title: LRRK2 Expression Is Deregulated in Fibroblasts and Neurons from Parkinson Patients with Mutations in PINK1.
abstract: Mutations in PINK1 (PARK6), a serine/threonine kinase involved in mitochondrial homeostasis, are associated with early onset Parkinson's disease. Fibroblasts from Parkinson's disease patients with compound heterozygous mutations in exon 7 (c.1488 + 1G > A; c.1252_1488del) showed no apparent signs of mitochondrial impairment. To elucidate changes primarily caused by lack of functional PINK1, we over-expressed wild-type PINK1, which induced a significant downregulation of LRRK2 (PARK8). Indeed, we found that LRRK2 protein basal levels were significantly higher in the mutant PINK1 fibroblasts. To examine the interaction between the two PARK genes in a disease-relevant cell context, we generated induced pluripotent stem cell (iPSC) lines from mutant, carrier and control fibroblasts by lentiviral-mediated re-programming. Efficiency of neural induction and dopamine differentiation using a floor-plate induction protocol was similar in all genotypes. As observed in fibroblasts, PINK1 mutant neurons showed increased LRRK2 expression both at the RNA and protein level and transient over-expression of wild-type PINK1 efficiently downregulated LRRK2 levels. Additionally, we confirmed a dysregulation of LRRK2 expression in fibroblasts from patients with a different homozygous mutation in PINK1 exon 4, c.926G > A (G309D). Thus, our results identify a novel role of PINK1 modulating the levels of LRRK2 in Parkinson's disease fibroblasts and neurons, suggest a convergent pathway for these PARK genes, and broaden the role of LRRK2 in the pathogenesis of Parkinson's disease.
authors.affiliation: Laboratory of Stem Cells and Neural Repair, Inbiomed, Paseo Mikeletegi, 81, 20009, San Sebastian, Spain.|Animal Model Unit, Inbiomed, San Sebastian, Spain.|Animal Research Facility, Scientific and Technological Centers, University of Barcelona, Barcelona, Spain.
authors.forename: Garikoitz
authors.initials: G
authors.lastname: Azkona
authors.affiliation: Laboratory of Stem Cells and Neural Repair, Inbiomed, Paseo Mikeletegi, 81, 20009, San Sebastian, Spain.
authors.forename: Rakel
authors.initials: R
authors.lastname: López de Maturana
authors.affiliation: Laboratory of Stem Cells and Neural Repair, Inbiomed, Paseo Mikeletegi, 81, 20009, San Sebastian, Spain.
authors.forename: Patricia
authors.initials: P
authors.lastname: Del Rio
authors.affiliation: Laboratory of Stem Cells and Neural Repair, Inbiomed, Paseo Mikeletegi, 81, 20009, San Sebastian, Spain.
authors.forename: Amaya
authors.initials: A
authors.lastname: Sousa
authors.affiliation: Laboratory of Stem Cells and Neural Repair, Inbiomed, Paseo Mikeletegi, 81, 20009, San Sebastian, Spain.
authors.forename: Nerea
authors.initials: N
authors.lastname: Vazquez
authors.affiliation: Laboratory of Stem Cells and Neural Repair, Inbiomed, Paseo Mikeletegi, 81, 20009, San Sebastian, Spain.
authors.forename: Amaia
authors.initials: A
authors.lastname: Zubiarrain
authors.affiliation: Institute of Functional Biology and Genomics (IBFG), University of Salamanca-CSIC, Salamanca, Spain.|Institute of Biomedical Research of Salamanca (IBSAL), University Hospital of Salamanca, Salamanca, Spain.
authors.forename: Daniel
authors.initials: D
authors.lastname: Jimenez-Blasco
authors.affiliation: Institute of Functional Biology and Genomics (IBFG), University of Salamanca-CSIC, Salamanca, Spain.|Institute of Biomedical Research of Salamanca (IBSAL), University Hospital of Salamanca, Salamanca, Spain.
authors.forename: Juan P
authors.initials: JP
authors.lastname: Bolaños
authors.affiliation: Department of Neurology, University Hospital San Cecilio, Granada, Spain.
authors.forename: Blas
authors.initials: B
authors.lastname: Morales
authors.affiliation: Experimental Neurology, Goethe University Medical School, Frankfurt am Main, Germany.
authors.forename: Georg
authors.initials: G
authors.lastname: Auburger
authors.affiliation: Parkinson's and Movement Disorders Unit, Department of Neurology, Hospital Universitario Insular de Gran Canaria, Las Palmas de Gran Canaria, Spain.
authors.forename: José Matias
authors.initials: JM
authors.lastname: Arbelo
authors.affiliation: Laboratory of Stem Cells and Neural Repair, Inbiomed, Paseo Mikeletegi, 81, 20009, San Sebastian, Spain. rpernaute@inbiomed.org.|Andalusian Initiative for Advanced Therapies, Junta de Andalucia, Sevilla, Spain. rpernaute@inbiomed.org.
authors.forename: Rosario
authors.initials: R
authors.lastname: Sánchez-Pernaute
mesh_terms: D015536:Down-Regulation; D005260:Female; D005347:Fibroblasts / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 / Q000235:genetics* / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D009154:Mutation / Q000235:genetics*; D009474:Neurons / Q000378:metabolism* / Q000473:pathology; D010300:Parkinson Disease / Q000235:genetics* / Q000473:pathology*; D011494:Protein Kinases / Q000235:genetics*; D000099202:PTEN-Induced Putative Kinase
chemical_list: D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D011494:Protein Kinases; D000099202:PTEN-Induced Putative Kinase
keywords: LRRK2; PINK1; Parkinson disease; iPSC