Glial Cultures Differentiated from iPSCs of Patients with PARK2-Associated Parkinson’s Disease Demonstrate a Pro-Inflammatory Shift and Reduced Response to TNFα Stimulation

Abstract

Parkinson’s disease (PD) is the second most common neurodegenerative diseases characterized by progressive loss of midbrain dopaminergic neurons in the substantia nigra. Mutations in the PARK2 gene are a frequent cause of familial forms of PD. Sustained chronic neuroinflammation in the central nervous system makes a significant contribution to neurodegeneration events. In response to inflammatory factors produced by activated microglia, astrocytes change their transcriptional programs and secretion profiles, thus acting as immunocompetent cells. Here, we investigated iPSC-derived glial cell cultures obtained from healthy donors (HD) and from PD patients with PARK2 mutations in resting state and upon stimulation by TNFα. The non-stimulated glia of PD patients demonstrated higher IL1B and IL6 expression levels and increased IL6 protein synthesis, while BDNF and GDNF expression was down-regulated when compared to that of the glial cells of HDs. In the presence of TNFα, all of the glial cultures displayed a multiplied expression of genes encoding inflammatory cytokines: TNFA, IL1B, and IL6, as well as IL6 protein synthesis, although PD glia responded to TNFα stimulation less strongly than HD glia. Our results demonstrated a pro-inflammatory shift, a suppression of the neuroprotective gene program, and some depletion of reactivity to TNFα in PARK2-deficient glia compared to glial cells of HDs.

Genetically Encoded and Modular SubCellular Organelle Probes (GEM-SCOPe) reveal lysosomal and mitochondrial dysfunction driven byPRKNknockout

“…Gliosis has been widely reported in human post-mortem brain studies of patients with PRKN loss-of-function mutations [94-97]. Similar pro-inflammatory phenotypes have also been observed in mouse models and stem-cell-derived glial models of PRKN deficiency [98, 99]. However, there is still a limited understanding of how genetic mutations related to PD modify astrocytes and their role in neurodegeneration.…”

IPSC-Derived Astrocytes Contribute to In Vitro Modeling of Parkinson’s Disease Caused by the GBA1 N370S Mutation

“…Studies on the generation of PD models based on astrocytes derived from iPSCs and their successful application to study various aspects of PD pathogenesis are increasingly appearing in the literature [ 29 , 30 , 31 ]. However, it is important to note that the model has some peculiarities because of astroglial cell ontogenesis.…”

Human Glial Cells as Innovative Targets for the Therapy of Central Nervous System Pathologies

“…Conversely, TNFα-stimulated glial cultures from both PD patients and healthy donors displayed an increased expression of genes encoding for pro-inflammatory cytokines, although PD glia responded to TNF-α stimulation less strongly than healthy glia. Authors assumed that glial cells in PARK2-associated PD have a "more inflammatory" status in the resting state but respond less strongly than healthy glia to inflammatory challenges, suggesting a reduced activation capacity [83]. Human iPSC-derived neurons and astrocytes were exposed to pro-inflammatory cytokines (i.e., TNF-α and IL-17A) typically associated with progressive multiple sclerosis (PMS).…”