Long Non-Coding RNAs Gene Variants as Molecular Markers for Diabetic Retinopathy Risk and Response to Anti-VEGF Therapy

Abstract

Background: Long non-coding RNAs (lncRNAs) play essential roles in molecular diagnosis and therapeutic response in several diseases. Purpose: For the first time, we aimed to evaluate the association of four lncRNAs TUG1 (rs7284767G/A), MIAT (rs1061540T/C), MALAT1 (rs3200401C/T), and SENCR (rs12420823C/T) variants with susceptibility to diabetic retinopathy (DR), disease severity, and early therapeutic response to intravitreous anti-vascular endothelial growth factor aflibercept therapy. Patients and Methods: This case-control study enrolled 126 adult patients with type 2 diabetes. TaqMan assays using Real-Time PCR were run for genotyping. Multivariable regression analyses were applied to assess the role of each polymorphism after the adjustment of covariates. Results: Carriers of TUG1 A/G and MIAT T/C and C/C genotypes were more likely to develop DR [OR=3.15 (95% CI=1.15-8.64), and OR=4.31 (95% CI=1.78-10.47)], while MALAT1 T/C conferred protection (OR=0.40, 95% CI=0.16-0.99). For TUG1, MALAT1, MIAT, and SENCR genotype combinations, GTCT and GCCC had a higher disease risk (P=0.012). For disease severity, MIAT T/T homozygosity was associated with higher DR grade [33.3% (T/T) vs 10% (C/C) and 4.2% (C/T) carriers, P=0.012]. Otherwise, patients with the SENCR T variant exhibited better pre-treatment best-corrected visual acuity level (p=0.021). Following aflibercept administration, carrying the TUG1 A or MIAT T/C was associated with a poor therapeutic response (OR=5.02, 95% CI=1. and OR=10.23, respectively). Conclusion:The lncRNAs TUG1 (rs7284767G/A) and MIAT (rs1061540T/C) were associated with increased DR susceptibility and poor response to aflibercept treatment, while MALAT1 (rs3200401C/T) conferred protection to DR. These genetic determinants could be useful in DR risk stratification and pharmacogenetics after validation in large-scale studies.

Association of Angio-LncRNAs MIAT rs1061540/MALAT1 rs3200401 Molecular Variants with Gensini Score in Coronary Artery Disease Patients Undergoing Angiography

“…The study demonstrated that the MIAT rs1061540 T/T genotype was associated with a more severe Gensini score under codominant and recessive models, although no association with disease susceptibility was observed. In agreement with our outcome, TT homozygosity of the same MIAT variant was associated with a more advanced grade of diabetic retinopathy compared with CC and TC genotypes [ 27 ]. Ishii et al conducted a large-scale association study and reported that six MIAT SNPs (i.e., rs2331291, rs2301523, exon 3 (8813), and exon 5 (11093, 11741, and 12311)) were associated with myocardial infarction (MI) risk; however, the current study polymorphism was not found to be a risk variant [ 31 ], which could support in part our finding of inability to identify an association of this variant with the disease risk.…”

“…Real-time polymerase chain reaction allelic discrimination was applied in a StepOne™ Real-Time PCR System (Applied Biosystems, Thermo Fisher Scientific, Foster City, CA, USA) using TaqMan assays (Assay IDs: C__30952613_10 (A/C) for PUNISHER rs12318065, C__11783392_10 (C/T) for SENCR rs12420823, C___2467719_10 (C/T) for MIAT rs1061540, C___3246069_10 (T/C) for MALAT1 rs3200401, and C__98039038_10 (A/G) for GATA6-AS1 rs73390820, Applied Biosystems). The details of PCR contents/concentrations and PCR programming were detailed in our previous work [ 27 ]. We believe that the no template control (NTC) samples used for every assay that were run on a plate were enough to ensure no contamination.…”

LncRNAs and the Angiogenic Switch in Cancer: Clinical Significance and Therapeutic Opportunities

“…The reported findings may underscore the importance of these oxygen sensitive lncRNAs in tumour angiogenesis as this process is largely triggered by hypoxia. One of the highly expressed lncRNAs in ECs, MALAT1, functions to protect ECs from the effects of oxygen deprivation and nutrient deprivation by stimulating moderate autophagy [ 13 ]. MALAT1 is active during the initiation stage of autophagy and during autolysosomal fusion [ 13 , 14 ].…”

“…One of the highly expressed lncRNAs in ECs, MALAT1, functions to protect ECs from the effects of oxygen deprivation and nutrient deprivation by stimulating moderate autophagy [ 13 ]. MALAT1 is active during the initiation stage of autophagy and during autolysosomal fusion [ 13 , 14 ]. Autophagy is a process of cellular degradation in response to various stresses such as nutrient deprivation and it plays an important role in the maintenance of homeostasis.…”

“…Of note is that the silencing of ROR in vitro results in the downregulation of VEGF [ 16 ]. Several lncRNAs were reportedly expressed in ECs under conditions of hypoxia, including MEG8 and 9, as well as H19 [ 13 , 19 ]. Interestingly, the knockdown of H19 reduced the ability of ECs to form cords in an in-vitro assay of angiogenesis.…”