RESUMO
OBJECTIVES: Blood-brain barrier impairment is frequent in people living with human immunodeficiency virus (PLWHIV), affecting the penetration of target cells and antiretrovirals into the central nervous system, through transporters (e.g. ABCB1), leading to neuroinflammation. This study aimed to identify variants of genes encoding transporters able to predict neuroinflammation biomarker levels. METHODS: Cerebrospinal fluid (CSF) and plasma samples were obtained from PLWHIV. The CSF biomarkers were quantified by commercial assays. Genetic variants were evaluated through real-time polymerase chain reaction (PCR). RESULTS: A total of 107 PLWHIV (163 samples) were included in the study: 79% were male, median age was 48.5 years, CD4% was 25%, and HIV-associated neurocognitive disorder (HAND) was observed in 17.8%. The ABCB1 2677G>T genetic variant showed a different allelic distribution according to the clinical group (P = 0.026). In linear regression analyses, HIV-related central nervous system disorders, ABCG2 1194+928CC genotype, log viral load, CSF-to-serum albumin ratio, ß-1,42 levels, and CSF proteins were retained in the final model as factors independently associated with CSF neopterin levels; CSF proteins and integrase inhibitor use were associated with CSF tau level in the multivariate model. Phospho-tau regression analysis reported the ABCB1 2677GT/TT genotype and CSF proteins as predictors in the final model; sex, protease inhibitors, neopterin, and ABCB1 2677 GT/ TT genotype were predictors in the multivariate regression for ß-1,42. CONCLUSIONS: For the first time, pharmacogenetic and clinical features were found to be predictors of neuro-inflammation biomarkers.
Assuntos
Subfamília B de Transportador de Cassetes de Ligação de ATP , Biomarcadores , Infecções por HIV , Polimorfismo de Nucleotídeo Único , Humanos , Masculino , Feminino , Pessoa de Meia-Idade , Biomarcadores/líquido cefalorraquidiano , Biomarcadores/sangue , Infecções por HIV/complicações , Adulto , Subfamília B de Transportador de Cassetes de Ligação de ATP/genética , Barreira Hematoencefálica , Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/genética , Inflamação/líquido cefalorraquidiano , Carga Viral , Genótipo , Complexo AIDS Demência/líquido cefalorraquidiano , Complexo AIDS Demência/patologia , Complexo AIDS Demência/genética , Proteínas de NeoplasiasRESUMO
Globally, 43 million people are living with HIV, 90% in developing countries. Increasing life expectancy with combination antiretroviral therapy (cART) results in chronic complications, including HIV-associated neurocognitive disorders (HAND) and eye diseases. HAND screening is currently challenging. Our aim was to evaluate clinical utility of retinopathy as a screening measure of HAND in older cART-treated individuals in Tanzania and feasibility of smartphone-based retinal screening in this low-resource setting. A cross-sectional systematic sample aged ≥ 50-years attending routine HIV follow-up in Tanzania were comprehensively assessed for HAND by American Academy of Neurology criteria and received ophthalmic assessment including smartphone-based retinal imaging. HAND and ophthalmic assessments were independent and blinded. Diagnostic accuracy was evaluated by AUROC curves. Of 129 individuals assessed, 69.8% were visually impaired. Thirteen had retinopathy. HAND prevalence was 66.7%. Retinopathy was significantly associated with HAND but HIV-disease factors (CD4, viral load) were not. Diagnostic accuracy of retinopathy for HAND was poor (AUROC 0.545-0.617) but specificity and positive predictive value were high. We conclude that ocular pathology and HAND appear highly prevalent in this low-resource setting. Although retinal screening cannot be used alone identify HAND, prioritization of individuals with abnormal retinal screening is a potential strategy in low-resource settings.
Assuntos
Complexo AIDS Demência/diagnóstico por imagem , Fármacos Anti-HIV/uso terapêutico , Programas de Rastreamento/métodos , Retina/diagnóstico por imagem , Retinoscopia/métodos , Complexo AIDS Demência/patologia , Terapia Antirretroviral de Alta Atividade , Contagem de Linfócito CD4 , Estudos Transversais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Aplicativos Móveis , Valor Preditivo dos Testes , Curva ROC , Retina/efeitos dos fármacos , Retina/patologia , Tanzânia , Carga ViralRESUMO
People over the age of 50 are the fastest growing segment of the HIV-infected population in the USA. Although antiretroviral therapy has remarkable success controlling the systemic HIV infection, HIV-associated neurocognitive disorder (HAND) prevalence has increased or remained the same among this group, and cognitive deficits appear more severe in aged patients with HIV. The mechanisms of HAND in the aged population are not completely understood; a leading hypothesis is that aged individuals with HIV might be at higher risk of developing Alzheimer's disease (AD) or one of the AD-related dementias (ADRD). There are a number of mechanisms through which chronic HIV disease alone or in combination with antiretroviral therapy and other comorbidities (e.g., drug use, hepatitis C virus (HCV)) might be contributing to HAND in individuals over the age of 50 years, including (1) overlapping pathogenic mechanisms between HIV and aging (e.g., decreased proteostasis, DNA damage, chronic inflammation, epigenetics, vascular), which could lead to accelerated cellular aging and neurodegeneration and/or (2) by promoting pathways involved in AD/ADRD neuropathogenesis (e.g., triggering amyloid ß, Tau, or α-synuclein accumulation). In this manuscript, we will review some of the potential common mechanisms involved and evidence in favor and against a role of AD/ADRD in HAND.
Assuntos
Complexo AIDS Demência/etiologia , Complexo AIDS Demência/epidemiologia , Complexo AIDS Demência/patologia , Idade de Início , Idoso , Idoso de 80 Anos ou mais , Proteínas Amiloidogênicas/metabolismo , Fármacos Anti-HIV/uso terapêutico , Autofagia , Encéfalo/metabolismo , Encéfalo/patologia , Encéfalo/virologia , Comorbidade , Epigênese Genética , Feminino , HIV/isolamento & purificação , HIV/fisiologia , Humanos , Macrófagos/virologia , Masculino , Microglia/virologia , Pessoa de Meia-Idade , Células-Tronco Neurais/patologia , Transtornos Neurocognitivos/epidemiologia , Agregação Patológica de Proteínas , Proteostase , Tropismo Viral , Substância Branca/patologiaRESUMO
The marked increase in life expectancy for HIV-1 seropositive individuals, following the great success of combination antiretroviral therapy (cART), heralds an examination of the progression of HIV-1 associated neurocognitive disorders (HAND). However, since the seminal call for animal models of HIV-1/AIDS in 1988, there has been no extant in vivo animal model system available to provide a truly longitudinal study of HAND. Here, we demonstrate that the HIV-1 transgenic (Tg) rat, resembling HIV-1 seropositive individuals on lifelong cART, exhibits age-related, progressive neurocognitive impairments (NCI), including alterations in learning, sustained attention, flexibility, and inhibition; deficits commonly observed in HIV-1 seropositive individuals. Pyramidal neurons from layers II-III of the medial prefrontal cortex (mPFC) displayed profound synaptic dysfunction in HIV-1 Tg animals relative to controls; dysfunction that was characterized by alterations in dendritic branching complexity, synaptic connectivity, and dendritic spine morphology. NCI and synaptic dysfunction in pyramidal neurons from layers II-III of the mPFC independently identified the presence of the HIV-1 transgene with at least 78.5% accuracy. Thus, even in the absence of sensory or motor system deficits and comorbidities, HAND is a neurodegenerative disease characterized by age-related disease progression; impairments which may be due, at least partly, to synaptic dysfunction in the mPFC. Further, the progression of HAND with age in the HIV-1 Tg rat and associated synaptic dysfunction affords an instrumental model system for the development of therapeutics and functional cure strategies.
Assuntos
Complexo AIDS Demência/patologia , Doenças Neurodegenerativas/patologia , Células Piramidais/patologia , Complexo AIDS Demência/virologia , Animais , Modelos Animais de Doenças , Feminino , HIV-1 , Masculino , Doenças Neurodegenerativas/virologia , Córtex Pré-Frontal/citologia , Córtex Pré-Frontal/patologia , Córtex Pré-Frontal/virologia , Células Piramidais/virologia , Ratos , Ratos TransgênicosRESUMO
Internal RNA modifications have been known for decades, however their roles in mRNA regulation have only recently started to be elucidated. Here we investigated the most abundant mRNA modification, N6-methyladenosine (m6A) in transcripts from the hippocampus of HIV transgenic (Tg) rats. The distribution of m6A peaks within HIV transcripts in HIV Tg rats largely corresponded to the ones observed for HIV transcripts in cell lines and T cells. Host transcripts were found to be differentially m6A methylated in HIV Tg rats. The functional roles of the differentially m6A methylated pathways in HIV Tg rats is consistent with a key role of RNA methylation in the regulation of the brain transcriptome in chronic HIV disease. In particular, host transcripts show significant differential m6A methylation of genes involved in several pathways related to neural function, suggestive of synaptodendritic injury and neurodegeneration, inflammation and immune response, as well as RNA processing and metabolism, such as splicing. Changes in m6A methylation were usually positively correlated with differential expression, while differential m6A methylation of pathways involved in RNA processing were more likely to be negatively correlated with gene expression changes. Thus, sets of differentially m6A methylated, functionally-related transcripts appear to be involved in coordinated transcriptional responses in the context of chronic HIV. Altogether, our results support that m6A methylation represents an additional layer of regulation of HIV and host gene expression in vivo that contributes significantly to the transcriptional effects of chronic HIV.
Assuntos
Complexo AIDS Demência/genética , HIV-1/genética , Hipocampo/patologia , RNA Mensageiro/genética , Transcriptoma/genética , Complexo AIDS Demência/imunologia , Complexo AIDS Demência/patologia , Complexo AIDS Demência/virologia , Adenosina/análogos & derivados , Adenosina/metabolismo , Animais , Modelos Animais de Doenças , Epigênese Genética/genética , Epigênese Genética/imunologia , Regulação da Expressão Gênica/imunologia , HIV-1/imunologia , Interações Hospedeiro-Patógeno/genética , Interações Hospedeiro-Patógeno/imunologia , Humanos , Masculino , Metilação , Splicing de RNA/genética , RNA Mensageiro/isolamento & purificação , RNA Viral/genética , Ratos , Ratos Sprague-Dawley , Ratos Transgênicos , Análise de Sequência de RNA , Transcriptoma/imunologiaRESUMO
OBJECTIVE: The relationship of cerebrospinal fluid (CSF) extracellular vesicles to neurocognitive impairment (NCI) in HIV-infected individuals is unclear. Here, we characterize CSF extracellular vesicles and their association with central nervous system (CNS) injury related biomarkers [neurofilament light (NFL), S100B, neopterin] and NCI in HIV-positive individuals on combination antiretroviral therapy (cART). DESIGN: A cross-sectional and longitudinal study of CSF samples from HIV-positive individuals on cART. METHODS: NFL, S100B and neopterin were measured by ELISA in 190 CSF samples from 112 individuals (67 HIV-positive and 45 HIV-negative). CSF extracellular vesicles were isolated and characterized by electron microscopy, nanoparticle tracking analysis, immunoblotting for exosome markers (CD9, CD63, CD81, FLOT-1) and ELISA for HLA-DR. RESULTS: HIV-positive individuals had median age 52 years, 67% with suppressed plasma viral load (< 50âcopies/ml), median CD4 nadir 66âcells/µl and CD4 cell count 313âcells/µl. CSF NFL, S100B and neopterin levels were higher in HIV-positive vs. HIV-negative individuals, and nonsuppressed vs. suppressed HIV-positive individuals. Although CSF NFL and S100B levels were higher in NCI vs. unimpaired HIV-positive individuals (Pâ<â0.05), only NFL was associated with NCI in adjusted models (Pâ<â0.05). CSF extracellular vesicles were increased in HIV-positive vs. HIV-negative individuals, and NCI vs. unimpaired HIV-positive individuals (Pâ<â0.05), and correlated positively with NFL (Pâ<â0.001). HLA-DR was enriched in CSF extracellular vesicles from HIV-positive individuals with NCI (Pâ<â0.05), suggesting that myeloid cells are a potential source of CSF extracellular vesicles during HIV infection. CONCLUSION: Increased CSF extracellular vesicles correlate with neuronal injury biomarker NFL in cART-treated HIV-positive individuals with neurocognitive impairment, suggesting potential applications as novel biomarkers of CNS injury.
Assuntos
Complexo AIDS Demência/patologia , Biomarcadores/líquido cefalorraquidiano , Líquido Cefalorraquidiano/química , Vesículas Extracelulares , Infecções por HIV/complicações , Infecções por HIV/patologia , Proteínas de Neurofilamentos/análise , Antirretrovirais/uso terapêutico , Terapia Antirretroviral de Alta Atividade , Sistema Nervoso Central , Estudos Transversais , Feminino , Infecções por HIV/tratamento farmacológico , Humanos , Estudos Longitudinais , Masculino , Pessoa de Meia-IdadeRESUMO
HIV-associated neurocognitive impairment (NCI) is a term established to capture a wide spectrum of HIV related neurocognitive deficits ranging in severity from asymptomatic to dementia. The genetic underpinnings of this complex phenotype are incompletely understood. Mitochondrial function has long been thought to play a role in neurodegeneration, along with iron metabolism and transport. In this work, we aimed to characterize the interplay of mitochondrial DNA (mtDNA) haplogroup and nuclear genetic associations to NCI phenotypes in the CHARTER cohort, encompassing 1025 individuals of European-descent, African-descent, or admixed Hispanic. We first employed a polygenic modeling approach to investigate the global effect of previous marginally associated nuclear SNPs, and to examine how the polygenic effect of these SNPs is influenced by mtDNA haplogroups. We see evidence of a significant interaction between nuclear SNPs en masse and mtDNA haplogroups within European-descent and African-descent individuals. Subsequently, we performed an analysis of each SNP by mtDNA haplogroup, and detected significant interactions between two nuclear SNPs (rs17160128 and rs12460243) and European haplogroups. These findings, which require validation in larger cohorts, indicate a potential new role for nuclear-mitochondrial DNA interactions in susceptibility to NCI and shed light onto the pathophysiology of this neurocognitive phenotype.
Assuntos
Complexo AIDS Demência/genética , Núcleo Celular/genética , Estudos de Associação Genética , Infecções por HIV/complicações , Mitocôndrias/genética , Polimorfismo de Nucleotídeo Único , Complexo AIDS Demência/patologia , Haplótipos , Humanos , Proteínas Mitocondriais/genética , Proteínas Nucleares/genética , Estudos Prospectivos , Grupos RaciaisRESUMO
OBJECTIVE: We investigated whether HIV brain latency was associated with brain injury in virally suppressed HIV infection. DESIGN: Observational cross-sectional and longitudinal study. METHODS: The study included 26 virally suppressed HIV-infected men (61.5% with HIV-associated neurocognitive disorder) who undertook cerebrospinal fluid (CSF) analyses at baseline. They also completed a proton magnetic resonance spectroscopy (1H MRS) and neuropsychological assessments at baseline and 18 months. To quantify whether there was residual brain HIV transcription, we measured CSF HIV-tat. As an HIV brain latency biomarker, we used concentrations of CSF BcL11b - a microglia transcription factor that inhibits HIV transcription. Concurrently, we assessed neuroinflammation with CSF neopterin, neuronal injury with CSF neurofilament light-chain (NFL), and in-vivo neurochemistry with 1H MRS of N-acetyl aspartate (NAA), choline (Cho), creatine, myo-inositol (MI), glutamine/glutamate (Glx) in the frontal white matter (FWM), posterior cingulate cortex (PCC), and caudate nucleus area. RESULTS: Baseline adjusted regression models for neopterin, NFL, and tat showed that a higher CSF BcL11b was consistently associated with lower FWM creatine (when adjusted for neopterin: ßâ=â-0.30, Pâ=â0.15; when adjusted for NFL: ßâ=â-0.47, Pâ=â0.04; and when adjusted for tat: ßâ=â-0.47, Pâ=â0.02). In longitudinal analyses, we found no time effect, but a consistent BcL11b altering effect on FWM creatine. The effect reached a significant moderate effect size range when corrected for CSF NFL (ßâ=â-0.36, Pâ=â0.02) and CSF tat (ßâ=â-0.34, Pâ=â0.02). CONCLUSIONS: Reduced frontal white matter total creatine may indicate subclinical HIV brain latency-related injury. H MRS may offer a noninvasive option to measure HIV brain latency.
Assuntos
Complexo AIDS Demência/patologia , Encéfalo/virologia , Infecções por HIV/complicações , Proteínas Repressoras/líquido cefalorraquidiano , Proteínas Supressoras de Tumor/líquido cefalorraquidiano , Complexo AIDS Demência/diagnóstico por imagem , Idoso , Biomarcadores/líquido cefalorraquidiano , Encéfalo/diagnóstico por imagem , Estudos Transversais , Humanos , Estudos Longitudinais , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Testes NeuropsicológicosRESUMO
HIV-associated neurocognitive disorders (HAND) are prevalent despite combined antiretroviral therapy, affecting nearly half of HIV-infected patients worldwide. During HIV infection of macrophages secretion of the lysosomal protein, cathepsin B, is increased. Secreted cathepsin B has been shown to induce neurotoxicity. Oxidative stress is increased in HIV-infected patients, while antioxidants are decreased in monocytes from patients with HIV-associated dementia (HAD). Dimethyl fumarate (DMF), an antioxidant, has been reported to decrease HIV replication and neurotoxicity mediated by HIV-infected macrophages. Thus, we hypothesized that DMF will decrease cathepsin B release from HIV-infected macrophages by preventing oxidative stress and enhancing lysosomal function. Monocyte-derived macrophages (MDM) were isolated from healthy donors, inoculated with HIV-1ADA, and treated with DMF following virus removal. After 12 days post-infection, HIV-1 p24 and total cathepsin B levels were measured from HIV-infected MDM supernatants using ELISA; intracellular reactive oxygen and nitrogen species (ROS/RNS) were measured from MDM lysates, and functional lysosomes were assessed using a pH-dependent lysosomal dye. Neurons were incubated with serum-free conditioned media from DMF-treated MDM and neurotoxicity was determined using TUNEL assay. Results indicate that DMF reduced HIV-1 replication and cathepsin B secretion from HIV-infected macrophages in a dose-dependent manner. Also, DMF decreased intracellular ROS/RNS levels, and prevented HIV-induced lysosomal dysfunction and neuronal apoptosis. In conclusion, the improvement in lysosomal function with DMF treatment may represent the possible mechanism to reduce HIV-1 replication and cathepsin B secretion. DMF represents a potential therapeutic strategy against HAND.
Assuntos
Antioxidantes/uso terapêutico , Catepsina B/metabolismo , Fumarato de Dimetilo/uso terapêutico , Infecções por HIV/prevenção & controle , Macrófagos/metabolismo , Complexo AIDS Demência/patologia , Apoptose/efeitos dos fármacos , Proteína do Núcleo p24 do HIV/metabolismo , Infecções por HIV/patologia , HIV-1/efeitos dos fármacos , Humanos , Marcação In Situ das Extremidades Cortadas , Macrófagos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Nitrogênio/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Replicação Viral/efeitos dos fármacosAssuntos
Complexo AIDS Demência/epidemiologia , Fármacos Anti-HIV/uso terapêutico , Suspensão da Respiração , Doenças Arteriais Cerebrais/diagnóstico , Testes Diagnósticos de Rotina/métodos , Infecções por HIV/complicações , Programas de Rastreamento/métodos , Complexo AIDS Demência/patologia , Adulto , Doenças Arteriais Cerebrais/complicações , Doenças Arteriais Cerebrais/patologia , China , Feminino , Infecções por HIV/tratamento farmacológico , Humanos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , População UrbanaRESUMO
The human immunodeficiency virus-1 (HIV-1) regulatory protein Tat plays an important role during HIV-1-associated neurocognitive disorders (HAND) by inducing neuronal autophagy. In this study, we used immunohistochemistry, immunofluorescence, western blot, qRT-PCR, and RNA interference to elucidate the involvement of Bcl-2-associated athanogene 3 (BAG3) in the pathogenesis of HIV-1 Tat-induced autophagy during HAND. We found that BAG3 expression is elevated in astrocytes in frontal cortex of macaques infected with simian immunodeficiency virus-human immunodeficiency chimeric virus (SHIV). In addition, in human primary glioblastoma cells (U87), HIV-1 Tat upregulated BAG3 in an NF-κB-dependent manner to induce autophagy. Importantly, suppression of BAG3 or inhibition of NF-κB activity reversed the HIV-1 Tat-induced autophagy. These results indicate that HIV-1 Tat induces autophagy by upregulating BAG3 via NF-κB signaling, which suggests BAG3 and NF-κB could potentially serve as novel targets for HAND therapies.
Assuntos
Complexo AIDS Demência/patologia , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Proteínas Reguladoras de Apoptose/metabolismo , Autofagia , HIV-1/metabolismo , NF-kappa B/metabolismo , Transdução de Sinais , Produtos do Gene tat do Vírus da Imunodeficiência Humana/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/genética , Animais , Proteínas Reguladoras de Apoptose/genética , Astrócitos/metabolismo , Astrócitos/virologia , Linhagem Celular Tumoral , Feminino , Macaca , Masculino , RNA Viral/metabolismo , Regulação para Cima/genéticaRESUMO
OBJECTIVE: To compare retinal layer thickness in HIV-infected subjects with (CI-HIV) and without (NCI-HIV) cognitive impairment, with a control population and to correlate this with the cognitive status of the patient and other clinical parameters. DESIGN: Single-center cross-sectional study. METHODS: Participants with controlled HIV infection aged between 40 and 70 years and sex-matched and age-matched controls were enrolled. Retinal nerve fiber layer (RNFL), ganglion cell layer (GCL) and inner plexiform layer (IPL) thickness were assessed using optical coherence tomography. These measurements in HIV patients were compared with those in controls. Age-related and sex-related changes were compared in both groups. Other variables studied in HIV patients included: duration of HIV infection, CD4 cell count nadir, antiretroviral therapy regimen and cognitive status using the Montreal Cognitive Assessment (MoCA) test. RESULTS: Sixty-nine individuals, 34 with and 35 without cognitive impairment, and 70 controls were enrolled. GCL was significantly thinner in CI-HIV patients compared with NCI-HIV patients and controls (Pâ=â0.01 and Pâ=â0.02, respectively). GCL and IPL thickness significantly decreased with age in patients with HIV (Pâ=â0.0003, Pâ=â0.02, respectively, for the entire cohort). This change was not seen in controls. MoCA test score significantly decreased with age in HIV patients and controls. GCL thickness positively correlated with cognitive function across the entire HIV cohort (Pâ=â0.02). CONCLUSION: GCL was thinner in HIV patients with cognitive impairment. GCL thickness correlated positively with cognitive function and negatively with age in HIV patients. GCL thickness may reflect accelerated cognitive aging in HIV.
Assuntos
Complexo AIDS Demência/patologia , Cognição , Retina/patologia , Adulto , Idoso , Animais , Estudos Transversais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Retina/diagnóstico por imagem , Tomografia de Coerência ÓpticaRESUMO
HIV-associated neurocognitive disorders (HANDs) share common symptoms with Alzheimer's disease (AD), which is characterized by amyloid-ß (Aß) plaques. Plaques are formed by aggregation of Aß oligomers, which may be the toxic species in AD pathogenesis, and oligomers are generated by cleavage of amyloid precursor protein (APP) by ß-site amyloid precursor protein cleaving enzyme 1 (BACE1). BACE1 inhibitors reverse neuronal loss and cognitive decline in animal models of AD. Although studies have also found evidence of altered APP processing in HIV+ patients, it is unknown whether increased BACE1 expression or Aß oligomer production is a common neuropathological feature of HAND. Moreover, it is unknown whether BACE1 or APP is involved in the excitotoxic, NMDAR-dependent component of HIV-associated neurotoxicity in vitro Herein, we hypothesize that HIV-associated neurotoxicity is mediated by NMDAR-dependent elevation of BACE1 and subsequent altered processing of APP. Supporting this, we observed elevated levels of BACE1 and Aß oligomers in CNS of male and female HIV+ patients. In a model of HIV-associated neurotoxicity in which rat neurons are treated with supernatants from HIV-infected human monocyte-derived macrophages, we observed NMDAR-dependent elevation of BACE1 protein. NMDA treatment also increased BACE1 and both pharmacological BACE1 inhibition and genetic loss of APP were partially neuroprotective. Moreover, in APP knock-out (APP-/-) mouse neurons, NMDA-induced toxicity was BACE1 independent, indicating that cytotoxicity of BACE1 is dependent upon APP cleavage. Our findings suggest that increased BACE1 and the resultant Aß oligomer production may contribute to HIV-associated neuropathogenesis and inhibition of BACE1 could have therapeutic potential in HANDs.SIGNIFICANCE STATEMENT HIV-associated neurocognitive disorders (HANDs) represent a range of cognitive impairments affecting â¼50% of HIV+ individuals. The specific causes of HAND are unknown, but evidence suggests that HIV-infected macrophage infiltration into the brain may cause neuronal damage. Herein, we show that neurons treated with conditioned media from HIV-infected macrophages have increased expression of ß-site amyloid precursor protein cleaving enzyme 1 (BACE1), a protein implicated in Alzheimer's disease pathogenesis. Moreover, inhibition of BACE1 prevented neuronal loss after conditioned media exposure, but had no effect on HIV-associated neurotoxicity in neurons lacking its cleavage target amyloid precursor protein. We also observed increased BACE1 expression in HIV+ patient brain tissue, confirming the potential relevance of BACE1 as a therapeutic target in HANDs.
Assuntos
Complexo AIDS Demência/genética , Complexo AIDS Demência/patologia , Secretases da Proteína Precursora do Amiloide/genética , Precursor de Proteína beta-Amiloide/genética , Ácido Aspártico Endopeptidases/genética , Infecções por HIV/patologia , Neurônios/patologia , Adulto , Secretases da Proteína Precursora do Amiloide/antagonistas & inibidores , Animais , Ácido Aspártico Endopeptidases/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Agonistas de Aminoácidos Excitatórios/toxicidade , Feminino , Hipocampo/metabolismo , Humanos , Macrófagos/química , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Pessoa de Meia-Idade , N-Metilaspartato/toxicidade , Ratos , Ratos Sprague-Dawley , Receptores de N-Metil-D-Aspartato/genéticaRESUMO
BACKGROUND: Human immunodeficiency virus type 1 (HIV-1)-associated neurocognitive disorder (HAND) is a common outcome of a majority of HIV-1-infected subjects and is associated with synaptodendritic damage. Neurogranin (Ng), a postsynaptic protein, and calmodulin (CaM) are two important players of synaptic integrity/functions. The biological role of Ng in the context of HAND is unknown. METHODS: We compared the expression of Ng in frontal cortex (FC) tissues from control and HIV-1-positive subjects with and without HAND by immunohistochemistry, western blot, and qRT-PCR. The interaction between Ng and CaM was analyzed by co-immunoprecipitation. Ng, microtubule-associated protein 2 (MAP2), CaM, CaM-dependent protein kinase II (CaMKII), CREB, synaptophysin (Syp), and synapsin I (Syn I) expressions were evaluated by western blot using FC tissue lysates and differentiated SH-SY5Y (dSH-SY5Y) cells. Identification of inflammatory factors related to Ng loss was accomplished by exposing dSH-SY5Y cells to HIV-1 and mock-infected monocyte-derived macrophage (MDM) supernatants or HIV-1 NLYU2 pseudotyped with VSV-G-Env. Levels of interleukin (IL)-1ß, IL-8, tumor necrosis factor (TNF)-α, monocyte chemoattractant protein (MCP)-1, MCP-2, and CXCL5 in MDM supernatants were measured by ELISA. Association of IL-1ß and IL-8 to Ng expression in context of HIV-1 infection was evaluated in the presence or absence of neutralizing antibodies against these cytokines. RESULTS: Expression level of Ng was reduced significantly in FC of HAND-positive (HAND+) patients compared to uninfected individuals. Although no difference was found in CaM expression, interaction between Ng and CaM was reduced in HAND+ patients, which was associated with decreased level of CaMKII, a downstream signaling molecule of CaM pathway. This in turn resulted in reduction of synaptic markers, Syp and Syn I. HIV-1 infection directly had no considerable effect on dysregulation of Ng expression in dSH-SY5Y cells, whereas high amount of pro-inflammatory IL-1ß and IL-8 in HIV-1-infected MDM supernatants was associated with significant reduction in Ng expression. CONCLUSIONS: Synaptic damage in HAND+ patients could be a result of abrogation of Ng through HIV-1-induced inflammation that dysregulates Ng-CaM interaction and downstream signaling cascades associated with synaptodendritic functions. This is the first study evaluating the potential role of Ng in the context of HIV-1 neuropathogenesis.
Assuntos
Complexo AIDS Demência/metabolismo , Dendritos/metabolismo , Lobo Frontal/metabolismo , HIV-1 , Neurogranina/biossíntese , Sinapses/metabolismo , Complexo AIDS Demência/patologia , Adulto , Idoso , Dendritos/patologia , Feminino , Lobo Frontal/patologia , Humanos , Inflamação/metabolismo , Inflamação/patologia , Masculino , Pessoa de Meia-Idade , Sinapses/patologiaRESUMO
Despite achieving human immunodeficiency virus type 1 (HIV-1) RNA suppression below levels of detection and, for most, improved CD4+ T-cell counts, those aging with HIV experience excess low-level inflammation, hypercoagulability, and immune dysfunction (chronic inflammation), compared with demographically and behaviorally similar uninfected individuals. A host of biomarkers that are linked to chronic inflammation are also associated with HIV-associated non-AIDS-defining events, including cardiovascular disease, many forms of cancer, liver disease, renal disease, neurocognitive decline, and osteoporosis. Furthermore, chronic HIV infection may interact with long-term treatment toxicity and weight gain after ART initiation. These observations suggest that future biomarker-guided discovery and treatment may require attention to multiple biomarkers and, possibly, weighted indices. We are clinical trialists, epidemiologists, pragmatic trialists, and translational scientists. Together, we offer an operational definition of a biomarker and consider how biomarkers might facilitate progress along the translational pathway from therapeutic discovery to intervention trials and clinical management among people aging with or without HIV infection.
Assuntos
Antirretrovirais/uso terapêutico , Biomarcadores/análise , Pesquisa Biomédica/tendências , Descoberta de Drogas/tendências , Infecções por HIV/complicações , Infecções por HIV/tratamento farmacológico , Resposta Viral Sustentada , Complexo AIDS Demência/tratamento farmacológico , Complexo AIDS Demência/patologia , Nefropatia Associada a AIDS/tratamento farmacológico , Nefropatia Associada a AIDS/patologia , Doenças Cardiovasculares/tratamento farmacológico , Doenças Cardiovasculares/patologia , Síndrome de Lipodistrofia Associada ao HIV/tratamento farmacológico , Síndrome de Lipodistrofia Associada ao HIV/patologia , HumanosRESUMO
The human immunodeficiency virus (HIV) envelope protein gp120 promotes axonal damage and neurite pruning, similar to that observed in HIV-positive subjects with neurocognitive disorders. Thus, gp120 has been used to examine molecular and cellular pathways underlying HIV-mediated neuronal dysfunction. Gp120 binds to tubulin beta III, a component of neuronal microtubules. Microtubule function, which modulates the homeostasis of neurons, is regulated by polymerization and post-translational modifications. Based on these considerations, we tested the hypothesis that gp120 induces dynamic instability of neuronal microtubules. We first observed that gp120 prevents the normal polymerization of tubulin in vitro. We then tested whether gp120 alters the post-translational modifications in tubulin by examining the ability of gp120 to change the levels of acetylated tubulin in primary rat neuronal cultures. Gp120 elicited a time-dependent decrease in tubulin acetylation that was reversed by Helix-A peptide, a compound that competitively displaces the binding of gp120 to neuronal microtubules. To determine whether post-translational modifications in tubulin also occur in vivo, we measured acetylated tubulin in the cerebral cortex of HIV transgenic rats (HIV-tg). We observed a decrease in tubulin acetylation in 5- and 9-month-old HIV-tg rats when compared to age-matched wild type. Neither changes in microglia morphology nor alterations in mRNA levels for interleukin-1ß and tumor necrosis factor α were detected in 5-month-old animals. Our findings propose neuronal microtubule instability as a novel mechanism of HIV neurotoxicity, without evidence of enhanced inflammation.
Assuntos
Proteína gp120 do Envelope de HIV/farmacologia , Proteína gp120 do Envelope de HIV/toxicidade , Neurônios/metabolismo , Tubulina (Proteína)/metabolismo , Complexo AIDS Demência/patologia , Acetilação , Animais , Células Cultivadas , Humanos , Interleucina-1beta/biossíntese , Interleucina-1beta/genética , Masculino , Microtúbulos/efeitos dos fármacos , Processamento de Proteína Pós-Traducional , Ratos , Ratos Sprague-Dawley , Ratos Transgênicos , Fator de Necrose Tumoral alfa/biossíntese , Fator de Necrose Tumoral alfa/genéticaRESUMO
OBJECTIVE: Dickkopf-related protein 1 (DKK1) is a soluble antagonist of the Wningless (Wnt) pathway. It binds to and sequesters low-density lipoprotein receptor-related proteins 5/6 away from Wnts. Because the Wnt pathway regulates synaptic transmission and plasticity, we hypothesized that increased DKK1 would increase the risk for neurocognitive impairment (NCI) in HIV-positive (HIV) individuals. We evaluated, here, the relationship between plasma DKK1 and global NCI. METHODS: Plasma samples and data from 41 HIV to 42 HIV adults were obtained from the University of California, San Diego, California, USA. Concentrations of DKK1 and a comparator protein, monocyte chemoattractant protein-1 (MCP-1), were quantified in plasma by immunoassay. All study participants completed a standardized comprehensive neuropsychological test battery and their performance was summarized using the global deficit score method. RESULTS: A higher DKK1 level was associated with NCI among HIV participants (dâ=â0.63, Pâ=â0.05), particularly among the 26 participants whose plasma HIV RNA level was suppressed (dâ=â0.74, Pâ=â0.08). DKK1 level was not associated with NCI among HIV participants (Pâ=â0.98). was not associated with NCI in either group. In HIV adults with suppressed plasma HIV RNA, a receiver operator characteristic curve identified that a DKK1 level of at least 735âpg/ml had a positive predictive value of 83.3% for a diagnosis of NCI. This association did not weaken after accounting for the effect of AIDS, nadir CD4 T-cell count, addictive drug use, or demographic characteristics. CONCLUSION: DKK1 is a specific biomarker for NCI in HIV adults, implicating the Wnt pathway in HIV neuropathogenesis.
Assuntos
Complexo AIDS Demência/patologia , Biomarcadores/sangue , Infecções por HIV/complicações , Infecções por HIV/patologia , Peptídeos e Proteínas de Sinalização Intercelular/sangue , Plasma/química , Adolescente , Adulto , Idoso , California , Feminino , Humanos , Imunoensaio , Masculino , Pessoa de Meia-Idade , Testes Neuropsicológicos , Adulto JovemRESUMO
OBJECTIVE: Higher plasma soluble cluster of differentiation (CD)163 (sCD163), shed by monocytes and macrophages, correlates with neurocognitive impairment in HIV infection. We hypothesized that higher antemortem plasma or cerebrospinal fluid (CSF) sCD163 would be associated with greater postmortem neurodegeneration and/or microgliosis. DESIGN: Retrospective, postmortem observational study. METHODS: We measured sCD163 levels in antemortem plasma (nâ=â54) and CSF (nâ=â32) samples from 74 HIV-seropositive participants (median 5 months before death) who donated their brains to research at autopsy. Postmortem, we quantified markers of synaptodendritic damage (microtubule-associated protein 2, synaptophysin), microgliosis [human leukocyte antigen DR (HLA-DR), ionized calcium-binding adaptor molecule 1], astrocytosis (glial fibrillary acidic protein), and impaired protein clearance (ß-amyloid) in frontal cortex, hippocampus, putamen, and internal capsule. Multivariable least-squares regression was used to evaluate the association between plasma or CSF sCD163 and histological measures, correcting for multiple comparisons. RESULTS: Higher plasma sCD163 was associated with lower microtubule-associated protein 2 in frontal cortex [Bâ=â-0.23, 95% confidence interval (CI) -0.41 to -0.06, Pâ=â0.04], putamen (Bâ=â0.32, 95% CI -0.52 to -0.12, Pâ=â0.02), and hippocampus (Bâ=â-0.23, 95% CI -0.35 to -0.10, Pâ=â0.01), and with lower synaptophysin in hippocampus (Bâ=â-0.25, 95% CI -0.42 to -0.03, Pâ=â0.02) but not putamen or frontal cortex (Pâ>â0.05). Higher plasma sCD163 was associated with higher HLA-DR in putamen (Bâ=â0.17, 95% CI 0.08 to 0.26, Pâ=â0.008). CSF sCD163 was not associated with any histological measure (Pâ>â0.05). CONCLUSION: Higher plasma sCD163 in life is associated with greater synaptodendritic damage and microglial activation in cortical and subcortical brain regions.
Assuntos
Complexo AIDS Demência/patologia , Antígenos CD/sangue , Antígenos de Diferenciação Mielomonocítica/sangue , Encéfalo/patologia , Infecções por HIV/complicações , Infecções por HIV/patologia , Receptores de Superfície Celular/sangue , Adulto , Idoso , Antígenos CD/líquido cefalorraquidiano , Antígenos de Diferenciação Mielomonocítica/líquido cefalorraquidiano , Líquido Cefalorraquidiano/química , Feminino , HIV , Hipocampo , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Plasma/química , Estudos RetrospectivosRESUMO
BACKGROUND: HIV+ patients on highly active antiretroviral therapy (HAART) with suppressed viral loads have a low incidence of HIV-associated dementia, but increased prevalence of milder forms of HIV-associated neurocognitive disorders (HAND). These milder forms of HAND are often associated with minimal histological abnormalities, and their pathophysiology is unclear. Comorbidities, altered amyloid metabolism, accelerated brain aging, and activated interferon responses are suspected to play a role in HAND pathogenesis in HAART-treated persons. METHODS: To investigate associations between liver disease, accelerated brain aging, and HAND in HIV+ patients in the late HAART era (2002-2015), we studied liver and brain autopsy tissues from 53 older subjects evaluated at UCLA and BWH using histopathological stains, a sensitive fluorescent amyloid stain (AmyloGlo), and targeted gene expression profiling (NanoString). RESULTS: The majority of HIV+ subjects (median age 56) were on HAART (89.3%) with last pre-mortem plasma viral load <400 copies/mL (81.5%); 50% had CD4+ counts <200 cells/µL. Compared to HIV- controls (median age 65), HIV+ subjects had more cancer (p = 0.04), illicit drug use (p <0.00001), and HCV co-infection (p = 0.002), less cardiovascular disease (p = 0.03), and similar prevalence of cerebrovascular disease (~40%), hypertension, hyperlipidemia, and diabetes. Deep frontal white matter showed increased gliosis in HIV+ subjects vs. HIV- controls (p = 0.09), but no significant differences in myelin loss, blood vessel thickening, or inflammation. Liver showed more severe fibrosis/cirrhosis (p = 0.02) and less steatosis (p = 0.03) in HIV+ subjects, but no significant differences in inflammation, blood vessel thickness, or pigment deposition. There were no significant associations between liver and brain pathologies. AmyloGlo staining detected large amyloid deposits in only one HIV+ case (age 69 with Alzheimer's disease pathology) and two HIV- controls (ages 66 and 74). White matter from HIV+ cases vs. HIV- seronegative controls showed a trend (p = 0.06) towards increased interferon response gene expression (ISG15, MX1, IFIT1, IFIT2, and IFITM1). CONCLUSIONS: Gliosis and cerebrovascular disease, but not accelerated amyloid deposition, are common brain pathologies among older HIV+ patients in the late HAART era. Although HIV+ subjects had more cirrhosis, liver pathology was not associated with any consistent pattern of brain pathology. Cerebrovascular disease, interferon responses, and neuroinflammation are likely factors contributing to brain aging and HAND in older HIV+ patients on current HAART regimens.
Assuntos
Complexo AIDS Demência/metabolismo , Complexo AIDS Demência/patologia , Amiloide/metabolismo , Terapia Antirretroviral de Alta Atividade , Encéfalo/patologia , Interferons/imunologia , Cirrose Hepática/patologia , Complexo AIDS Demência/imunologia , Idoso , Encéfalo/metabolismo , Contagem de Linfócito CD4 , Transtornos Cerebrovasculares/metabolismo , Transtornos Cerebrovasculares/patologia , Feminino , Gliose/metabolismo , Gliose/patologia , Humanos , Interferons/metabolismo , Cirrose Hepática/metabolismo , Masculino , Pessoa de Meia-Idade , Transcriptoma , Carga ViralRESUMO
Astrocyte activation is a hallmark of HIV infection and aging in the CNS. In chronically infected HIV patients, prolonged activation of astrocytes has been linked to accelerated aging including but not limited to neurocognitive impairment and frailty. The current study addresses the role of HIV protein Tat in inducing a set of small noncoding microRNAs (miRNA) that play critical role in astrogliosis. In our efforts to link astrocyte activation as an indicator of aging, we assessed the brains of both wild type and HIV transgenic rats for the expression of glial fibrillary acidic protein (GFAP). As expected, in the WT animals we observed age-dependent increase in astrogliosis in the older animals compared to the younger group. Interestingly, compared to the young WT group, young HIV Tg rats exhibited higher levels of GFAP in this trend was also observed in the older HIV Tg rats compared to the older WT group. Based on the role of SIRT1 in aging and the regulation of SIRT1 by miRNAs-34a and -138, we next assessed the expression levels of these miRs in the brains of both the young an old WT and HIV Tg rats. While there were no significant differences in the young WT versus the HIV Tg rats, in the older HIV Tg rats there was a significant upregulation in the expression of miRs-34a & -138 in the brains. Furthermore, increased expression of miRs-34a & -138 in the older Tg rats, correlated with a concomitant decrease in their common anti-aging target protein SIRT1, in the brains of these animals. To delineate the mechanism of action we assessed the role of HIV-Tat (present in the Tg rats) in inducing miRs-34a & -138 in both the primary astrocytes and the astrocytoma cell line A172, thereby leading to posttranscriptional suppression of SIRT1 with a concomitant up regulation of NF-kB driven expression of GFAP.