title: Transcriptional targeting of primary and metastatic tumor neovasculature by an adenoviral type 5 roundabout4 vector in mice.
abstract: New approaches targeting metastatic neovasculature are needed. Payload capacity, cellular transduction efficiency, and first-pass cellular uptake following systemic vector administration, motivates persistent interest in tumor vascular endothelial cell (EC) adenoviral (Ad) vector targeting. While EC transductional and transcriptional targeting has been accomplished, vector administration approaches of limited clinical utility, lack of tumor-wide EC expression quantification, and failure to address avid liver sequestration, challenged prior work. Here, we intravenously injected an Ad vector containing 3 kb of the human roundabout4 (ROBO4) enhancer/promoter transcriptionally regulating an enhanced green fluorescent protein (EGFP) reporter into immunodeficient mice bearing 786-O renal cell carcinoma subcutaneous (SC) xenografts and kidney orthotopic (KO) tumors. Initial experiments performed in human coxsackie virus and adenovirus receptor (hCAR) transgenic:Rag2 knockout mice revealed multiple ECs with high-level Ad5ROBO4-EGFP expression throughout KO and SC tumors. In contrast, Ad5CMV-EGFP was sporadically expressed in a few tumor vascular ECs and stromal cells. As the hCAR transgene also facilitated Ad5ROBO4 and control Ad5CMV vector EC expression in multiple host organs, follow-on experiments engaged warfarin-mediated liver vector detargeting in hCAR non-transgenic mice. Ad5ROBO4-mediated EC expression was undetectable in most host organs, while the frequencies of vector expressing intratumoral vessels and whole tumor EGFP protein levels remained elevated. In contrast, AdCMV vector expression was only detectable in one or two stromal cells throughout the whole tumor. The Ad5ROBO4 vector, in conjunction with liver detargeting, provides tractable genetic access for in-vivo EC genetic engineering in malignancies.
authors.affiliation: Urology Division and Department of Surgery, Washington University in St. Louis Medical School, St. Louis, Missouri, United States of America.
authors.forename: Zhi Hong
authors.initials: ZH
authors.lastname: Lu
authors.affiliation: Department of Radiation Oncology and Biological Therapeutics Center, Washington University in St. Louis Medical School, St. Louis, Missouri, United States of America.
authors.forename: Sergey
authors.initials: S
authors.lastname: Kaliberov
authors.affiliation: Urology Division and Department of Surgery, Washington University in St. Louis Medical School, St. Louis, Missouri, United States of America.
authors.forename: Rebecca E
authors.initials: RE
authors.lastname: Sohn
authors.affiliation: Department of Radiation Oncology and Biological Therapeutics Center, Washington University in St. Louis Medical School, St. Louis, Missouri, United States of America.
authors.forename: Lyudmila
authors.initials: L
authors.lastname: Kaliberova
authors.affiliation: Department of Radiation Oncology and Biological Therapeutics Center, Washington University in St. Louis Medical School, St. Louis, Missouri, United States of America ; Siteman Cancer Center, Washington University in St. Louis Medical School, St. Louis, Missouri, United States of America.
authors.forename: David T
authors.initials: DT
authors.lastname: Curiel
authors.affiliation: Urology Division and Department of Surgery, Washington University in St. Louis Medical School, St. Louis, Missouri, United States of America ; Siteman Cancer Center, Washington University in St. Louis Medical School, St. Louis, Missouri, United States of America ; Cell Biology Department, Washington University in St. Louis Medical School, St. Louis, Missouri, United States of America.
authors.forename: Jeffrey M
authors.initials: JM
authors.lastname: Arbeit
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D045744:Cell Line, Tumor; D002471:Cell Transformation, Neoplastic; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein / Q000235:genetics; D042783:Endothelial Cells / Q000378:metabolism / Q000473:pathology; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D007680:Kidney Neoplasms / Q000098:blood supply / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D009362:Neoplasm Metastasis; D009389:Neovascularization, Pathologic; D011956:Receptors, Cell Surface / Q000235:genetics*; D014158:Transcription, Genetic / Q000235:genetics*; D019076:Transgenes / Q000235:genetics; D014859:Warfarin / Q000494:pharmacology; D000097905:Roundabout Proteins
chemical_list: D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D011956:Receptors, Cell Surface; D014859:Warfarin; C568771:CLMP protein, human; C462951:ROBO4 protein, human; D000097905:Roundabout Proteins