Dataset Information

N⁶-methyladenosine modification of ITGA6 mRNA promotes the development and progression of bladder cancer.

ABSTRACT:

Background

Accumulating evidence has revealed the critical roles of N⁶-methyladenosine (m⁶A) modification of mRNA in various cancers. However, the biological function and regulation of m⁶A in bladder cancer (BC) are not yet fully understood.

Methods

We performed cell phenotype analysis and established in vivo mouse xenograft models to assess the effects of m⁶A-modified ITGA6 on BC growth and progression. Methylated RNA immunoprecipitation (MeRIP), RNA immunoprecipitation and luciferase reporter and mutagenesis assays were used to define the mechanism of m⁶A-modified ITGA6. Immunohistochemical analysis was performed to assess the correlation between METTL3 and ITGA6 expression in bladder cancer patients.

Findings

We show that the m⁶A writer METTL3 and eraser ALKBH5 altered cell adhesion by regulating ITGA6 expression in bladder cancer cells. Moreover, upregulation of ITGA6 is correlated with the increase in METTL3 expression in human BC tissues, and higher expression of ITGA6 in patients indicates a lower survival rate. Mechanistically, m⁶A is highly enriched within the ITGA6 transcripts, and increased m⁶A methylations of the ITGA6 mRNA 3'UTR promotes the translation of ITGA6 mRNA via binding of the m⁶A readers YTHDF1 and YTHDF3. Inhibition of ITGA6 results in decreased growth and progression of bladder cancer cells in vitro and in vivo. Furthermore, overexpression of ITGA6 in METTL3-depleted cells partially restores the BC adhesion, migration and invasion phenotypes.

Interpretation

Our results demonstrate an oncogenic role of m⁶A-modified ITGA6 and show its regulatory mechanisms in BC development and progression, thus identifying a potential therapeutic target for BC. FUND: This work was supported by National Natural Science Foundation of China (81772699, 81472999).

SUBMITTER: Jin H

PROVIDER: S-EPMC6796523 | biostudies-literature | 2019 Sep

REPOSITORIES: biostudies-literature

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Publications

N<sup>6</sup>-methyladenosine modification of ITGA6 mRNA promotes the development and progression of bladder cancer.

Jin Huan H Ying Xiaoling X Que Biao B Wang Xiaoxue X Chao Yinghui Y Zhang Haiqing H Yuan Zusen Z Qi Defeng D Lin Shuibin S Min Wang W Yang Mei M Ji Weidong W

EBioMedicine 20190810

<h4>Background</h4>Accumulating evidence has revealed the critical roles of N<sup>6</sup>-methyladenosine (m<sup>6</sup>A) modification of mRNA in various cancers. However, the biological function and regulation of m<sup>6</sup>A in bladder cancer (BC) are not yet fully understood.<h4>Methods</h4>We performed cell phenotype analysis and established in vivo mouse xenograft models to assess the effects of m<sup>6</sup>A-modified ITGA6 on BC growth and progression. Methylated RNA immunoprecipitatio ...[more]

PMID: 31409574

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Project description:IntroductionN6-methyladenosine (m6A) modification contributes to the pathogenesis and development of various cancers, including bladder cancer (BCa). In particular, integrin α6 (ITGA6) promotes BCa progression by cooperatively regulating multisite m6A modification. However, the therapeutic effect of targeting ITGA6 multisite m6A modifications in BCa remains unknown.ObjectivesWe aim to develop a multisite dCasRx- m6A editor for assessing the effects of the multisite dCasRx-m6A editor targeted m6A demethylation of ITGA6 mRNA in BC growth and progression.MethodsThe multisite dCasRx- m6A editor was generated by cloning. m6A-methylated RNA immunoprecipitation (meRIP), luciferase reporter, a single-base T3 ligase-based qPCR-amplification, Polysome profiling and meRIP-seq experiments were performed to determine the targeting specificity of the multisite dCasRx-m6A editor. We performed cell phenotype analysis and used in vivo mouse xenograft models to assess the effects of the multisite dCasRx-m6A editor in BC growth and progression.ResultsWe designed a targeted ITGA6 multi-locus guide (g)RNA and established a bidirectional deactivated RfxCas13d (dCasRx)-based m6A-editing platform, comprising a nucleus-localized dCasRx fused with the catalytic domains of methyltransferase-like 3 (METTL3-CD) or α-ketoglutarate-dependent dioxygenase alkB homolog 5 (ALKBH5-CD), to simultaneously manipulate the methylation of ITGA6 mRNA at four m6A sites. The results confirmed the dCasRx-m6A editor modified m6A at multiple sites in ITGA6 mRNA, with low off-target effects. Moreover, targeted m6A demethylation of ITGA6 mRNA by the multisite dCasRx-m6A editor significantly reduced BCa cell proliferation and migration in vitro and in vivo. Furthermore, the dCasRx-ALKBH5-CD and ITGA6 multi-site gRNA delivered to 5-week-old BALB/cJNju-Foxn1nu/Nju nude mice via adeno-associated viral vectors significantly inhibited BCa cell growth.ConclusionOur study proposes a novel therapeutic tool for the treatment of BC by applying the multisite dCasRx-m6A editor while highlighting its potential efficacy for treating other diseases associated with abnormal m6A modifications.

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Dataset Information

N⁶-methyladenosine modification of ITGA6 mRNA promotes the development and progression of bladder cancer.

Background

Methods

Findings

Interpretation

Publications

N<sup>6</sup>-methyladenosine modification of ITGA6 mRNA promotes the development and progression of bladder cancer.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Dataset Information

N6-methyladenosine modification of ITGA6 mRNA promotes the development and progression of bladder cancer.

Background

Methods

Findings

Interpretation

Publications

N<sup>6</sup>-methyladenosine modification of ITGA6 mRNA promotes the development and progression of bladder cancer.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

N⁶-methyladenosine modification of ITGA6 mRNA promotes the development and progression of bladder cancer.