Potential relationship and clinical significance of miRNAs and Th17 cytokines in patients with multiple myeloma

doi:10.1016/j.leukres.2014.07.005

Leukemia Research

Volume 38, Issue 9, September 2014, Pages 1130-1135

https://doi.org/10.1016/j.leukres.2014.07.005 Get rights and content

Highlights

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MiRNAs and Th17 cytokines were aberrantly expressed in MM patients.
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The expression of them was differentially present according to the ISS.
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Upregulated IL-17, IL-21 and IL-27 may potentially down-regulate miRNAs in MM.
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The relationship may be useful for understanding the pathogenesis of MM.

Abstract

We evaluated the potential relationship between miRNAs and Th17 cytokines in multiple myeloma (MM) patients. Twenty-seven newly diagnosed myeloma patients and eight normal donors were studied. We determined that the relative expression levels of miR-15a/16, miR-34a, miR-194 in MM patients were significantly lower than those in the healthy controls with exception for miR-181a/b, which showed significantly higher in MM patients (P < 0.05). In contrast, the levels of IL-17, IL-21 and IL-27 were up-regulated in MM patients compared to healthy controls while IL-22 was down-regulated (P < 0.05). The expression patterns of them were differentially present in various groups according to the International Staging System (ISS) criteria. Up-regulated IL-17, IL-21 and IL-27 may potentially down-regulate the expression of several miRNAs in MM patients. Establishment of the relationship may be useful for understanding the pathogenesis of MM and for clinical diagnosis of the disease.

Introduction

Multiple myeloma (MM), which accounts for approximately 10% of all hematologic malignancies, is a mature B-cell neoplasm characterized by clonal expansion and proliferation of plasma cells (PCs) in the bone marrow (BM). The clinical characteristics of MM are bone destruction, high calcium levels, renal failure and altered immunity, which is commonly ascribed to over-production of monoclonal immunoglobins from the tumor cells. Several studies have previously shown that MM is an immunologically relevant disease which subverts and suppresses immunity, owing to the perturbation of immuno-regulatory responses [1]. Elevated IL-17 produced by Th17 cells can promote myeloma cell growth but inhibit immune functions in MM [2], suggesting that Th17 cells and IL-17 may serve as the potential therapeutic targets for patients with MM.miRNAs are a class of small non-coding single stranded RNAs of approximately 22 nucleotides in length, which regulate gene expression by targeting specific messenger RNAs (mRNAs) for translational repression and degradation through base pairing to partially or fully complementary sites. Recent studies have also found abnormal expression patterns of miRNAs in MM. Alterations in certain miRNAs expression are associated with gene subtypes and chromosome abnormalities. More than 50% of miRNAs genes are found to locate within regions of loss of heterozygosity, amplification, fragile sites, viral integration sites and other cancer-related genomic regions [3]. Several miRNAs have been found to be involved in the activation, differentiation and function of immune cells by affecting key transcripts and play a significant role in modulating the differentiation of Th17 cells [4], [5], [6], [7].

Although the development of new drugs and stem cell transplantation prolong survival rate of MM patients in recent years, it remains a major clinical challenge for many patients with recurrence, resistance and refractory. Recently, there are published researches concerning miR-15a/16, miR-34a, miR-192/194 and miR-181a/b deregulation in MM, and they have been deeply discussed in preclinical and clinical research because of their diverse functions. Based on previous observations showing the involvement of miRNAs, Th17 cells and inflammation in MM, we sought to investigate the expression of these miRNAs and Th17 cytokines in patients with MM and their potential roles in the pathogenesis of MM.

Section snippets

Patient samples

Informed consents were obtained from patients and healthy donors and the study was complied with the Declaration of Helsinki and its amendments. A cohort of 35 subjects from the Affiliated Hospital of Xuzhou Medical College was enrolled in this study. 27 patients with MM were newly diagnosed in the Department of Hematology during the period June 2012 through January 2013. All patients conformed to the diagnostic criteria of multiple myeloma according to the International Staging System (ISS).

Selected miRNAs profiling in patients with MM before treatment

Based on previous investigation on the involvement of miRNAs, we examined in BMMCs, if the selected miR-15a/16、miR-34a and miR-194 levels are altered as compared to normal marrow donors. As shown in Fig. 1A, the expression levels of the selected miRNAs were more than 2–4 folds lower except miR-181a/b, which was increased in patients with MM as compared to the control group. There were statistically significant differences between the two groups, but miR-192 was an exception. There was no

Discussion

MM is a currently incurable B-cell malignancy whose cell of origin mediates human humoral immunity. Abnormal secretion of related cytokines, activation of oncogenes and many other molecular genetic abnormalities play important roles in the initiation and progression of the disease. Th17 cells are important immune cells in tumor immunity. Studies have shown that the percentage of Th17 cells in BM in monoclonal gammopathy of undetermined significance (MGUS) patients was higher than that in MM

Conflict of interest statement

The authors have declared no conflicts of interest.

Acknowledgments

The authors thank Professor Lingyu Zeng for permission to use the instrument and we gratefully acknowledge the assistance of Weiwei Xing and Ning Li for sample preps.

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Li et al. have shown that the promoter region of miR-34a is hypermethylated and epigenetically silenced in MM patients. This abnormal expression of miR-34a and on the other side, increased IL-21 may lead to the initiation of Th17-induced inflammatory responses and increase the myeloma cell proliferation via raising the vascular endothelial growth factor (VEGF) level and angiogenesis [60] Moreover, Misso and his colleagues indicated that the interleukin-6 receptor (IL-6R)/signal transducer and activator of transcription 3 (STAT3)/miR-34a act as a loop and actively participate in transforming plasma cells and so play a prominent role in the biology of MM cells. IL-6R is a direct target of miR-34a.
miRNAs are a class of non-coding RNAs and very conserve molecules that negatively regulate the expression of many genes by targeting the 3′ UTR of mRNAs. miRNAs are involved in the modulation of T-cell biology during the earliest and last stages and key controllers of T-cell differentiation and function. The miR-34a, as a major hub of the regulatory network of T cells, plays an important role in T cell activation. miR-34a is widely expressed in immune cells (dendritic cells, macrophages, mast cells, B cells, and T cells) and regulates their development, function, and survival. This miRNA, by targeting over 30 genes across different cellular pathways controls immune response. miR-34a expression is controlled by p53 in transcription level. As well as, miR-34a by activating dendritic cells mediates innate immune response and increases tumor-infiltrating CD8 expression T lymphocytes. In various types of cancers and autoimmune diseases, miR-34a can regulate T cell function and become a possible significant target of microRNA-based therapy. Therefore, in this review, we focus on miR-34a-related regulatory mechanisms in T cell function and understanding mechanisms and molecules involved in this network.
c-Myc, RMRP, and miR-34a-5p form a positive-feedback loop to regulate cell proliferation and apoptosis in multiple myeloma
2019, International Journal of Biological Macromolecules
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miR-34a-5p, located at chromosome 1, is a member of the miR-34 family and participates in the occurrence and development of human cancers by modulating the expression of target genes implicated in cell proliferation, cell cycle and cell apoptosis [31–33]. miR-34a has been postulated to be underexpressed in numerous cancers, including pancreatic cancer, ovarian cancer, nasopharyngeal carcinoma and MM [34–37]. Previously, miR-34a has been documented to enhance the anti-tumor effect of γ-secretase inhibitor, Sirtinol or zoledronic acid in RPMI-8226 cells, which can be considered as a therapeutic approach for MM therapy [38].
Long non-coding RNA (lncRNA) component of mitochondrial RNA processing endoribonuclease (RMRP) has been demonstrated to be implicated in human cancer processes. However, the role of lncRNA RMRP in multiple myeloma (MM) remains unknown. In this paper, we proved that RMRP and c-Myc were upregulated, while miR-34a-5p was downregulated in MM cell lines and bone marrows of MM patients. High RMRP expression significantly correlated with worse disease-free survival and overall survival in MM patients. c-Myc promoted RMRP transcription by directly binding to its promoter region. Knockdown of RMRP inhibited proliferation and promoted apoptosis of OPM2 and RPMI-8226 cells. Negative correlation between RMRP, and miR-34a-5p was discovered in bone marrows of MM patients. c-Myc expression was inversely correlated with miR-34a-5p in bone marrows of MM patients. Additionally, silencing of RMRP led to a marked reduction in c-Myc expression in OPM2 and RPMI-8226 cells, and this action was obviously blocked by miR-34a-5p knockdown. Moreover, upregulation of miR-34a-5p repressed proliferation and promoted apoptosis of OPM2 and RPMI-8226 cells. However, RMRP overexpression blocked these changes triggered by miR-34a-5p mimic. Besides, RMRP knockdown repressed MM tumor growth in vivo. Conclusions, RMRP functions as a miR-34a-5p sponge to promote cell proliferation and repress cell apoptosis through upregulation of c-Myc in MM.
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Chromosome 13q14 deletion is one of the most common cytogenetic abnormalities which was accounted for about 54% of MM patients. miR-15a and miR-16 which were previously indicated to be dysregulated in MM are located at the chromosome 13q14 [3–5]. Previous studies have found that miR-15a/16 took an important part in the pathogenesis of MM, and it has been reported that inhibition of miR-15a/16 can promote cell proliferation and angiogenesis in MM [6,7].
miRNAs have been reported to be involved in the pathogenesis of many cancers. In this article, we investigated the role and the mechanisms of miR-15a/16 in the pathogenesis of multiple myeloma (MM). We found that miR-15a/16 was down-regulated in bone marrow-derived mononuclear cells (BM-MNCs) of newly diagnosed patients with MM and the downregulation of miR-15a/16 was correlated with International Staging System (ISS) stage. We then demonstrated miR-15a/16 inhibited myeloma cells proliferation, and increased apoptosis rate of U266 cells by suppressing the expression of anti-apoptosis protein Bcl-2. We also found miR-15a/16 could decrease VEGF-A and IL-17 levels in the supernatant of myeloma cells. These results indicate that miR-15a/16 may function as a tumor suppressor in MM through multiple regulatory mechanisms and they may be potential targets for the therapy of MM.
Dysregulation of Non-Coding RNAs: Roles of miRNAs and lncRNAs in the Pathogenesis of Multiple Myeloma
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¹: Both authors contributed to this work equally.

View full text

Potential relationship and clinical significance of miRNAs and Th17 cytokines in patients with multiple myeloma

Highlights

Abstract

Introduction

Section snippets

Patient samples

Selected miRNAs profiling in patients with MM before treatment

Discussion

Conflict of interest statement

Acknowledgments

Blood

Methods

Blood

Cancer Cell

Blood

Blood

Cancer Cell

Drug-mediated and cellular immunotherapy in multiple myeloma

Immunotherapy

Lack of BIC and microRNA miR-155 expression in cases of Burkitt lymphoma

Genes Chromosomes Cancer

microRNAs: critical regulators in Th17 cells and players in diseases

Cell Mol Immunol

MicroRNA miR-326 regulates TH-17 differentiation and is associated with the pathogenesis of multiple sclerosis

Nat Immunol