Characterisation of the patient-specific induced pluripotent stem cell line UKEi075-A carrying the heterozygous PRKAG2 frameshift variant c.769_770del (p.E257Kfs*15)

Q: What is the clinical evidence from this study?

Population: Severe dilated cardiomyopathy carrying heterozygous PRKAG2 frameshift variant c.769_770del (p.E257Kfs*15) (n=1). Intervention: Sendai virus-mediated reprogramming of dermal fibroblasts to hiPSCs (UKEi075-A) vs. Control hiPSC-line UKEi001-A (for RT-qPCR pluripotency marker comparison). Primary outcome: Characterization of hiPSC line pluripotency (SSEA-3 positivity) and 2D-cardiac differentiation potential (cTnT-positive cells).

Q: What does this research mean for the field?

Sendai virus-mediated reprogramming of dermal fibroblasts from a dilated cardiomyopathy patient carrying the heterozygous PRKAG2 frameshift variant c.769_770del produces a stable, pluripotent stem cell line (UKEi075-A) capable of tri-lineage differentiation. Novelty: ClaimNovelty.METHODOLOGICAL. Consensus alignment: ConsensusAlignment.NEUTRAL.

Benedict Hoffmann; Tim Knochenhauer; Julian Schlobohm; Κωνσταντίνα Σταθοπούλου; Bärbel Maria Ulmer; 柴宮綾; Sandra D. Laufer; Sarina Schäfer; Elisabeth Krämer; Sigrid Fuchs; Christa Augustin; Evaldas Girdauskas; Till Joscha Demal; Sabine Klaassen; Tanja Zeller; Stefan Blankenberg; Thomas Eschenhagen; Friederike Cuello

doi:10.1016/j.scr.2026.104123

October 27, 2026Stem Cell ResearchOpen Access

Characterisation of the patient-specific induced pluripotent stem cell line UKEi075-A carrying the heterozygous PRKAG2 frameshift variant c.769_770del (p.E257Kfs*15)

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Key result

Reprogramming PRKAG2-mutant fibroblasts yields patient-derived hiPSCs with ~99% pluripotency marker expression.

n=1

Why the study?

Missense variants in PRKAG2 can cause hypertrophic cardiomyopathy, cardiac conduction defects, and glycogen storage disorders, but the clinical consequences of truncating PRKAG2 variants are poorly understood.

Key Points

To generate and thoroughly characterise a patient-derived human induced pluripotent stem cell line carrying the heterozygous PRKAG2 truncating variant c.769_770del (p.E257Kfs*15).
Reprogrammed dermal fibroblasts from a 43-year-old male dilated cardiomyopathy patient carrying the heterozygous PRKAG2 c.769_770del variant using non-integrating Sendai virus vectors delivering OCT-4, SOX2, c-Myc, and KLF4.
Assessed pluripotency marker expression via flow cytometry (SSEA-3) and RT-qPCR (NANOG, SOX2), tested for Sendai virus clearance and mycoplasma contamination, and verified the variant by Sanger sequencing.
Evaluated genomic stability using G-banding karyotyping and copy number variation arrays, and verified differentiation potential into the three primary germ layers.
HiPSCs displayed normal colony morphology, absence of mycoplasma, and an intact male 46,XY karyotype with no detectable copy number alterations.
Flow-cytometric analysis showed > 99 % SSEA-3 positive cells, alongside robust gene expression of pluripotency markers NANOG and SOX2 confirmed by RT-qPCR.
Master cell bank at passage 20 tested negative for Sendai virus vectors by RT-qPCR and successfully demonstrated in vitro differentiation into all three primary germ layers.

Structured PICO

P

Population

Dermal fibroblasts from 1 male Caucasian patient (age 43) with severe dilated cardiomyopathy carrying the heterozygous PRKAG2 frameshift variant c.769_770del (p.E257Kfs*15).

I

Intervention

Sendai virus-mediated reprogramming of dermal fibroblasts expressing OCT-4, SOX2, c-Myc, and KLF4 (CytoTune-iPS 2.0 Reprogramming Kit), followed by directed trilineage differentiation (over 5 days) and 2D-cardiac differentiation.

C

Comparator

Control hiPSC-line UKEi001-A (for pluripotency analysis) and isotype control antibodies (for flow cytometry).

O

Outcome

Generation and quality control characterization of the hiPSC line UKEi075-A, including colony morphology, genomic stability (G-banding and NanoString karyotype panel), pluripotency marker expression (SSEA-3, NANOG, SOX2), Sendai virus clearance, mycoplasma absence, and trilineage/cardiac differentiation potential (cTnT, SOX17, HNF-3b, PAX6, Nestin, Brachyury).surrogate

The successfully generated and characterized patient-specific hiPSC line UKEi075-A provides a versatile preclinical model to investigate the molecular pathomechanisms of dilated cardiomyopathy caused by truncating PRKAG2 variants.

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Implication

Patient-specific hiPSC model enables mechanistic studies of PRKAG2 truncating variants; hypothesis-generating with no immediate clinical implications.

Abstract

Missense variants in PRKAG2, the gene encoding for the γ2-subunit of adenosine monophosphate-activated protein kinase (AMPK), can cause hypertrophic cardiomyopathy, cardiac conduction defects and glycogen storage disorders. Clinical consequences evoked by truncating PRKAG2 variants are poorly understood. Dermal fibroblasts of a male dilated cardiomyopathy-patient carrying the truncating PRKAG2 variant c.769_770del (p.E257Kfs*15) at a heterozygous state were reprogrammed by Sendai virus-mediated expression of OCT-4, SOX2, c-Myc and KLF4 to human induced pluripotent stem cells (hiPSCs). HiPSCs displayed a normal karyotype, high pluripotency, mycoplasma-absence and were successfully differentiated into three primary germ layers. This hiPSC-line will enable analysing molecular pathomechanisms underlying PRKAG2-cardiomyopathy.

Discussion

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Cite This Study

Hoffmann et al. (2026) studied Severe dilated cardiomyopathy carrying heterozygous PRKAG2 frameshift variant c.769_770del (p.E257Kfs*15) (n=1). Sendai virus-mediated reprogramming of dermal fibroblasts to hiPSCs (UKEi075-A) vs. Control hiPSC-line UKEi001-A (for RT-qPCR pluripotency marker comparison) was evaluated on Characterization of hiPSC line pluripotency (SSEA-3 positivity) and 2D-cardiac differentiation potential (cTnT-positive cells). Sendai virus reprogramming of dermal fibroblasts from a PRKAG2 c.769_770del dilated cardiomyopathy patient generated hiPSC line UKEi075-A with >99% SSEA-3 positivity and 26-81% cTnT+ cardiomyocytes.

synapsesocial.com/papers/6af10ed2a61b6e74a6de158d https://doi.org/10.1016/j.scr.2026.104123

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Characterisation of the patient-specific induced pluripotent stem cell line UKEi075-A carrying the heterozygous PRKAG2 frameshift variant c.769_770del (p.E257Kfs*15)

Key Points

Structured PICO

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Implication

Abstract

Discussion

Cite This Study

Characterisation of the patient-specific induced pluripotent stem cell line UKEi075-A carrying the heterozygous PRKAG2 frameshift variant c.769_770del (p.E257Kfs*15)

Key Points

Structured PICO

Ask about this paper

Implication

Abstract

Discussion

Cite This Study

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