Human Huntington's disease pluripotent stem cell-derived microglia develop normally but are abnormally hyper-reactive and release elevated levels of reactive oxygen species

• Published in: Journal of neuroinflammation Vol. 18; no. 1; pp. 94 - 17
• Main Authors: O'Regan, Grace C, Farag, Sahar H, Casey, Caroline S, Wood-Kaczmar, Alison, Pocock, Jennifer M, Tabrizi, Sarah J, Andre, Ralph
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 19.04.2021; BioMed Central; BMC
• Subjects: Apoptosis; Biopsy; Cell Differentiation; Cell Line; Corpus Striatum - metabolism; Cytokines; Development and progression; Fibroblasts; Genetic aspects; Health aspects; Humans; Huntingtin Protein - genetics; Huntingtin Protein - metabolism; Huntington Disease - genetics; Huntington Disease - metabolism; Huntington Disease - pathology; Huntington Disease - physiopathology; Huntington's chorea; Huntingtons disease; Huntington’s disease; Immune system; Immunofluorescence; Induced Pluripotent Stem Cells - metabolism; Induced Pluripotent Stem Cells - pathology; Inflammation; Interleukin 6; Microglia; Microglia - metabolism; Microglia - pathology; Monocytes; Mutants; Mutation; Neostriatum; Neurodegeneration; Neurons - metabolism; Neurotoxicity; Pathology; Penicillin; Phenotypes; Pluripotency; Pluripotent stem cells; Polyglutamine; Reactive oxygen species; Reactive Oxygen Species - metabolism; Stem cells; Striatal neurons; Trinucleotide repeat diseases; Tumor necrosis factor-α; United Kingdom; Reactive oxygen species; Cytokines; Neurodegeneration; Striatal neurons; Huntington’s disease; Pluripotent stem cells; Microglia
• ISSN: 1742-2094; 1742-2094
• DOI: 10.1186/s12974-021-02147-6

Neuroinflammation may contribute to the pathogenesis of Huntington's disease, given evidence of activated microglia and elevated levels of inflammatory molecules in disease gene carriers, even those many years from symptom onset. We have shown previously that monocytes from Huntington's disease patients are hyper-reactive to stimulation in a manner dependent on their autonomous expression of the disease-causing mutant HTT protein. To date, however, whether human microglia are similarly hyper-responsive in a cell-autonomous manner has not been determined. Microglial-like cells were derived from human pluripotent stem cells (PSCs) expressing mutant HTT containing varying polyglutamine lengths. These included lines that are otherwise isogenic, such that any observed differences can be attributed with certainty to the disease mutation itself. Analyses by quantitative PCR and immunofluorescence microscopy respectively of key genes and protein markers were undertaken to determine whether Huntington's disease PSCs differentiated normally to a microglial fate. The resultant cultures and their supernatants were then assessed by various biochemical assays and multiplex ELISAs for viability and responses to stimulation, including the release of pro-inflammatory cytokines and reactive oxygen species. Conditioned media were applied to PSC-derived striatal neurons, and vice versa, to determine the effects that the secretomes of each cell type might have on the other. Human PSCs generated microglia successfully irrespective of the expression of mutant HTT. These cells, however, were hyper-reactive to stimulation in the production of pro-inflammatory cytokines such as IL-6 and TNFα. They also released elevated levels of reactive oxygen species that have neurotoxic potential. Accompanying such phenotypes, human Huntington's disease PSC-derived microglia showed increased levels of apoptosis and were more susceptible to exogenous stress. Such stress appeared to be induced by supernatants from human PSC-derived striatal neurons expressing mutant HTT with a long polyglutamine tract. These studies show, for the first time, that human Huntington's disease PSC-derived microglia are hyper-reactive due to their autonomous expression of mutant HTT. This provides a cellular basis for the contribution that neuroinflammation might make to Huntington's disease pathogenesis.