Mitochondria interaction networks show altered topological patterns in Parkinson's disease.

in NPJ systems biology and applications (2020), 6(1), 38

Mitochondrial dysfunction is linked to pathogenesis of Parkinson's disease (PD). However, individual mitochondria-based analyses do not show a uniform feature in PD patients. Since mitochondria interact ... [more ▼]

Mitochondrial dysfunction is linked to pathogenesis of Parkinson's disease (PD). However, individual mitochondria-based analyses do not show a uniform feature in PD patients. Since mitochondria interact with each other, we hypothesize that PD-related features might exist in topological patterns of mitochondria interaction networks (MINs). Here we show that MINs formed nonclassical scale-free supernetworks in colonic ganglia both from healthy controls and PD patients; however, altered network topological patterns were observed in PD patients. These patterns were highly correlated with PD clinical scores and a machine-learning approach based on the MIN features alone accurately distinguished between patients and controls with an area-under-curve value of 0.989. The MINs of midbrain dopaminergic neurons (mDANs) derived from several genetic PD patients also displayed specific changes. CRISPR/CAS9-based genome correction of alpha-synuclein point mutations reversed the changes in MINs of mDANs. Our organelle-interaction network analysis opens another critical dimension for a deeper characterization of various complex diseases with mitochondrial dysregulation. [less ▲]

Fibroblast mitochondria in idiopathic Parkinson’s disease display morphological changes and enhanced resistance to depolarization

in Scientific Reports (2020)

Mitochondrial dysfunction is a hallmark in idiopathic Parkinson’s disease (IPD). Here, we established screenable phenotypes of mitochondrial morphology and function in primary fibroblasts derived from ... [more ▼]

Mitochondrial dysfunction is a hallmark in idiopathic Parkinson’s disease (IPD). Here, we established screenable phenotypes of mitochondrial morphology and function in primary fibroblasts derived from patients with IPD. Upper arm punch skin biopsy was performed in 41 patients with mid-stage IPD and 21 age-matched healthy controls. At the single-cell level, the basal mitochondrial membrane potential (Ψm) was higher in patients with IPD than in controls. Similarly, under carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) stress, the remaining Ψm was increased in patients with IPD. Analysis of mitochondrial morphometric parameters revealed significantly decreased mitochondrial connectivity in patients with IPD, with 9 of 14 morphometric mitochondrial parameters differing from those in controls. Significant morphometric mitochondrial changes included the node degree, mean volume, skeleton size, perimeter, form factor, node count, erosion body count, endpoints, and mitochondria count (all P-values < 0.05). These functional data reveal that resistance to depolarization was increased by treatment with the protonophore FCCP in patients with IPD, whereas morphometric data revealed decreased mitochondrial connectivity and increased mitochondrial fragmentation. [less ▲]

Functional characterization of novel RhoT1 variants, which are associated with Parkinson's disease.

Doctoral thesis (2016)

Parkinson’s disease (PD) is a common neurodegenerative disease affecting up to 2 % of the population older than 65 years. Most PD cases are sporadic with unknown cause, and about 10 % are familial ... [more ▼]

Parkinson’s disease (PD) is a common neurodegenerative disease affecting up to 2 % of the population older than 65 years. Most PD cases are sporadic with unknown cause, and about 10 % are familial inherited. PD is a progressive neurodegenerative disease characterized by loss of predominantly dopaminergic neurons, leading to typical symptoms like rigidity and tremor. Commonly involved pathogenic pathways are linked to mitochondrial dysfunction, e.g. increased oxidative stress, disruption of calcium homeostasis, decreased energy supply and mitochondrial-controlled apoptosis. The mitochondrial outer membrane protein Miro1 is important for mitochondrial distribution, quality control and maintenance. To date Miro1 is not established as risk factor for PD. Using a comprehensive mutation screening of RhoT1 in German PD patients we dissected the role of the first PD-associated mutations in RhoT1, the gene encoding for Miro1. Three mutations in RhoT1 have been identified in three PD patients with positive family history for PD. For analysis of mitochondrial phenotypes patient-derived fibroblasts from two of the three patients were available. As independent cell model served the neuroblastoma cell line M17 with stable knockdown of endogenous RhoT1 and transiently overexpression of the RhoT1 mutant variants. Investigation of yeast with knockout of endogenous Gem1 (the yeast orthologue of Miro1) and overexpression of mutant Gem1 revealed that growth on non-fermentable carbon source was impaired. These findings suggest that Miro1-mutant1 is a loss of function mutation. Interestingly, the Miro1 protein amount was significantly reduced in Miro1-mutant1 and Miro1-mutant2 fibroblast lines compared to controls. Functional analysis revealed that mitochondrial mass was decreased in Miro1-mutant2, but not in Miro1-mutant1 fibroblasts, whereas mitochondrial biogenesis was increased in Miro1-mutant2 fibroblasts, as indicated by elevation of PGC1α. A similar phenotype with reduction of mitochondrial mass was also observed in M17 cells overexpressing Miro1-mutant1 or Miro1-mutant2. Additionally, spare respiratory capacity was reduced in Miro1-mutant1 fibroblasts compared to Ctrl 1 fibroblasts. In contrast, Miro1-mutant2 fibroblasts showed increased respiratory activity compared to Ctrl 1, despite citrate synthase activity was significantly reduced. Both alterations of respiratory activity lead to mitochondrial membrane hyperpolarization in Miro1-mutant1 and Miro1-mutant2 fibroblasts, a phenotype which was also found in M17 cells with knockdown of RhoT1. Both Miro1 mutant fibroblasts lines displayed different problems with cytosolic calcium buffering: in Miro1-mutant1 fibroblasts histamine treatment increased cytosolic calcium concentration significantly compared to Ctrl 1 fibroblasts, indicating that calcium homeostasis was impaired, whereas in Miro1-mutant2 fibroblasts the buffering capacity for cytosolic calcium was impaired. The results indicate that mutations in Miro1 cause significant mitochondrial dysfunction, which are likely contributing to neurodegeneration in PD and underline the importance of Miro1 for mitochondrial maintenance. [less ▲]