12 research outputs found
Generation of isogenic control DJ-1-delP GC13 for the genetic Parkinson's disease-patient derived iPSC line DJ-1-delP (LCSBi008-A-1)
Generation and characterization of a genetic Parkinson`s disease-patient derived iPSC line DJ-1-delP (LCSBi008-A)
GDAP1 loss of function inhibits the mitochondrial pyruvate dehydrogenase complex by altering the actin cytoskeleton
Mitochondrial and Clearance Impairment in p.D620N VPS35 Patient-Derived Neurons
Background: VPS35 is part of the
retromer complex and is responsible for the trafficking
and recycling of proteins implicated in autophagy and
lysosomal degradation, but also takes part in the degradation
of mitochondrial proteins via mitochondria-derived
vesicles. The p.D620N mutation of VPS35 causes an
autosomal-dominant form of Parkinson’s disease (PD),
clinically representing typical PD.
Objective: Most of the studies on p.D620N VPS35 were
performed on human tumor cell lines, rodent models
overexpressing mutant VPS35, or in patient-derived
fibroblasts. Here, based on identified target proteins, we
investigated the implication of mutant VPS35 in
autophagy, lysosomal degradation, and mitochondrial
function in induced pluripotent stem cell-derived neurons
from a patient harboring the p.D620N mutation.
Methods: We reprogrammed fibroblasts from a PD
patient carrying the p.D620N mutation in the VPS35
gene and from two healthy donors in induced pluripotent
stem cells. These were subsequently differentiated into
neuronal precursor cells to finally generate midbrain
dopaminergic neurons.
Results: We observed a decreased autophagic flux and
lysosomal mass associated with an accumulation of
α-synuclein in patient-derived neurons compared to controls.
Moreover, patient-derived neurons presented a mitochondrial
dysfunction with decreased membrane potential,
impaired mitochondrial respiration, and increased production
of reactive oxygen species associated with a defect in
mitochondrial quality control via mitophagy.
Conclusion: We describe for the first time the impact of the
p.D620N VPS35 mutation on autophago-lysosome pathway
and mitochondrial function in stem cell-derived neurons
from an affected p.D620N carrier and define neuronal phenotypes
for future pharmacological intervention
The Role of DJ-1 in Cellular Metabolism and Pathophysiological Implications for Parkinson’s Disease
The Role of DJ-1 in Cellular Metabolism and Pathophysiological Implications for Parkinson’s Disease
DJ-1 is a multifunctional protein associated with pathomechanisms implicated in different chronic diseases including neurodegeneration, cancer and diabetes. Several of the physiological functions of DJ-1 are not yet fully understood; however, in the last years, there has been increasing evidence for a potential role of DJ-1 in the regulation of cellular metabolism. Here, we summarize the current knowledge on specific functions of DJ-1 relevant to cellular metabolism and their role in modulating metabolic pathways. Further, we illustrate pathophysiological implications of the metabolic effects of DJ-1 in the context of neurodegeneration in Parkinson´s disease
Generation and characterization of a genetic Parkinson's disease-patient derived iPSC line DJ-1-delP (LCSBi008-A)
Here, we describe an induced pluripotent stem cell (iPSC) line that was derived from fibroblasts obtained from a monogenic Parkinson's disease (PD) patient. The disease was caused by a c.634-636delGCC mutation in the PARK7 gene leading to p.158P deletion in the protein DJ-1. iPSCs were generated via electroporation using three episomal plasmids encoding human Oct3/4, Sox2, Klf4, Lin28, L-Myc combined with a short hairpin RNA for p53. The presence of the c.471_473delGCC mutation in exon 7 of PARK7 was confirmed by Sanger sequencing. The iPSCs express pluripotency markers, are capable of in vitro differentiation into the three germ layers and obtain karyotypic integrity
Generation of isogenic control DJ-1-delP GC13 for the genetic Parkinson's disease-patient derived iPSC line DJ-1-delP (LCSBi008-A-1)
We describe the generation of an isogenic control cell line DJ-1-delP GC13 from an induced pluripotent stem cell (iPSC) line DJ-1-delP LCSBi008-A that was derived from fibroblasts obtained from a Parkinson’s disease (PD) patient. Using CRISPR/Cas9 technology, we corrected the disease causing c.471_473delGCC homozygous mutation in the PARK7 gene leading to p.158P deletion in the encoded protein DJ-1. The generated isogenic pair will be used for phenotypic analysis of PD-patient derived neurons and astrocytes