title: Human anti-nucleolin recombinant immunoagent for cancer therapy.
abstract: Nucleolin (NCL) is a nucleocytoplasmic protein involved in many biological processes, such as ribosomal assembly, rRNA processing, and mRNA stabilization. NCL also regulates the biogenesis of specific microRNAs (miRNAs) involved in tumor development and aggressiveness. Interestingly, NCL is expressed on the surface of actively proliferating cancer cells, but not on their normal counterparts. Therefore, NCL is an attractive target for antineoplastic treatments. Taking advantage of phage-display technology, we engineered a fully human single-chain fragment variable, named 4LB5. This immunoagent binds NCL on the cell surface, it is translocated into the cytoplasm of target cells, and it abrogates the biogenesis of NCL-dependent miRNAs. Binding of 4LB5 to NCL on the cell surface of a variety of breast cancer and hepatocellular carcinoma cell lines, but not to normal-like MCF-10a breast cells, dramatically reduces cancer cell viability and proliferation. Finally, in orthotopic breast cancer mouse models, 4LB5 administration results in a significant reduction of the tumor volume without evident side effects. In summary, here we describe, to our knowledge, the first anti-NCL single-chain fragment variable displaying antineoplastic activity against established solid tumors, which could represent the prototype of novel immune-based NCL-targeting drugs with clinical potential as diagnostic and therapeutic tools in a wide variety of human cancers.
authors.affiliation: Department of Molecular Virology, Immunology and Medical Genetics, The Ohio State University, Columbus, OH 43210;
authors.forename: Dario
authors.initials: D
authors.lastname: Palmieri
authors.affiliation: Department of Molecular Virology, Immunology and Medical Genetics, The Ohio State University, Columbus, OH 43210;
authors.forename: Timothy
authors.initials: T
authors.lastname: Richmond
authors.affiliation: Department of Molecular Virology, Immunology and Medical Genetics, The Ohio State University, Columbus, OH 43210;
authors.forename: Claudia
authors.initials: C
authors.lastname: Piovan
authors.affiliation: Department of Molecular Virology, Immunology and Medical Genetics, The Ohio State University, Columbus, OH 43210;
authors.forename: Tyler
authors.initials: T
authors.lastname: Sheetz
authors.affiliation: Department of Molecular Virology, Immunology and Medical Genetics, The Ohio State University, Columbus, OH 43210;
authors.forename: Nicola
authors.initials: N
authors.lastname: Zanesi
authors.affiliation: Centro di Ingegneria Genetica (CEINGE) Biotecnologie Avanzate, 80145 Naples, Italy;
authors.forename: Fulvia
authors.initials: F
authors.lastname: Troise
authors.affiliation: Department of Mass Spectroscopy and Proteomics, The Ohio State University, Columbus, OH 43210;
authors.forename: Cindy
authors.initials: C
authors.lastname: James
authors.affiliation: Department of Molecular Virology, Immunology and Medical Genetics, The Ohio State University, Columbus, OH 43210;
authors.forename: Dorothee
authors.initials: D
authors.lastname: Wernicke
authors.affiliation: Department of Molecular Virology, Immunology and Medical Genetics, The Ohio State University, Columbus, OH 43210;
authors.forename: Fata
authors.initials: F
authors.lastname: Nyei
authors.affiliation: Department of Veterinary Biosciences, The Ohio State University, Columbus, OH 43210;
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Gordon
authors.affiliation: Department of Molecular Virology, Immunology and Medical Genetics, The Ohio State University, Columbus, OH 43210;
authors.forename: Jessica
authors.initials: J
authors.lastname: Consiglio
authors.affiliation: Centro di Ingegneria Genetica (CEINGE) Biotecnologie Avanzate, 80145 Naples, Italy;
authors.forename: Francesco
authors.initials: F
authors.lastname: Salvatore
authors.affiliation: Department of Molecular Virology, Immunology and Medical Genetics, The Ohio State University, Columbus, OH 43210;
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Coppola
authors.affiliation: Department of Internal Medicine, Division of Hematology, The Ohio State University, Columbus, OH 43210;
authors.forename: Flavia
authors.initials: F
authors.lastname: Pichiorri
authors.affiliation: Centro di Ingegneria Genetica (CEINGE) Biotecnologie Avanzate, 80145 Naples, Italy; Department of Molecular Medicine and Medical Biotechnologies, University of Napoli "Federico II", 80131 Naples, Italy carlo.croce@osumc.edu cladelor@unina.it.
authors.forename: Claudia
authors.initials: C
authors.lastname: De Lorenzo
authors.affiliation: Department of Molecular Virology, Immunology and Medical Genetics, The Ohio State University, Columbus, OH 43210; carlo.croce@osumc.edu cladelor@unina.it.
authors.forename: Carlo M
authors.initials: CM
authors.lastname: Croce
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry*; D017209:Apoptosis; D006528:Carcinoma, Hepatocellular / Q000378:metabolism; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002465:Cell Movement; D049109:Cell Proliferation; D002470:Cell Survival; D003593:Cytoplasm / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism; D051379:Mice; D016513:Mice, SCID; D009368:Neoplasm Transplantation; D009369:Neoplasms / Q000276:immunology* / Q000378:metabolism / Q000628:therapy*; D019151:Peptide Library; D010750:Phosphoproteins / Q000737:chemistry*; D015202:Protein Engineering; D016601:RNA-Binding Proteins / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D057127:Single-Chain Antibodies / Q000737:chemistry*; D000097796:Nucleolin
chemical_list: D000970:Antineoplastic Agents; D019151:Peptide Library; D010750:Phosphoproteins; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D000097796:Nucleolin
keywords: cancer immunotherapy; microRNA; nucleolin; phage-display; scFv