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Obesity (Poster Session)

(P20-030-25) A 70 Amino Acid Fragment of Insparin Retains Its Potential to Upregulate Distal Insulin Signaling

Sunday, June 1, 2025
12:45 PM - 1:45 PM ET
Location: Poster Board D-19
Disclosure(s):
Tanvi Kale, MS Biotechnology: No relevant financial relationship(s) with ineligible companies to disclose.
Vijay Hegde, PhD: No relevant financial relationship(s) with ineligible companies to disclose.


Objectives: E4orf1 (Insparin), a 125 amino acid protein derived from Adenovirus 36 upregulates pAKT in cellular insulin signaling pathway and increases Glut4 abundance and membrane translocation to promote cellular glucose uptake independent of insulin. A four amino acid protein binding motif (PBM) at the C-terminal of Insparin is necessary for its therapeutic action. Preclinical studies show strong anti-diabetic potential of subcutaneous administration of Insparin. We hypothesized that a smaller fragment of Insparin that retains the PBM may provide better safety and efficacy profile. Here we synthesized a 70 aa fragment of Insparin and screened for its Insparin-like potential by determining upregulation of pAKT and GLUT 4 abundance in vitro.

Methods: Structures of Insparin peptide for 50, 70, and 125 aa with its conserved PBM were generated using bioinformatics software Phyre2 and Geneious Prime. Since the 70 aa peptide retained its secondary structure, it was synthesized and screened for its effect on pAKT and Glut 4 abundance. 3T3 L1- MBX cells were treated with either 23.8 µM or 0 µM (control) of 70 aa peptide using a Pulsin-mediated delivery and incubated for 5 or 7 h. Next, cell lysates were used to determine protein expression of pAKT and GLUT4 via western blot.

Results: Cells transfected with the 70 aa peptide for 5h increased pAKT abundance (normalized to AKT; p=0.025) and GLUT 4 abundance (normalized to GAPDH; p=0.067) compared with control. There was no significant difference in pAKT or Glut4 abundance for cells transfected for 7h with 70 aa peptide compared with control.

Conclusions: The 70 aa fragment of Insparin appears to capture the ability of Insparin to promote glucose uptake, suggesting its potential use as an anti-diabetic agent. Further research is needed to determine its efficacy in improving diabetes in vivo and to compare with that of Insparin.

Funding Sources: Insparin Therapeutics