title: Rationally designed chemokine-based toxin targeting the viral G protein-coupled receptor US28 potently inhibits cytomegalovirus infection in vivo.
abstract: The use of receptor-ligand interactions to direct toxins to kill diseased cells selectively has shown considerable promise for treatment of a number of cancers and, more recently, autoimmune disease. Here we move the fusion toxin protein (FTP) technology beyond cancer/autoimmune therapeutics to target the human viral pathogen, human cytomegalovirus (HCMV), on the basis of its expression of the 7TM G protein-coupled chemokine receptor US28. The virus origin of US28 provides an exceptional chemokine-binding profile with high selectivity and improved binding for the CX3C chemokine, CX3CL1. Moreover, US28 is constitutively internalizing by nature, providing highly effective FTP delivery. We designed a synthetic CX3CL1 variant engineered to have ultra-high affinity for US28 and greater specificity for US28 than the natural sole receptor for CX3CL1, CX3CR1, and we fused the synthetic variant with the cytotoxic domain of Pseudomonas Exotoxin A. This novel strategy of a rationally designed FTP provided unparalleled anti-HCMV efficacy and potency in vitro and in vivo.
authors.affiliation: INAGEN ApS., DK-2800 Kongens Lyngby, Denmark; Laboratory for Molecular Pharmacology, Department of Neuroscience and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, 2200 Copenhagen, Denmark;
authors.forename: Katja
authors.initials: K
authors.lastname: Spiess
authors.affiliation: INAGEN ApS., DK-2800 Kongens Lyngby, Denmark;
authors.forename: Mads G
authors.initials: MG
authors.lastname: Jeppesen
authors.affiliation: INAGEN ApS., DK-2800 Kongens Lyngby, Denmark;
authors.forename: Mikkel
authors.initials: M
authors.lastname: Malmgaard-Clausen
authors.affiliation: INAGEN ApS., DK-2800 Kongens Lyngby, Denmark;
authors.forename: Karen
authors.initials: K
authors.lastname: Krzywkowski
authors.affiliation: Department of Microbiology and Molecular Genetics, Rutgers-New Jersey Medical School, Newark, NJ 07184;
authors.forename: Kalpana
authors.initials: K
authors.lastname: Dulal
authors.affiliation: National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Sciences, Xiamen University, 361100 Fujian Province, China;
authors.forename: Tong
authors.initials: T
authors.lastname: Cheng
authors.affiliation: Laboratory for Molecular Pharmacology, Department of Neuroscience and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, 2200 Copenhagen, Denmark;
authors.forename: Gertrud M
authors.initials: GM
authors.lastname: Hjortø
authors.affiliation: Laboratory for Molecular Pharmacology, Department of Neuroscience and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, 2200 Copenhagen, Denmark;
authors.forename: Olav
authors.initials: O
authors.lastname: Larsen
authors.affiliation: Stanford University School of Medicine, Stanford, CA 94305; Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, CA 94305;
authors.forename: John S
authors.initials: JS
authors.lastname: Burg
authors.affiliation: School of Biomedical and Healthcare Sciences, University of Plymouth, Plymouth PL4 8AA, United Kingdom.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Jarvis
authors.affiliation: Stanford University School of Medicine, Stanford, CA 94305; Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, CA 94305; kcgarcia@stanford.edu tnkl@vet.dtu.dk rosenkilde@sund.ku.dk.
authors.forename: K Christopher
authors.initials: KC
authors.lastname: Garcia
authors.affiliation: Department of Microbiology and Molecular Genetics, Rutgers-New Jersey Medical School, Newark, NJ 07184;
authors.forename: Hua
authors.initials: H
authors.lastname: Zhu
authors.affiliation: INAGEN ApS., DK-2800 Kongens Lyngby, Denmark; kcgarcia@stanford.edu tnkl@vet.dtu.dk rosenkilde@sund.ku.dk.
authors.forename: Thomas N
authors.initials: TN
authors.lastname: Kledal
authors.affiliation: Laboratory for Molecular Pharmacology, Department of Neuroscience and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, 2200 Copenhagen, Denmark; kcgarcia@stanford.edu tnkl@vet.dtu.dk rosenkilde@sund.ku.dk.
authors.forename: Mette M
authors.initials: MM
authors.lastname: Rosenkilde
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D054428:Chemokine CX3CL1 / Q000235:genetics / Q000378:metabolism*; D003587:Cytomegalovirus / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D003586:Cytomegalovirus Infections / Q000517:prevention & control* / Q000821:virology; D004305:Dose-Response Relationship, Drug; D015195:Drug Design; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism / Q000821:virology; D057809:HEK293 Cells; D054884:Host-Pathogen Interactions / Q000187:drug effects; D006801:Humans; D008168:Lung / Q000166:cytology; D011485:Protein Binding / Q000187:drug effects; D019707:Receptors, Chemokine / Q000037:antagonists & inhibitors* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D013997:Time Factors; D014764:Viral Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D054428:Chemokine CX3CL1; D019707:Receptors, Chemokine; D011993:Recombinant Fusion Proteins; C411191:US28 receptor, Cytomegalovirus; D014764:Viral Proteins; C111749:pseudomonas exoprotein A protein, Pseudomonas aeruginosa
keywords: 7TM GPCR; HCMV; antiviral drug; chemokine; protein engineering