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Published on: May 4, 2021
MicroRNA-615-3p decreases apo B expression in human liver cells
Abulaish Ansari1, Pradeep Kumar Yadav2, Swati Valmiki3
1Department of Foundations of Medicine, New York University Grossman Long Island School of Medicine, Mineola, NY, USA; Department of Cell and Developmental Biology, Weill Cornell Medicine, New York, NY, USA; Department of Research, Weill Cornell Medicine-Qatar, Qatar Foundation, Education City, Doha, Qatar.
Abstract:
Plasma lipids are mainly carried in apolipoprotein B (apoB) containing lipoproteins. High levels of these lipoproteins are associated with several metabolic diseases and lowering their plasma levels is associated with reduced incidence of atherosclerotic cardiovascular disease. MicroRNAs (miRs) are small non-coding RNAs that reduce the protein expression of their target mRNAs and are potential therapeutic agents. Here, we identified a novel miR-615-3p that interacts with human 3'-UTR of apoB mRNA, induces post-transcriptional mRNA degradation, and reduces cellular and secreted apoB100 in human hepatoma Huh-7 cells. Reducing cellular miR-615-3p levels by CRISPR-sgRNA increased cellular and secreted apoB100 indicating endogenous miR regulates apoB expression. Overexpression of miR-615-3p along with or without palmitic acid treatment decreased cellular and media apoB and increased cellular triglyceride levels without inducing endoplasmic reticulum stress. These studies have identified miR-615-3p as a negative regulator of apoB expression in human liver-derived cells. It is likely that there are more miRs that regulate apoB-containing lipoprotein assembly and secretion. Discovery of additional miRs may uncover novel mechanisms that control lipoprotein assembly and secretion.
Insights
Researchers discovered a new microRNA (miR-615-3p) that lowers apolipoprotein B (apoB) levels, a key factor in metabolic diseases. This finding offers potential new strategies for treating cardiovascular disease by targeting apoB production.
Area of Science:
- Biochemistry
- Molecular Biology
- Genetics
Background:
- Plasma lipids are transported by apolipoprotein B (apoB)-containing lipoproteins.
- Elevated apoB levels are linked to metabolic diseases and atherosclerotic cardiovascular disease.
- MicroRNAs (miRs) are regulators of gene expression with therapeutic potential.
Purpose of the Study:
- To identify novel microRNAs regulating apolipoprotein B (apoB) expression.
- To investigate the role of miR-615-3p in apoB regulation and lipoprotein metabolism.
- To explore the therapeutic potential of targeting miR-615-3p for metabolic and cardiovascular diseases.
Main Methods:
- Identification of miR-615-3p using bioinformatics and cell-based assays.
- Luciferase reporter assays to confirm interaction between miR-615-3p and apoB mRNA 3'-UTR.
- CRISPR-sgRNA technology to modulate endogenous miR-615-3p levels.
- Overexpression studies of miR-615-3p in human hepatoma Huh-7 cells.
- Measurement of cellular and secreted apoB100, triglycerides, and endoplasmic reticulum stress markers.
Main Results:
- miR-615-3p directly targets the 3'-UTR of apoB mRNA, inducing degradation and reducing apoB100 levels.
- Downregulation of endogenous miR-615-3p using CRISPR-sgRNA increased apoB100 expression.
- Overexpression of miR-615-3p decreased cellular and secreted apoB, and increased cellular triglycerides without ER stress.
- miR-615-3p acts as a negative regulator of apoB expression in human liver cells.
Conclusions:
- miR-615-3p is a novel regulator of apolipoprotein B (apoB) expression in human liver-derived cells.
- Modulating miR-615-3p levels presents a potential therapeutic strategy for managing apoB-related metabolic and cardiovascular diseases.
- Further discovery of miRs regulating apoB may reveal new mechanisms in lipoprotein assembly and secretion.
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