CONICET | Buscador de Institutos y Recursos Humanos

Neuroblastoma (NB) is the most common extracranial pediatric cancer, originating fromcells of the sympathoadrenal lineage. ASCL1 is a critical transcription factor in thislineage's development whose expression should be embryonic and transient, however,it is overexpressed in NB and has been linked to a worse clinical prognosis. The aimwas to investigate the consequences of its knockdown (KD) in NB cells. We conductedASCL1 KD in SK-N-SH cells. Additionally, we analyzed publicly available single-cellRNA sequencing (scRNA-seq) data obtained from tumors of NB patients, to gaininsights into the heterogeneity of NB. ASCL1 loss of function results in the induction ofdifferentiation of these cells. NB cells lacking ASCL1 acquire neuronal morphology anddecrease their proliferative and migratory capacities, without affecting their viability. Forour scRNA-seq analysis, we integrated the data into a single dataset, and clustered bydimensional reduction. We characterized the derived clusters by performing functionalenrichment analysis and evaluating the expression patterns of ASCL1 with non-negative matrix factorization. Our findings revealed that NB cells share a commonorigin, specifically deriving from sympathoblasts of the adrenal medulla and that theobserved heterogeneity can be explained by the grade of cellular differentiation and thepredicted phase of the cell cycle. We discovered that ASCL1 is expressed acrossvarious clusters, and co-expresses with genes linked to neurogenesis and cell cycle.Finally, for future testing of ASCL1 KD, we optimized the implantation of SK-N-SH spheroids onto the chorioallantoic membrane of chicken.The results obtained led to two significant points: a) ASCL1 plays a role in blockingterminal neuronal differentiation in NB, thereby promoting tumor progression; b) theobserved effects of reducing ASCL1 function in vitro that correlate with the roleassessed in silico suggest that ASCL1 is a potential target for NB therapy.

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