Research Projects

Demyelination is one of the cardinal features that can cause disruption in motor coordination during viral infection. Japanese encephalitis (JE), an acute and uncontrolled inflammatory disease of the CNS in humans, is caused by the Japanese encephalitis virus (JEV), a member of the family of Flaviviridae. The endemic also considered a neglected tropical disease, is highly prevalent among acute encephalitis in India and the rate of incidence is increasing exponentially, especially in the Northern regions of India. Understanding the intrinsic mechanisms of demyelination disorder caused by neurotropic viruses may help to discover novel drugs to combat neurological disorders. However, current existing models have technical limitations and are not suitable for high-throughput drug screening studies. Thus, there is a need for developing a novel tool that can be utilized for understanding mechanisms as well as can be used as a tool to screen novel drugs to treat demyelination. The organotypic brain culture is a suitable model for the neurovirology field to better understand CNS infections. However, a promising organotypic brain slice culture model of demyelination is yet to be established for JEV infection. Here, we hypothesize that the development of organotypic cultures of the neurogenic niches will be a valid alternative method to study virus-induced demyelination in the CNS, which complements in vivo models and neurosphere cultures. The proposal also assumes that the ex-vivo model is suitable for evaluating the anti-demyelinating effect of drugs. The role of the ceramide biosynthetic pathway is well-established in the pathophysiology of demyelination and virus-induced demyelination. In addition, it has been documented that ceramide plays a significant role in the propagation of JEV in the cell culture study. However, the effect of JEV infection on the ceramide biosynthetic pathway remains to be elaborated in demyelination. It is to be emphasized that a promising drug candidate is yet to be established in the management of JEV-induced demyelination. Hence, an organotypic cerebellar culture model will be developed and characterized to assess demyelination. Subsequently, the extent of alteration in the ceramide biosynthetic pathway will be investigated in the standardized ex-vivo model. Thereafter, the therapeutic effectiveness of teriflunomide, dimethyl furoate, and fingolimod will be evaluated on the activity of the selected enzyme(s) and the extent of demyelination in the ex-vivo model. In the last objective, the therapeutic effectiveness of teriflunomide, dimethyl furoate, and fingolimod will be evaluated on the activity of the selected enzyme(s) and the extent of demyelination in the standardized animal model of JE. If the proposed project works, then a better-optimized JEV-challenged organotypic cerebellar slice culture model of demyelination will be developed that could be effective in the screening of anti-demyelinating drugs.