Chemotrap-1: an engineered soluble receptor that blocks chemokine-induced migration of metastatic cancer cells in vivo.
Cancer and dendritic cells recognize and migrate toward chemokines secreted from lymphatics and use this mechanism to invade the lymphatic system, and cancer cells metastasize through it. The lymphatic-secreted chemokine ligand CCL21 has been identified as a key regulatory molecule in the switch to a metastatic phenotype in melanoma and breast cancer cells. However, it is not known whether CCL21 inhibition is a potential therapeutic strategy for inhibition of metastasis. Here, we describe an engineered CCL21-soluble inhibitor, Chemotrap-1, which inhibits migration of metastatic melanoma cells in vivo. Two-hybrid, pull-down, and coimmunoprecipitation assays allowed us to identify a naturally occurring human zinc finger protein with CCL21 chemokine-binding properties. Further analyses revealed a short peptide (∼70 amino acids), with a predicted coiled-coil structure, which is sufficient for association with CCL21. This CCL21 chemokine-binding peptide was then fused to the Fc region of human IgG1 to generate Chemotrap-1, a human chemokine-binding Fc fusion protein. Surface plasmon resonance and chemotaxis assays showed that Chemotrap-1 binds CCL21 and inhibits CCL21-induced migration of melanoma cells in vitro with subnanomolar affinity. In addition, Chemotrap-1 blocked migration of melanoma cells toward lymphatic endothelial cells in vitro and in vivo. Finally, Chemotrap-1 strongly reduced lymphatic invasion, tracking, and metastasis of CCR7-expressing melanoma cells in vivo. Together, these results show that CCL21 chemokine inhibition by Chemotrap-1 is a potential therapeutic strategy for metastasis and provide further support for the hypothesis that lymphatic-mediated metastasis is a chemokine-dependent process.
Lanati
Silvia
S
Microvascular Research Laboratories, Bristol Heart Institute, Department of Physiology and Pharmacology, School of Veterinary Sciences, University of Bristol, Bristol, United Kingdom.
Dunn
Darryl B
DB
Roussigné
Myriam
M
Emmett
Maxine S
MS
Carriere
Virginie
V
Jullien
Denis
D
Budge
Jessica
J
Fryer
Justin
J
Erard
Monique
M
Cailler
Françoise
F
Girard
Jean-Phillippe
JP
Bates
David O
DO
D001665:Binding Sites; D002465:Cell Movement; D018932:Chemokine CCL2 / Q000378:metabolism; D054421:Chemokine CCL21 / Q000235:genetics / Q000378:metabolism; D018925:Chemokines / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002633:Chemotaxis / Q000502:physiology; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics / Q000378:metabolism*; D008207:Lymphatic Metastasis / Q000517:prevention & control; D008545:Melanoma / Q000378:metabolism / Q000473:pathology*; D009369:Neoplasms / Q000473:pathology; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000378:metabolism / Q000494:pharmacology*
C515478:CCL21 protein, human; D018932:Chemokine CCL2; D054421:Chemokine CCL21; D018925:Chemokines; D007141:Immunoglobulin Fc Fragments; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase