Banca de QUALIFICAÇÃO: LUKAS IOHAN DA CRUZ CARVALHO

Uma banca de QUALIFICAÇÃO de DOUTORADO foi cadastrada pelo programa.
STUDENT : LUKAS IOHAN DA CRUZ CARVALHO
DATE: 29/09/2023
TIME: 09:00
LOCAL: Google Meet, https://meet.google.com/bgb-nhev-jbe
TITLE:

Bioinformatics Molecular Characterization of Induced Human Neurons

KEY WORDS:

HiNPCs; ASCL1; SMAD; scRNA-Seq

PAGES: 56
BIG AREA: Ciências Biológicas
AREA: Biologia Geral
SUMMARY:

Various protocols have been developed in recent years for generating neuronal cells from human induced pluripotent stem cells (hiNPCs), revolutionizing neuroscience research and enhancing our comprehension of human disease pathogenesis. These protocols encompass approaches such as SMAD inhibition or the forced conversion of hiNPCs into human induced neurons (hINs) through the expression of pivotal transcription factors (TFs) associated with neuronal differentiation. In this study, we undertook a comparative analysis of a novel hIN generation method, which combines SMAD inhibition with the overexpression of ASCL1, a well-known TF implicated in neuronal induction, alongside two differentiation strategies: NEUROG2-induced differentiation and spontaneous differentiation of hiNPCs. Following a 4-6 week differentiation period, cells from these distinct protocols were isolated, subjected to sequencing, and subsequently analyzed using single-cell analytical tools. Our findings from differential expression and Gene Set Enrichment Analysis (GSEA) unveiled that ASCL1-induced hINs, encompassing both glutamatergic and GABAergic subtypes, exhibited significant upregulation of genes associated with synaptic maturation and neuronal development, distinguishing them from hiNPCs_4w, hiNPCs_6w, and NEUROG2-induced hINs. Furthermore, regional identity analysis revealed a prevalent cortex identity among glutamatergic cells, in contrast to the diverse regional identities observed among GABAergic neurons. Additionally, regulon analysis identified candidate transcription factors that may elucidate the mechanism behind the enhanced maturation properties of the novel ASCL1 protocol. Further exploration of the downstream targets of these regulons and the associated regulatory networks presents an intriguing avenue for future investigations into this innovative hIN differentiation approach.

COMMITTEE MEMBERS:
Presidente - 1674643 - MARCOS ROMUALDO COSTA
Externo ao Programa - 2183828 - TARCISO ANDRE FERREIRA VELHO - UFRN