidative phosphorylation processes were consistently dysregulated and confirm these findings in previously published iPSC and murine models. Discussion These results implicate perturbed gene expression and pathways that may contribute to the severe phenotypes in LMNA-R541C. Informatic analysis of pathways suggests several drug classes including multiple cardiac glycosides as potential targeted therapeutic candidates to be explored.
Thomas E. Keller, Ci Koehring, Brett W. Higgins et al.· bioRxiv· 0 citations
The YARS2 variant, harboring the compound heterozygous pathogenic mutations F185L/E264del, was identified in the gene for mitochondrial tyrosyl-tRNA synthetase in a proband that suffered a neonatal phenotype. To facilitate studies to better understand the severity of the mutations, we created a patient-derived inducible pluripotent stem cell (iPSC) model. We first derived iPSCs from fibroblasts of the patient Q1818, which contain two mutations, c.553T>C (p.F185L) and c.792_794delAGA (p.E264del) in the YARS2 gene. We then generated three isogenic control iPSC lines with one or both mutations corrected by using CRISPR-Cas9 technology. The correction of mutations in YARS2 was confirmed by Sanger sequencing. The stemness of iPSC lines was demonstrated by the expression of stem cell markers in the iPSCs, as determined using qPCR, immunostaining, and trilineage differentiation. Moreover, three positive clones of each iPSC line were extensively characterized, confirming that they originated from Q1818 fibroblasts, had normal karyotypes, and did not contain off-targets in the YARS2 coding sequence; genome wide off target effects were not a major concern. Subsequently, Q1818 iPSCs and the three isogenic control iPSCs were differentiated into clinically relevant motor neurons. In addition, we demonstrated that the patient fibroblasts and the derived iPSCs are heterozygous for either c. 553T>C or c.792_794delAGA, and that the two mutations are located on different alleles of the YARS2 gene, providing critical information for studying the mutation-associated disease. In conclusion, we have generated a set of four iPSC lines, which can be used as a model to study a clinically severe case of YARS2 disease.
Chen-Bo Zeng, Andrew Gray, R. Ganetzky et al.· International Journal of Ste...· 0 citations
