Development of Myelinating Human Brain Organoids for Modelling Multiple Sclerosis: A Platform For Drug Screening Studies and Mechanistic Investigations
Abstract
Multiple sclerosis (MS) is a neurodegenerative disease that is characterized mainly by demyelination and inflammation. The disadvantages of the current two-dimensional cell culture models and experimental animals imply the need for the development of new disease models. Brain organoids are emerging and powerful tools for recapitulating the central nervous system. Although the models have been generated for many years, time-consuming processes and less cellular diversity are important challenges in defined protocols. In this study, we aimed to generate an optimized brain organoid protocol and model organoids for MS. In these processes, human embryonic stem cells were directed to neurons via the extracellular matrix (ECM) and growth factors to generate brain organoids with diverse neural cells. The presence of myelinating cells, astrocytes, microglia and excitatory neurons in the organoids was verified by immunostaining. MS was then used to induce inflammation and demyelination via lipopolysaccharide (LPS). The model was also confirmed by immunostaining, which revealed an ~50% increase in GFAP and an ~60% decrease in CNPase-positive cells. Finally, the use of organoids in the drug screening field was tested via fingolimod treatment of LPS-induced organoids. A comparison between fingolimod-treated and untreated organoids revealed that fingolimod decreased GFAP by more than 80% and increased the percentage of CNPase-positive cells by 90%. Additionally, the relative expression levels of inflammation-related transcripts (FOXP3 and GFAP) after fingolimod treatment were significantly decreased. In conclusion, a human brain organoid model for MS studies was successfully generated for use in drug screening and mechanistic studies.
Keywords
brain organoids , myelination , inflammation , multiple sclerosis
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