Adipocyte and Cell Biology

Nanoparticle-mediated in vitro delivery of E4orf1 to preadipocytes is a clinically relevant delivery system to improve glucose uptake

Abstract

Objective

Impaired glycemic control is a common comorbidity of obesity. E4orf1(E4), an adenovirus-derived protein, reduces the activity of insulin receptor substrate (IRS), yet activates Akt and promotes the membrane translocation of GLUT4, resulting in better glycemic control in mice. To develop a clinically suitable delivery system, here we constructed and tested liposome nanoparticles (NP), to deliver E4 to preadipocytes.

Methods

Glutathione-S-transferase (GST)-tagged E4 was encapsulated in Rhodamine-phosphatidylethanolamine (PE)-tagged soy-phosphatidylcholine-NP. The NP were characterized. Preadipocytes were treated with free E4, E4 containing NP (E4 NP) or E4-free NP (void NP).

Results

For void and E4 NP, the average size was ~150 and 130 nm, PDI was ~0.25 and 0.27, and Zeta potential was −23 and −25, respectively. The average encapsulation efficiency (EE) was ~50%. Cells treated with E4 showed maximum GST expression and Rhodamine signals at 24 h. The presence of E4 in cells was confirmed at 24, 48, and 72 h. At 72 h after exposure, E4 NP significantly decreased pTyr-IRS, yet increased pAkt protein abundance, membrane translocation of GLUT4, and glucose uptake, compared with cells treated with void NP. Free E4 (without NP) had no effect.

Conclusions

NP-mediated delivery of E4 promotes glucose uptake in preadipocytes. The next step is to test the efficacy of this clinically compatible delivery approach in vivo.

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Fig. 1: Morphology of NP.
Fig. 2: NP are able to encapsulate E4 protein and deliver it to the cells.
Fig. 3: Encapsulated E4 downregulates proximal insulin signaling but upregulates distal insulin signaling in preadipocytes.
Fig. 4: E4 NP enhance cellular glucose uptake in preadipocytes.
Fig. 5: NP are stable and functional after one week of storage at 4 °C.

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Acknowledgements

We would like to thank Mary Catherine Hastert for assistance with the transmission electron microscopy and Dr Peter Keyel for his help with flow cytometry (both at Texas Tech University).

Author information

Correspondence to Nikhil V. Dhurandhar.

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Conflict of interest

NVD has received several United States and international patents that protect intellectual property about the use of adenoviruses and E4 protein in obesity, diabetes, and related areas.

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Feizy, Z., Peddibhotla, S., Khan, S. et al. Nanoparticle-mediated in vitro delivery of E4orf1 to preadipocytes is a clinically relevant delivery system to improve glucose uptake. Int J Obes (2020). https://doi.org/10.1038/s41366-020-0526-6

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