scholarly journals Structural basis for strand-transfer inhibitor binding to HIV intasomes

Science ◽  
2020 ◽  
Vol 367 (6479) ◽  
pp. 810-814 ◽  
Author(s):  
Dario Oliveira Passos ◽  
Min Li ◽  
Ilona K. Jóźwik ◽  
Xue Zhi Zhao ◽  
Diogo Santos-Martins ◽  
...  
Keyword(s):  
High Resolution ◽  
Drug Target ◽  
Antiretroviral Drugs ◽  
Drug Binding ◽  
Structural Basis ◽  
Strand Transfer ◽  
Cryo Electron Microscopy ◽  
Integrase Strand Transfer Inhibitors ◽  
Transfer Inhibitor ◽  
Host Chromatin

The HIV intasome is a large nucleoprotein assembly that mediates the integration of a DNA copy of the viral genome into host chromatin. Intasomes are targeted by the latest generation of antiretroviral drugs, integrase strand-transfer inhibitors (INSTIs). Challenges associated with lentiviral intasome biochemistry have hindered high-resolution structural studies of how INSTIs bind to their native drug target. Here, we present high-resolution cryo–electron microscopy structures of HIV intasomes bound to the latest generation of INSTIs. These structures highlight how small changes in the integrase active site can have notable implications for drug binding and design and provide mechanistic insights into why a leading INSTI retains efficacy against a broad spectrum of drug-resistant variants. The data have implications for expanding effective treatments available for HIV-infected individuals.

Increase in visceral adipose tissue in a woman living with HIV after introduction of integrase strand transfer inhibitor

2020 ◽  
Vol 31 (14) ◽  
pp. 1407-1410
Author(s):  
Daniela Piacentini ◽  
Massimiliano Lanzafame ◽  
Sebastiano Rizzardo ◽  
Sheila Chiesi ◽  
Emanuela Lattuada ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Antiretroviral Drugs ◽  
Strand Transfer ◽  
Metabolic Abnormalities ◽  
Tissue Density ◽  
Integrase Strand Transfer Inhibitors ◽  
Transfer Inhibitor ◽  
Living With Hiv ◽  
Visceral Adipose ◽  
Integrase Strand Transfer Inhibitor

Integrase strand transfer inhibitors (INSTIs) are a class of antiretroviral drugs with high virologic efficacy and excellent tolerability. Recent evidence showed a possible link of dolutegravir-based regimens with weight gain, and a relationship between raltegravir use and changes in adipose tissue density and metabolic abnormalities, with an increased cardiovascular risk, has been suggested. We describe a case where dolutegravir monotherapy led to a decrease in adipose tissue density.

scholarly journals Cryo-EM structures and atomic model of the HIV-1 strand transfer complex intasome

Science ◽  
2017 ◽  
Vol 355 (6320) ◽  
pp. 89-92 ◽  
Author(s):  
Dario Oliveira Passos ◽  
Min Li ◽  
Renbin Yang ◽  
Stephanie V. Rebensburg ◽  
Rodolfo Ghirlando ◽  
...  
Keyword(s):  
High Resolution ◽  
Higher Order ◽  
Strand Transfer ◽  
Structural Studies ◽  
Cryo Electron Microscopy ◽  
Target Dna ◽  
Carboxyl Terminal ◽  
Host Target ◽  
Hiv 1 ◽  
Host Chromatin

Like all retroviruses, HIV-1 irreversibly inserts a viral DNA (vDNA) copy of its RNA genome into host target DNA (tDNA). The intasome, a higher-order nucleoprotein complex composed of viral integrase (IN) and the ends of linear vDNA, mediates integration. Productive integration into host chromatin results in the formation of the strand transfer complex (STC) containing catalytically joined vDNA and tDNA. HIV-1 intasomes have been refractory to high-resolution structural studies. We used a soluble IN fusion protein to facilitate structural studies, through which we present a high-resolution cryo–electron microscopy (cryo-EM) structure of the core tetrameric HIV-1 STC and a higher-order form that adopts carboxyl-terminal domain rearrangements. The distinct STC structures highlight how HIV-1 can use the common retroviral intasome core architecture to accommodate different IN domain modules for assembly.

How Good Can Single-Particle Cryo-EM Become? What Remains Before It Approaches Its Physical Limits?

2019 ◽  
Vol 48 (1) ◽  
pp. 45-61 ◽  
Author(s):  
Robert M. Glaeser
Keyword(s):  
Electron Microscopy ◽  
High Resolution ◽  
Single Particle ◽  
Energy Use ◽  
Optimal Choice ◽  
Structural Basis ◽  
Detective Quantum Efficiency ◽  
Quantum Metrology ◽  
Cryo Electron Microscopy ◽  
Induced Motion

Impressive though the achievements of single-particle cryo–electron microscopy are today, a substantial gap still remains between what is currently accomplished and what is theoretically possible. As is reviewed here, twofold or more improvements are possible as regards ( a) the detective quantum efficiency of cameras at high resolution, ( b) converting phase modulations to intensity modulations in the image, and ( c) recovering the full amount of high-resolution signal in the presence of beam-induced motion of the specimen. In addition, potential for improvement is reviewed for other topics such as optimal choice of electron energy, use of aberration correctors, and quantum metrology. With the help of such improvements, it does not seem to be too much to imagine that determining the structural basis for every aspect of catalytic control, signaling, and regulation, in any type of cell of interest, could easily be accelerated fivefold or more.

scholarly journals High-resolution structure determination of sub-100 kilodalton complexes using conventional cryo-EM

2018 ◽  
Author(s):  
Mark A. Herzik ◽  
Mengyu Wu ◽  
Gabriel C. Lander
Keyword(s):  
High Resolution ◽  
Structure Determination ◽  
Drug Target ◽  
Phase Plate ◽  
Biological Macromolecules ◽  
Cryo Electron Microscopy ◽  
Transmission Electron ◽  
High Resolution Structure ◽  
Resolution Structure

Determining high-resolution structures of biological macromolecules with masses of less than 100 kilodaltons (kDa) has long been a goal of the cryo-electron microscopy (cryo-EM) community. While the Volta Phase Plate has enabled cryo-EM structure determination of biological specimens of this size range, use of this instrumentation is not yet fully automated and can present technical challenges. Here, we show that conventional defocus-based cryo-EM methodologies can be used to determine the high-resolution structures of specimens amassing less than 100 kDa using a transmission electron microscope operating at 200 keV coupled with a direct electron detector. Our ~2.9 Å structure of alcohol dehydrogenase (82 kDa) proves that bound ligands can be resolved with high fidelity, indicating that these methodologies can be used to investigate the molecular details of drug-target interactions. Our ~2.8 Å and ~3.2 Å resolution structures of methemoglobin demonstrate that distinct conformational states can be identified within a dataset for proteins as small as 64 kDa. Furthermore, we provide the first sub-nanometer cryo-EM structure of a protein smaller than 50 kDa.

scholarly journals Nonhuman Primates and Humanized Mice for Studies of HIV-1 Integrase Inhibitors: A Review

2016 ◽  
Vol 1 (1) ◽  
pp. 41 ◽  
Author(s):  
Said A. Hassounah ◽  
Thibault Mesplède ◽  
Mark A. Wainberg
Keyword(s):  
Nonhuman Primates ◽  
Antiretroviral Drugs ◽  
Strand Transfer ◽  
Resistance Mutations ◽  
Humanized Mouse ◽  
Hiv Replication ◽  
Hiv Therapy ◽  
Integrase Strand Transfer Inhibitors ◽  
Hiv 1

Since the discovery of the first inhibitors of HIV replication, drug resistance has been a major problem in HIV therapy, due, in part, to the high mutation rate of HIV. Therefore, the development of a predictive animal model is important to identify impending resistance mutations and to possibly inform treatment decisions. Significant advances have been made possible through use of nonhuman primates infected by SIV, SHIV, and stHIV-1, and use of humanized mouse models of HIV-1 infections. In this review, we describe some of the findings from animal models used for the preclinical testing of integrase strand transfer inhibitors as well as other antiretroviral drugs. These models have led to important findings about the potential role of integrase strand transfer inhibitors in both the prevention and treatment of HIV-1 infection.

scholarly journals Potential drug–drug interactions in the era of integrase strand transfer inhibitors: a cross-sectional single-center study in Japan

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Yusuke Kunimoto ◽  
Ryosuke Matamura ◽  
Hiroshi Ikeda ◽  
Satoshi Fujii ◽  
Tomoko Kimyo ◽  
...  
Keyword(s):  
Drug Interactions ◽  
Health Care Providers ◽  
Median Number ◽  
Antiretroviral Drugs ◽  
Strand Transfer ◽  
Potential Drug ◽  
Care Providers ◽  
Cross Sectional ◽  
Single Center Study ◽  
Integrase Strand Transfer Inhibitors

Abstract Background Potential drug–drug interactions (PDDIs) commonly occur because of aging and comorbidities in people living with human immunodeficiency virus (HIV; PLWH). Protease inhibitors and non-nucleoside reverse transcriptase inhibitors have been reported to cause PDDIs in these patients. However, there are few reports of PDDIs in the era of treatment using integrase strand transfer inhibitors. Therefore, we investigated PDDIs in Japanese PLWH receiving antiretroviral drugs (ARVs). Methods This was a cross-sectional observational study conducted in Japanese outpatients. All eligible patients who had received ARV therapy for at least 48 weeks were enrolled. The primary endpoint was the incidence of PDDIs detected using the Lexicomp® interface. Results Of the 71 eligible patients, 51 (71.8%) were prescribed concomitant non-ARV medications. In 21 patients (29.6%), PDDIs with the potential to reduce the effects of ARVs occurred, although the HIV load was suppressed in all cases. Polypharmacy (the use of ≥5 non-ARVs) was observed in 25 patients (35.2%). There was a significantly higher median number of non-ARV medications in the PDDI group than in the non-PDDI group (6 vs. 3, P <  0.001). Furthermore, the proportion of patients on polypharmacy was significantly higher in those with PDDIs than in those without PDDIs (81.0% vs. 26.7%, P <  0.001). Conclusions The incidence of PDDIs is relatively high in Japanese PLWH, even in the era of treatment using integrase strand transfer inhibitors. Therefore, it is important for patients and health care providers to be constantly aware of PDDIs associated with ARV treatment.

scholarly journals Molecular insights into ago-allosteric modulation of the human glucagon-like peptide-1 receptor

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Zhaotong Cong ◽  
Li-Nan Chen ◽  
Honglei Ma ◽  
Qingtong Zhou ◽  
Xinyu Zou ◽  
...  
Keyword(s):  
Drug Target ◽  
Metabolic Disorders ◽  
Allosteric Modulation ◽  
Structural Basis ◽  
Allosteric Modulators ◽  
Cryo Electron Microscopy ◽  
Covalently Bonded ◽  
Glucagon Like Peptide ◽  
Glucagon Like Peptide 1 ◽  
Orthosteric Ligands

AbstractThe glucagon-like peptide-1 (GLP-1) receptor is a validated drug target for metabolic disorders. Ago-allosteric modulators are capable of acting both as agonists on their own and as efficacy enhancers of orthosteric ligands. However, the molecular details of ago-allosterism remain elusive. Here, we report three cryo-electron microscopy structures of GLP-1R bound to (i) compound 2 (an ago-allosteric modulator); (ii) compound 2 and GLP-1; and (iii) compound 2 and LY3502970 (a small molecule agonist), all in complex with heterotrimeric Gs. The structures reveal that compound 2 is covalently bonded to C347 at the cytoplasmic end of TM6 and triggers its outward movement in cooperation with the ECD whose N terminus penetrates into the GLP-1 binding site. This allows compound 2 to execute positive allosteric modulation through enhancement of both agonist binding and G protein coupling. Our findings offer insights into the structural basis of ago-allosterism at GLP-1R and may aid the design of better therapeutics.

Integrase strand transfer inhibitor-associated diabetes mellitus: A case report

2016 ◽  
Vol 28 (6) ◽  
pp. 626-628 ◽  
Author(s):  
Peter S Fong ◽  
Devon M Flynn ◽  
Christopher D Evans ◽  
P Todd Korthuis
Keyword(s):  
Diabetes Mellitus ◽  
Case Report ◽  
Hemoglobin A1c ◽  
Hemophilia A ◽  
Strand Transfer ◽  
Exogenous Insulin ◽  
Integrase Strand Transfer Inhibitors ◽  
History Of ◽  
Transfer Inhibitor ◽  
Integrase Strand Transfer Inhibitor

Integrase strand transfer inhibitors (INSTIs) have become integral antiretroviral therapy (ART) agents for treating HIV infection. We report the case of a 44-year-old male with a history of hemophilia A who developed diabetes mellitus four months after switching from abacavir, lamivudine, and efavirenz to abacavir, lamivudine, and raltegravir. Hemoglobin A1C normalized without further need for exogenous insulin after raltegravir was switched back to efavirenz. In this case report, we will review a possible mechanism for INSTI-induced hyperglycemia and/or diabetes mellitus.

scholarly journals Resistance of human immunodeficiency virus type 1 to integrase strand transfer inhibitors in Croatia

2019 ◽  
Vol 2 (1) ◽  
pp. 29-33
Author(s):  
Ana Planinic ◽  
Maja Oroz ◽  
Josip Begovac ◽  
Snjezana Zidovec Lepej
Keyword(s):  
Population Based ◽  
Antiretroviral Drugs ◽  
Protease Gene ◽  
Strand Transfer ◽  
Resistance Mutations ◽  
Integrase Gene ◽  
Integrase Strand Transfer Inhibitors ◽  
Clinically Significant ◽  
First Time ◽  
Hiv 1

Integrase strand transfer inhibitors (INSTIs) are the latest class of antiretroviral drugs that prevent the integration of proviral DNA into the host genome. The aim of this study was to describe, for the first time, INSTI resistance mutations observed in Croatian HIV-infected patients. Methods: The study was conducted between March 2016 and September 2018 and included 4 previously untreated patients (antiretroviral, ARV-naive) as well as 18 unsuccessfully treated HIV-infected patients (ARV-experienced) that have been tested for INSTI resistance. The genetic data on INSTI resistance was obtained by population-based sequencing of the integrase gene. Resistance analysis to other classes of antiretroviral drugs has been performed in some patients by sequencing the protease gene and a part of the reverse transcriptase HIV-1 gene. Results: INSTI resistance mutations were not found in ARV-naive patients. Mutations associated with resistance to INSTIs have been observed in 5 of 18 (27.8%) patients failing INSTI-based ARV regiment. Resistance to INSTIs in ARV-experienced patients was attributed to major resistance mutations Q148R, N155H and E92Q that confer resistance to two INSTIs (raltegravir and elvitegravir). Conclusions: The results of this study describe the first 5 cases of ARV-experienced HIV-1 infected patients with clinically significant resistance to INSTIs, and emphasize the need for continuous surveillance of INSTI resistance in patients experiencing virological failure to antiretroviral treatment in Croatia.

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