No difference in monocyte cholesterol metabolism gene expression in people living with HIV and matched uninfected controls: the HIV UBPEAT CAD Substudy
D Alalwan2, P McGettrick1,2, W Tinago2, J O'Halloran2, J O'Brien3, N Mahon4, L Lawler5, A Cotter1,2, J Lambert1,2, G Sheehan1,2, C Sabin8, P Mallon1,2
1Department of Infectious Diseases, Mater Misericordiae University Hospital, Dublin, Ireland; 2HIV Molecular Research Group, School of Medicine, University College Dublin, Dublin, Ireland; 3Department of Radiology, University Hospital Limerick, Limerick, Ireland; 4Department of Cardiology, Mater Misericordiae University Hospital, Dublin, Ireland; 5Department of Radiology, Mater Misericordiae University Hospital, Dublin, Ireland; 6School of Microbiology & Alimentary Pharmabiotic Centre, University Cork, Cork, Ireland; 7Rush University Medical Center, Chicago, USA; 8Research Department of Infection and Population Health, University College London, London, United Kingdom
Objectives/aims: People living with HIV (PLWH) have an increased risk of coronary artery disease (CAD) with previous studies reporting monocyte gene expression consistent with high intracellular cholesterol in treatment-naive PLWH which improves but does not normalize with ART. We aimed to examine differences in cholesterol metabolism gene expression in a cohort of treated PLWH with CAD risk factor (CADRF) matched uninfected controls.
Methods: The UPBEAT CAD substudy enrolled 100 participants of the HIV UPBEAT study, a prospective longitudinal cohort of PLWH and uninfected controls with over 10 years follow-up. Participants were over the age of 40 years, had no known history of CAD and were closely matched for HIV serostatus and traditional CADRF.
Quantitative polymerase chain reaction used to assess the expression of 17 cholesterol metabolism genes in monocytes isolated from fasting fresh blood samples. Gene expression was reported relative to 3 housekeeping genes. Between group differences was assessed using Mann–Whitney U test and analysis of covariance to adjust for confounders (SPSS vers 24).
Results: 99 participants were included in the analysis. Median age was 50.81 (46.29, 56.24) years, 71.71% male, 76.76% Caucasian and 48.48% were current smokers. PLWH had lower HDL cholesterol (HIV+ 1.27 [1.0,1.3]; HIV- 1.4 [1.1,1.6]; P=0.017) and more likely to be on statin therapy (HIV+ 49%, HIV- 12%; P<0.01). Other demographics and CADRF were similar between groups.
After adjustment for HDL and statin use, there remained no significant association between HIV serostatus and cholesterol metabolism gene expression (Table 1).
(Abstract P16)
Gene
HIV Negative Estimated Mean (95% CI)
HIV Positive Estimated Mean (95% CI)
P
Cholesterol Sensing
SCAP
0.0906 (0.0766, 0.1046)
0.0779 (0.0641, 0.0918)
0.227
SREBF1
0.0131 (0.0112, 0.0150)
0.0117 (0.0099, 0.0136)
0.34
SREBF2
0.0196 (0.0163, 0.0229)
0.0193 (0.0163, 0.0223)
0.901
MBTPS1
0.0308 (0.0253, 0.0363)
0.0328 (0.0274, 0.0382)
0.62
MBTPS2
0.0048 (0.0033, 0.0064)
0.0056 (0.0041, 0.0071)
0.513
PPARA
0.0095 (0.0072, 0.0117)
0.0092 (0.0070, 0.0115)
0.893
NR1H3
0.0053 (0.0042, 0.0064)
0.0053 (0.0042, 0.0064)
0.995
LPL
0.0028 (0.0018, 0.0038)
0.0031 (0.0021, 0.0041)
0.635
Cholesterol Uptake
LDLR
0.0433 (0.0363, 0.0503)
0.0458 (0.0383, 0.0533)
0.643
CD36
0.0348 (0.0186, 0.0510)
0.0515 (0.0353, 0.0677)
0.178
De Novo Cholesterol Synthesis
HMGCR
0.0275 (0.0173, 0.0377)
0.0367 (0.0264, 0.0470)
0.233
PMVK
0.0227 (0.0179, 0.0275)
0.0181 (0.0134, 0.0229)
0.204
ACAT2
0.0108 (0.0090, 0.0126)
0.0114 (0.0096, 0.0131)
0.684
Cholesterol Efflux
ABCA1
0.0009 (0.0004, 0.0014)
0.0017 (0.0012, 0.0023)
0.053
ABCG1
0.0058 (0.0045, 0.0072)
0.0074 (0.0060, 0.0087)
0.143
SCARB1
0.0259 (0.0218, 0.0300)
0.0232 (0.0191, 0.0273)
0.369
Model adjusted for HDL and Statin use, CI; Confidence Interval (lower bound, upper bound)
Conclusions/discussion: In a cohort of treatment-experienced PLWH and CADRF matched controls, there was no significant difference in monocyte cholesterol gene expression suggesting a persistent dysfunctional intracellular cholesterol metabolism may not contribute to the increased risk of CAD observed in stable, ART-treated PLWH.
Abstract P17
Antiviral Therapy 2020; 25 Suppl 1:A53
Prevalence of silent atherosclerosis and other comorbidities in an outpatient cohort of adults living with HIV: associations with HIV parameters and biomarkers
J Ghosn1, H Abdoul1, S Fellahi2, A Merler1, D Salmon1, JP Morini1, J Deleuze1, J Blacher1, J Capeau2, JP Bastard2, JP Viard1
1Assistance Publique-Hôpitaux de Paris. Centre-Université de Paris; 2Assistance Publique-Hôpitaux de Paris. Sorbonne Université, Paris, France
Background: People living with HIV (PLWH) are at risk of non-infectious comorbidities. It is important to individualize those at higher risk.
Methods: In a single-centre cohort of PLWH, we performed a cross-sectional analysis of comorbidities, diagnosed according to standard procedures. The primary end point was the prevalence of subclinical carotid/ coronary atherosclerosis. Secondary end points were its association with selected inflammatory/immune activation biomarkers and with other comorbidities. Associations were examined using Chi-2 or Fisher's exact test for categorical variables and Student or Wilcoxon tests for quantitative variables, and a stepwise multivariate logistical model was performed for further exploration.
Results: Among 790 participants (median age: 49.8 years [IQR: 44.5–55.6], 77.1% males, median CD4: 536/mm3 [IQR: 390–754], 83.6% with undetectable viral load), asymptomatic atherosclerosis was found in 26% and was associated in multivariate analysis with older age, longer known duration of infection, longer exposure to antiretroviral drugs and ever exposure to stavudine, higher sCD14 and lower adiponectin levels. Hypertension was found in 33.5% of participants, diabetes in 19.4%, renal impairment in 14.6%, elevated LDL-cholesterol in 13.3%, elevated triglyceride/HDL-cholesterol ratio in 6.6% and osteoporosis in 7.9%. The presence of two or more comorbidities was found in 42.1% of participants and was associated in mul-tivariate analysis with older age, longer exposure to antiretrovirals, ever exposure to stavudine and higher sCD14 levels. Comorbidities were diversely associated with biomarkers: osteoporosis with higher IL-6, renal impairment with higher sCD14, hypertension with higher D-dimer, diabetes and elevated triglyceride/ HDL-cholesterol ratio both with lower adiponectin and lower 25hydroxy-vitamin D.
Conclusions: Asymptomatic atherosclerosis and multimorbidity were frequent in a cohort of middle-aged, well-controlled, PLWH and were associated with traditional and HIV-specific factors. Associations between morbidities and inflammatory/immune activation biomarkers were diverse.
Abstract P18
Antiviral Therapy 2020; 25 Suppl 1:A54
Higher interactions of leukocytes and platelets with the endothelium as a potential hint of cardiovascular risk in ABC-treated HIV patients
MA Blanch-Ruiz1, A Sanchez-Lopez1, R Ortega-Luna1, P García-Martínez1, S Orden1, MA Martinez-Cuesta1, R Ferrando-Vilalta2, MJ Galindo2, JV Esplugues1, A Alvarez1
1University of Valencia, Valencia, Spain; 2Hospital Clinic of Valencia, Valencia, Spain
Aims: Abacavir (ABC) has been linked with a higher risk of myocardial infarction. We have demonstrated in vitro, in cells from healthy donors, that clinical concentrations of ABC, but not of tenofovir diso-proxil fumarate (TDF), have pro-inflammatory effects, inducing leukocyte–endothelium interactions and prothrombotic actions, causing the interplay of platelets with endothelial cells and leukocytes (specifically with neutrophils). Furthermore, ABC promoted thrombus formation in a well-established murine in vivo model. Thus, given that platelets and their interactions with other vascular cells play an important role in thrombus formation that can lead finally to myocardial infarction, the aim of the present study was to test the pro-inflammatory and pro-thrombotic status of HIV patients undergoing ABC versus tenofovir (TFV), including TDF or tenofovir alafenamide (TAF), treatment by analysing polymorphonuclear (PMN)- and/or platelet–endothelium interactions in cells isolated from blood of these two groups of HIV patients.
Methods: This is a non-randomized prospective observational study in which we used leukocytes and platelets from blood withdrawn from HIV-patients at Hospital Clínico Universitario de Valencia who had been receiving treatment, for at least 6 months, with an ART regime that included either ABC or TFV. Interactions of isolated PMNs, rolling and adhesion, and isolated platelets, adhesion, with a non-infected endothelium monolayer were evaluated by means of a parallel-plate flow chamber system. Platelets were labelled with an anti-CD41 (specific platelet marker) antibody linked to Alexa-Fluor®488 in order to visualize them by Epi-fluorescence microscopy.
Results: 50 patients were included in the study, 25 of whom were receiving ABC and 25 of whom were receiving TFV. There were no significant differences in demographic and cardiovascular risk parameters between the two groups. PMN rolling (Figure 1A) and adhesion (Figure 1B) along the endothelium were significantly higher in the ABC group than in the TFV group. Moreover, the number of platelets adhering to endothelial cells was higher in the ABC versus TFV group (Figure 1C).
(Abstract P18)
Conclusions: Treatment with ABC enhances PMN-endothelium interactions, thus promoting the initial phases of the inflammatory process. Furthermore, it induces platelet adhesion to endothelial cells, which is an important step in thrombus formation. Our results give support to the increased risk of myocardial infarction observed in ABC-treated HIV patients.
Abstract P19
Antiviral Therapy 2020; 25 Suppl 1:A55
Protective effects of lamivudine, emtricitabine, didanosine, raltegravir and atazanavir on platelet aggregation
R Ortega-Luna1, S Tiracorrendo1, MA Blanch-Ruiz1, S Orden1, MA Martinez-Cuesta1, JV Esplugues1, A Alvarez1
1University of Valencia, Valencia, Spain
Objectives/aims: Cardiovascular toxicity associated with combined antiretroviral therapy has been attributed mainly to nucleoside reverse transcriptase inhibitors (NRTI; specifically abacavir), though particular drugs from other families (non-nucleoside reverse tran-scriptase inhibitors [NNRTI], chemokine co-receptor 5 antagonist [aCCR5], protease inhibitors [PI] and integrase strand transfer inhibitors [INSTI]) may also produce this side effect as they are administered in combination. This study evaluates the actions of clinically relevant concentrations of antiretrovirals belonging to the different families on platelet aggregation – key step in the transition from inflammation to thrombus generation.
Methods: We employed platelet-rich plasma (PRP) from healthy donors from Hospital Clínico Universitario de Valencia, who had not been receiving drugs for at least 10 days. Platelet aggregation in response to positive stimulus such as ADP (1 mM) or collagen (2 μg/ml) was assessed by an impedance aggregometer (Chrono-Log Model 590-2D) in the presence of selected antiretroviral drugs (4 min treatment): the NRTIs abacavir (5 μg/ml), tenofovir (0.3 μg/ml), didanosine (1.8 μg/ml), emtricitabine (2.5 μg/ml) and lamivudine (2.3 μg/ml); the INSTIs raltegravir (5 μg/ml) and elvitegravir (10 μg/ml); the aCCR5 maroviroc (5 μg/ml); the PIs lopinavir (25 μg/ml) and atazanavir (25 μg/ml) and the NNRTIs efavirenz (25 μg/ml) and rilpivirine (1 μg/ml). AUC (area under curve) is the parameter shown as it includes both amplitude and slope of the curve.
Results: In contrast to what was expected, none of the drugs potentiated platelet aggregation induced by ADP or collagen. However, some of them had a protective action. Particularly lamivudine and raltegravir reduced platelet aggregation induced by ADP (Figure 1A) whereas emtricitabine, didanosine and atazanavir inhibited that caused by collagen (Figure 1B). On the other hand, the rest of antiretroviral drugs analysed did not induce significant changes in platelet aggregation stimulated by ADP or collagen.
(Abstract P19)
Conclusions/discussion: Our results suggest that lamivudine, emtricitabine, didanosine, atazanavir and raltegravir could be safer alternatives than other antiretrovi-ral drugs used in acquired immune deficiency syndrome treatment such as abacavir which have been correlated with the appearance of cardiovascular side effects.
Abstract P20
Antiviral Therapy 2020; 25 Suppl 1:A56
Changes in inflammatory markers after switching to elvitegravir/cobicistat/emtricitabine/tenofovir alafenamide in virologically suppressed older people living with HIV
N Funderburg1, E Bowman1, A Kettlehut1, J Gabriel1, S Navadeh2, A Weinberg2, C Blair2, I McNicholl2, S Chuck2, R Haubrich2
1Ohio State University, Columbus, OH, USA; 2Gilead Sciences, Foster City, CA, USA
Aim: Antiretroviral therapy reduces both immune activation and systemic inflammation, yet, little data exist on these indices in older people living with HIV. We examined the effect of age on plasma biomarkers of monocyte activation (sCD14, sCD163), and systemic (IL-6, hsCRP, sTNFR-I, TNFR-2 and D-dimer) and vascular (Lp-PLA2) inflammation in virologically suppressed adults living with HIV who switched from a stable regimen to elvitegravir/cobicistat/emtricitabine/ tenofovir alafenamide in two randomized, controlled trials (GS-US-292-1823, GS-US-292-1826).
Methods: Samples were included from participants who were on abacavir/lamivudine or emtricitabine/ tenofovir disoproxil fumarate plus a 3rd agent for ≥6 months and switched to elvitegravir/cobicistat/emtricitabine/tenofovir alafenamide, received at least one dose of study medication, and had both baseline and at least one post-baseline value at weeks 12 or 24 for any biomarker. Samples from participants who had an HIV-1 RNA ≥50 copies/ml at any time point were excluded. Biomarker levels were measured at baseline and at weeks 12 and 24. We compared baseline levels and change from baseline between two age groups (<60 years, ≥60 years) using the Wilcoxon rank sum test. We used a multivariate, stepwise regression model to identify factors associated with percent change in bio-markers. Demographics, disease characteristics, prior HIV treatment duration, CD4 cell count, eGFR, lipid profiles, CVD risk score, diabetes, hypertension, hyperlipidaemia, current smoking status, baseline statin use and baseline third agent were included in the stepwise regression models.
Results: 235 participants were included (<60 years, 119; ≥60 years, 116): 14% women, 10% Black, 30% current smokers, 29% statin users and 45% with 10-year cardiovascular risk ≥10%. Median baseline CD4 count (651 cells/μl) was similar in both groups. Baseline levels IL-6, TNFRs 1 and 2, D-dimer and sCD163 were significantly higher in the ≥60 years old group compared with levels in the <60 years. In the total population, median levels of IL-6, D-dimer, TNFRs 1 and 2, and Lp-PlA2 increased from baseline through week 24. There were no differences in the median change in biomarkers between age groups at weeks 12 and 24 (P>0.05) except for sCD163 (P=0.007 and 0.03, respectively). By stepwise regression model, age was not a significant predictor for percent change in any of the biomarkers through week 24. See Table 1.
Conclusions: Although there were differences in biomarkers between age groups at baseline, age was not associated with longitudinal biomarker changes in virologically suppressed adults after switching to elvitegravir/cobicistat/emtricitabine/tenofovir alafenamide.
Baseline and change in biomarkers by age group (Abstract P20)
P-value for difference between age groups by 2-sided Wilcoxon rank sum test
Abstract P21
Antiviral Therapy 2020; 25 Suppl 1:A57
Attitudes toward COVID-19 among people living with HIV attending a multiethnic outpatient centre in Houston, Texas
P Debroy1, S Lupo1, M Reimer-McAtee1, JE Gioia, JE Lake1
1University of Texas Health Sciences Center, Houston, TX, USA
Objectives: The evolving outbreak of coronavirus 2019 (COVID-19) may impact people with HIV (PWH) in unique ways, beyond the increased risk of medical complications. Social distancing measures resulting in social isolation and interruptions in medical care may disproportionately affect PWH. The purpose of this study is to capture and understand how COVID-19 is affecting PWH in a low-income, multiethnic urban health centre in Houston, Texas.
Methods: Cross-sectional survey of adult PWH who attend Thomas Street Health Center for outpatient care and volunteered to complete a short telephone questionnaire. Descriptive statistics were calculated for all patient characteristics and survey responses.
Results: Of the 188 participants, 26% were women, 3.9% were transgender women, 33% were non-Hispanic Black and 57.4% Hispanic. The median (interquartile range) age was 49 (40–56) years; 73% had 1 or more comorbidities. Overall, most (82%) people had not experienced difficulty accessing HIV medicines; 25% of patients reported difficulty accessing medical care and failing to attend clinic due to fear of exposure to COVID-19. Most (82.8%) reported being affected by COVID-19 in their daily life; 14% reported being extremely affected. 64% feared getting COVID-19, though a minority feared they were at increased risk of becoming sick, a finding especially true among Black (22.4%) and Hispanic individuals (33.9%) compared with White participants (15.4%). Nearly half (48.4%) reported increased anxiety. More than a third (37.8%) reported feeling depressed, more frequently reported among Black (46.6%) compared with Hispanic individuals (34.8%) and White participants (23%); 31% felt more alone; 90% reported not seeking behavioural health resources. Only 10% reported increased use in alcohol or illegal substances. One in four did not have access to an electronic device for a telemedicine visit. Of the 14% who had lost their job after the pandemic started, 61% were Hispanic.
Conclusions: PWH experienced a wide range of effects from the COVID-19 pandemic. It is important to recognize the socioeconomical and psychological implications that will be seen in disadvantaged communities of PWH as a consequence of COVID-19.
