TY  - JOUR
AU  - Knab, Felix
AU  - Lee, Jun-Hoe
AU  - Nirujogi, Raja
AU  - Menden, Kevin
AU  - Braunger, Luca
AU  - Logarnudi, Lambrianna
AU  - Riebenbauer, Benjamin
AU  - Isik, Fatma Busra
AU  - Rajkumar, Anto Praveen
AU  - Czemmel, Stefan
AU  - Fitzgerald, Julia
AU  - Gasser, Thomas
AU  - Gloeckner, Christian Johannes
TI  - Cellular and Extracellular MicroRNA Dysregulation in LRRK2-Linked Parkinson's Disease.
JO  - Molecular neurobiology
VL  - 63
IS  - 1
SN  - 0893-7648
CY  - Totowa, NJ
PB  - Humana Press
M1  - DZNE-2025-01329
SP  - 189
PY  - 2025
AB  - Cell-free microRNAs in body fluids have emerged as promising biomarker candidates in neurodegenerative diseases. While several studies have identified dysregulated miRNAs in sporadic Parkinson's disease, it remains unclear whether distinguishable alterations of cell-free miRNAs occur in genetic forms of the disease, such as those associated with the LRRK2 G2019S mutation. In this proof-of-concept study, we used a human induced pluripotent stem cell-derived dopaminergic neuron model to investigate whether the LRRK2 G2019S mutation induces detectable changes in the intra- and extracellular miRNAome, and whether miRNA signatures identified in vitro can be validated in patient-derived cerebrospinal fluid. We differentiated dopaminergic neurons from induced pluripotent stem cells carrying the LRRK2 G2019S mutation and an isogenic gene-corrected control. Extracellular vesicles were isolated from the culture medium and used as a source of cell-free miRNA. Next, small RNA libraries were generated and analyzed. Differentially expressed microRNAs were validated in an independent batch using RT-qPCR. We further quantified candidate microRNAs in cerebrospinal fluid samples from five LRRK2 G2019S patients and matching healthy controls. The patient cohort included the fibroblast donor from whom the stem cells were originally derived. We successfully isolated extracellular vesicles from induced pluripotent stem cell-derived human dopaminergic neurons. We identified a distinct set of differentially expressed miRNAs in cellular and cell-free RNA, among which let-7g-5p and miR-21-5p were consistently upregulated and validated across independent replicates. These alterations were reflected in the cerebrospinal fluid of the original donor and partially reproduced in additional LRRK2 patients, supporting the concept of patient-specific signatures. A strong correlation between intra- and extracellular miRNA expression was observed. Our findings demonstrate that induced pluripotent stem cell-derived dopaminergic neurons can serve as a model to identify individualized, cell-free microRNA signatures associated with the LRRK2 G2019S mutation. The dysregulated miRNAs detected in vitro were mirrored in patient cerebrospinal fluid, supporting their potential as accessible molecular readouts. These results lay the groundwork for personalized biomarker strategies in genetic forms of Parkinson's disease and warrant further validation in larger patient cohorts.
KW  - Humans
KW  - Leucine-Rich Repeat Serine-Threonine Protein Kinase-2: genetics
KW  - Leucine-Rich Repeat Serine-Threonine Protein Kinase-2: metabolism
KW  - MicroRNAs: genetics
KW  - MicroRNAs: metabolism
KW  - MicroRNAs: cerebrospinal fluid
KW  - Parkinson Disease: genetics
KW  - Parkinson Disease: cerebrospinal fluid
KW  - Induced Pluripotent Stem Cells: metabolism
KW  - Dopaminergic Neurons: metabolism
KW  - Dopaminergic Neurons: pathology
KW  - Extracellular Vesicles: metabolism
KW  - Mutation: genetics
KW  - Cell Differentiation
KW  - Middle Aged
KW  - Male
KW  - Female
KW  - Biomarker (Other)
KW  - IPSCs (Other)
KW  - LRRK2 (Other)
KW  - Micro-RNA (Other)
KW  - Parkinson’s disease (Other)
KW  - Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 (NLM Chemicals)
KW  - MicroRNAs (NLM Chemicals)
KW  - LRRK2 protein, human (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:41298996
C2  - pmc:PMC12657546
DO  - DOI:10.1007/s12035-025-05379-2
UR  - https://pub.dzne.de/record/282566
ER  -