LEDGIN-mediated Inhibition of Integrase–LEDGF/p75 Interaction Reduces Reactivation of Residual Latent HIV

Vranckx, Lenard; Demeulemeester, Jonas; Saleh, Suha; Boll, Annegret; Vansant, Greet; Schrijvers, R.; Weydert, Caroline; Battivelli, Emilie; Verdin, Eric; Cereseto, Anna; Christ, Frauke; Gijsbers, Rik; Debyser, Zeger

doi:10.1016/j.ebiom.2016.04.039

Cited by 97 publications

(169 citation statements)

References 81 publications

Supporting

Mentioning

140

Contrasting

Order By: Relevance

“…Second, since integrase is produced by protease-mediated cleavage of the Pr160 Gag-Pol precursor protein, PIs might inhibit HIV-1 integration by preventing the formation of a functional integrase19. Impaired integrase activity and/or impairment of the integrase-LEDGF/p75 interaction could result in a decreased number of integrated provirus in transcriptionally active sites, but also could favor the viral integration in less transcriptionally active sites which will result in lower viral reactivation22. Third, HIV PIs have been reported to block Akt activation23, and we recently observed that PIs, by blocking Akt activation especially triggered by Nef, limit HIV-1 recovery from latently-infected T cells24.…”

Section: Discussionmentioning

confidence: 99%

Limited HIV-1 Reactivation in Resting CD4+ T cells from Aviremic Patients under Protease Inhibitors

Kumar

Abbas

Bouchat

et al. 2016

Sci Rep

View full text Add to dashboard Cite

A latent viral reservoir that resides in resting CD4+ T cells represents a major barrier for eradication of HIV infection. We test here the impact of HIV protease inhibitor (PI) based combination anti-retroviral therapy (cART) over nonnucleoside reverse transcriptase inhibitor (NNRTI)-based cART on HIV-1 reactivation and integration in resting CD4+ T cells. This is a prospective cohort study of patients with chronic HIV-1 infection treated with conventional cART with an undetectable viremia. We performed a seven-year study of 47 patients with chronic HIV-infection treated with cART regimens and with undetectable plasma HIV-1 RNA levels for at least 1 year. Of these 47 patients treated with cART, 24 were treated with a PI-based regimen and 23 with a NNRTI-based regimen as their most recent treatment for more than one year. We evaluated the HIV-1 reservoir using reactivation assay and integrated HIV-1 DNA, respectively, in resting CD4+ T cells. Resting CD4+ T cells isolated from PI-treated patients compared to NNRTI-treated patients showed a limited HIV-1 reactivation upon T-cell stimulation (p = 0·024) and a lower level of HIV-1 integration (p = 0·024). Our study indicates that PI-based cART could be more efficient than NNRTI-based cART for limiting HIV-1 reactivation in aviremic chronically infected patients.

show abstract

Section: Discussionmentioning

confidence: 99%

Limited HIV-1 Reactivation in Resting CD4+ T cells from Aviremic Patients under Protease Inhibitors

Kumar

Abbas

Bouchat

et al. 2016

Sci Rep

View full text Add to dashboard Cite

show abstract

“…Vranckx et al have shown that LEDGF/p75 depletion hampers HIV-1 reactivation in cell culture, and they demonstrated that LEDGINs relocate and retarget HIV integration, resulting in a HIV reservoir that is refractory to reactivation by different latency-reversing agents (20).…”

Section: Introductionmentioning

confidence: 99%

“…Initially designed to prevent the IN-LEDGF/p75 interaction, it was later evidenced that these molecules have an additional biochemical activity based on the induction of allosteric conformational changes of IN that ultimately trigger its aberrant multimerization (10)(11)(12)(15)(16)(17)(18)(19). This effect is independent of the presence of LEDGF/p75 and the viral DNA and results from the binding of the inhibitors to the IN dimer interface that is also part of the LEDGF-binding pocket.Given their dual biochemical activities, INLAIs were shown to block HIV-1 replication at two different steps: the inhibition of IN-LEDGF/p75 binding accounts for an "early" block at integration, while INLAI-promoted IN multimerization results in a "late" effect during virus maturation (11,12,17,18).Vranckx et al have shown that LEDGF/p75 depletion hampers HIV-1 reactivation in cell culture, and they demonstrated that LEDGINs relocate and retarget HIV integration, resulting in a HIV reservoir that is refractory to reactivation by different latency-reversing agents (20).HIV-1 virions produced in the presence of INLAIs are non-infectious because they are unable to complete reverse transcription upon target cell infection (12,17,18). Investigating the molecular bases of the observed infectivity defects, we found that HIV-1 virions produced in the presence of the quinoline INLAI compound BI-D (developed by Boehringer Ingelheim) package normal levels of genomic RNA dimer and harbor a properly placed tRNA lys3 primer that could be extended ex vivo.…”

mentioning

confidence: 99%

Structure-function analyses unravel distinct effects of allosteric inhibitors of HIV-1 integrase on viral maturation and integration

Bonnard

Rouzic

Eiler

et al. 2018

Journal of Biological Chemistry

View full text Add to dashboard Cite

Recently, a new class of HIV-1 integrase (IN) inhibitors with a dual mode of action, called IN-LEDGF/p75 allosteric inhibitors (INLAIs), was described. Designed to interfere with the IN-LEDGF/p75 interaction during viral integration, unexpectedly, their major impact was on virus maturation. This activity has been linked to induction of aberrant IN multimerization, while inhibition of the IN-LEDGF/p75 interaction accounts for weaker antiretroviral effect at integration. Since these dual activities result from INLAI binding to IN at a single binding site, we expected that these activities co-evolved together, driven by the affinity for IN. Using an original INLAI, MUT-A, and its activity on an Ala-125 (A125) IN variant, we found that these two activities on A125 IN can be fully dissociated: MUT-A-induced IN multimerization and the formation of eccentric condensates in viral particles, that are responsible for inhibition of virus maturation, were lost, while inhibition of the IN-LEDGF/p75 interaction and consequently integration, was fully retained. Hence the mere binding of INLAI to A125 IN is insufficient to promote the conformational changes of IN required for aberrant multimerization. By analyzing the Xray structures of MUT-A bound to the IN catalytic core domain (CCD) with or without the A125 polymorphism, we discovered that the loss of IN multimerization is due to stabilization of the A125 IN variant CCD dimer, highlighting the importance of the CCD dimerization energy for IN multimerization. Our study reveals that affinity for the LEDGF/p75-binding pocket is not sufficient to induce INLAI-dependent IN multimerization and the associated inhibition of viral maturation.The integrase (IN) protein of Human Immunodeficiency Virus type 1 (HIV-1) catalyzes the stable insertion of the viral cDNA genome into the host cell chromatin, a step of the viral life cycle that is required for efficient viral gene expression. Integration occurs via a two-step reaction where IN initially cleaves after a conserved CA dinucleotide at the 3' end of the viral cDNA genome to free a 3'-OH group (3' processing), which is next used to carry out a nucleophilic attack on cellular chromosomal DNA (strand transfer).IN is one of the preferred targets for the development of antiretroviral (ARV) drugs. However, given the high genetic variability of HIV-1, IN mutations conferring cross-resistance to the first generation INSTIs, RAL and EVG, were described in patients receiving INSTI-containing regimens (2). The second generation INSTI DTG has a higher genetic barrier and conserves good ARV activity against a number of RAL-and EVGresistant strains. Recent reports showed that Bictegravir, a second generation INSTI still in development from Gilead Sciences, has a resistance profile similar to DTG (3). Nevertheless, DTG and Bictegravir are sensitive to the most detrimental INSTI resistant mutations albeit at lower levels than first generation INSTIs (4). Therefore, the development of small molecule inhibitors impairing IN functions with dis...

show abstract

“…In addition to decreasing integration levels, the R263K substitution may also alter integration sites. Changes in integration sites were observed when infections are performed in the presence of suboptimal concentrations of RAL or noncatalytic integrase inhibitors but also with the D116N catalytically inactive integrase mutant (27–29). In the case of the R263K substitution, however, these changes are unlikely to alter the establishment of or reactivation from latency, since we previously reported these processes to be unaffected by this substitution (30).…”

Section: Discussionmentioning

confidence: 99%

The R263K Dolutegravir Resistance-Associated Substitution Progressively Decreases HIV-1 Integration

Mésplède

Leng

Pham

et al. 2017

mBio

View full text Add to dashboard Cite

Human immunodeficiency virus (HIV) infection persists despite decades of active antiretroviral therapy (ART), effectively preventing viral eradication. Treatment decreases plasma viral RNA, but viral DNA persists, mostly integrated within the genome of nucleated blood cells. Viral DNA blood levels correlate with comorbidities and the rapidity of viral rebound following treatment interruption. To date, no intervention aiming at decreasing HIV DNA levels below those attained through ART has been successful. This includes use of some integrase inhibitors either as part of ART or in treatment intensification studies. We have argued that using the integrase inhibitor dolutegravir (DTG) in similar studies may yield better results, but this remains to be studied. In treatment-experienced individuals, the most frequent substitution associated with failure with dolutegravir is R263K in integrase. R263K decreases integration both in cell-free and tissue culture assays. We investigated here how integrated DNA levels evolve over time during prolonged infections with R263K viruses. To investigate a potential defect in reverse transcription with R263K, the levels of reverse transcripts were measured by quantitative PCR. We measured HIV type 1 (HIV-1) integration in Jurkat cells over the course of 4-week infections using Alu-mediated quantitative PCR. The results show that R263K did not decrease reverse transcription. Prolonged infections with R263K mutant viruses led to less HIV-1 integrated DNA over time compared to wild-type viruses. These tissue culture results help to explain the absence of the R263K substitution in most individuals experiencing failure with DTG and support studies aiming at longitudinally measuring the levels of integrated DNA in individuals treated with this drug.

show abstract

LEDGIN-mediated Inhibition of Integrase–LEDGF/p75 Interaction Reduces Reactivation of Residual Latent HIV

Cited by 97 publications

References 81 publications

Limited HIV-1 Reactivation in Resting CD4+ T cells from Aviremic Patients under Protease Inhibitors

Limited HIV-1 Reactivation in Resting CD4+ T cells from Aviremic Patients under Protease Inhibitors

Structure-function analyses unravel distinct effects of allosteric inhibitors of HIV-1 integrase on viral maturation and integration

The R263K Dolutegravir Resistance-Associated Substitution Progressively Decreases HIV-1 Integration

Contact Info

Product

Resources

About