CXCR4-targeting nanobodies differentially inhibit CXCR4 function and HIV entry

Hout, Anneleen Van; Klarenbeek, A.; Bobkov, Vladimir; Doijen, Jordi; Arimont, Marta; Zhao, Chao; Heukers, Raimond; Rimkunas, Rebecca; Graaf, Chris de; Verrips, Theo; Woning, Bas van der; Haard, Hans de; Rucker, Joseph; Vermeire, Kurt; Handel, Tracy M.; Loy, Tom Van; Smit, Martine J.; Schols, Dominique

doi:10.1016/j.bcp.2018.10.015

Cited by 39 publications

(62 citation statements)

References 56 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Nanobodies have been used previously to stabilise GPCRs and investigate membrane protein conformations to elucidate new aspects of GPCR function (Zimmermann et al, 2018;Heukers et al, 2018;De Groof et al, 2019a;Heukers et al, 2019). For example, nanobodies recognizing active GPCRs have been used to monitor G protein-mediated signalling after receptor internalization (Irranejad et al, 2013, Stoeber et al, 2018.…”

Section: Discussionmentioning

confidence: 99%

“…Next, a nanobody-HiBiT fusion construct was generated. We used the previously described nanobody, VUN400, as this is known to bind to key residues in ECL2 of the CXCR4 receptor Van Hout et al, 2018). VUN400-HiBiT was expected to bind in close proximity to the N-terminal LgBiT on the LgBiT-CXCR4 receptor, increasing the likelihood of HiBiT-LgBiT complementation upon nanobody binding to the LgBiT-CXCR4.…”

Section: Characterisation Of Lgbit-cxcr4 With Nanobitmentioning

confidence: 99%

“…The improved selectivity and extended half-lives of antibodies compared to small molecules has meant there has been much interest in using antibody-based approaches to target CXCR4 therapeutically (Hutchings et al, 2017;Bobkov et al, 2019). This has included the recent development of a panel of single domain antibody fragments, called nanobodies, which are able to bind CXCR4 (Jahnichen et al, 2010;de Wit et al, 2017;Bobkov et al, 2018;Van Hout et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

“…Nanobodies which that bind to the extracellular domains of GPCRs are able to modulate receptor activity and organization (De Groof et al, 2019a). Several studies have investigated the therapeutic potential of extracellular nanobodies which target chemokine GPCRs, including CXCR2 (Bradley et al, 2015), CXCR4 (Jahnichen et al, 2010;de Wit et al, 2017;Bobkov et al, 2018;Van Hout et al, 2018), ACKR3 (Maussang et al, 2013), and US28 (De Groof et al, 2019b). Given their relatively large N-terminus compared to the other Class A GPCRs, and the fact that their endogenous ligands are peptides, chemokine GPCRs are ideal candidates to target with extracellular nanobodies.…”

Section: Introductionmentioning

confidence: 99%

“…Given their relatively large N-terminus compared to the other Class A GPCRs, and the fact that their endogenous ligands are peptides, chemokine GPCRs are ideal candidates to target with extracellular nanobodies. Most recently, several nanobodies binding to the N-terminus and second extracellular loop (ECL2) of CXCR4 were generated Van Hout et al, 2018). For example, VUN400 was one of these nanobodies that acted as an antagonist and inhibited CXCL12-induced signalling by CXCR4, as well as internalization.…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Monitoring ligand-induced changes in receptor conformation with NanoBiT conjugated nanobodies

Soave

Heukers

Kellam

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

max 150 words)Camelid single-domain antibody fragments (nanobodies) offer the specificity of an antibody in a single 15kDa immunoglobulin domain. Their small size allows for easy genetic manipulation of the nanobody sequence to incorporate protein tags, facilitating their use as biochemical probes. The nanobody VUN400, which recognises the second extracellular loop of the human CXCR4 chemokine receptor, was used as a probe to monitor specific CXCR4 conformations. VUN400 was fused via its C-terminus to the 11-amino acid HiBiT tag (VUN400-HiBiT) which complements to LgBiT protein, forming a full length functional NanoLuc luciferase. Here, complemented luminescence was used to detect VUN400-HiBiT binding to CXCR4 receptors expressed in living HEK293 cells. VUN400-HiBiT binding to CXCR4 could be prevented by orthosteric and allosteric ligands, allowing VUN400-HiBiT to be used as a probe to detect specific conformations of CXCR4. These data demonstrate that the high specificity offered by extracellular-targeted nanobodies can be utilised to probe receptor pharmacology.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Characterisation Of Lgbit-cxcr4 With Nanobitmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Monitoring ligand-induced changes in receptor conformation with NanoBiT conjugated nanobodies

Soave

Heukers

Kellam

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

Nanosome‐Mediated Delivery Of Hdac Inhibitors and Oxygen Molecules for the Transcriptional Reactivation of Latent Hiv‐Infected Cd4⁺ T Cells

Hong

Choi

Lee

et al. 2023

Small

View full text Add to dashboard Cite

The treatment of human immunodeficiency virus (HIV) infection is notoriously difficult due to the ability of this virus to remain latent in the host's CD4+ T cells. Histone deacetylases (HDACs) interfere with DNA transcription in HIV‐infected hosts, resulting in viral latency. Therefore, HDAC inhibitors can be used to activate viral transcription in latently infected cells, after which the virus can be eliminated through a shock‐and‐kill strategy. Here, a drug delivery system is developed to effectively deliver HDAC inhibitors to latent HIV‐infected cells. Given that the efficacy of HDAC inhibitors is reduced under hypoxic conditions, oxygen‐containing nanosomes are used as drug carriers. Oxygen‐containing nanosomes can improve the efficiency of chemotherapy by delivering essential oxygen to cells. Additionally, their phospholipid bilayer structure makes them uniquely well‐suited for drug delivery. In this study, a novel drug delivery system is developed by taking advantage of the oxygen carriers in these oxygen nanosomes, incorporating a multi‐drug strategy consisting of HDAC inhibitors and PKA activators, and introducing CXCR4 binding peptides to specifically target CD4+ T cells. Oxygen nanosomes with enhanced targeting capability through the introduction of the CXCR4 binding peptide mitigate drug toxicity and slow down drug release. The observed changes in the expression of p24, a capsid protein of HIV, indirectly confirm that the proposed drug delivery system can effectively induce transcriptional reactivation of HIV in latent HIV‐infected cells.

show abstract

Nanobodies in the fight against infectious diseases: repurposing nature's tiny weapons

Rizk,

Moustafa,

ElBanna

et al. 2024

World J Microbiol Biotechnol

View full text Add to dashboard Cite

Nanobodies are the smallest known antigen-binding molecules to date. Their small size, good tissue penetration, high stability and solubility, ease of expression, refolding ability, and negligible immunogenicity in the human body have granted them excellence over conventional antibodies. Those exceptional attributes of nanobodies make them promising candidates for various applications in biotechnology, medicine, protein engineering, structural biology, food, and agriculture. This review presents an overview of their structure, development methods, advantages, possible challenges, and applications with special emphasis on infectious diseases-related ones. A showcase of how nanobodies can be harnessed for applications including neutralization of viruses and combating antibiotic-resistant bacteria is detailed. Overall, the impact of nanobodies in vaccine design, rapid diagnostics, and targeted therapies, besides exploring their role in deciphering microbial structures and virulence mechanisms are highlighted. Indeed, nanobodies are reshaping the future of infectious disease prevention and treatment.

show abstract

CXCR4-targeting nanobodies differentially inhibit CXCR4 function and HIV entry

Cited by 39 publications

References 56 publications

Monitoring ligand-induced changes in receptor conformation with NanoBiT conjugated nanobodies

Monitoring ligand-induced changes in receptor conformation with NanoBiT conjugated nanobodies

Nanosome‐Mediated Delivery Of Hdac Inhibitors and Oxygen Molecules for the Transcriptional Reactivation of Latent Hiv‐Infected Cd4⁺ T Cells

Nanobodies in the fight against infectious diseases: repurposing nature's tiny weapons

Contact Info

Product

Resources

About

CXCR4-targeting nanobodies differentially inhibit CXCR4 function and HIV entry

Cited by 39 publications

References 56 publications

Monitoring ligand-induced changes in receptor conformation with NanoBiT conjugated nanobodies

Monitoring ligand-induced changes in receptor conformation with NanoBiT conjugated nanobodies

Nanosome‐Mediated Delivery Of Hdac Inhibitors and Oxygen Molecules for the Transcriptional Reactivation of Latent Hiv‐Infected Cd4+ T Cells

Nanobodies in the fight against infectious diseases: repurposing nature's tiny weapons

Contact Info

Product

Resources

About

Nanosome‐Mediated Delivery Of Hdac Inhibitors and Oxygen Molecules for the Transcriptional Reactivation of Latent Hiv‐Infected Cd4⁺ T Cells