Polymorphism in ANRIL is associated with relapse in patients with multiple myeloma after autologous stem cell transplant

Abstract

Multiple myeloma (MM) is a hematologic malignancy characterized by clonal proliferation of plasma cells and overproduction of monoclonal immunoglobins. Treatment with melphalan is currently standard of care for younger and fit patients when followed by hematopoietic stem cell transplantation (HSCT), and in transplant ineligible patients when used in combination regimens. It has been previously shown that changes in the p53 pathway are associated with melphalan efficacy, but the regulatory role of the p14ARF-MDM2-p53 axis has yet to be fully explored. Recently, a non-coding RNA, ANRIL (antisense non-coding RNA in the INK4-ARF locus) has been shown to negatively regulate the transcription of the entire INK4-ARF locus and simultaneously modulate the p53 and pRb pathways. Moreover, some single nucleotide polymorphisms (SNPs) in ANRIL have previously been associated with susceptibility to several malignancies. Here we investigated select ANRIL SNPs in DNA from patient-derived peripheral blood mononuclear cells obtained from 108 MM patients treated with high-dose melphalan followed by HSCT. Our results show that the rs2151280 (CàT) SNP in ANRIL was associated with worse progression-free survival (TC/CC vs TT: HR = 0.53, 95%CI, [0.26, 1.07], P = 0.07; adjusted HR = 0.39, 95%CI, [0.18, 0.84], P = 0.016), and the TT variant had higher ANRIL expression and lower p15, p14ARF, and p16 expression compared to the TC/CC variants. Our results indicate that ANRIL may be involved in melphalan-mediated apoptosis via down-regulating p14ARF and subsequent p53, and that the rs2151280 polymorphism may be a potential prognostic biomarker for relapse in melphalan-treated MM patients.

Impact of four lncRNA polymorphisms (rs2151280, rs7763881, rs1136410, and rs3787016) on glioma risk and prognosis: A case‐control study

“…Another study found that ANRIL was upregulated in glioma, promoting cell proliferation, migration and invasion, and inhibiting apoptosis by downregulating mir‐34a . In addition, ANRIL rs2151280 polymorphism was found to be a potential prognostic biomarker for relapse in patients with multiple myeloma treated with melphalan by downregulating p14ARF and subsequent p53 . LncRNA ANRIL was positively correlated with glioma grade and negatively correlated with mir‐203a.…”

XRCC1‐mediated DNA repair is associated with progression‐free survival of multiple myeloma patients after autologous stem cell transplant

“…Studies in our laboratory also demonstrated that rs2151280 in noncoding ANRIL RNA gene (antisense noncoding RNA in proximity to the INK4‐ARF locus, also designated as CDKN2BAS ) was significantly associated with PFS in the same cohort of MM patients . ANRIL is able to transcriptionally repress the transcription of the entire INK4‐ARF locus (including p15 , p14ARF , and p16 tumor suppressor genes), thus simultaneously modulating both the p53 and pRb pathways, both of which are directly/indirectly involved in DNA repair, apoptosis, and other cellular processes .…”

“…The genotype of XRCC1 rs25487 was determined on a QuantStudio 7 Flex System (Thermo Fisher Scientific, Waltham, MA) using a prevalidated Taqman genotyping assay (Life Technologies), C__622564_10. Experiments were conducted in duplicate …”

“…The relative expression level (REL) of a target was determined using a comparative Cq method in which REL was defined as 2 − δ Cq . Another reference gene, HPRT1 gene (assay ID: 4333768‐0803017; hypoxanthine phosphoribosyltransferase 1) was also used in this study for the following purpose: if both GAPDH and HPRT1 messenger RNAs (mRNAs) were undetectable in a sample, the sample was excluded from further gene expression analyses due to the lack of adequate amounts of total RNA . Of note, the expression level of a target gene in PBMC presented in this study should be regarded as the baseline expression level of that specific target gene as PBMCs were separated from patients’ blood samples before melphalan treatment and RNA and DNA samples were extracted from these PBMCs in the absence of ex vivo melphalan treatment.…”

“…Gene expression data were subjected to log transformation to fulfill the normality requirement . Group‐wise differences in gene expression were analyzed using the unpaired/paired two‐sample Student's t tests where appropriate.…”