MicroRNA-15a-5p acts as a tumor suppressor in histiocytosis by mediating CXCL10-ERK-LIN28a-let-7 axis
Erdheim–Chester disease (ECD) is a rare hematological malignancy. Here we describe that miR-15a function as a tumor suppressor in ECD through the CXCL10-ERK-LIN28a-let7 axis, highlighting a novel layer of regulation and suggesting that upregulation of miR-15a may have a therapeutic role.
MicroRNAs (miRNAs) are short, non-coding RNAs that play a pivotal role in cancer initiation, progression, and treatment response via regulation of post-transcriptional gene expression (1).
Erdheim–Chester disease (ECD) is a rare histiocytic disorder with diverse clinical manifestations, ranging from indolent, localized presentations to a life-threatening, multi-system disease (2). Recurrent activating kinase mutations and fusions involving the ERK cascade and PI3K/AKT pathways and specifically, mutations in BRAF V600E and its downstream gene, MAP2K1 have been discovered in a large proportion of ECD patients (2-4). While genetic alterations driving ECD pathogenesis have been well studied (2), to date, only a few studies, have investigated transcriptional and post-transcriptional alterations in this disease.
We hypothesized that ECD relies on miRNA-mRNA interactions. Understanding the miRNA landscape in ECD may help to identify signaling pathways involved in this disease, and improve our understanding about disease pathogenesis.
We have recently showed that the miRNA profile of ECD patients differs from that of healthy controls (5). The most prominent and significant change detected between ECD and healthy controls was downregulation of miR-15a-5p. The functions of miR-15a-5p have been investigated in hematological malignancies (6-8), but whether the dysregulation of this miRNA contributes to the pathogenesis and development of ECD has not yet been elucidated.
In our manuscript, we describe evidence for a novel molecular mechanism which is mediated by miR-15a-5p and the chemokine CXCL10 [also named interferon- inducible protein-10 (IP-10)]. This chemokine is associated with a variety of human diseases including chronic inflammation, immune dysfunction, and tumor development, and metastasis (9).
By luciferase assay, we showed that CXCL10 is a bona fide target of miR-15a-5p. Analysis of plasma samples from ECD patients revealed elevated levels of CXCL10, associated with downregulation of miR-15a-5p in 70% (24/36) of the patients. This was further evaluated in tissue biopsies of ECD lesions as compared to matched healthy tissues from post-mortem autopsy. As observed in the plasma samples, miR-15a-5p was downregulated in ECD tissues and CXCL10 levels were upregulated compared with healthy controls, strengthening our hypothesis that miR-15a-5p mediates CXCL10 expression.
As several lines of evidence suggest that ECD originates from a myeloid progenitor (10-15), we investigated miR-15a-5p levels in the human myeloid cell lines KG-1a and OCI-AML3, which over express the MAPK-ERK pathway due to RAS mutations. We also used the cytokine-dependent murine lymphoid pro-B Ba/F3 line that stably expresses the MIGII-BRAFV600E vector.
We found that restoring miR-15a-5p levels by transfection with a miR-15a-5p mimic, resulted in downregulation of CXCL10 mRNA and protein levels. Since high levels of CXCL10 were previously shown to activate the MAPK-ERK pathway in solid tumors (9, 16, 17) we examined whether downregulation of this chemokine, following miR-15a-5p overexpression, affects the MAPK-ERK cascade in myeloid and lymphoid cells. As expected, expression of p-ERK was downregulated in all cell lines overexpressing miR-15a-5p, this was accompanied by downregulation of ERK target genes (DUSP6, SPRY2 and LIN28a), indicating a true biological effect of p-ERK downregulation. Furthermore, miR-15a-5p inhibited cell growth and induced apoptosis in all cell lines. Finally, analysis of sequential samples from 7 ECD patients treated with MAPK inhibitor (cobimetinib) for 4 months resulted in upregulation of miR-15a-5p and downregulation of CXCL10.
Our findings show that miR-15a-5p may function as a tumor suppressor in ECD patients through the CXCL10-ERK-LIN28a-let7 axis, highlighting an additional layer of post-transcriptional regulation in ECD patients. Upregulation of miR-15a-5p in ECD patients or in other myeloid malignancies with overexpression of the ERK cascade may have a potential therapeutic role.
- Peng Y, Croce CM. The role of MicroRNAs in human cancer. Signal Transduct Target Ther. 2016;1:15004.
- Goyal G, Heaney ML, Collin M, Cohen-Aubart F, Vaglio A, Durham BH, et al. Erdheim-Chester disease: consensus recommendations for evaluation, diagnosis, and treatment in the molecular era. Blood. 2020 May 28;135(22):1929-45.
- Janku F, Vibat CR, Kosco K, Holley VR, Cabrilo G, Meric-Bernstam F, et al. BRAF V600E mutations in urine and plasma cell-free DNA from patients with Erdheim-Chester disease. Oncotarget. 2014 Jun 15;5(11):3607-10.
- Diamond EL, Durham BH, Haroche J, Yao Z, Ma J, Parikh SA, et al. Diverse and Targetable Kinase Alterations Drive Histiocytic Neoplasms. Cancer Discov. 2016 Feb;6(2):154-65.
- Weissman R, Diamond EL, Haroche J, Pillar N, Shapira G, Durham BH, et al. The Contribution of MicroRNAs to the Inflammatory and Neoplastic Characteristics of Erdheim-Chester Disease. Cancers (Basel). 2020 Nov 3;12(11).
- Veronese A, Pepe F, Chiacchia J, Pagotto S, Lanuti P, Veschi S, et al. Allele-specific loss and transcription of the miR-15a/16-1 cluster in chronic lymphocytic leukemia. Leukemia. 2015 Jan;29(1):86-95.
- Calin GA, Sevignani C, Dumitru CD, Hyslop T, Noch E, Yendamuri S, et al. Human microRNA genes are frequently located at fragile sites and genomic regions involved in cancers. Proc Natl Acad Sci U S A. 2004 Mar 2;101(9):2999-3004.
- Handa H, Murakami Y, Ishihara R, Kimura-Masuda K, Masuda Y. The Role and Function of microRNA in the Pathogenesis of Multiple Myeloma. Cancers (Basel). 2019 Nov 6;11(11).
- Liu M, Guo S, Stiles JK. The emerging role of CXCL10 in cancer (Review). Oncol Lett. 2011 Jul;2(4):583-9.
- Haroche J, Cohen-Aubart F, Charlotte F, Maksud P, Grenier PA, Cluzel P, et al. The histiocytosis Erdheim-Chester disease is an inflammatory myeloid neoplasm. Expert Rev Clin Immunol. 2015;11(9):1033-42.
- Milne P, Bigley V, Bacon CM, Neel A, McGovern N, Bomken S, et al. Hematopoietic origin of Langerhans cell histiocytosis and Erdheim-Chester disease in adults. Blood. 2017 Jul 13;130(2):167-75.
- Durham BH, Roos-Weil D, Baillou C, Cohen-Aubart F, Yoshimi A, Miyara M, et al. Functional evidence for derivation of systemic histiocytic neoplasms from hematopoietic stem/progenitor cells. Blood. 2017 Jul 13;130(2):176-80.
- Papo M, Diamond EL, Cohen-Aubart F, Emile JF, Roos-Weil D, Gupta N, et al. High prevalence of myeloid neoplasms in adults with non-Langerhans cell histiocytosis. Blood. 2017 Aug 24;130(8):1007-13.
- Ghobadi A, Miller CA, Li T, O'Laughlin M, Lee YS, Ali M, et al. Shared cell of origin in a patient with Erdheim-Chester disease and acute myeloid leukemia. Haematologica. 2019 Aug;104(8):e373-e5.
- Cohen Aubart F, Roos-Weil D, Armand M, Marceau-Renaut A, Emile JF, Duployez N, et al. High frequency of clonal hematopoiesis in Erdheim-Chester disease. Blood. 2020 Oct 16.
- Maru SV, Holloway KA, Flynn G, Lancashire CL, Loughlin AJ, Male DK, et al. Chemokine production and chemokine receptor expression by human glioma cells: role of CXCL10 in tumour cell proliferation. J Neuroimmunol. 2008 Aug 13;199(1-2):35-45.
- Datta D, Flaxenburg JA, Laxmanan S, Geehan C, Grimm M, Waaga-Gasser AM, et al. Ras-induced modulation of CXCL10 and its receptor splice variant CXCR3-B in MDA-MB-435 and MCF-7 cells: relevance for the development of human breast cancer. Cancer Res. 2006 Oct 1;66(19):9509-18.