Llama-derived Single Variable Domains (Nanobodies) Directed against Chemokine Receptor CXCR7 Reduce Head and Neck Cancer Cell Growth in Vivo

Maussang, David; Mujić-Delić, Azra; Descamps, Francis J; Stortelers, Catelijne; Vanlandschoot, Peter; Walsum, Marijke Stigter-van; Vischer, Henry F.; Roy, Maarten Van; Vosjan, Maria J. W. D.; González-Pajuelo, María; Dongen, Guus A.M.S. van; Merchiers, Pascal; Rompaey, Philippe van; Smit, Martine J.

doi:10.1074/jbc.m113.498436

Cited by 126 publications

(90 citation statements)

References 48 publications

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“…Camelid V H Hs generated against the Kv1.3 ion channel targeted extracellular loops, not the channel cavity, 85 and the epitopes of V H Hs against the P2X7 ion channel were not defined. 86 Similarly, camelid V H Hs developed as potential therapeutics against the chemokine receptors CXCR4, 84 CXCR7 87 and ChemR23, 88 as well as V H Hs used as crystallization chaperones for several GPCRs, channels and transporters, 89–95 all appear to bind solvent-exposed extracellular or intracellular loops of these receptors in a manner similar to conventional antibodies and their fragments. By contrast, a synthetic CXCR4-binding “i-body” engineered from an Ig-like NCAM domain was found to penetrate deep into the receptor’s ligand-binding pocket to occupy a truly cryptic, partially transmembrane epitope.…”

Section: Single-domain Antibodies Directed Against Folded Proteinsmentioning

confidence: 99%

Antigen recognition by single-domain antibodies: structural latitudes and constraints

Henry

MacKenzie

2018

mAbs

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Single-domain antibodies (sdAbs), the autonomous variable domains of heavy chain-only antibodies produced naturally by camelid ungulates and cartilaginous fishes, have evolved to bind antigen using only three complementarity-determining region (CDR) loops rather than the six present in conventional VH:VL antibodies. It has been suggested, based on limited evidence, that sdAbs may adopt paratope structures that predispose them to preferential recognition of recessed protein epitopes, but poor or non-recognition of protuberant epitopes and small molecules. Here, we comprehensively surveyed the evidence in support of this hypothesis. We found some support for a global structural difference in the paratope shapes of sdAbs compared with those of conventional antibodies: sdAb paratopes have smaller molecular surface areas and diameters, more commonly have non-canonical CDR1 and CDR2 structures, and have elongated CDR3 length distributions, but have similar amino acid compositions and are no more extended (interatomic distance measured from CDR base to tip) than conventional antibody paratopes. Comparison of X-ray crystal structures of sdAbs and conventional antibodies in complex with cognate antigens showed that sdAbs and conventional antibodies bury similar solvent-exposed surface areas on proteins and form similar types of non-covalent interactions, although these are more concentrated in the compact sdAb paratope. Thus, sdAbs likely have privileged access to distinct antigenic regions on proteins, but only owing to their small molecular size and not to general differences in molecular recognition mechanism. The evidence surrounding the purported inability of sdAbs to bind small molecules was less clear. The available data provide a structural framework for understanding the evolutionary emergence and function of autonomous heavy chain-only antibodies.

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Section: Single-domain Antibodies Directed Against Folded Proteinsmentioning

confidence: 99%

Antigen recognition by single-domain antibodies: structural latitudes and constraints

Henry

MacKenzie

2018

mAbs

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“…Productions and purifications of monovalent, bivalent, and biparatopic Nanobodies were performed essentially as described elsewhere (10). The flexible Gly-Ser linkers used to combine the monovalent building blocks in the bivalent and biparatopic constructs are referred to as 40GS ((GGGGS) 8 ).…”

Section: Methodsmentioning

confidence: 99%

High Throughput Combinatorial Formatting of PcrV Nanobodies for Efficient Potency Improvement

Tavernier

Detalle

Morizzo

et al. 2016

Journal of Biological Chemistry

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Improving potencies through concomitant blockage of multiple epitopes and avid binding by fusing multiple (different) monovalent Nanobody building blocks via linker sequences into one multivalent polypeptide chain is an elegant alternative to affinity maturation. We explored a large and random formatting library of bivalent (combinations of two identical) and biparatopic (combinations of two different) Nanobodies for functional blockade of Pseudomonas aeruginosa PcrV. PcrV is an essential part of the P. aeruginosa type III secretion system (T3SS), and its oligomeric nature allows for multiple complex binding and blocking options. The library screening yielded a large number of promising biparatopic lead candidates, revealing significant (and non-trivial) preferences in terms of Nanobody building block and epitope bin combinations and orientations. Excellent potencies were confirmed upon further characterization in two different P. aeruginosa T3SS-mediated cytotoxicity assays. Three biparatopic Nanobodies were evaluated in a lethal mouse P. aeruginosa challenge pneumonia model, conferring 100% survival upon prophylactic administration and reducing lung P. aeruginosa burden by up to 2 logs. At very low doses, they protected the mice from P. aeruginosa infection-related changes in lung histology, myeloperoxidase production, and lung weight. Importantly, the most potent Nanobody still conferred protection after therapeutic administration up to 24 h post-infection. The concept of screening such formatting libraries for potency improvement is applicable to other targets and biological therapeutic platforms.

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“…Nbs targeting CXCR7 possess antagonistic properties inhibiting CXCL12 binding and recruitment of b-arrestin. 19 Biparatopic Nbs, targeting the N-terminus and extracellular loops of CXCR7 showed enhanced potencies. Extending these biparatopic Nbs with a Nb targeting albumin resulted in Nbs with increased half-life in vivo.…”

Section: Friday November 25 2016mentioning

confidence: 99%

“…These trivalent Nbs reduced tumor growth in a patient-derived, CXCR7-expressing head and neck cancer xenograft model, through inhibition of angiogenesis. 19 Cancer types showing high expression of CXCR7 might thus be effectively targeted by CXCR7 Nbs.…”

Section: Friday November 25 2016mentioning

confidence: 99%

Antibodies targeting G protein-coupled receptors: Recent advances and therapeutic challenges

Ayoub

Crépieux

Koglin

et al. 2017

mAbs

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Le STUDIUM conference was held November 24-25, 2016 in Tours, France as a satellite workshop of the 5(th) meeting of the French GDR 3545 on "G Protein-Coupled Receptors (GPCRs) - From Physiology to Drugs", which was held in Tours during November 22-24, 2016. The conference gathered speakers from academia and industry considered to be world leaders in the molecular pharmacology and signaling of GPCRs, with a particular interest in the development of therapeutic GPCR antibodies (Abs). The main topics were new advances and challenges in the development of antibodies targeting GPCRs and their potential applications to the study of the structure and function of GPCRs, as well as their implication in physiology and pathophysiology. The conference included two sessions, with the first dedicated to the recent advances in methodological strategies used for GPCR immunization using thermo-stabilized and purified GPCRs, and the development of various formats of Abs such as monoclonal IgG, single-chain variable fragments and nanobodies (Nbs) by in vitro and in silico approaches. The second session focused on GPCR Nbs as a "hot" field of research on GPCRs. This session started with discussion of the pioneering Nbs developed against GPCRs and their application to structural studies, then transitioned to talks on original ex vivo and in vivo studies on GPCR-selective Nbs showing promising therapeutic applications of Nbs in important physiological systems, such as the central nervous and the immune systems, as well as in cancer. The conference ended with the consensus that Abs and especially Nbs are opening a new era of research on GPCR structure, pharmacology and pathophysiology

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Llama-derived Single Variable Domains (Nanobodies) Directed against Chemokine Receptor CXCR7 Reduce Head and Neck Cancer Cell Growth in Vivo

Cited by 126 publications

References 48 publications

Antigen recognition by single-domain antibodies: structural latitudes and constraints

Antigen recognition by single-domain antibodies: structural latitudes and constraints

High Throughput Combinatorial Formatting of PcrV Nanobodies for Efficient Potency Improvement

Antibodies targeting G protein-coupled receptors: Recent advances and therapeutic challenges

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