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An Adipocyte Cell Culture Model to Study the Impact of Protein and Micro-RNA Modulation on Adipocyte Function
Published on: May 4, 2021
MicroRNA-192* impairs adipocyte triglyceride storage
Raghavendra Mysore1, You Zhou1, Sanja Sädevirta2
1Minerva Foundation Institute for Medical Research, Biomedicum 2U, FI-00290 Helsinki, Finland.
Abstract:
We investigated the expression of miR-192* (miR-192-3p) in the visceral adipose tissue (VAT) of obese subjects and its function in cultured human adipocytes. This miRNA is a 3' arm derived from the same pre-miRNA as miR-192 (miR-192-5p) implicated in type 2 diabetes, liver disease and cancers, and is predicted to target key genes in lipid metabolism. In morbidly obese subjects undergoing bariatric surgery preceded by a very low calorie diet, miR-192* in VAT correlated negatively (r=-0.387; p=0.046) with serum triglyceride (TG) and positively with high-density lipoprotein (HDL) concentration (r=0.396; p=0.041). In a less obese patient cohort, the miRNA correlated negatively with the body mass index (r=-0.537; p=0.026). To characterize the function of miR-192*, we overexpressed it in cultured adipocytes and analyzed the expression of adipogenic differentiation markers as well as cellular TG content. Reduced TG and expression of the adipocyte marker proteins aP2 (adipocyte protein 2) and perilipin 1 were observed. The function of miR-192* was further investigated by transcriptomic profiling of adipocytes expressing this miRNA, revealing impacts on key lipogenic genes. A number of the mRNA alterations were validated by qPCR. Western analysis confirmed a marked reduction of the lipogenic enzyme SCD (stearoyl coenzyme A desaturase-1), the fatty aldehyde dehydrogenase ALDH3A2 (aldehyde dehydrogenase 3 family member A2) and the high-density lipoprotein receptor SCARB1 (scavenger receptor B, type I). SCD and ALDH3A2 were demonstrated to be direct targets of miR-192*. To conclude, the present data identify miR-192* as a novel controller of adipocyte differentiation and lipid homeostasis.
Insights
MicroRNA miR-192* controls adipocyte differentiation and lipid homeostasis. This study found miR-192* expression in visceral adipose tissue correlates with obesity and lipid levels, impacting key metabolic genes.
Area of Science:
- Metabolic research
- Molecular biology
- Adipocyte biology
Background:
- MicroRNAs (miRNAs) play crucial roles in metabolic regulation.
- miR-192* (miR-192-3p), a less-studied miRNA, is derived from the same precursor as miR-192 (miR-192-5p), which is linked to metabolic diseases.
- Understanding miR-192*'s function in adipose tissue is essential for metabolic health.
Purpose of the Study:
- To investigate the expression of miR-192* in visceral adipose tissue (VAT) of obese individuals.
- To elucidate the functional role of miR-192* in human adipocyte differentiation and lipid metabolism.
- To identify the molecular targets of miR-192* involved in lipogenesis.
Main Methods:
- Expression analysis of miR-192* in VAT from obese and less obese subjects.
- Overexpression of miR-192* in cultured human adipocytes.
- Analysis of adipogenic differentiation markers and cellular triglyceride content.
- Transcriptomic profiling and validation of target genes using qPCR and Western blot.
Main Results:
- miR-192* expression in VAT negatively correlated with serum triglyceride and positively with HDL in morbidly obese subjects.
- miR-192* expression correlated negatively with body mass index in a less obese cohort.
- Overexpression of miR-192* in adipocytes reduced cellular triglyceride content and expression of adipocyte markers aP2 and perilipin 1.
- Transcriptomic analysis revealed miR-192* impacts lipogenic genes, including direct targeting of SCD and ALDH3A2, and reduced expression of SCARB1.
Conclusions:
- miR-192* is a novel regulator of adipocyte differentiation and lipid homeostasis.
- The findings suggest miR-192* plays a significant role in managing lipid metabolism within adipose tissue.
- miR-192* represents a potential therapeutic target for obesity and related metabolic disorders.
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