Early Antiretroviral Therapy at High CD4 Counts Does Not Improve Arterial Elasticity: A Substudy of the Strategic Timing of AntiRetroviral Treatment (START) Trial

BACKGROUND: Both human immunodeficiency virus (HIV) infection and antiretroviral therapy (ART) may increase cardiovascular disease (CVD) risk. Vascular function assessments can be used to study CVD pathogenesis. We compared the effect of immediate versus deferred ART initiation at CD4 counts >500 cells/mm(3) on small arterial elasticity (SAE) and large artery elasticity (LAE). METHODS: Radial artery blood pressure waveforms were recorded noninvasively. Small arterial elasticity and LAE were derived from analysis of the diastolic pulse waveform. Randomized treatment groups were compared with linear models at each visit and longitudinal mixed models. RESULTS: Study visits involved 332 participants in 8 countries: mean (standard deviation [SD]) age 35 (10), 70% male, 66% nonwhite, 30% smokers, and median CD4 count 625 cells/mm(3) and 10-year Framingham risk score for CVD 1.7%. Mean (SD) SAE and LAE values at baseline were 7.3 (2.9) mL/mmHg × 100 and 16.6 (4.1) mL/mmHg × 10, respectively. Median time on ART was 47 and 12 months in the immediate and deferred ART groups, respectively. The treatment groups did not demonstrate significant within-person changes in SAE or LAE during the follow-up period, and there was no difference in mean change from baseline between treatment groups. The lack of significant differences persisted after adjustment, when restricted to early or late changes, after censoring participants in deferred group who started ART, and among subgroups defined by CVD and HIV risk factors. CONCLUSIONS: Among a diverse global population of HIV-positive persons with high CD4 counts, these randomized data suggest that ART treatment does not have a substantial influence on vascular function among younger HIV-positive individuals with preserved immunity

Combination antiretroviral therapy -associated lipodystrophy : insights into pathogenesis and treatment

Introduction: Combination antiretroviral therapy (cART) has decreased morbidity and mortality of individuals infected with human immunodeficiency virus type 1 (HIV-1). Its use, however, is associated with adverse effects which increase the patients risk of conditions such as diabetes and coronary heart disease. Perhaps the most stigmatizing side effect is lipodystrophy, i.e., the loss of subcutaneous adipose tissue (SAT) in the face, limbs and trunk while fat accumulates intra-abdominally and dorsocervically. The pathogenesis of cART-associated lipodystrophy is obscure. Nucleoside reverse transcriptase inhibitors (NRTI) have been implicated to cause lipoatrophy via mitochondrial toxicity. There is no known effective treatment for cART-associated lipodystrophy during unchanged antiretroviral regimen in humans, but in vitro data have shown uridine to abrogate NRTI-induced toxicity in adipocytes. Aims: To investigate whether i) cART or lipodystrophy associated with its use affect arterial stiffness; ii) lipoatrophic SAT is inflamed compared to non-lipoatrophic SAT; iii) abdominal SAT from patients with compared to those without cART-associated lipoatrophy differs with respect to mitochondrial DNA (mtDNA) content, adipose tissue inflammation and gene expression, and if NRTIs stavudine and zidovudine are associated with different degree of changes; iv) lipoatrophic abdominal SAT differs from preserved dorsocervical SAT with respect to mtDNA content, adipose tissue inflammation and gene expression in patients with cART-associated lipodystrophy and v) whether uridine can revert lipoatrophy and the associated metabolic disturbances in patients on stavudine or zidovudine based cART. Subjects and methods: 64 cART-treated patients with (n=45) and without lipodystrophy/-atrophy (n=19) were compared cross-sectionally. A marker of arterial stiffness, heart rate corrected augmentation index (AgIHR), was measured by pulse wave analysis. Body composition was measured by magnetic resonance imaging and dual-energy X-ray absorptiometry, and liver fat content by proton magnetic resonance spectroscopy. Gene expression and mtDNA content in SAT were assessed by real-time polymerase chain reaction and microarray. Adipose tissue composition and inflammation were assessed by histology and immunohistochemistry. Dorsocervical and abdominal SAT were studied. The efficacy and safety of uridine for the treatment of cART-associated lipoatrophy were evaluated in a randomized, double-blind, placebo-controlled 3-month trial in 20 lipoatrophic cART-treated patients. Results: Duration of antiretroviral treatment and cumulative exposure to NRTIs and protease inhibitors, but not the presence of cART-associated lipodystrophy, predicted AgIHR independent of age and blood pressure. Gene expression of inflammatory markers was increased in SAT of lipodystrophic as compared to non-lipodystrophic patients. Expression of genes involved in adipogenesis, triglyceride synthesis and glucose disposal was lower and of those involved in mitochondrial biogenesis, apoptosis and oxidative stress higher in SAT of patients with than without cART-associated lipoatrophy. Most changes were more pronounced in stavudine-treated than in zidovudine-treated individuals. Lipoatrophic SAT had lower mtDNA than SAT of non-lipoatrophic patients. Expression of inflammatory genes was lower in dorsocervical than in abdominal SAT. Neither depot had characteristics of brown adipose tissue. Despite being spared from lipoatrophy, dorsocervical SAT of lipodystrophic patients had lower mtDNA than the phenotypically similar corresponding depot of non-lipodystrophic patients. The greatest difference in gene expression between dorsocervical and abdominal SAT, irrespective of lipodystrophy status, was in expression of homeobox genes that regulate transcription and regionalization of organs during embryonal development. Uridine increased limb fat and its proportion of total fat, but had no effect on liver fat content and markers of insulin resistance. Conclusions: Long-term cART is associated with increased arterial stiffness and, thus, with higher cardiovascular risk. Lipoatrophic abdominal SAT is characterized by inflammation, apoptosis and mtDNA depletion. As mtDNA is depleted even in non-lipoatrophic dorsocervical SAT, lipoatrophy is unlikely to be caused directly by mtDNA depletion. Preserved dorsocervical SAT of patients with cART-associated lipodystrophy is less inflamed than their lipoatrophic abdominal SAT, and does not resemble brown adipose tissue. The greatest difference in gene expression between dorsocervical and abdominal SAT is in expression of transcriptional regulators, homeobox genes, which might explain the differential susceptibility of these adipose tissue depots to cART-induced toxicity. Uridine is able to increase peripheral SAT in lipoatrophic patients during unchanged cART.Johdanto: Ihmisen immuunikatoviruksen (HIV) hoitoon käytetyt lääkeyhdistelmät ovat vähentäneet HIV-positiivisten henkilöiden sairastuvuutta ja kuolleisuutta. Yhdistelmähoitoon liittyy kuitenkin vakavia sivuvaikutuksia, jotka lisäävät potilaiden riskiä sairastua mm. diabetekseen ja sepelvaltimotautiin. Yksi leimaavimpia sivuvaikutuksia on lipodystrofia eli ihonalaisen rasvakudoksen häviäminen (lipoatrofia) kasvoista, raajoista ja vatsalta samalla kun rasvaa kertyy ylen määrin vatsaonteloon ja niskaan. Ilmiön syyt ovat epäselvät. Useiden HIV:ta vastaan suunnattujen lääkeaineiden on epäilty aiheuttavan lipodystrofiaa mm. tuhoamalla mitokondrioita, solujen energiatehtaita . Lipodystrofiaan ei ole tehokasta hoitoa, ellei HIV-lääkitystä muuteta, mutta esim. uridiini on ollut lupaava apu solumallitutkimusten valossa. Tavoitteet: Tutkia liittyykö yhdistelmähoitoon tai sen käyttöön liittyvään lipodystrofiaan verisuonien jäykistymistä, onko lipoatrofinen rasvakudos tulehtunutta verrattuna ei-lipoatrofiseen rasvakudokseen, eroaako lipoatrofinen ei-lipoatrofisesta rasvakudoksesta mm. mitokondriomäärän ja aineenvaihduntaan vaikuttavien geenien ilmentymisen suhteen sekä poikkeaako lipodystrofiassa paremmin säilyvä niskan rasva häviävästä vatsan ihonalaisrasvasta ja onko se mahdollisesti ruskeata rasvaa. Lisäksi tutkimme, voiko ravintolisänä käytetty uridiini parantaa lipoatrofiaa ja siihen liittyviä aineenvaihduntahäiriöitä, kuten rasvamaksaa ja heikentynyttä insuliiniherkkyyttä. Menetelmät: Tutkimuksiin osallistui 64 HIV-positiivista yhdistelmähoidettua potilasta, joista 45:lla oli ja 19:lla ei ollut kehittynyt lääkitykseen liittyviä rasvakudoksen muutoksia. Verisuonijäykkyys tutkittiin pulssiaaltoanalyysilla, kehon koostumus mitattiin kaksienergisella röntgenabsorptiometria- sekä magneettikuvaantamisella ja maksan rasvapitoisuus protonispektroskopialla. Rasvakudosnäytteet otettiin potilaiden vatsan ja niskan ihoalaisrasvasta ja niistä mitattiin eri geenien ilmentymistä sekä mitokondrioiden ja tulehdussolujen määrää mm. DNA:n monistustekniikalla ja kudosleikevärjäyksin. Uridiinin tehoa lipoatrofian hoidossa arvioitiin 3kk satunnaistetussa lumelääkekontrolloidussa tutkimuksessa, johon osallistui 20 HIV-positiivista yhdistelmähoidettua lipoatrofista henkilöä. Tulokset: HIV-lääkityksen kesto, mutta ei lipodystrofia, altistaa verisuonien jäykistymiselle iästä ja verenpainetasosta riippumatta. Lipoatrofisessa rasvakudoksessa tulehdukseen liittyvien geenien ilmentyminen ja tulehdussolujen määrä ovat lisääntyneet, kun taas mitokondriomäärä sekä rasvasolujen muodostumiseen ja toimintaan liittyvien geenien ilmentyminen vähentyneet verrattuna ei-lipoatrofiseen rasvakudokseen. Lipodystrofiassa säilyvä/lisääntyvä niskan rasva on vähemmän tulehtunutta kuin herkemmin häviävä vatsan ihonalaisrasva eikä se ole ruskeata rasvaa. Lipodystrofisten henkilöiden niskan rasvassa on vähemmän mitokondrioita kuin ei-lipodystrofisten henkilöiden niskan rasvassa, vaikka kudokset ovat ulkoisesti samannäköisiä. Niskan ja vatsan alueen ihonalaisrasva eroaa eniten ns. homeobox-geenien ilmentymisessä eli sellaisten geenien, jotka määrittelevät kudosten sijainnin ja ominaisuudet sikiökehityksen varhaisvaiheessa. Uridiini lisää ihonalaisrasvan määrää lipoatrofisilla potilailla, mutta ei vaikuta maksan rasvapitoisuuteen tai insuliiniherkkyyteen. Johtopäätökset: HIV:n hoitoon käytettyjen lääkkeiden pitkäaikaiskäyttö lisää verisuonien jäykkyyttä ja siten potilaiden riskiä sairastua sydän- ja verisuonitauteihin. Lipoatrofinen rasva on tulehtunut ja sen mitokondriovarannot vähentyneet. Koska mitokondrioiden vähyys on todettavissa niskarasvassa myös sellaisilla lipodystrofisilla henkilöillä, joilla se on säilynyt atrofialta, mitokondriokatoa ei voida pitää lipoatrofiaa suoraan aiheuttavana tekijänä. Niskan ja vatsan ihonalaisrasvan merkittävin ero on elinkehitystä ohjaavissa geeneissä, mikä voi selittää kudosten erilaisen alttiuden lääkkeiden sivuvaikutuksille. Uridiini on tehokas hoito HIV-potilaiden lipodystrofiaan muuttumattoman yhdistelmähoidon aikana

Carotid artery biomechanical parameters as measured with ultrasound elastography in HIV individuals – an assessment of the association to coronary atherosclerosis and comparison to traditional cardiovascular risk factors

Aim: This study aims to assess the association of biomechanical characteristics of carotid walls and carotid intima-media thickness (IMT), as assessed by ultrasound, when incorporated into prediction models for coronary CT plaque burden in both people living with HIV (PLWH) and HIV-negative control individuals. Methods: In this cross-sectional study, 164 participants (mean age 57 years ± 8 years; 134 males) with low to intermediate cardiovascular risk were recruited from the ongoing prospective Canadian HIV and Aging Cohort Study (CHACS). Among the 164 recruited participants, a total of 154 individuals (mean age, 56.5 years ± 7.55 years; 83 PLWH, 54%; 137 males; 88%) were evaluated. Ten participants were excluded due to unavailable coronary plaque data. The mean time interval between coronary CT and carotid ultrasound per participant was 7.69 ± 20.1 months. Using ultrasound, cumulated axial strain, cumulated shear strain, cumulated axial translation, cumulated lateral translation, and IMT of the common and internal carotid arteries were measured. Participants also underwent cardiac CT for coronary plaque assessment. Univariate and multivariate Poisson regression analyses with robust variance were performed to identify independent associations of cardiovascular risk factors, IMT, and elastography parameters with coronary plaque presence. Receiver operating characteristic (ROC) curve analysis and the area under the curve (AUC) were used to compare different prediction models for coronary plaque presence. Results: The study included 83 PLWH and 71 controls (N=154). The median 10-year Framingham risk score was 12% [IQR, 8 - 16] in PLWH and 9% [IQR, 7 - 15] in controls (p = 0.045). In the PLWH group, coronary plaques were observed in 55 participants (61.1%) compared to 42 (56.8%) in the non-HIV control group (p = .46). Carotid IMT and all elastography features for both the internal and common carotid arteries were similar between PLWH and healthy volunteers. 4 After adjusting for cardiovascular risk using multivariate Poisson regression, smoking exposure was significantly associated with coronary plaque presence on CT (prevalence ratio 1.10, 95% CI 1.04 – 1.13, p < 0.001). No significant associations were found with other coronary artery disease risk factors or HIV status in multivariate analysis. Carotid elastography parameters and carotid intima-media thickness were not associated with coronary atherosclerosis after adjustment. AUC analyses did not reveal any significant differences in predictive accuracy between models when adding either elastography parameters, IMT, or both elastography parameters and IMT results to the cardiovascular risk factor model, with AUC ranging from 0.647 to 0.681 in all models. Conclusion: In our study, models incorporating carotid elastography and IMT did not enhance the prediction of coronary plaque presence in PLWH or controls, compared to models including only traditional cardiovascular risk factors. Key words: HIV, computed tomography, angiography, us elastography, atherosclerosisObjectif: Évaluer l’association des caractéristiques biomécaniques des parois carotidiennes et de l'épaisseur intima-média (EIM) « Intima-media Thickness » (IMT) carotidienne, telles qu'évaluées par échographie, lorsque celles-ci sont incluses dans les modèles de prédiction de la charge de plaque coronarienne évaluée par tomodensitométrie (CT) chez les personnes vivant avec le VIH (PVVIH) et les personnes contrôles séronégatives. Méthodes : Dans notre étude transversale, 164 participants (âge moyen 57 ans ± 8 ans ; 134 hommes) présentant un risque cardiovasculaire faible/intermédiaire ont été recrutés, provenant tous de l’étude prospective Cohorte canadienne VIH et vieillissement (CHACS, Canadian HIV an Aging Cohort Study). Parmi les 164 participants recrutés, un total de 154 participants (âge moyen, 56.5 ans ±7.55 ans; 83 PPLWH, 54 %; 137 hommes; 88%) ont été évalués. Dix participants ont été exclus en raison de données de plaques non disponibles. L’intervalle de temps moyen entre le CT et l’élastographie carotidienne était de 7.7 ± 20.1 mois Avec l’imagerie par ultrasons, nous avons mesuré la déformation axiale cumulée, la déformation de cisaillement cumulée, la translation axiale cumulée, la translation latérale cumulée et l'IMT des artères carotides commune et interne. Les participants ont également subi une tomodensitométrie cardiaque pour l'évaluation de la plaque coronarienne. Des analyses de régression de Poisson univariées et multivariées avec une variance robuste ont été réalisées pour identifier comment les facteurs de risque cardiovasculaire, les paramètres IMT et élastographie sont indépendamment associés à la présence de plaque coronarienne. La fonction d’efficacité du récepteur (« caractéristique de fonctionnement du récepteur ») (ROC, receiver operating characteristic) et l'analyse de l'aire sous la courbe (AUC, area under the curve) ont également été utilisées pour comparer différents modèles de prédiction de la présence de plaque coronarienne. Résultats: Il y avait 83 PVVIH et 71 contrôles (N=154). Le score médian de risque de Framingham sur 10 ans était de 12% [IQR, 8 - 16] chez les PLWH and de 9% [IQR,7 -15] chez les témoins (p = 0.045). Dans le groupe PVVIH, des plaques coronaires étaient présentes chez 55 participants (61,1 %) contre 42 (56,8 %) dans le groupe contrôle non VIH (p = 0,46). 2 Après ajustement pour les facteurs de risque cardiovasculaire, on note que le tabagisme est associé à la présence de plaque coronarienne (ratio de prévalence 1.10, 95% CI 1.04 – 1.13, p < 0.001). Aucune autre association significative n’a été démontré avec d’autres facteurs de risque cardiovasculaire, ou avec le statut VIH, dans les analyses multivariées. L’analyse multivariée démontre que l'ajout des données d’IMT ou d’élastographie n'augmente pas la précision des modèles, au-delà du modèle n’incluant que les facteurs de risque traditionnels. Les analyses des courbes ROC et AUC n'ont montré aucune différence significative dans la précision prédictive entre les modèles qui incluent les paramètres d'élastographie, d'IMT et les facteurs de risque cardiovasculaire, versus les modèles qui n’incluent que les facteurs de risque cardiovasculaire, avec des AUC allant de 0,65 à 0,68. Conclusion: Dans notre étude, les modèles incluant l'élastographie carotidienne et l'IMT n'ont pas montré d’augmentation de la prédiction de la présence de plaque coronarienne chez les PVVIH ou les contrôles, en comparaison aux modèles incluant uniquement les facteurs de risque traditionnels

Functional changes of the vasculature in HIV/AIDS patients on Haart and Haart Naïve HIV participants

The present study sought to explore the functional changes that occur in the vasculature of HIV positive participants of African origin in Mthatha district of South africa which might lead to increased risk in their cardiovascular system. Available literature shows that arterial stiffness plays an important role in cardiovascular events such as stroke, vasculitis and myocardial infarction. Measurement of (aortic pulse wave velocity; PWV) provides some of the strongest evidence concerning the prognostic significance of large artery stiffening. This study was aimed at investigating the relationship between anthropometry, age, E-Selectin level, cytokine levels, haemodynamic variables, blood counts and blood lipid profile with pulse wave velocity. Some traditional cardiovascular risk factors such as alcohol, and smoking were also taken into account. This was a cross-sectional study comprising of 169 participants (62 males and 107 females). 63 were HIV negative (group A), 54 HIV positive on treatment (group B), and 52 were HIV positive not on treatment (group C). Pulse wave velocity (PWV) was assessed using the Sphygmocor Vx. Statistically, ANOVA was used for variables with normal distribution and non parametric tests were used for variables with skewed distribution. Notable significant differences were seen in the means of the following variables across all the 3 groups. Conclusion: This study showed that HIV infected patients with or without antiretroviral therapy have increase arterial stiffness which is associated with an increased cardiovascular risk. The sphygmocor is an accurate, non invassive and useful tool in the evaluation of arterial stiffness and its use in clinical practice should be encouraged. PWV and the augmentation index (AIx) are the two major non- iv invasive methods of assessing arterial stiffness. Life style modification should be incorporated into the management of HIV patients so as the continuous monitoring of their haematological and lipid profile

Ankle-brachial index in HIV infection

Prognosis for patients with the human immunodeficiency virus (HIV) has improved with the introduction of highly active antiretroviral therapy (HAART). Evidence over recent years suggests that the incidence of cardiovascular disease is increasing in HIV patients. The ankle-brachial index (ABI) is a cheap and easy test that has been validated in the general population. Abnormal ABI values are associated with increased cardiovascular mortality. To date, six series of ABI values in persons with HIV have been published, but none was a prospective study. No agreement exists concerning the risk factors for an abnormal ABI, though its prevalence is clearly higher in these patients than in the general population. Whether this higher prevalence of an abnormal ABI is associated with a higher incidence of vascular events remains to be determined

A Review of Arterial Stiffness and HIV Infection in Adult Africans

Aim: To review the impact of the human immunodeficiency virus and antiretroviral therapy on the vasculature. Objectives: This review seeks to identify the burden which the human immunodeficiency virus and antiretroviral therapy have on the vasculature. Method: Medline/PubMed and Google scholar were searched. There were over 100 publications reviewed. Some people who worked in similar fields were also contacted. The present review summarized current understanding of Human immunodeficiency virus, antiretroviral therapy and effect on the vasculature such as arterial stiffness. Atherosclerosis, endothelial dysfunction, the strengths and weaknesses of current testing strategies, and their potential applications in clinical research and patient care. The association of inflammatory biomarkers, blood pressure and ageing with arterial stiffness were also reviewed. Conclusion: Available literature shows that HIV and antiretroviral agents have a great impact on the vasculature and its progression

Circulating microparticles are increased amongst people presenting with HIV and advanced immune suppression in Malawi and correlate closely with arterial stiffness: a nested case control study

Background: We aimed to investigate whether circulating microparticle (CMPs) subsets were raised amongst people presenting with human immunodeficiency virus (HIV) and advanced immune suppression in Malawi, and whether they associated with arterial stiffness. Methods: Antiretroviral therapy (ART)-naïve adults with a new HIV diagnosis and CD4 <100 cells/µL had microparticle characterisation and carotid femoral Pulse Wave Velocity (cfPWV) at 2 weeks post ART initiation. HIV uninfected controls were matched on age, systolic blood pressure (BP) and diastolic BP in a 1:1 ratio.  Circulating microparticles were identified from platelet poor plasma and stained for endothelial, leucocyte, monocyte and platelet markers. Results: The median (IQ) total CMP count for 71 participants was 1 log higher in HIV compared to those without (p<0.0001) and was associated with arterial stiffness (spearman rho 0.47, p<0.001). In adjusted analysis, every log increase in circulating particles showed a 20% increase in cfPWV (95% confidence interval [CI] 4 – 40%, p=0.02). In terms of subsets, endothelial and platelet derived microparticles were most strongly associated with HIV. Endothelial derived E-selectin+ CMPs were 1.3log-fold higher and platelet derived CD42a+ CMPs were 1.4log-fold higher (both p<0.0001). Endothelial and platelet derived CMPs also correlated most closely with arterial stiffness (spearman rho: E-selectin+ 0.57 and CD42a 0.56, both p<0.0001). Conclusions: Circulating microparticles associate strongly with arterial stiffness among people living with HIV in Malawi. Endothelial damage and platelet microparticles are the predominant cell origin types and future translational studies could consider prioritising these pathways

Cardiovascular risks associated with protease inhibitors for the treatment of HIV

Introduction: Cumulative use of some first-generation protease inhibitors has been associated with higher rates of dyslipidemia and increased risk of cardiovascular disease. The protease inhibitors most commonly in use are atazanavir and darunavir, which have fewer detrimental lipid effects and greater tolerability. This paper aims to review the evidence of a potential association of these contemporary protease inhibitors with the risk of ischemic CVD and atherosclerotic markers.Areas covered: We searched for publications of randomized trials and observational studies on PubMed from 1st January 2000 onwards, using search terms including: protease inhibitors; darunavir; atazanavir; cardiovascular disease; cardiovascular events; dyslipidemia; mortality; carotid intima media thickness; arterial elasticity; arterial stiffness and drug discontinuation. Ongoing studies registered on clinicaltrials.gov as well as conference abstracts from major HIV conferences from 2015-2020 were also searched.Expert opinion: Atazanavir and darunavir are no longer part of first-line HIV treatment, but continue to be recommended as alternative first line, second- and third-line regimens, as part of two drug regimens, and darunavir is used as salvage therapy. Although these drugs will likely remain in use globally for several years to come, baseline CVD risk should be considered when considering their use, especially as the population with HIV ages

A Review of Arterial Stiffness and HIV Infection in Adult Africans

Aim: To review the impact of the human immunodeficiency virus and antiretroviral therapy on the vasculature. Objectives: This review seeks to identify the burden which the human immunodeficiency virus and antiretroviral therapy have on the vasculature. Method: Medline/PubMed and Google scholar were searched. There were over 100 publications reviewed. Some people who worked in similar fields were also contacted. The present review summarized current understanding of Human immunodeficiency virus, antiretroviral therapy and effect on the vasculature such as arterial stiffness. Atherosclerosis, endothelial dysfunction, the strengths and weaknesses of current testing strategies, and their potential applications in clinical research and patient care. The association of inflammatory biomarkers, blood pressure and ageing with arterial stiffness were also reviewed. Conclusion: Available literature shows that HIV and antiretroviral agents have a great impact on the vasculature and its progression

Mechanisms of HIV-Nef Induced Endothelial Cell Stress: Implications of HIV-Nef Protein Persistence in Aviremic HIV Patients

Indiana University-Purdue University Indianapolis (IUPUI)HIV-associated cardio-pulmonary vascular pathologies such as coronary artery disease, pulmonary hypertension and emphysema remain a major issue in the HIVinfected population even in the era of antiretroviral therapy (ART). The continued production of HIV encoded pro-apoptotic protein, such as Nef in latently HIV-infected cells is a possible mechanism for vascular dysfunction underlying these diseases. HIVNef persists in two compartments in these patients: (i) extracellular vesicles (EV) of plasma and bronchoalveolar lavage (BAL) fluid and (ii) PBMC and BAL derived cells. Here I demonstrate that the presence of HIV-Nef protein in cells and EV is capable of stressing endothelial cells by inducing ROS production leading to endothelial cell apoptosis. HIV-Nef protein hijacks host cell signaling by interacting with small GTP binding protein Rac1 which activates PAK2 to promote the release of pro-apoptotic cargo containing EV and surface expression of pro-apoptotic protein Endothelial Monocyte Activating Polypeptide II (EMAPII). Using this mechanism, Nef protein robustly induces apoptosis in Human Coronary Artery Endothelial Cells and Human Lung microvascular endothelial cells. Endothelial specific expression of HIV-Nef protein in transgenic mice was sufficient to induce vascular pathologies as evidenced by impaired endothelium mediated vasodilation of the aorta and vascular remodeling and emphysema like alveolar rarefaction in the lung. Furthermore, EV isolated from HIV patients on ART was capable of inducing endothelial apoptosis in a Nef dependent fashion. Of therapeutic interest, EMAPII neutralizing antibodies to block EMAPII mediated apoptosis and statin treatment to ameliorate Nef induced Rac1 signaling was capable of blocking Nef induced endothelial stress in both in vivo and in vitro. In conclusion, HIV-Nef protein uses a Rac1-Pak2 signaling axis to promote its dissemination in EV, which in turn induces endothelial cell stress after its uptake