Ghrelin intronic lncRNAs, lnc-GHRL-3:2 and lnc-GHRL-3:3, as novel biomarkers in type 2 diabetes mellitus.

BACKGROUND: We aim to identify circulating lncRNAs located in the region of the ghrelin (GHRL) gene that play a role in the development of T2DM. METHODS: Bioinformatic tool was used to identify candidates GHRL-lncRNAs. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to compare the expression levels of selected lncRNAs on diabetic patients and non-diabetic controls.Receiver operating characteristic (ROC) curve analysis was performed to evaluate the discriminatory power of selected GHRL-lncRNAs. RESULTS: The bioinformatic analysis predicted three antisense and eight sense-intronic GHRL- lncRNAs. Two differentially expressed GHRL-lncRNAs were detected in diabetic patients. The expression levels of lnc-GHRL-3:2, lnc-GHRL-3:3, and the GHRL mRNA were significantly (p ≤ .0001) lower in the diabetic patients. ROC analysis showed that the area under the curve (AUC) value was 0.93 for lnc-GHRL-3:2 and 0.90 for lnc-GHRL-3:3. CONCLUSION: lnc-GHRL-3:2 and lnc-GHRL-3:3 are novel biomarkers and might play a regulatory role in T2DM pathogenesis.

Global analysis of biogenesis, stability and sub-cellular localization of lncRNAs mapping to intragenic regions of the human genome.

Long noncoding RNAs (lncRNAs) that map to intragenic regions of the human genome with the same (intronic lncRNAs) or opposite orientation (antisense lncRNAs) relative to protein-coding mRNAs have been largely dismissed from biochemical and functional characterization due to the belief that they are mRNA precursors, byproducts of RNA splicing or simply transcriptional noise. In this work, we used a custom microarray to investigate aspects of the biogenesis, processing, stability, evolutionary conservation, and cellular localization of ∼ 6,000 intronic lncRNAs and ∼ 10,000 antisense lncRNAs. Most intronic (2,903 of 3,427, 85%) and antisense lncRNAs (4,945 of 5,214, 95%) expressed in HeLa cells showed evidence of 5' cap modification, compatible with their transcription by RNAP II. Antisense lncRNAs (median t1/2 = 3.9 h) were significantly (p < 0.0001) more stable than mRNAs (median t1/2 = 3.2 h), whereas intronic lncRNAs (median t1/2 = 2.1 h) comprised a more heterogeneous class that included both stable (t1/2 > 3 h) and unstable (t1/2 < 1 h) transcripts. Intragenic lncRNAs display evidence of evolutionary conservation, have little/no coding potential and were ubiquitously detected in the cytoplasm. Notably, a fraction of the intronic and antisense lncRNAs (13 and 15%, respectively) were expressed from loci at which the corresponding host mRNA was not detected. The abundances of a subset of intronic/antisense lncRNAs were correlated (r ≥ |0.8|) with those of genes encoding proteins involved in cell division and DNA replication. Taken together, the findings of this study contribute novel biochemical and genomic information regarding intronic and antisense lncRNAs, supporting the notion that these classes include independently transcribed RNAs with potentials for exerting regulatory functions in the cell.