title: Llama-derived single variable domains (nanobodies) directed against chemokine receptor CXCR7 reduce head and neck cancer cell growth in vivo.
abstract: The chemokine receptor CXCR7, belonging to the membrane-bound G protein-coupled receptor superfamily, is expressed in several tumor types. Inhibition of CXCR7 with either small molecules or small interference (si)RNA has shown promising therapeutic benefits in several tumor models. With the increased interest and effectiveness of biologicals inhibiting membrane-bound receptors we made use of the "Nanobody platform" to target CXCR7. Previously we showed that Nanobodies, i.e. immunoglobulin single variable domains derived from naturally occurring heavy chain-only camelids antibodies, represent new biological tools to efficiently tackle difficult drug targets such as G protein-coupled receptors. In this study we developed and characterized highly selective and potent Nanobodies against CXCR7. Interestingly, the CXCR7-targeting Nanobodies displayed antagonistic properties in contrast with previously reported CXCR7-targeting agents. Several high affinity CXCR7-specific Nanobodies potently inhibited CXCL12-induced β-arrestin2 recruitment in vitro. A wide variety of tumor biopsies was profiled, showing for the first time high expression of CXCR7 in head and neck cancer. Using a patient-derived CXCR7-expressing head and neck cancer xenograft model in nude mice, tumor growth was inhibited by CXCR7-targeting Nanobody therapy. Mechanistically, CXCR7-targeting Nanobodies did not inhibit cell cycle progression but instead reduced secretion of the angiogenic chemokine CXCL1 from head and neck cancer cells in vitro, thus acting here as inverse agonists, and subsequent angiogenesis in vivo. Hence, with this novel class of CXCR7 inhibitors, we further substantiate the therapeutic relevance of targeting CXCR7 in head and neck cancer.
authors: From the Amsterdam Institute for Molecules Medicines and Systems, Division of Medicinal Chemistry, Faculty of Sciences, VU University Amsterdam, 1081 HV Amsterdam, The Netherlands.
authors: David
authors: D
authors: Maussang
authors: Azra
authors: A
authors: Mujić-Delić
authors: Francis J
authors: FJ
authors: Descamps
authors: Catelijne
authors: C
authors: Stortelers
authors: Peter
authors: P
authors: Vanlandschoot
authors: Marijke
authors: M
authors: Stigter-van Walsum
authors: Henry F
authors: HF
authors: Vischer
authors: Maarten
authors: M
authors: van Roy
authors: Maria
authors: M
authors: Vosjan
authors: Maria
authors: M
authors: Gonzalez-Pajuelo
authors: Guus A M S
authors: GA
authors: van Dongen
authors: Pascal
authors: P
authors: Merchiers
authors: Philippe
authors: P
authors: van Rompaey
authors: Martine J
authors: MJ
authors: Smit
mesh_terms: D000818:Animals; D019390:Arrestins / Q000276:immunology / Q000378:metabolism; D001667:Binding, Competitive / Q000276:immunology; D002161:Camelids, New World / Q000276:immunology; D045744:Cell Line, Tumor; D054377:Chemokine CXCL12 / Q000494:pharmacology; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D057809:HEK293 Cells; D006258:Head and Neck Neoplasms / Q000276:immunology* / Q000473:pathology / Q000517:prevention & control; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D041681:NIH 3T3 Cells; D011869:Radioligand Assay; D054387:Receptors, CXCR / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects / Q000276:immunology; D061905:Single-Domain Antibodies / Q000276:immunology* / Q000494:pharmacology; D047368:Tumor Burden / Q000187:drug effects / Q000276:immunology; D023041:Xenograft Model Antitumor Assays; D000071557:beta-Arrestins
chemical_list: C501896:ACKR3 protein, human; D019390:Arrestins; D054377:Chemokine CXCL12; D054387:Receptors, CXCR; D061905:Single-Domain Antibodies; D000071557:beta-Arrestins
keywords: Antibody Engineering; CXCR7; Chemokines; G Protein-coupled Receptors (GPCR); Head and Neck Cancer; Nanobodies; Signaling
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