title: The myeloid-binding peptide adenoviral vector enables multi-organ vascular endothelial gene targeting.
abstract: Vascular endothelial cells (ECs) are ideal gene therapy targets as they provide widespread tissue access and are the first contact surfaces following intravenous vector administration. Human recombinant adenovirus serotype 5 (Ad5) is the most frequently used gene transfer system because of its appreciable transgene payload capacity and lack of somatic mutation risk. However, standard Ad5 vectors predominantly transduce liver but not the vasculature following intravenous administration. We recently developed an Ad5 vector with a myeloid cell-binding peptide (MBP) incorporated into the knob-deleted, T4 fibritin chimeric fiber (Ad.MBP). This vector was shown to transduce pulmonary ECs presumably via a vector handoff mechanism. Here we tested the body-wide tropism of the Ad.MBP vector, its myeloid cell necessity, and vector-EC expression dose response. Using comprehensive multi-organ co-immunofluorescence analysis, we discovered that Ad.MBP produced widespread EC transduction in the lung, heart, kidney, skeletal muscle, pancreas, small bowel, and brain. Surprisingly, Ad.MBP retained hepatocyte tropism albeit at a reduced frequency compared with the standard Ad5. While binding specifically to myeloid cells ex vivo, multi-organ Ad.MBP expression was not dependent on circulating monocytes or macrophages. Ad.MBP dose de-escalation maintained full lung-targeting capacity but drastically reduced transgene expression in other organs. Swapping the EC-specific ROBO4 for the CMV promoter/enhancer abrogated hepatocyte expression but also reduced gene expression in other organs. Collectively, our multilevel targeting strategy could enable therapeutic biological production in previously inaccessible organs that pertain to the most debilitating or lethal human diseases.
authors: Urology Division, Department of Surgery, Washington University School of Medicine, St Louis, MO, USA.
authors: Zhi Hong
authors: ZH
authors: Lu
authors: Department of Radiation Oncology and Biological Therapeutics Center, Washington University School of Medicine, St Louis, MO, USA.
authors: Sergey
authors: S
authors: Kaliberov
authors: Urology Division, Department of Surgery, Washington University School of Medicine, St Louis, MO, USA.
authors: Jingzhu
authors: J
authors: Zhang
authors: Cancer Biology Division, Department of Radiation Oncology, Washington University School of Medicine, St Louis, MO, USA.
authors: Barbara
authors: B
authors: Muz
authors: Cancer Biology Division, Department of Radiation Oncology, Washington University School of Medicine, St Louis, MO, USA.
authors: Abdel K
authors: AK
authors: Azab
authors: Urology Division, Department of Surgery, Washington University School of Medicine, St Louis, MO, USA.
authors: Rebecca E
authors: RE
authors: Sohn
authors: Department of Radiation Oncology and Biological Therapeutics Center, Washington University School of Medicine, St Louis, MO, USA.
authors: Lyudmila
authors: L
authors: Kaliberova
authors: Urology Division, Department of Surgery, Washington University School of Medicine, St Louis, MO, USA.
authors: Yingqiu
authors: Y
authors: Du
authors: 1] Department of Radiation Oncology and Biological Therapeutics Center, Washington University School of Medicine, St Louis, MO, USA [2] Siteman Cancer Center, Washington University School of Medicine, St Louis, MO, USA.
authors: David T
authors: DT
authors: Curiel
authors: 1] Urology Division, Department of Surgery, Washington University School of Medicine, St Louis, MO, USA [2] Siteman Cancer Center, Washington University School of Medicine, St Louis, MO, USA [3] Cell Biology Department, Washington University School of Medicine, St Louis, MO, USA.
authors: Jeffrey M
authors: JM
authors: Arbeit
mesh_terms: D000256:Adenoviridae / Q000235:genetics* / Q000502:physiology; D000818:Animals; D003587:Cytomegalovirus / Q000235:genetics; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism* / Q000821:virology; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage / Q000502:physiology*; D049452:Green Fluorescent Proteins / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism / Q000821:virology; D006801:Humans; D007275:Injections, Intravenous; D051379:Mice; D008810:Mice, Inbred C57BL; D022423:Myeloid Cells / Q000166:cytology / Q000378:metabolism / Q000821:virology; D010455:Peptides / Q000008:administration & dosage / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011956:Receptors, Cell Surface / Q000008:administration & dosage* / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000378:metabolism; D014764:Viral Proteins / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism; D056189:Viral Tropism; D014771:Virion / Q000502:physiology; D000097905:Roundabout Proteins
chemical_list: D049452:Green Fluorescent Proteins; D010455:Peptides; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; C462951:ROBO4 protein, human; D000097905:Roundabout Proteins; C485184:enhanced green fluorescent protein; C057839:fibritin protein, Enterobacteria phage T4