Inhibition of OCT4 binding at the MYCN locus induces neuroblastoma cell death accompanied by downregulation of transcripts with high-open reading frame dominance
Amplification of MYCN is observed in high-risk neuroblastomas (NBs) and is associated with a poor prognosis. MYCN 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 MYCN locus...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2024-02-01
|
| Series: | Frontiers in Oncology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fonc.2024.1237378/full |
| _version_ | 1797320779821481984 |
|---|---|
| author | Kazuma Nakatani Kazuma Nakatani Kazuma Nakatani Kazuma Nakatani Hiroyuki Kogashi Hiroyuki Kogashi Takanori Miyamoto Taiki Setoguchi Tetsushi Sakuma Kazuto Kugou Yoshinori Hasegawa Takashi Yamamoto Yoshitaka Hippo Yoshitaka Hippo Yoshitaka Hippo Yusuke Suenaga |
| author_facet | Kazuma Nakatani Kazuma Nakatani Kazuma Nakatani Kazuma Nakatani Hiroyuki Kogashi Hiroyuki Kogashi Takanori Miyamoto Taiki Setoguchi Tetsushi Sakuma Kazuto Kugou Yoshinori Hasegawa Takashi Yamamoto Yoshitaka Hippo Yoshitaka Hippo Yoshitaka Hippo Yusuke Suenaga |
| author_sort | Kazuma Nakatani |
| collection | DOAJ |
| description | Amplification of MYCN is observed in high-risk neuroblastomas (NBs) and is associated with a poor prognosis. MYCN 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 MYCN 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 MYCN locus, a critical site for the human-specific OCT4–MYCN positive feedback loop, induces caspase-2-mediated cell death in MYCN-amplified NB. We used the CRISPR/deactivated Cas9 (dCas9) technology to specifically inhibit transcription factors from binding to the MYCN locus in the MYCN-amplified NB cell lines CHP134 and IMR32. In both cell lines, the inhibition of OCT4 binding at the MYCN 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 HNRNPA1 and PTBP1. 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 MYCN 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 MYCN-amplified NB cells. Therefore, disruption of the OCT4 binding at the MYCN locus may serve as an effective therapeutic strategy for MYCN-amplified NB. |
| first_indexed | 2024-03-08T04:47:55Z |
| format | Article |
| id | doaj.art-acc0491d59654de7b7070657dd1c0837 |
| institution | Directory Open Access Journal |
| issn | 2234-943X |
| language | English |
| last_indexed | 2024-03-08T04:47:55Z |
| publishDate | 2024-02-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Oncology |
| spelling | doaj.art-acc0491d59654de7b7070657dd1c08372024-02-08T08:23:00ZengFrontiers Media S.A.Frontiers in Oncology2234-943X2024-02-011410.3389/fonc.2024.12373781237378Inhibition of OCT4 binding at the MYCN locus induces neuroblastoma cell death accompanied by downregulation of transcripts with high-open reading frame dominanceKazuma Nakatani0Kazuma Nakatani1Kazuma Nakatani2Kazuma Nakatani3Hiroyuki Kogashi4Hiroyuki Kogashi5Takanori Miyamoto6Taiki Setoguchi7Tetsushi Sakuma8Kazuto Kugou9Yoshinori Hasegawa10Takashi Yamamoto11Yoshitaka Hippo12Yoshitaka Hippo13Yoshitaka Hippo14Yusuke Suenaga15Laboratory of Evolutionary Oncology, Chiba Cancer Center Research Institute, Chiba, JapanGraduate School of Medical and Pharmaceutical Sciences, Chiba University, Chiba, JapanInnovative Medicine CHIBA Doctoral WISE Program, Chiba University, Chiba, JapanAll Directional Innovation Creator Ph.D. Project, Chiba University, Chiba, JapanLaboratory of Evolutionary Oncology, Chiba Cancer Center Research Institute, Chiba, JapanGraduate School of Medical and Pharmaceutical Sciences, Chiba University, Chiba, JapanLaboratory of Evolutionary Oncology, Chiba Cancer Center Research Institute, Chiba, JapanDepartment of Neurosurgery, Chiba Cancer Center, Chiba, JapanGraduate School of Integrated Sciences for Life, Hiroshima University, Hiroshima, JapanDepartment of Applied Genomics, Kazusa DNA Research Institute, Chiba, JapanDepartment of Applied Genomics, Kazusa DNA Research Institute, Chiba, JapanGraduate School of Integrated Sciences for Life, Hiroshima University, Hiroshima, JapanLaboratory of Evolutionary Oncology, Chiba Cancer Center Research Institute, Chiba, JapanGraduate School of Medical and Pharmaceutical Sciences, Chiba University, Chiba, JapanLaboratory of Precision Tumor Model Systems, Chiba Cancer Center Research Institute, Chiba, JapanLaboratory of Evolutionary Oncology, Chiba Cancer Center Research Institute, Chiba, JapanAmplification of MYCN is observed in high-risk neuroblastomas (NBs) and is associated with a poor prognosis. MYCN 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 MYCN 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 MYCN locus, a critical site for the human-specific OCT4–MYCN positive feedback loop, induces caspase-2-mediated cell death in MYCN-amplified NB. We used the CRISPR/deactivated Cas9 (dCas9) technology to specifically inhibit transcription factors from binding to the MYCN locus in the MYCN-amplified NB cell lines CHP134 and IMR32. In both cell lines, the inhibition of OCT4 binding at the MYCN 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 HNRNPA1 and PTBP1. 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 MYCN 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 MYCN-amplified NB cells. Therefore, disruption of the OCT4 binding at the MYCN locus may serve as an effective therapeutic strategy for MYCN-amplified NB.https://www.frontiersin.org/articles/10.3389/fonc.2024.1237378/fullneuroblastomaMYCNoct4open reading frame dominanceCRISPR/dCas9p53 |
| spellingShingle | Kazuma Nakatani Kazuma Nakatani Kazuma Nakatani Kazuma Nakatani Hiroyuki Kogashi Hiroyuki Kogashi Takanori Miyamoto Taiki Setoguchi Tetsushi Sakuma Kazuto Kugou Yoshinori Hasegawa Takashi Yamamoto Yoshitaka Hippo Yoshitaka Hippo Yoshitaka Hippo Yusuke Suenaga Inhibition of OCT4 binding at the MYCN locus induces neuroblastoma cell death accompanied by downregulation of transcripts with high-open reading frame dominance Frontiers in Oncology neuroblastoma MYCN oct4 open reading frame dominance CRISPR/dCas9 p53 |
| title | Inhibition of OCT4 binding at the MYCN locus induces neuroblastoma cell death accompanied by downregulation of transcripts with high-open reading frame dominance |
| title_full | Inhibition of OCT4 binding at the MYCN locus induces neuroblastoma cell death accompanied by downregulation of transcripts with high-open reading frame dominance |
| title_fullStr | Inhibition of OCT4 binding at the MYCN locus induces neuroblastoma cell death accompanied by downregulation of transcripts with high-open reading frame dominance |
| title_full_unstemmed | Inhibition of OCT4 binding at the MYCN locus induces neuroblastoma cell death accompanied by downregulation of transcripts with high-open reading frame dominance |
| title_short | Inhibition of OCT4 binding at the MYCN locus induces neuroblastoma cell death accompanied by downregulation of transcripts with high-open reading frame dominance |
| title_sort | inhibition of oct4 binding at the mycn locus induces neuroblastoma cell death accompanied by downregulation of transcripts with high open reading frame dominance |
| topic | neuroblastoma MYCN oct4 open reading frame dominance CRISPR/dCas9 p53 |
| url | https://www.frontiersin.org/articles/10.3389/fonc.2024.1237378/full |
| work_keys_str_mv | AT kazumanakatani inhibitionofoct4bindingatthemycnlocusinducesneuroblastomacelldeathaccompaniedbydownregulationoftranscriptswithhighopenreadingframedominance AT kazumanakatani inhibitionofoct4bindingatthemycnlocusinducesneuroblastomacelldeathaccompaniedbydownregulationoftranscriptswithhighopenreadingframedominance AT kazumanakatani inhibitionofoct4bindingatthemycnlocusinducesneuroblastomacelldeathaccompaniedbydownregulationoftranscriptswithhighopenreadingframedominance AT kazumanakatani inhibitionofoct4bindingatthemycnlocusinducesneuroblastomacelldeathaccompaniedbydownregulationoftranscriptswithhighopenreadingframedominance AT hiroyukikogashi inhibitionofoct4bindingatthemycnlocusinducesneuroblastomacelldeathaccompaniedbydownregulationoftranscriptswithhighopenreadingframedominance AT hiroyukikogashi inhibitionofoct4bindingatthemycnlocusinducesneuroblastomacelldeathaccompaniedbydownregulationoftranscriptswithhighopenreadingframedominance AT takanorimiyamoto inhibitionofoct4bindingatthemycnlocusinducesneuroblastomacelldeathaccompaniedbydownregulationoftranscriptswithhighopenreadingframedominance AT taikisetoguchi inhibitionofoct4bindingatthemycnlocusinducesneuroblastomacelldeathaccompaniedbydownregulationoftranscriptswithhighopenreadingframedominance AT tetsushisakuma inhibitionofoct4bindingatthemycnlocusinducesneuroblastomacelldeathaccompaniedbydownregulationoftranscriptswithhighopenreadingframedominance AT kazutokugou inhibitionofoct4bindingatthemycnlocusinducesneuroblastomacelldeathaccompaniedbydownregulationoftranscriptswithhighopenreadingframedominance AT yoshinorihasegawa inhibitionofoct4bindingatthemycnlocusinducesneuroblastomacelldeathaccompaniedbydownregulationoftranscriptswithhighopenreadingframedominance AT takashiyamamoto inhibitionofoct4bindingatthemycnlocusinducesneuroblastomacelldeathaccompaniedbydownregulationoftranscriptswithhighopenreadingframedominance AT yoshitakahippo inhibitionofoct4bindingatthemycnlocusinducesneuroblastomacelldeathaccompaniedbydownregulationoftranscriptswithhighopenreadingframedominance AT yoshitakahippo inhibitionofoct4bindingatthemycnlocusinducesneuroblastomacelldeathaccompaniedbydownregulationoftranscriptswithhighopenreadingframedominance AT yoshitakahippo inhibitionofoct4bindingatthemycnlocusinducesneuroblastomacelldeathaccompaniedbydownregulationoftranscriptswithhighopenreadingframedominance AT yusukesuenaga inhibitionofoct4bindingatthemycnlocusinducesneuroblastomacelldeathaccompaniedbydownregulationoftranscriptswithhighopenreadingframedominance |