Mutation-Specific Cardiomyocyte Lines from Patients with Fabry Disease: A Sustainable In Vitro Model to Investigate Structure, Function, and Disease Mechanisms

International Journal of Translational Medicine, Journal Year: 2025, Volume and Issue: 5(2), P. 15 - 15

Published: April 15, 2025

Background: Fabry disease (FD) results from pathogenic GLA variants, causing lysosomal α-galactosidase A (α-GalA) deficiency and sphingolipid ceramide trihexoside (Gb3 or THC) accumulation. Disease phenotype varies widely but cardiomyopathy is commonly life-limiting. As a multisystemic disorder, FD initiates at the cellular level; however, mechanism/s underlying Gb3-induced cell dysfunction remains largely unknown. This study established an in vitro mutation-specific model of using human-induced pluripotent stem (iPSC)-derived cardiomyocytes to explore pathology. Methods: Skin biopsies consenting patients normal control subjects were reprogrammed iPSCs then differentiated into cardiomyocytes. The mutations lines corrected CRISP-Cas9. Phenotypic characteristics, α-Gal activity, Gb3 accumulation, functional status, lipid analysis assessed. Cardiomyocytes derived two with severe clinical genotypes, GLAc.851T>C, GLAc.1193_1196del, their respective lines, GLAcorr c.851T>C, c.1193_1196del, selected for further studies. Results: individuals exhibited stable expression cardiomyocyte markers spontaneous contraction, morphological features FD, reduced accumulation Gb3. Lipidomic profiling revealed differences isoform profile between patient iPSC-derived Contraction strength was unchanged relaxation after contraction delayed, mimicking diastolic typical cardiomyopathy. Conclusions: provide useful aspects cardiomyopathy, including disruptions pathways, proteomics, multigene that together link genotype phenotype. platform potentially offers broad applicability across many genetic diseases prospect testing implementation individualised therapies.

Language: Английский

Investigating lysosomal dysfunction in Fabry disease using induced pluripotent stem cell-derived podocytes

Journal of Translational Genetics and Genomics, Journal Year: 2025, Volume and Issue: 9(1), P. 48 - 51

Published: Feb. 26, 2025

Aims: This study used induced pluripotent stem cell-derived podocytes from a Fabry disease (FD) patient carrying the p.Met284Thr pathogenic variant as an in vitro model to investigate lysosomal abnormalities driving cell pathology. Proteomic analysis was assess changes protein abundance FD compared controls. Additionally, temporal lysosome number were analyzed using automated live-cell imaging. Methods: Label-free mass spectrometry proteomics performed on at day 10 of differentiation For imaging, cultured transfected with CellLight Lysosomes-GFP and Plasma Membrane-CFP, then visualized quantified days 20 post-differentiation Perkin Elmer Phenix High Content Screening Microscope. Results: showed dysregulation glycosphingolipid metabolism proteins, including decreased galactosidase alpha (GLA; P < 0.01) increased galactosylceramidase glucosylceramidase (P podocytes. Lysosomal proteins enriched, significant increase cathepsin B 0.001) decrease lipase A 0.01). Furthermore, involved cycle regulation growth signaling pathways, such polo-like kinase 1 (PLK1; 0.0001) proto-oncogene tyrosine-protein Src (SRC; 0.01), suggested broader impacts cellular processes. Temporal imaging revealed controls Conclusions: These findings collectively suggest that undergo progressive impairment, which may contribute dysfunction progression. proof-of-concept lay foundation for future research targeted therapies high-throughput screening advanced analytical techniques.

Language: Английский