The N6-methyladenosine, also known as m6A, is the most common post-transcriptional modification in mRNAs and long non-coding RNAs and that profoundly influences mRNA biology, from early processing in the nucleus to final steps of translation and decay in the cytoplasm. Taking into consideration the importance of RNA in shaping cell fate, m6A is widely recognized as an additional layer in the regulation of gene expression, also thanks to its dynamic and reversible nature. Therefore, it is not surprising that any misregulation in m6A content might lead to the loss of cellular homeostasis. This effect is particularly evident when it comes to stem cells differentiation, embryo development and cancer. In a tumorigenic context, the m6A could affect the development, progression, cancer stem cells (CSCs) renewal and drug resistance of solid and liquid tumours. So, the m6A is consistently becoming a new attractive pharmacological target. Neuroblastoma (NB) is a neuroendocrine tumour of early childhood that derives from undifferentiated cells of the sympathoadrenal lineage of the neural crest. About 50% of patients have a very aggressive form of NB, with an overall survival rate of less than 30% despite heavy treatments. Moreover, NB is a challenging druggable tumour due to a low rate of somatic mutations. Somatic mutations at significant frequency have been identified in only five genes that also show detectable expression. Among these, only one is currently a directly validated druggable target. Two m6A regulators (METTL14 and ALKBH5) are aberrantly expressed in high-risk NB patients, and their alteration in NB cell lines affects tumour aggressiveness. Specifically, the overexpression of the methyltransferase METTL14 increases cell proliferation and invasion in vitro and tumour growth in mice acting as an oncogene, while ALKBH5 restoration affects cell proliferation, apoptosis and invasion in an opposite fashion. Importantly, the demethylase ALKBH5 impaired tumour formation in vivo when costitutively expressed and dramatically slows down tumor progression in mice when is induced by causing massive apoptosis. These data suggest that ALKBH5 acts as a potent tumour suppressor in NB. We discovered that METTL14 and ALKBH5 exert their effect on different levels by affecting mRNA stability or translation, respectively. Although the contribution to NB of the altered stability of transcripts related to mRNA processing in METTL14-overexpressing cells is less understand, the increase translation of pro-apoptotic genes in the ALKBH5-overexpression condition leaves little doubts. Our results unveil the m6A and its regulators as potential therapeutic targets for treating NB. Indeed, in collaboration with the Laboratory of Genomic Screening of Professor Alessandro Provenzani, we presented an encouraging proof-of-concept of the reader YTHDF1 as a possible pharmacological target.

The m6A RNA modification sustains neuroblastoma tumour aggressiveness / Montuori, Giulia. - (2020 Oct 19), pp. 1-117. [10.15168/11572_276989]

The m6A RNA modification sustains neuroblastoma tumour aggressiveness

Montuori, Giulia
2020-10-19

Abstract

The N6-methyladenosine, also known as m6A, is the most common post-transcriptional modification in mRNAs and long non-coding RNAs and that profoundly influences mRNA biology, from early processing in the nucleus to final steps of translation and decay in the cytoplasm. Taking into consideration the importance of RNA in shaping cell fate, m6A is widely recognized as an additional layer in the regulation of gene expression, also thanks to its dynamic and reversible nature. Therefore, it is not surprising that any misregulation in m6A content might lead to the loss of cellular homeostasis. This effect is particularly evident when it comes to stem cells differentiation, embryo development and cancer. In a tumorigenic context, the m6A could affect the development, progression, cancer stem cells (CSCs) renewal and drug resistance of solid and liquid tumours. So, the m6A is consistently becoming a new attractive pharmacological target. Neuroblastoma (NB) is a neuroendocrine tumour of early childhood that derives from undifferentiated cells of the sympathoadrenal lineage of the neural crest. About 50% of patients have a very aggressive form of NB, with an overall survival rate of less than 30% despite heavy treatments. Moreover, NB is a challenging druggable tumour due to a low rate of somatic mutations. Somatic mutations at significant frequency have been identified in only five genes that also show detectable expression. Among these, only one is currently a directly validated druggable target. Two m6A regulators (METTL14 and ALKBH5) are aberrantly expressed in high-risk NB patients, and their alteration in NB cell lines affects tumour aggressiveness. Specifically, the overexpression of the methyltransferase METTL14 increases cell proliferation and invasion in vitro and tumour growth in mice acting as an oncogene, while ALKBH5 restoration affects cell proliferation, apoptosis and invasion in an opposite fashion. Importantly, the demethylase ALKBH5 impaired tumour formation in vivo when costitutively expressed and dramatically slows down tumor progression in mice when is induced by causing massive apoptosis. These data suggest that ALKBH5 acts as a potent tumour suppressor in NB. We discovered that METTL14 and ALKBH5 exert their effect on different levels by affecting mRNA stability or translation, respectively. Although the contribution to NB of the altered stability of transcripts related to mRNA processing in METTL14-overexpressing cells is less understand, the increase translation of pro-apoptotic genes in the ALKBH5-overexpression condition leaves little doubts. Our results unveil the m6A and its regulators as potential therapeutic targets for treating NB. Indeed, in collaboration with the Laboratory of Genomic Screening of Professor Alessandro Provenzani, we presented an encouraging proof-of-concept of the reader YTHDF1 as a possible pharmacological target.
XXXII
2018-2019
CIBIO (29/10/12-)
Biomolecular Sciences
Quattrone, Alessandro
no
Inglese
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11572/276989
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