Objective: This study aimed to generate and characterize human induced pluripotent stem cell (iPSC) lines derived from two Parkinson’s Disease (PD) patients carrying combinations of pathogenic mutations. One iPSC line harbors an SNCA duplication together with the GBA1 T369M variant, while the other carries a PRKN deletion alongside the LRRK2 G2019S mutation.

Background: PD is increasingly recognized as a multifactorial disorder in which the convergence of multiple genetic and environmental risk factors can accelerate the course of neurodegeneration. While monogenic forms of PD have been extensively studied, the pathogenic interactions between co-occurring mutations, such as SNCA duplication with a GBA1 risk variant or PRKN deletion combined with a LRRK2 G2019S mutation, remain poorly understood in human iPSC-derived models. Patient-specific iPSC lines with these “double-hit” genotypes are essential tools to investigate synergistic molecular disturbances that drive complex PD phenotypes.

Method: Fibroblasts from both patients were reprogrammed using Sendai virus (SeV)-based non-integrative vectors encoding the Yamanaka factors (OCT4, SOX2, KLF4, and L‑MYC). Pluripotency was assessed by immunofluorescence, RT‑PCR, and flow cytometry using standard pluripotency markers, including SSEA4, TRA‑1‑80, SOX2, OCT4, and NANOG. Trilineage differentiation potential was evaluated through embryoid body formation. Karyotyping and genetic fingerprinting were used to verify genomic stability and identity. Presence of the patient‑specific mutations in the iPSCs was confirmed by MLPA and Sanger sequencing.

Results: Stable iPSC lines were successfully generated for both genotypes. These lines exhibited robust expression of pluripotency markers, maintained normal karyotypes, and preserved the parental genetic fingerprint and genotypes. Furthermore, both iPSC lines demonstrated the capacity to differentiate into derivatives of the three embryonic germ layers.

Conclusion: These patient-specific iPSC models provide valuable tools for investigating synergistic interactions between PD-related genetic alterations. By modeling the molecular interplay among key pathways disrupted in PD, these lines offer a novel platform to study how these mechanisms converge to drive PD pathophysiology and to support the development of targeted, disease-modifying therapeutic strategies.

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MDS Abstracts - https://www.mdsabstracts.org/abstract/modeling-the-genetic-complexity-of-parkinsons-disease-generation-of-patient-specific-ipscs-with-co-occurring-snca-gba1-and-prkn-lrrk2-variants/