Browsing by Subject "MS"

Multiple sclerosis (MS) is an inflammatory neurodegenerative disease of the central nervous system, that affects over 2 million people worldwide. In MS, there is an autoimmune reaction against the myelin surrounding axons, causing neurological deficits and progressive neurological deterioration over time. Currently, there is no curative treatment for MS; only disease-modifying treatments are available. In this study, the effect of C-MANF (C-terminal mesencephalic astrocyte-derived neurotrophic factor) on unfolded protein response (UPR), neuroinflammation and myelination was investigated in different models of myelination by analyzing gene expression with quantitative polymerase chain reaction (qPCR). C-MANF is a C-terminal fragment of endoplasmic reticulum (ER) located protein MANF, which has been shown to protect cells against ER-stress induced apoptosis. UPR is a cellular defense mechanism against ER stress, but when prolonged, it can direct cells into apoptosis. The effect of C-MANF was investigated in ER-stressed Oli-neu cell cultures, both undifferentiated and differentiated. Cells were treated for 5 hours with thapsigargin to induce ER stress and simultaneously with C-MANF. In differentiated cells, a longer 24 hour exposure to ER stress and C-MANF was also investigated. Organotypic cerebellar brain slices from C57BL/6 P10 mice were demyelinated with lysophosphatidylcholine (LPC) for 17 hours, after which C-MANF was given during 5 day remyelination period. After the experiments were conducted, RNA was isolated and converted to cDNA for quantitative polymerase chain reaction (qPCR) using SYBR green. In addition, spinal cords from experimental autoimmune encephalomyelitis (EAE) animal experiment, where mice were given C-MANF subcutaneously, were included in this study, and qPCR was performed on them. This study showed significant changes in UPR gene expression in differentiated Oli-neu cells and EAE mice, indicating that C-MANF could alleviate UPR signaling and thus protect cells. Neuroinflammation was assessed with cerebellar brain slices and EAE mice; both models showed significant changes in gene expression related to neuroinflammation, indicating reduced neuroinflammation in response to C-MANF treatment. C-MANF did not seem to have an effect on myelination in this study. Large variation in between different groups in cerebellar brain slices may be due to challenges in RNA isolation an may have affected the results. However, these findings suggest the potential of C-MANF as a therapeutic for diseases associated with upregulated UPR signaling and a high degree neuroinflammation, such as MS. Further research and additional methodologies are needed to confirm these findings.