Improved adipose tissue function with initiation of protease inhibitor-only ART

Robert T Maughan, Eoin R Feeney, Emilie Capel, Jacqueline Capeau, Pere Domingo, Marta Giralt, Joep M A Lange, Praphan Phanuphak, David A Cooper, Peter Reiss, Patrick W G Mallon, HIVNAT-019 Study Group, Saskia R Autar, Jasper van der Lugt, Anchalee Avihingsanon, Supalak Klungklang, Sasiwimol Ubolyam, Robert T Maughan, Eoin R Feeney, Emilie Capel, Jacqueline Capeau, Pere Domingo, Marta Giralt, Joep M A Lange, Praphan Phanuphak, David A Cooper, Peter Reiss, Patrick W G Mallon, HIVNAT-019 Study Group, Saskia R Autar, Jasper van der Lugt, Anchalee Avihingsanon, Supalak Klungklang, Sasiwimol Ubolyam

Abstract

Objectives: Use of ART containing HIV PIs has previously been associated with toxicity in subcutaneous adipose tissue (SAT), potentially contributing to the development of lipodystrophy and insulin resistance. However, the effect of PIs on SAT function in ART-naive patients independent of other ART classes is unknown. This study aimed to elucidate the effect of initiating PI-only ART on SAT function in ART-naive subjects.

Methods: In the HIVNAT-019 study, 48 HIV-infected, ART-naive Thai adults commencing PI-only ART comprising lopinavir/ritonavir/saquinavir for 24 weeks underwent assessments of fasting metabolic parameters and body composition. In a molecular substudy, 20 subjects underwent SAT biopsies at weeks 0, 2 and 24 for transcriptional, protein, mitochondrial DNA (mtDNA) and histological analyses. ClinicalTrials.gov registration number: NCT00400738.

Results: Over 24 weeks, limb fat increased (+416.4 g, P = 0.023), coinciding with larger adipocytes as indicated by decreased adipocyte density in biopsies (-32.3 cells/mm2, P = 0.047) and increased mRNA expression of adipogenesis regulator PPARG at week 2 (+58.1%, P = 0.003). Increases in mtDNA over 24 weeks (+600 copies/cell, P = 0.041), decreased NRF1 mRNA expression at week 2 (-33.7%, P < 0.001) and increased COX2/COX4 protein ratio at week 24 (+288%, P = 0.038) indicated improved mitochondrial function. Despite decreased AKT2 mRNA at week 2 (-28.6%, P = 0.002) and increased PTPN1 mRNA at week 24 (+50.3%, P = 0.016) suggesting insulin resistance, clinical insulin sensitivity [by homeostasis model assessment (HOMA-IR)] was unchanged.

Conclusions: Initiation of PI-only ART showed little evidence of SAT toxicity, the changes observed being consistent with a return to health rather than contributing to lipodystrophy.

© The Author 2016. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy.

Figures

Figure 1.
Figure 1.
Gene expression changes in the adipose tissue of subjects initiating HIV PI-only ART. Changes in the expression of genes related to (a) adipogenesis and lipid metabolism, (b) insulin signalling, (c) markers of stress and adipocyte-secreted hormones and (d) mitochondrial function were measured by qPCR array. Target gene expression was normalized to the average crossing point of the reference genes: actin β (ACTB), ribosomal protein L13a (RPL13A) and TATA box binding protein (TBP). The bars represent the median (IQR) percentage change from week 0 to week 2 and to week 24; n = 13 at week 2 and n = 12 at week 24. Statistically significant differences from baseline are labelled *P ≤ 0.05 and ***P ≤ 0.005 (Wilcoxon signed rank test).
Figure 2.
Figure 2.
Changes in adipose tissue mtDNA content over 24 weeks in subjects initiating HIV PI-only ART. mtDNA was measured at two regions of the mitochondrial genome by qPCR: mitochondrially encoded cytochrome b (MT-CYB; region 1) and mitochondrially encoded cytochrome c oxidase I (MT-CO1; region 2). mtDNA levels were normalized to nuclear genome DNA copy number (primers targeting peroxisome proliferator-activated receptor γ), with mtDNA copies/cell calculated as copy number of mtDNA/(copy number of nuclear DNA/2). Data are presented as mean (SEM) and P values correspond to the output of marginal model analyses. The number of samples (n) included at each timepoint is indicated under each graph. Wk, week.
Figure 3.
Figure 3.
Changes in the expression of proteins in the adipose tissue of subjects initiating HIV PI-only ART. Quantification was by immunoblotting with subsequent densitometric analysis. The bars represent the median (IQR) percentage change from week 0 to week 2 and to week 24. P values correspond to the Wilcoxon signed rank test. (a) PPARG, peroxisome proliferator-activated receptor γ. (b) SREBP1, sterol regulatory element binding protein 1. (c) COX4, cytochrome c oxidase subunit 4. (d) COX2, mitochondrially encoded cytochrome c oxidase subunit 2. (e) B2M, β2 microglobulin. The number of samples (n) included at each timepoint is given under each graph. Wk, week.
Figure 4.
Figure 4.
Histological analysis of the adipose tissue biopsies of subjects initiating HIV PI-only ART. (a) Change in adipose tissue adipocyte density over 24 weeks. The number of adipocytes per field (×10 magnification) was quantified using viewing software with adipocyte density expressed as the number of adipocytes/mm2. Data are presented as mean (SEM) and P values correspond to the output of marginal model analyses; n = 18, 18 and 14 at weeks 0, 2 and 24, respectively. Wk, week. (b) Representative micrographs of adipose tissue biopsy sections at week 0 and at week 24 taken at ×10 magnification, stained with haematoxylin phloxine saffron.

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