Generation of induced pluripotent stem cells from epithelial cells in urine from individuals with Down Syndrome

Abstract

Down syndrome (DS), which is the consequence of the trisomy of chromosome 21 (T21), has an approximate incidence of 1:660 live births. DS is the most prevalent cause of intellectual disability of genetic origin and results in general cognitive impairment, which is typically in the mild-to-moderate range. Although DS has been known for over 160 years, our knowledge regarding the mechanisms responsible for its phenotype, as well as effective treatments, are still incipient. The differentiation of human cells into induced pluripotent stem cells (iPSCs) has become an essential tool for modeling genetic diseases. In 2011, Zhou et al. showed that epithelial cells in urine samples could efficiently be isolated and reprogrammed into iPSCs. In 2013, Xue et al. described a detailed protocol for iPSCs generation using cells isolated from urine from donors with different ages and various clinical histories, including disease genetic backgrounds. In 2017, as part of my postdoctoral training, I was part of the research team that optimized these techniques and produced for the first time T21-iPSC lines from epithelial cells in urine samples from individuals with DS. In the present project, we propose to bring this new technology to Brazil and not only generate T21-iPSC lines, but also to differentiate them into glutamatergic neurons and astrocytes and assess their nmethyldaspartate (NMDA) receptor and complement system associated cellular phenotypes. The generation of T21-iPSCs through a completely noninvasive method should help the advance of future basic and translational studies in DS. The investigation of NMDA receptors and complement system in neuronal cells derived from T21-iPSCs are unpublished, will be relevant for the understanding of the DS pathology, and could make a powerful resource for the assessment and development of candidate therapeutic strategies. (AU)