title: Sindbis Virus-Pseudotyped Lentiviral Vectors Carrying VEGFR2-Specific Nanobody for Potential Transductional Targeting of Tumor Vasculature.
abstract: Introduction of selectivity/specificity into viral-based gene delivery systems, such as lentiviral vectors (LVs), is crucial in their systemic administration for cancer gene therapy. The pivotal role of tumor-associated endothelial cells (TAECs) in tumor angiogenesis and overexpression of vascular endothelial growth factor receptor-2 (VEGFR2 or KDR) in TAECs makes them a potent target in cancer treatment. Herein, we report the development of VEGFR2-targeted LVs pseudotyped with chimeric sindbis virus E2 glycoprotein (cSVE2s). For this purpose, either sequence of a VEGFR2-specific nanobody or its natural ligand (VEGF121) was inserted into the binding site of sindbis virus E2 glycoprotein. In silico modeling data suggested that the inserted targeting motifs were exposed in the context of cSVE2s. Western blot analysis of LVs indicated the incorporation of cSVE2s into viral particles. Capture ELISA demonstrated the specificity/functionality of the incorporated cSVE2s. Transduction of 293/KDR (expressing VEGFR2) or 293T cells (negative control) by constructed LVs followed by fluorescent microscopy and flow cytometric analyses indicated selective transduction of 293/KDR cells (30 %) by both targeting motifs compared to 293T control cells (1-2 %). These results implied similar targeting properties of VEGFR2-specific nanobody compared to the VEGF121and indicated the potential for transductional targeting of tumor vasculature by the nanobody displaying LVs.
authors.affiliation: Department of Virology, Pasteur Institute of Iran, 69 Pasteur Avenue, Kargar Avenue, Tehran, 1316943551, Iran.
authors.forename: Roshank
authors.initials: R
authors.lastname: Ahani
authors.affiliation: Department of Virology, Pasteur Institute of Iran, 69 Pasteur Avenue, Kargar Avenue, Tehran, 1316943551, Iran. rfarzin@pasteur.ac.ir.
authors.forename: Farzin
authors.initials: F
authors.lastname: Roohvand
authors.affiliation: New Technologies Research Group, Department of Pilot Nanobiotechnology, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Reza Ahangari
authors.initials: RA
authors.lastname: Cohan
authors.affiliation: Department of Virology, Pasteur Institute of Iran, 69 Pasteur Avenue, Kargar Avenue, Tehran, 1316943551, Iran.
authors.forename: Mohammad Hossein
authors.initials: MH
authors.lastname: Etemadzadeh
authors.affiliation: Department of Virology, Pasteur Institute of Iran, 69 Pasteur Avenue, Kargar Avenue, Tehran, 1316943551, Iran.
authors.forename: Nasir
authors.initials: N
authors.lastname: Mohajel
authors.affiliation: Biotechnology Research Center, Venom & Biotherapeutics Molecules Lab, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Mahdi
authors.initials: M
authors.lastname: Behdani
authors.affiliation: Department of Virology, Pasteur Institute of Iran, 69 Pasteur Avenue, Kargar Avenue, Tehran, 1316943551, Iran.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Shahosseini
authors.affiliation: Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA, 02215, USA.|Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA, USA.
authors.forename: Navid
authors.initials: N
authors.lastname: Madani
authors.affiliation: Department of Virology, Pasteur Institute of Iran, 69 Pasteur Avenue, Kargar Avenue, Tehran, 1316943551, Iran. k.azadmanesh@gmail.com.
authors.forename: Kayhan
authors.initials: K
authors.lastname: Azadmanesh
mesh_terms: D003198:Computer Simulation; D018390:Gene Targeting; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008958:Models, Molecular; D012845:Sindbis Virus / Q000235:genetics / Q000378:metabolism*; D061905:Single-Domain Antibodies / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014161:Transduction, Genetic; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D061905:Single-Domain Antibodies; D014759:Viral Envelope Proteins; C064950:glycoprotein E2, Sindbis virus; C000588866:KDR protein, human; D040301:Vascular Endothelial Growth Factor Receptor-2
keywords: Angiogenesis; Cancer gene therapy; Lentiviral vectors; Nanobody; Pseudotyping; Transductional targeting; VEGFR2