Methylation-mediated miR-155-FAM133A axis contributes to the attenuated invasion and migration of IDH mutant gliomas

doi:10.1016/j.canlet.2018.06.007

Cancer Letters

Volume 432, 28 September 2018, Pages 93-102

https://doi.org/10.1016/j.canlet.2018.06.007 Get rights and content

Highlights

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MiR-155 is down-regulated in IDH^MT gliomas and is an independent risk factor for glioma patients.
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MiR-155 is down-regulated via IDH mutation-induced promoter CpG island methylation.
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Cancer testis antigen FAM133A is a direct downstream target of miR-155.
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FAM133A predicts favorable prognosis for glioma patients.
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Down-regulation of FAM133A promotes glioma invasion/migration by regulating MMP14.

Abstract

Gliomas with isocitrate dehydrogenases gene mutations (IDH^MT) were found to be less aggressive than their wildtype (IDH^WT) counterparts. However, the mechanism remains unclear. The current study aims to investigate the role of silenced oncogenic microRNAs in IDH^MT gliomas, which were largely ignored and may contribute to the less aggressive behavior of IDH^MT gliomas. Microarrays, bioinformatics analysis of the data from TCGA and qPCR analysis of samples from our experimental cohort (LGG: IDH^WT = 10, IDH^MT = 31; GBM: IDH^WT = 34, IDH^MT = 9) were performed. The results show that miR-155 was consistently down-regulated in IDH^MT gliomas. Establishment of IDH1^R132H overexpressing glioma cell line and bisulfite sequencing PCR suggested that miR-155 down-regulation was associated with IDH1^R132H mutation induced promoter CpG islands methylation. The cancer testis antigen FAM133A is a direct downstream target of miR-155 and is a negative regulator of glioma invasion and migration possibly by regulating matrix metallopeptidase 14 (MMP14). Together, we found that methylation-regulated miR-155-FAM133A axis may contribute to the attenuated invasion and migration of IDH^MT gliomas by targeting MMP14.

Introduction

In the last decade, somatic mutations in isocitrate dehydrogenases (IDH^MT) were found to be widespread in lower grade gliomas (LGG, WHO grade II and III) and secondary glioblastomas (GBM) [1], and were thought to be the earliest event in glioma development [2,3]. Meanwhile, IDH^MT gliomas were found to be less aggressive than the WHO grade-matched wild-type counterparts [4]. The possible mechanisms include the attenuated invasion and migration [5], decreased proliferation [6], altered metabolic pathways [7,8] and sensitization to chemo-/radio-therapy [9,10]. Based on the profound influence of IDH mutation on gliomas, the current WHO classification of tumors of the central nervous system (2016) has placed IDH mutation as a standard prognostic molecular marker to help better define the specific entities of gliomas [11].

IDH mutations are located specifically within the enzymatically active sites at the amino acid residue arginine 132th (for IDH1) and 172th (for IDH2), with IDH1^R132H and IDH2^R172K substitutions being most common [12]. These mutations give IDH a neomorphic enzymatic activity catalyzing α-KG to a new metabolite (R)-enantiomer of 2-hydroxyglutarate (R-2HG), an onco-metabolite inhibiting α-KG-dependent enzymes, resulting in widespread genomic DNA hypermethylation [13]. The causal role of IDH mutation to the G-CIMP [14,15] had challenged researchers to find specific promoter methylation-mediated down-regulated gene signatures which may explain the tumor biology of IDH^MT gliomas. Tumor suppressor genes such as RBP1 (retinol-binding protein 1) [16], miR-148a [17] and MTSS1 [18] have been found to be silenced by promoter CpG island methylation and can contribute to glioma formation, linking vital roles of IDH mutation in gliomagenesis. On the other hand, the role of silenced oncogenes or oncogenic miRNAs, which may play roles in the less malignant behaviors of IDH^MT gliomas, has been largely under-investigated.

MicroRNAs (miRNAs) are small noncoding RNAs regulating gene expression by binding to the 3′-untranslated region (3′UTR) of target mRNAs, leading to mRNA cleavage and degradative repression [19]. It has been widely demonstrated that miRNAs play extremely important roles in regulating the malignant phenotypes of gliomas [20]. In this study, we identified a methylation-mediated miR-155-FAM133A axis in regulating the invasion/migration of IDH^MT gliomas, which provides new insight into understanding the less aggressive malignant behaviors of IDH^MT gliomas.

Section snippets

Glioma cohorts and clinical samples

A total of 41 LGG and 43 GBM tissues from our cohort (XQ cohort) with complete clinical follow-up information were recruited in this study. Another validation cohort from the Cancer Genome Atlas (TCGA) database was investigated in silico. Characteristics of patients are detailed in Supplementary Table 1.

Cell culture

Human anaplastic astrocytoma cell line SF268 (WHO grade III) [21] and GBM cell lines U251, SNB19, SF295 (WHO grade IV) [21] with less than 5 total passage times were directly selected from the

Oncogenic miR-155 is down-regulated in IDH-mutant gliomas

We initially isolated primary GBMs cells (IDH^WT = 4 and IDH^MT = 2) for miRNA-array analysis to determine miRNAs differentially expressed (DE-miRs) between IDH^WT and IDH^MT gliomas (Fig. 1A). To validate the list of candidates, we sought for clinical analysis either from publicly available database or from our own hospital. We first investigated an independent cohort of glioma patients from TCGA, including 121 cases of lower grade gliomas (LGG, IDH^MT = 87) and early glioblastomas (eGBM, IDH^WT

Discussion

Migration and invasion, key features of glioma aggressiveness, are largely responsible for difficulty of achieving surgical total resection and poor patients' prognosis [33]. Interestingly, it has been repeatedly reported that the migration and invasion are inhibited in IDH^MT gliomas [5,10,34,35], this may in part explain the less aggressive behaviors of IDH^MT gliomas. In this study, we provide new evidence that down-regulation of oncogenic miR-155 can contribute to the attenuated

Disclosure of potential conflicts of interest

The authors declare no potential conflicts of interest.

Acknowledgement

We thank Dr. Clive Da Costa (Francis Crick Institute, London, UK) for providing useful suggestions and carefully commenting on this manuscript. This work was supported by the National Natural Science Foundation of China (NSFC:81272783).

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Original ArticlesMethylation-mediated miR-155-FAM133A axis contributes to the attenuated invasion and migration of IDH mutant gliomas

Highlights

Abstract

Introduction

Section snippets

Glioma cohorts and clinical samples

Cell culture

Oncogenic miR-155 is down-regulated in IDH-mutant gliomas

Discussion

Disclosure of potential conflicts of interest

Acknowledgement

Ann. Oncol.

Canc. Cell

Canc. Lett.

Radiother. Oncol.

Biochim. Biophys. Acta

Canc. Cell

Transl Oncol

Cell

Exp. Cell Res.

Biomed. Pharmacother.

Biochim. Biophys. Acta

Int. J. Biochem. Cell Biol.

Mol. Cell

Evidence for sequenced molecular evolution of IDH1 mutant glioblastoma from a distinct cell of origin

J. Clin. Oncol.

Isocitrate dehydrogenase 1 codon 132 mutation is an important prognostic biomarker in gliomas

J. Clin. Oncol.

IDH1 R132H decreases proliferation of glioma cell lines in vitro and in vivo

Ann. Neurol.

IDH1 R132H mutation generates a distinct phospholipid metabolite profile in glioma

Canc. Res.

Inhibition of glutaminase preferentially slows growth of glioma cells with mutant IDH1

Canc. Res.

IDH mutations predict longer survival and response to temozolomide in secondary glioblastoma

Canc. Sci.

The 2016 world health organization classification of tumors of the central nervous system: a summary

Acta Neuropathol.

A heterozygous IDH1R132H/WT mutation induces genome-wide alterations in DNA methylation

Genome Res.

IDH1 mutation is sufficient to establish the glioma hypermethylator phenotype

Nature

Original Articles
Methylation-mediated miR-155-FAM133A axis contributes to the attenuated invasion and migration of IDH mutant gliomas