Research Article: MicroRNA signature constituted of miR-30d, miR-93, and miR-181b is a promising prognostic marker in primary central nervous system lymphoma

Date Published: January 7, 2019

Publisher: Public Library of Science

Author(s): Yasuo Takashima, Atsushi Kawaguchi, Yasuo Iwadate, Hiroaki Hondoh, Junya Fukai, Koji Kajiwara, Azusa Hayano, Ryuya Yamanaka, Hiroyoshi Ariga.


MicroRNAs (miRNAs) are small RNA molecules that inhibit gene function by suppressing translation of target genes. However, in primary central nervous system lymphoma (PCNSL), the biological significance of miRNAs is largely unknown, although some miRNAs are known to be prognosis markers. Here, we analyzed 847 miRNAs expressed in 27 PCNSL specimens using microarray profiling and surveyed miRNA signature for prognostic prediction. Of these, 16 miRNAs were expressed in 27 PCNSL specimens at a frequency of 48%. Their variable importance measured by Random forest model revealed miR-192, miR-486, miR-28, miR-52, miR-181b, miR-194, miR-197, miR-93, miR-708, and let-7g as having positive effects; miR-29b-2*, miR-126, and miR-182 as having negative effects; and miR-18a*, miR-425, and miR-30d as neutral. After principal component analysis, the prediction formula for prognosis, consisting of the expression values of the above-mentioned miRNAs, clearly divided Kaplan-Meier survival curves by the calculated Z-score (HR = 6.4566, P = 0.0067). The 16 miRNAs were enriched by gene ontology terms including angiogenesis, cell migration and proliferation, and apoptosis, in addition to signaling pathways including TGF-β/SMAD, Notch, TNF, and MAPKinase. Their target genes included BCL2-related genes, HMGA2 oncogene, and LIN28B cancer stem cell marker. Furthermore, three miRNAs including miR-181b, miR-30d, and miR-93, selected from the 16 miRNAs, also showed comparable results for survival (HR = 8.9342, P = 0.0007), suggestive of a miRNA signature for prognostic prediction in PCNSL. These results indicate that this miRNA signature is useful for prognostic prediction in PCNSL and would help us understand target pathways for therapies in PCNSL.

Partial Text

Primary central nervous system lymphoma (PCNSL), a rare subgroup of diffuse large B-cell lymphoma (DLBCL) arising in the central nervous system (CNS), is an aggressive malignant variant of nodal non-Hodgkin lymphoma (NHL) [1,2]. PCNSLs account for 3% of all primary CNS tumors and 1% of NHLs in adults [3]. Most PCNSLs are immune privilege site-associated DLBCLs, according to the WHO diagnostic criteria [1]. Despite intensive treatments including high-dose methotrexate (HD-MTX) based polychemotherapy with whole brain radiotherapy, the median overall survival (OS) time is approximately 4 years for PCNSLs and shows a poorer prognosis than that of extracerebral DLBCLs [4–6].

PCNSL is a rare subtype of extra-nodal NHL, in which biological significance have been poorly understood, with only a few studies on the CNS signature of PCNSL being reported compared to non-CNS DLBCL [35–37]. Previous studies have reported that, in DLBCL, the up-regulated miRNAs are miR-155, miR-210, miR-17-5p, and miR-106, whereas the down-regulated miRNAs are miR-150, miR-145, miR-328, miR-139, miR-99a, miR-10a, miR-95, miR-149, miR-320, miR-151, and let-7e, compared to that in normal lymph nodes [38]. Reduced expression of miR-195 and let-7g also shows good EFS in DLBCL [38]. In this study as well, only the down-regulated miR-10a showed poorer survival in PCNSL (S2B Fig). There is a bias in miRNA expression pattern between PCNSL and nodal DLBCL; miR-9, miR-20a/b, miR-155, miR-340, miR-17-5p, miR-148a, miR-30b/c, miR-27b, miR-26b, miR-146b, and let-7g are expressed in PCNSL; whereas miR-199a, miR-214, miR-432, miR-193b, and miR-145 are expressed in nodal DLBCL [17]. This study also demonstrated that a subgroup associated with higher expression of let-7g, a suppressor of LIN28B stem cell marker [39], showed poor prognosis in PCNSL tumors (Fig 1D and 1E). A recent study has revealed that increased expression of miR-30c in patients with secondary CNS lymphoma (SCNSL), compared to those with PCNSL, allows the lymphomas to engraft into CNS, by binding to the cadherin EGF LAG seven-pass G-type receptor (CELSR) 3 gene [40]. In non-CNS DLBCL, exosomal RNAs are considered biomarkers in the blood, serum, and cerebrospinal fluid (CSF) [19,41]. Such serum biomarkers include high expression of miR-15, miR-16, miR-21, miR-29c, miR-155, and miR-210, but low expression of miR-34a within the serum [28,42]. The miR-30c derived from the CSF can serve as a biomarker to distinguish PCNSL from SCNSL [40]. We also examined the expression of 33 above-mentioned miRNAs and their correlation to the OS in this study. However, no differences were found except for let-7g and miR-10a (Fig 1D and 1B, and S2 Fig). Thus, we should also address comprehensive expression profiling of miRNAs in a larger population of PCNSL, as well as a non-CNS DLBCL study [43].