RESUMEN
Neuroinflammation plays a pathogenic role in neurodegenerative diseases and recent findings suggest that it may also be involved in X-linked Dystonia-Parkinsonism (XDP) pathogenesis. Previously, fibroblasts and neuronal stem cells derived from XDP patients demonstrated hypersensitivity to TNF-α, dysregulation in NFκB signaling, and an increase in several pro-inflammatory markers. However, the role of inflammatory processes in XDP patient brain remains unknown. Here we demonstrate that there is a significant increase in astrogliosis and microgliosis in human post-mortem XDP prefrontal cortex (PFC) compared to control. Furthermore, there is a significant increase in histone H3 citrullination (H3R2R8R17cit3) with a concomitant increase in peptidylarginine deaminase 2 (PAD2) and 4 (PAD4), the enzymes catalyzing citrullination, in XDP post-mortem PFC. While there is a significant increase in myeloperoxidase (MPO) levels in XDP PFC, neutrophil elastase (NE) levels are not altered, suggesting that MPO may be released by activated microglia or reactive astrocytes in the brain. Similarly, there was an increase in H3R2R8R17cit3, PAD2 and PAD4 levels in XDP-derived fibroblasts. Importantly, treatment of fibroblasts with Cl-amidine, a pan inhibitor of PAD enzymes, reduced histone H3 citrullination and pro-inflammatory chemokine expression, without affecting cell survival. Taken together, our results demonstrate that inflammation is increased in XDP post-mortem brain and fibroblasts and unveil a new epigenetic potential therapeutic target.
Asunto(s)
Citrulinación , Trastornos Distónicos/metabolismo , Enfermedades Genéticas Ligadas al Cromosoma X/metabolismo , Histonas/metabolismo , Inflamación/metabolismo , Corteza Prefrontal/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Astrocitos/metabolismo , Astrocitos/patología , Autopsia , Supervivencia Celular , Quimiocinas/efectos de los fármacos , Quimiocinas/metabolismo , Citrulinación/efectos de los fármacos , Trastornos Distónicos/patología , Femenino , Fibroblastos/efectos de los fármacos , Enfermedades Genéticas Ligadas al Cromosoma X/patología , Gliosis/metabolismo , Gliosis/patología , Histonas/efectos de los fármacos , Humanos , Inflamación/patología , Elastasa de Leucocito/metabolismo , Masculino , Microglía/metabolismo , Microglía/patología , Persona de Mediana Edad , Ornitina/análogos & derivados , Ornitina/farmacología , Peroxidasa/metabolismo , Corteza Prefrontal/patología , Arginina Deiminasa Proteína-Tipo 2/metabolismo , Arginina Deiminasa Proteína-Tipo 4/metabolismoRESUMEN
X-linked Dystonia-Parkinsonism (XDP) is a neurodegenerative disease linked to an insertion of a SINE-VNTR-Alu (SVA)-type retrotransposon within an intron of TAF1. This SVA insertion induces aberrant TAF1 splicing and partial intron retention, thereby decreasing levels of the full-length transcript. Here we sought to determine if these altered transcriptional dynamics caused by the SVA are also accompanied by local changes in histone acetylation, given that these modifications influence gene expression. Because TAF1 protein may itself exhibit histone acetyltransferase activity, we also examined whether decreased TAF1 expression in XDP cell lines and post-mortem brain affects global levels of acetylated histone H3 (AcH3). The results demonstrate that total AcH3 are not altered in XDP post-mortem prefrontal cortex or cell lines. We also did not detect local differences in AcH3 associated with TAF1 exons or intronic sites flanking the SVA insertion. There was, however, a decrease in AcH3 association with the exon immediately proximal to the intronic SVA, and this decrease was normalized by CRISPR/Cas-excision of the SVA. Collectively, these data suggest that the SVA insertion alters histone status in this region, which may contribute to the dysregulation of TAF1 expression.