The m6A writers regulated by the IL-6/STAT3 inflammatory pathway facilitate cancer cell stemness in cholangiocarcinoma

Abstract

Objective: Investigation of the regulatory mechanisms of cell stemness in cholangiocarcinoma (CCA) is essential for developing effective therapies to improve patient outcomes. The purpose of this study was to investigate the function and regulatory mechanism of m6A modifications in CCA cell stemness. Methods: Interleukin 6 (IL-6) treatment was used to induce an inflammatory response, and loss-of-function studies were conducted using mammosphere culture assays. Chromatin immunoprecipitation, polysome profiling, and methylated RNA immunoprecipitation analyses were used to identify signaling pathways. The in vitro findings were verified in a mice model. Results: We first identified that m6A writers were highly expressed in CCAs and further showed that STAT3 directly bound to the gene loci of m6A writers, showing that IL-6/STAT3 signaling regulated expressions of m6A writers. Downregulating m6A writers prevented cell proliferation and migration in vitro and suppressed CCA tumorigenesis in vivo. Notably, the knockdown of m6A writers inhibited CCA cell stemness that was triggered by IL-6 treatment. Mechanistically, IGF2BP2 was bound to CTNNB1 transcripts, significantly enhancing their stability and translation, and conferring stem-like properties. Finally, we confirmed that the combination of m6A writers, IGF2BP2, and CTNNB1 distinguished CCA tissues from normal tissues. Conclusions: Overall, this study showed that the IL-6-triggered inflammatory response facilitated the expressions of m6A writers and cell stemness in an m6A-IGF2BP2-dependent manner. Furthermore, the study showed that m6A modification was a targetable mediator of the response to inflammation factor exposure, was a potential diagnostic biomarker for CCA, and was critical to the progression of CCA.

Multi-Omics Binary Integration via Lasso Ensembles (MOBILE) for identification of context-specific networks and new regulatory mechanisms

“…Some were (i) shown to be part of prognostic markers (SLC12A4-CHEK2 (77)), (ii) differentially expressed together in response to perturbations (SEMA3C-TCEAL1 (78), IGFBP2-SIX1 (79), TINCR-POU2AF1 (80), NTSR1-TOMM6 (81)), and (iii) part of gene signatures for different classes of tumors (SNORA73A-MAPK (82), FDCSP-GJA1 (83)). A few of the associations had related mechanistic interactions as well (CDC42-MTOR (84,85), IGFBP2-SIX1 (79), WTAP-STAT3 (86,87), TINCR-POU2AF1 (80)). The rest of the associations are either Self: same gene, different data type, Unknown: non-curated gene(s), or Novel: no known interactions.…”

m5C and m6A modification of long noncoding NKILA accelerates cholangiocarcinoma progression via the miR‐582‐3p‐YAP1 axis

“…In CCA, IL‐6/STAT3 signalling regulates expressions of m 6 A writers. Knockdown of these m 6 A writers attenuates the CCA cell stemness that was triggered by IL‐6 treatment 27 . Moreover, m 6 A demethylase ALKBH5 interacts with PD‐L1 mRNA.…”

“…Knockdown of these m 6 A writers attenuates the CCA cell stemness that was triggered by IL-6 treatment. 27 Moreover, m 6 A demethylase ALKBH5 interacts with PD-L1 mRNA. ALKBH5 deficiency elevates m 6 A modification of PD-L1 mRNA, thereby promoting its degradation in a YTHDF2-dependent manner and enhancing the sensitivity of anti-PD-1 immunotherapy.…”

METTL3 promotes glycolysis and cholangiocarcinoma progression by mediating m6A modification of AKR1B10

“…CCA is a rare cancer but still affects a wide range of people and its morbidity and mortality increasing at an alarming rate [22]. Numerous studies reveal that m6A modi cation on RNA is tightly associated with the tumorigenesis and development of multiple cancers through various mechanisms, including CCA [23], bladder cancer [24], ovarian cancer [25], and liver cancer [26]. m6A methylation is catalyzed by a multicomponent methyltransferase complex that includes m6A writer METTL3 [27].…”