Functional integration of neurons derived from reprogrammed pericytes isolated from the adult human brain

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Cellular reprogramming of somatic cells into induced neurons could pave the way for new therapies of currently uncurable neurodegenerative diseases. We could previously show that pericytes isolated from the adult human brain can be reprogrammed into induced neurons by forced expression of the two transcription factors Sox2 and Ascl1. This proposal aims at investigating whether such reprogramming can also take place in vivo and whether thereby newly generated neurons can functionally integrate into a neuronal network. We will examine this by using a transplantation model established by Prof. Marcos R Costa at the Federal University of Rio Grande do Norte (Brazil). Hereby in vitro retrovirus-transduced human pericytes are transplanted into the lateral ventricle of mouse embryos. The retroviruses encode Sox2 and/or Ascl1 as well as a reporter gene (GFP or RFP). After different survival times, the transplanted cell will be located in the host tissue based on their reporter expression and examined for their marker expression (neuronal versus pericyte markers). If we can show that reprogramming can also occur in the in vivo context, we will next examine by means of a rabies virus-based tracing technique whether and which type of presynaptic inputs impinge on neurons derived from pericytes. This study will be complemented by classical patch-clamp electrophysiology for synaptic activity. The experiments described here will provide evidence whether neurons derived from reprogrammed pericytes of human origin can be recognized by authentic neurons as target cells and thus proof of principle evidence for or against their synaptic integration. This represents a major advancement in our understanding of the potential of this cell-based approach. This project will be conducted in close collaboration with Prof. Marcos R. Costa, who has applied for a grant at the CNPq to study the integration of neurons of reprogrammed mouse astrocytes. Both project will be highly complementary and require the expertise of both laboratories.