While human immunodeficiency virus 1(HIV-1) infectionis controlled through continuous, life-long use of a combination of drugs targeting different steps of the virus cycle, HIV-1 is never completely eradicated from th...While human immunodeficiency virus 1(HIV-1) infectionis controlled through continuous, life-long use of a combination of drugs targeting different steps of the virus cycle, HIV-1 is never completely eradicated from the body. Despite decades of research there is still no effective vaccine to prevent HIV-1 infection. Therefore, the possibility of an RNA interference(RNAi)-based cure has become an increasingly explored approach. Endogenous gene expression is controlled at both, transcriptional and post-transcriptional levels by noncoding RNAs, which act through diverse molecular mechanisms including RNAi. RNAi has the potential to control the turning on/off of specific genes through transcriptional gene silencing(TGS), as well as finetuning their expression through post-transcriptional gene silencing(PTGS). In this review we will describe in detail the canonical RNAi pathways for PTGS and TGS, the relationship of TGS with other silencing mechanisms and will discuss a variety of approaches developed to suppress HIV-1 via manipulation of RNAi. We will briefly compare RNAi strategies against other approaches developed to target the virus, highlighting their potential to overcome the major obstacle to finding a cure, which is the specific targeting of the HIV-1 reservoir within latently infected cells.展开更多
The human immunodeficiency virus （HIV） lentiviral vector is an ideal vector for gene therapy. In the present study, the wild-type HIV-1 genome was segregated into four plasmids, and an optimized novel HIV-1 lentivir...The human immunodeficiency virus （HIV） lentiviral vector is an ideal vector for gene therapy. In the present study, the wild-type HIV-1 genome was segregated into four plasmids, and an optimized novel HIV-1 lentiviral vector containing green fluorescent protein and vesicular stomatitis virus G pseudo-capsule was constructed. The plasmids were pHR-CMV-EGFP, pCMVΔ8.9, pRSV-Rev, pCMV-VSV-G. The four plasmid system was co-transfected into 293T cells, and green fluorescent protein expression was observed. The present study obtained lentiviral particles by high-speed centrifugation, and the lentiviral particle titer was 4 × 108 TU/mL after centrifugation. Thus, an optimized novel HIV-1 lentiviral vector was successfully constructed.展开更多
Eradication of human immunodeficiency virus(HIV) in infected individuals is currently not possible because of the presence of the persistent cellular reservoir of latent infection. The identification of HIV latency bi...Eradication of human immunodeficiency virus(HIV) in infected individuals is currently not possible because of the presence of the persistent cellular reservoir of latent infection. The identification of HIV latency biomarkers and a better understanding of the molecular mechanisms contributing to regulation of HIV expression might provide essential tools to eliminate these latently infected cells. This review aims at summarizing gene expression profiling and systems biology applications to studies of HIV latency and eradication. Studies comparing gene expression in latently infected and uninfected cells identify candidate latency biomarkers and novel mechanisms of latency control. Studies that profiled gene expression changes induced by existing latency reversing agents(LRAs) highlight uniting themes driving HIV reactivation and novel mechanisms that contribute to regulation of HIV expression by different LRAs. Among the reviewed gene expression studies, the common approaches included identification of differentially expressed genes and gene functional category assessment. Integration of transcriptomic data with other biological data types is presently scarce, and the field would benefit from increased adoption of these methods in future studies. In addition, designing prospective studies that use the same methods of data acquisition and statistical analyses will facilitate a more reliableidentification of latency biomarkers using different model systems and the comparison of the effects of different LRAs on host factors with a role in HIV reactivation. The results from such studies would have the potential to significantly impact the process by which candidate drugs are selected and combined for future evaluations and advancement to clinical trials.展开更多
Background: It is controversial whether the apolipoprotein E epsilon 4 allele (APOE ε4) is a risk gene for human immunodeficiency virus (HIV)-related neurocognitive impairment. This meta-analysis aimed to summarize e...Background: It is controversial whether the apolipoprotein E epsilon 4 allele (APOE ε4) is a risk gene for human immunodeficiency virus (HIV)-related neurocognitive impairment. This meta-analysis aimed to summarize evidence of the associations betweenAPOE ε4 and cognitive impairment in people living with HIV (PLWH).Methods: Our study conducted a systematic literature search of PubMed, Web of Science, Embase, Google Scholar, and ProQuest for studies published before April 11, 2022 that evaluated associations betweenAPOE ε4 and cognitive impairment in adult PLWH (aged ≥18 years). We calculated pooled odds ratios (ORs) of global cognitive impairment and 95% confidence intervals (CIs) and standardized mean differences (SMDs) for specific cognitive domains betweenAPOE ε4 carriers and non-carriers. Subgroup meta-analyses were used to evaluate the result profiles across different categorical variables.Results: Twenty studies met the inclusion criteria, including 19 that evaluated global cognitive impairment.APOE ε4 was significantly associated with global cognitive impairment in PLWH (OR = 1.36, 95% CI = [1.05, 1.78], number of estimates [k] = 19,P = 0.02, random effects). Subgroup meta-analysis based percentage of females showed evident intergroup differences in global cognitive performance between ε4 carriers and non-carriers (P = 0.015).APOE ε4 carriers had lower cognitive test scores than non-carriers in all seven cognitive domains, including fluency (SMD = -0.51, 95% CI = [-0.76, -0.25],P < 0.001,k = 4,I^(2)= 0%), learning (SMD = -0.52, 95% CI = [-0.75, -0.28],P < 0.001,k = 5,I^(2) = 0%), executive function (SMD = -0.41, 95% CI= [-0.59, -0.23],P < 0.001,k= 8,I^(2)= 0%), memory (SMD=-0.41, 95% CI= [-0.61, -0.20],P < 0.001,k= 10,I^(2)= 36%), attention/working memory (SMD=-0.34, 95% CI= [-0.54, -0.14],P= 0.001,k= 6,I^(2)= 0%), speed of information processing (SMD = -0.34, 95% CI = [-0.53, -0.16],P < 0.001,k = 8,I^(2) = 0%), and motor function (SMD = -0.19, 95% CI = [-0.38, -0.01],P = 0.04,k = 7,I^(2) = 0%).Conclusions: Our meta-analysis provides significant evidence thatAPOE ε4 is a risk genotype for HIV-associated cognitive impairment, especially in cognitive domains of fluency, learning, executive function, and memory. Moreover, the impairment is sex specific.Meta analysis registration: PROSPERO, CRD 42021257775.展开更多
AIM: To explore the possibility of the replacement of the gag gene between human immunodeficiency virus and bovine immunodeficiency virus, to achieve chimeric virions,and thereby gain a new kind of AIDS vaccine based ...AIM: To explore the possibility of the replacement of the gag gene between human immunodeficiency virus and bovine immunodeficiency virus, to achieve chimeric virions,and thereby gain a new kind of AIDS vaccine based on BHIV chimeric viruses.METHODS: A series of chimeric BHIV proviral DNAs differing in the replacement regions in gag gene were constructed, and then were transfected into 293T cells. The expression of chimeric viral genes was detected at the RNA and protein level. The supematant of 293T cell was ultra centrifuged to detect the probable chimeric virion. Once the chimeric virion was detected, its biological activities were also assayed by infecting HIV-sensitive MT4 cells.RESULTS: Four chimeric BHIV proviral DNAs were constructed. Genes in chimeric viruses expressed correctly in transfected 293T cells. All four constructs assembled chimeric virions with different degrees of efficiency. These virions had complete structures common to retroviruses and packaged genomic RNAs, but the cleavages of the precursor Gag proteins were abnormal to some extent. Three of these virions tested could attach and enter into MT4 cells, and one of them could complete the course of reverse transcription. Yet none of them could replicate in MT4 cells.CONCLUSION: The replacement of partial gag gene of HIV with BIV gaggene is feasible. Genes in chimeric BHIVs are accurately expressed, and virions are assembled. These chimeric BHIVs (proviral DNA together with virus particles) have the potential to become a new kind of HIV/AIDS vaccine.展开更多
In the last decade, RNA interference(RNAi) advanced to one of the most widely applied techniques in the biomedical research field and several RNAi therapeutic clinical trials have been launched. We focus on RNAibased ...In the last decade, RNA interference(RNAi) advanced to one of the most widely applied techniques in the biomedical research field and several RNAi therapeutic clinical trials have been launched. We focus on RNAibased inhibitors against the chronic infection with human immunodeficiency virus type 1(HIV-1). A lentiviral gene therapy is proposed for HIV-infected patients that will protect and reconstitute the vital immune cell pool. The RNAi-based inhibitors that have been developed are short hairpin RNA molecules(sh RNAs), of which multiple are needed to prevent viral escape. In ten distinct steps, we describe the selection process that started with 135 sh RNA candidates, from the initial design criteria, via testing of the in vitro and in vivo antiviral activity and cytotoxicity to the final design of a combinatorial therapy with three sh RNAs. These sh RNAs satisfied all 10 selection criteria such as targeting conserved regions of the HIV-1 RNA genome,exhibiting robust inhibition of HIV-1 replication and having no impact on cell physiology. This combinatorial sh RNA vector will soon move forward to the first clinical studies.展开更多
Viral protein U(Vpu) is an accessory protein associated with two main functions important in human immunodeficiency virus type 1(HIV-1) replication and dissemination; these are down-regulation of CD4 receptor through ...Viral protein U(Vpu) is an accessory protein associated with two main functions important in human immunodeficiency virus type 1(HIV-1) replication and dissemination; these are down-regulation of CD4 receptor through mediating its proteasomal degradation and enhancement of virion release by antagonizing tetherin/BST2. It is also well established that Vpu is one of the most highly variable proteins in the HIV-1 proteome. However it is still unclear what drives Vpu sequence variability, whether Vpu acquires polymorphisms as a means of immune escape, functional advantage, or otherwise. It is assumed that the host-pathogen interaction is a cause of polymorphic phenotype of Vpu and that the resulting functional heterogeneity of Vpu may have critical significance in vivo. In order to comprehensively understand Vpu variability, it is important to integrate at the population level the genetic associationapproaches to identify specific amino acid residues and the immune escape kinetics which may impose Vpu functional constraints in vivo. This review will focus on HIV-1 accessory protein Vpu in the context of its sequence variability at population level and also bring forward evidence on the role of the host immune responses in driving Vpu sequence variability; we will also highlight the recent findings that illustrate Vpu functional implication in HIV-1 pathogenesis.展开更多
目的了解北京市部分男男同性恋(men who have sex with men,MSM)人群HIV-1的分子流行情况,监测该地区人群HIV毒株的最新流行情况。方法提取17例男男同性恋者HIV-1抗体阳性样本全血基因组DNA,直接进行巢式PCR扩增HIVenv基因C2-V5区及gag...目的了解北京市部分男男同性恋(men who have sex with men,MSM)人群HIV-1的分子流行情况,监测该地区人群HIV毒株的最新流行情况。方法提取17例男男同性恋者HIV-1抗体阳性样本全血基因组DNA,直接进行巢式PCR扩增HIVenv基因C2-V5区及gag基因P17-P24区,对PCR产物进行核苷酸序列测定和分析,确定基因亚型。结果17例患者中,共存在CRF01-AE、B亚型和CRF07-BC3种亚型,其中CRF01-AE9例(52.94%),B亚型5例(29.41%),CRF07-BC3例(17.65%)。结论小样本量的流行病学调查显示,北京地区部分男男同性恋人群中主要存在CRF01-AE、B和CRF07-BC3种亚型;CRF01-AE已取代B亚型而成为北京市MSM人群中HIV主要流行亚型,CRF07-BC重组亚型在北京市MSM人群中增长迅速。展开更多
文摘While human immunodeficiency virus 1(HIV-1) infectionis controlled through continuous, life-long use of a combination of drugs targeting different steps of the virus cycle, HIV-1 is never completely eradicated from the body. Despite decades of research there is still no effective vaccine to prevent HIV-1 infection. Therefore, the possibility of an RNA interference(RNAi)-based cure has become an increasingly explored approach. Endogenous gene expression is controlled at both, transcriptional and post-transcriptional levels by noncoding RNAs, which act through diverse molecular mechanisms including RNAi. RNAi has the potential to control the turning on/off of specific genes through transcriptional gene silencing(TGS), as well as finetuning their expression through post-transcriptional gene silencing(PTGS). In this review we will describe in detail the canonical RNAi pathways for PTGS and TGS, the relationship of TGS with other silencing mechanisms and will discuss a variety of approaches developed to suppress HIV-1 via manipulation of RNAi. We will briefly compare RNAi strategies against other approaches developed to target the virus, highlighting their potential to overcome the major obstacle to finding a cure, which is the specific targeting of the HIV-1 reservoir within latently infected cells.
基金the National Natural Science Foundation of China, No. 30770755
文摘The human immunodeficiency virus （HIV） lentiviral vector is an ideal vector for gene therapy. In the present study, the wild-type HIV-1 genome was segregated into four plasmids, and an optimized novel HIV-1 lentiviral vector containing green fluorescent protein and vesicular stomatitis virus G pseudo-capsule was constructed. The plasmids were pHR-CMV-EGFP, pCMVΔ8.9, pRSV-Rev, pCMV-VSV-G. The four plasmid system was co-transfected into 293T cells, and green fluorescent protein expression was observed. The present study obtained lentiviral particles by high-speed centrifugation, and the lentiviral particle titer was 4 × 108 TU/mL after centrifugation. Thus, an optimized novel HIV-1 lentiviral vector was successfully constructed.
基金Supported by The grant from the National Institutes of Health,Martin Delaney Collaboratory of AIDS Researchers for Eradication(CARE,U19 AI 096113)the Swiss National Science Foundation(grant 31003A_146579)the University of California,San Diego Fellowships for Graduate Researchers,Frontiers of Innovation Scholars Program
文摘Eradication of human immunodeficiency virus(HIV) in infected individuals is currently not possible because of the presence of the persistent cellular reservoir of latent infection. The identification of HIV latency biomarkers and a better understanding of the molecular mechanisms contributing to regulation of HIV expression might provide essential tools to eliminate these latently infected cells. This review aims at summarizing gene expression profiling and systems biology applications to studies of HIV latency and eradication. Studies comparing gene expression in latently infected and uninfected cells identify candidate latency biomarkers and novel mechanisms of latency control. Studies that profiled gene expression changes induced by existing latency reversing agents(LRAs) highlight uniting themes driving HIV reactivation and novel mechanisms that contribute to regulation of HIV expression by different LRAs. Among the reviewed gene expression studies, the common approaches included identification of differentially expressed genes and gene functional category assessment. Integration of transcriptomic data with other biological data types is presently scarce, and the field would benefit from increased adoption of these methods in future studies. In addition, designing prospective studies that use the same methods of data acquisition and statistical analyses will facilitate a more reliableidentification of latency biomarkers using different model systems and the comparison of the effects of different LRAs on host factors with a role in HIV reactivation. The results from such studies would have the potential to significantly impact the process by which candidate drugs are selected and combined for future evaluations and advancement to clinical trials.
基金This work was supported by grants from the National Natural Science Foundation of China (No. NSFC, 81974303)the High-Level Public Health Specialized Talents Project of Beijing Municipal Health Commission (Nos. 2022-1-007, 2022-2-018)+2 种基金the "Climbing the peak (Dengfeng) " Talent Training Program of Beijing Hospitals Authority (No. DFL20191701)the Beijing Health Technologies Promotion Program (No. BHTPP2020)and the Beijing Key Laboratory for HIV/AIDS Research (No. BZ0089) .
文摘Background: It is controversial whether the apolipoprotein E epsilon 4 allele (APOE ε4) is a risk gene for human immunodeficiency virus (HIV)-related neurocognitive impairment. This meta-analysis aimed to summarize evidence of the associations betweenAPOE ε4 and cognitive impairment in people living with HIV (PLWH).Methods: Our study conducted a systematic literature search of PubMed, Web of Science, Embase, Google Scholar, and ProQuest for studies published before April 11, 2022 that evaluated associations betweenAPOE ε4 and cognitive impairment in adult PLWH (aged ≥18 years). We calculated pooled odds ratios (ORs) of global cognitive impairment and 95% confidence intervals (CIs) and standardized mean differences (SMDs) for specific cognitive domains betweenAPOE ε4 carriers and non-carriers. Subgroup meta-analyses were used to evaluate the result profiles across different categorical variables.Results: Twenty studies met the inclusion criteria, including 19 that evaluated global cognitive impairment.APOE ε4 was significantly associated with global cognitive impairment in PLWH (OR = 1.36, 95% CI = [1.05, 1.78], number of estimates [k] = 19,P = 0.02, random effects). Subgroup meta-analysis based percentage of females showed evident intergroup differences in global cognitive performance between ε4 carriers and non-carriers (P = 0.015).APOE ε4 carriers had lower cognitive test scores than non-carriers in all seven cognitive domains, including fluency (SMD = -0.51, 95% CI = [-0.76, -0.25],P < 0.001,k = 4,I^(2)= 0%), learning (SMD = -0.52, 95% CI = [-0.75, -0.28],P < 0.001,k = 5,I^(2) = 0%), executive function (SMD = -0.41, 95% CI= [-0.59, -0.23],P < 0.001,k= 8,I^(2)= 0%), memory (SMD=-0.41, 95% CI= [-0.61, -0.20],P < 0.001,k= 10,I^(2)= 36%), attention/working memory (SMD=-0.34, 95% CI= [-0.54, -0.14],P= 0.001,k= 6,I^(2)= 0%), speed of information processing (SMD = -0.34, 95% CI = [-0.53, -0.16],P < 0.001,k = 8,I^(2) = 0%), and motor function (SMD = -0.19, 95% CI = [-0.38, -0.01],P = 0.04,k = 7,I^(2) = 0%).Conclusions: Our meta-analysis provides significant evidence thatAPOE ε4 is a risk genotype for HIV-associated cognitive impairment, especially in cognitive domains of fluency, learning, executive function, and memory. Moreover, the impairment is sex specific.Meta analysis registration: PROSPERO, CRD 42021257775.
基金Supported by the National Basic Research Program (973 Program) of China, No. 01999054107
文摘AIM: To explore the possibility of the replacement of the gag gene between human immunodeficiency virus and bovine immunodeficiency virus, to achieve chimeric virions,and thereby gain a new kind of AIDS vaccine based on BHIV chimeric viruses.METHODS: A series of chimeric BHIV proviral DNAs differing in the replacement regions in gag gene were constructed, and then were transfected into 293T cells. The expression of chimeric viral genes was detected at the RNA and protein level. The supematant of 293T cell was ultra centrifuged to detect the probable chimeric virion. Once the chimeric virion was detected, its biological activities were also assayed by infecting HIV-sensitive MT4 cells.RESULTS: Four chimeric BHIV proviral DNAs were constructed. Genes in chimeric viruses expressed correctly in transfected 293T cells. All four constructs assembled chimeric virions with different degrees of efficiency. These virions had complete structures common to retroviruses and packaged genomic RNAs, but the cleavages of the precursor Gag proteins were abnormal to some extent. Three of these virions tested could attach and enter into MT4 cells, and one of them could complete the course of reverse transcription. Yet none of them could replicate in MT4 cells.CONCLUSION: The replacement of partial gag gene of HIV with BIV gaggene is feasible. Genes in chimeric BHIVs are accurately expressed, and virions are assembled. These chimeric BHIVs (proviral DNA together with virus particles) have the potential to become a new kind of HIV/AIDS vaccine.
基金Supported by The NWO-CW(Chemical Sciences),Zon Mw(Medical Sciences),the Dutch AIDS Fund(project 2006006)the DAAD(German Academic Exchange Service)the FRM(Fondation pour la Recherche Medicale)
文摘In the last decade, RNA interference(RNAi) advanced to one of the most widely applied techniques in the biomedical research field and several RNAi therapeutic clinical trials have been launched. We focus on RNAibased inhibitors against the chronic infection with human immunodeficiency virus type 1(HIV-1). A lentiviral gene therapy is proposed for HIV-infected patients that will protect and reconstitute the vital immune cell pool. The RNAi-based inhibitors that have been developed are short hairpin RNA molecules(sh RNAs), of which multiple are needed to prevent viral escape. In ten distinct steps, we describe the selection process that started with 135 sh RNA candidates, from the initial design criteria, via testing of the in vitro and in vivo antiviral activity and cytotoxicity to the final design of a combinatorial therapy with three sh RNAs. These sh RNAs satisfied all 10 selection criteria such as targeting conserved regions of the HIV-1 RNA genome,exhibiting robust inhibition of HIV-1 replication and having no impact on cell physiology. This combinatorial sh RNA vector will soon move forward to the first clinical studies.
基金Supported by A Grant-in-Aid for Scientific Research from the Ministry of Education,Science,Sports,and Culture(MEXT)of JapanA Grant-in-Aid for AIDS Research from the Ministry of Health,Labor,and Welfare of Japan+1 种基金The Scholarship for the International Priority Graduate Programs,to Hasan Z and Kamori DAdvanced Graduate Courses for International Students(Doctoral Course),MEXT,Japan,to Hasan Z and Kamori D
文摘Viral protein U(Vpu) is an accessory protein associated with two main functions important in human immunodeficiency virus type 1(HIV-1) replication and dissemination; these are down-regulation of CD4 receptor through mediating its proteasomal degradation and enhancement of virion release by antagonizing tetherin/BST2. It is also well established that Vpu is one of the most highly variable proteins in the HIV-1 proteome. However it is still unclear what drives Vpu sequence variability, whether Vpu acquires polymorphisms as a means of immune escape, functional advantage, or otherwise. It is assumed that the host-pathogen interaction is a cause of polymorphic phenotype of Vpu and that the resulting functional heterogeneity of Vpu may have critical significance in vivo. In order to comprehensively understand Vpu variability, it is important to integrate at the population level the genetic associationapproaches to identify specific amino acid residues and the immune escape kinetics which may impose Vpu functional constraints in vivo. This review will focus on HIV-1 accessory protein Vpu in the context of its sequence variability at population level and also bring forward evidence on the role of the host immune responses in driving Vpu sequence variability; we will also highlight the recent findings that illustrate Vpu functional implication in HIV-1 pathogenesis.
文摘目的了解北京市部分男男同性恋(men who have sex with men,MSM)人群HIV-1的分子流行情况,监测该地区人群HIV毒株的最新流行情况。方法提取17例男男同性恋者HIV-1抗体阳性样本全血基因组DNA,直接进行巢式PCR扩增HIVenv基因C2-V5区及gag基因P17-P24区,对PCR产物进行核苷酸序列测定和分析,确定基因亚型。结果17例患者中,共存在CRF01-AE、B亚型和CRF07-BC3种亚型,其中CRF01-AE9例(52.94%),B亚型5例(29.41%),CRF07-BC3例(17.65%)。结论小样本量的流行病学调查显示,北京地区部分男男同性恋人群中主要存在CRF01-AE、B和CRF07-BC3种亚型;CRF01-AE已取代B亚型而成为北京市MSM人群中HIV主要流行亚型,CRF07-BC重组亚型在北京市MSM人群中增长迅速。
