Background

Co-expression of heparin-binding EGF-like (HB-EGF) and a disintegrin and metallprotease (ADAM) 12S is capable of reprogramming a number of mammalian cells into brown adipose tissue-like cells as evidenced by up-regulation of BAT genes and down-regulation of white adipose tissue (WAT) genes. Furthermore, the metabolic activity of the reprogramed BAT-like cells is similar to that of BAT.

Methods

Human primary adipodyctes were infected with either mock or ADAM 12S high titer adenovirus, monitored for fluorescence, lipid accumulation, and qRT-PCR conducted for tl BAT genes, PRDM16, PGC-1α, and UCP-1. Metabolic analysis for functionality of BAT-like cells was determined using a Seahorse XF24 analyzer.

Results

ADAM 12S infected cells demonstrated lipid droplet accumulation and positive Oil Red O stainingin comparison to the mock infected cells.qRT-PCR demonstrated significant up-regulation ofPGC-1α in ADAM 12S infectedcells (p = 0.05) compared to mock infected cells. Although UCP-1 was up-regulated, is was not statistically significant (p = 0.05).The Seahorse metabolic assay demonstrated an increase rate of glycolysis for ADAM 12S cells after exposure to a stressor mix, composed of FCCP and Oligomycin, when compared to their basal rate. ECAR was significantly increased in ADAM 12S cells compared to MOCK cells after exposure to catecholamines and the stressor mix (FCCP and Oligomycin).

Conclusions

These results further support previous findings that co-expression of HB-EGF and ADAM 12S stimulates cellular reprogramming into brown adipose tissue (BAT)-like cells. We believe that ADAM 12S stimulates cellular reprogramming into BAT-like cells utilizing endogenous hHB-EGF. These novel insights may provide the first evidence demonstrating BAT-like cellular reprogramming occurs in vivo in humans. This research has possible therapeutic applications to combat obesity and type II diabetes.