Inhibition of OCT4 Binding at the Locus Induces Neuroblastoma Cell Death Accompanied by Downregulation of Transcripts with High-open Reading Frame Dominance

Overview

Journal Front Oncol

Specialty Oncology

Date 2024 Feb 23

PMID 38390263

Authors

Kazuma Nakatani

Hiroyuki Kogashi

Takanori Miyamoto

Taiki Setoguchi

Tetsushi Sakuma

Kazuto Kugou

Yoshinori Hasegawa

Takashi Yamamoto

Yoshitaka Hippo

Yusuke Suenaga

Affiliations

Soon will be listed here.

Abstract

Amplification of is observed in high-risk neuroblastomas (NBs) and is associated with a poor prognosis. expression is directly regulated by multiple transcription factors, including OCT4, MYCN, CTCF, and p53 in NB. Our previous study showed that inhibition of p53 binding at the locus induces NB cell death. However, it remains unclear whether inhibition of alternative transcription factor induces NB cell death. In this study, we revealed that the inhibition of OCT4 binding at the locus, a critical site for the human-specific OCT4-MYCN positive feedback loop, induces caspase-2-mediated cell death in -amplified NB. We used the CRISPR/deactivated Cas9 (dCas9) technology to specifically inhibit transcription factors from binding to the locus in the -amplified NB cell lines CHP134 and IMR32. In both cell lines, the inhibition of OCT4 binding at the locus reduced MYCN expression, thereby suppressing MYCN-target genes. After inhibition of OCT4 binding, differentially downregulated transcripts were associated with high-open reading frame (ORF) dominance score, which is associated with the translation efficiency of transcripts. These transcripts were enriched in splicing factors, including MYCN-target genes such as and . Furthermore, transcripts with a high-ORF dominance score were significantly associated with genes whose high expression is associated with a poor prognosis in NB. Because the ORF dominance score correlates with the translation efficiency of transcripts, our findings suggest that MYCN maintains the expression of transcripts with high translation efficiency, contributing to a poor prognosis in NB. In conclusion, the inhibition of OCT4 binding at the locus resulted in reduced MYCN activity, which in turn led to the downregulation of high-ORF dominance transcripts and subsequently induced caspase-2-mediated cell death in -amplified NB cells. Therefore, disruption of the OCT4 binding at the locus may serve as an effective therapeutic strategy for -amplified NB.

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