The Integrase Inhibitors Dolutegravir and Raltegravir Exert Proadipogenic and Profibrotic Effects and Induce Insulin Resistance in Human/Simian Adipose Tissue and Human Adipocytes

Abstract

Background Although some integrase strand transfer inhibitors (INSTIs) promote peripheral and central adipose tissue/weight gain in people with human immunodeficiency virus (PHIV), the underlying mechanism has not been identified. Here, we used human and simian models to assess the impact of INSTIs on adipose tissue phenotype and function. Methods Adipocyte size and fibrosis were determined in biopsies of subcutaneous and visceral adipose tissue (SCAT and VAT, respectively) from 14 noninfected macaques and 19 PHIV treated or not treated with an INSTI. Fibrosis, adipogenesis, oxidative stress, mitochondrial function, and insulin sensitivity were assessed in human proliferating or adipocyte-differentiated adipose stem cells after long-term exposure to dolutegravir or raltegravir. Results We observed elevated fibrosis, adipocyte size, and adipogenic marker expression in SCAT and VAT from INSTI-treated noninfected macaques. Adiponectin expression was low in SCAT. Accordingly, SCAT and VAT samples from INSTI-exposed patients displayed higher levels of fibrosis than those from nonexposed patients. In vitro, dolutegravir and, to a lesser extent, raltegravir were associated with greater extracellular matrix production and lipid accumulation in adipose stem cells and/or adipocytes as observed in vivo. Despite the INSTIs’ proadipogenic and prolipogenic effects, these drugs promoted oxidative stress, mitochondrial dysfunction, and insulin resistance. Conclusions Dolutegravir and raltegravir can directly impact adipocytes and adipose tissue. These INSTIs induced adipogenesis, lipogenesis, oxidative stress, fibrosis, and insulin resistance. The present study is the first to shed light on the fat modifications observed in INSTI-treated PHIV.

SIV Infection and the HIV Proteins Tat and Nef Induce Senescence in Adipose Tissue and Human Adipose Stem Cells, Resulting in Adipocyte Dysfunction

“…Several studies reported a higher level of fibrosis in adipose tissue of ART-treated HIV-infected patients compared to noninfected subjects [6,7,9]. Finally, integrase inhibitors (INSTIs), a recent widely used class of ART, have less impact on senescence of endothelial cells together with anti-inflammatory properties [58,59] but their use is associated with fat gain and higher AT fibrosis [60][61][62][63]). Taken as a whole, ART can also have a role in adipose tissue aging in HIV-infected patients.…”

Metformin alleviates aging-associated cellular senescence of human adipose stem cells and derived adipocytes

“…The study was performed in compliance with the principles of the Declaration of Helsinki, and was approved by the local independent ethics committee. The ASCs were isolated using a collagenase (Roche Diagnostics, Basel, Switzerland) digestion technique, as described previously (Gorwood, Bourgeois et al 2019, Gorwood, Ejlalmanesh et al 2020), after seeding in Eagle’s Minimum Essential Medium alpha (Thermo Fisher Scientific, Courtaboeuf, France) supplemented with 10% foetal bovine serum (FBS) (PAN-Biotech, Aidenbach, Germany), 2 mmol/L glutamine, 100 U/mL penicillin/streptomycin, 10 mmol/L HEPES (Thermo Fisher Scientific), and 2.5 ng/mL FGF2 (PeproTech, Rocky Hill, NJ, USA). Upon confluence, adherent cells were trypsinized (Thermo Fisher Scientific) and seeded at a density of between 2000 and 4000 cells/cm 2 .…”

“…Differentiation of ASCs was induced by the addition of proadipogenic Dulbecco Modified Eagle’s Medium (DMEM), 4.5 g/L glucose (Thermo Fisher Scientific), 10% FBS, 2 mmol/L glutamine, 100 U/mL penicillin/streptomycin, 10 mmol/L HEPES, 1 μmol/L dexamethasone, 250 μmol/L 3-Isobutyl-1-methylxanthine (IBMX), 1 μmol/L rosiglitazone, 1 μmol/L insulin (Merck, Sigma-Aldrich) for 5 days, and then maintained in DMEM with rosiglitazone and insulin up until day 14. Cells were then stained for Oil-Red-O (Merck, Sigma-Aldrich) and quantified at 520nm as described previously (Gorwood, Bourgeois et al 2019, Gorwood, Ejlalmanesh et al 2020).…”

“…Cellular senescence was evaluated in terms of the cell PDT at each cell passage, as described previously (Gorwood, Ejlalmanesh et al 2020). The positive blue staining of senescence-associated SA-β-galactosidase has been used as a biomarker of cellular senescence.…”

“…The positive blue staining of senescence-associated SA-β-galactosidase has been used as a biomarker of cellular senescence. To detect SA-β-galactosidase activity, cells were incubated in an appropriate buffer solution at pH6 containing bromo-4-chloro-3-indolyl-β-D-galactopyranoside (Euromedex, Souffelweyersheim, France), as described previously (Gorwood, Ejlalmanesh et al 2020). The percentage of blue SA-β-galactosidase-positive cells was estimated by cell counting in at least 3 random-selected fields at a magnification of 20X.…”