title: Preclinical Benefit of Hypoxia-Activated Intra-arterial Therapy with Evofosfamide in Liver Cancer.
abstract: OBJECTIVE
  To evaluate safety and characterize anticancer efficacy of hepatic hypoxia-activated intra-arterial therapy (HAIAT) with evofosfamide in a rabbit model.
  
  
  METHODS
  VX2-tumor-bearing rabbits were assigned to 4 intra-arterial therapy (IAT) groups (n = 7/group): (i) saline (control); (ii) evofosfamide (Evo); (iii) doxorubicin-lipiodol emulsion followed by embolization with 100-300 μm beads (conventional, cTACE); or (iv) cTACE and evofosfamide (cTACE + Evo). Blood samples were collected pre-IAT and 1, 2, 7, and 14 days post-IAT. A semiquantitative scoring system assessed hepatocellular damage. Tumor volumes were segmented on multidetector CT (baseline, 7/14 days post-IAT). Pathologic tumor necrosis was quantified using manual segmentation on whole-slide images. Hypoxic fraction (HF) and compartment (HC) were determined by pimonidazole staining. Tumor DNA damage, apoptosis, cell proliferation, endogenous hypoxia, and metabolism were quantified (γ-H2AX, Annexin V, caspase-3, Ki-67, HIF1α, VEGF, GAPDH, MCT4, and LDH).
  
  
  RESULTS
  cTACE + Evo showed a similar profile of liver enzymes elevation and pathologic scores compared with cTACE. Neither hematologic nor renal toxicity were observed. Animals treated with cTACE + Evo demonstrated smaller tumor volumes, lower tumor growth rates, and higher necrotic fractions compared with cTACE. cTACE + Evo resulted in a marked reduction in the HF and HC. Correlation was observed between decreases in HF or HC and tumor necrosis. cTACE + Evo promoted antitumor effects as evidenced by increased expression of γ-H2AX, apoptotic biomarkers, and decreased cell proliferation. Increased HIF1α/VEGF expression and tumor glycolysis supported HAIAT.
  
  
  CONCLUSIONS
  HAIAT achieved a promising step towards the locoregional targeting of tumor hypoxia. The favorable toxicity profile and enhanced anticancer effects of evofosfamide in combination with cTACE pave the way towards clinical trials in patients with liver cancer. Clin Cancer Res; 23(2); 536-48. ©2016 AACR.
authors.affiliation: Russell H. Morgan Department of Radiology and Radiological Science, Division of Vascular and Interventional Radiology, The Johns Hopkins Hospital, Baltimore, Maryland.|Department of Radiology and Biomedical Imaging, Yale University School of Medicine, New Haven, Connecticut.
authors.forename: Rafael
authors.initials: R
authors.lastname: Duran
authors.affiliation: Russell H. Morgan Department of Radiology and Radiological Science, Division of Vascular and Interventional Radiology, The Johns Hopkins Hospital, Baltimore, Maryland.
authors.forename: Sahar
authors.initials: S
authors.lastname: Mirpour
authors.affiliation: Russell H. Morgan Department of Radiology and Radiological Science, Division of Vascular and Interventional Radiology, The Johns Hopkins Hospital, Baltimore, Maryland.
authors.forename: Vasily
authors.initials: V
authors.lastname: Pekurovsky
authors.affiliation: Russell H. Morgan Department of Radiology and Radiological Science, Division of Vascular and Interventional Radiology, The Johns Hopkins Hospital, Baltimore, Maryland.
authors.forename: Shanmugasundaram
authors.initials: S
authors.lastname: Ganapathy-Kanniappan
authors.affiliation: Department of Molecular and Comparative Pathobiology, The Johns Hopkins University School of Medicine, Baltimore, Maryland.
authors.forename: Cory F
authors.initials: CF
authors.lastname: Brayton
authors.affiliation: Department of Pathology, Division of Gastrointestinal and Liver Pathology, The Johns Hopkins University School of Medicine, Baltimore, Maryland.
authors.forename: Toby C
authors.initials: TC
authors.lastname: Cornish
authors.affiliation: Russell H. Morgan Department of Radiology and Radiological Science, Division of Vascular and Interventional Radiology, The Johns Hopkins Hospital, Baltimore, Maryland.
authors.forename: Boris
authors.initials: B
authors.lastname: Gorodetski
authors.affiliation: Department of Radiation Oncology and Molecular Radiation Sciences, The Johns Hopkins Hospital, Baltimore, Maryland.
authors.forename: Juvenal
authors.initials: J
authors.lastname: Reyes
authors.affiliation: Russell H. Morgan Department of Radiology and Radiological Science, Division of Vascular and Interventional Radiology, The Johns Hopkins Hospital, Baltimore, Maryland.|Department of Radiology and Biomedical Imaging, Yale University School of Medicine, New Haven, Connecticut.
authors.forename: Julius
authors.initials: J
authors.lastname: Chapiro
authors.affiliation: Russell H. Morgan Department of Radiology and Radiological Science, Division of Vascular and Interventional Radiology, The Johns Hopkins Hospital, Baltimore, Maryland.|Department of Radiology and Biomedical Imaging, Yale University School of Medicine, New Haven, Connecticut.
authors.forename: Rüdiger E
authors.initials: RE
authors.lastname: Schernthaner
authors.affiliation: Department of Biostatistics, The Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland.
authors.forename: Constantine
authors.initials: C
authors.lastname: Frangakis
authors.affiliation: Department of Radiology and Biomedical Imaging, Yale University School of Medicine, New Haven, Connecticut.|U/S Imaging and Interventions (UII), Philips Research North America, Cambridge, Massachusetts.
authors.forename: MingDe
authors.initials: M
authors.lastname: Lin
authors.affiliation: Threshold Pharmaceuticals, South San Francisco, California.
authors.forename: Jessica D
authors.initials: JD
authors.lastname: Sun
authors.affiliation: Threshold Pharmaceuticals, South San Francisco, California.
authors.forename: Charles P
authors.initials: CP
authors.lastname: Hart
authors.affiliation: Department of Radiology and Biomedical Imaging, Yale University School of Medicine, New Haven, Connecticut. jeff.geschwind@yale.edu.
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Geschwind
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D014408:Biomarkers, Tumor / Q000235:genetics*; D049109:Cell Proliferation / Q000187:drug effects; D000074141:Circulating Tumor DNA / Q000235:genetics; D003131:Combined Modality Therapy; D004195:Disease Models, Animal; D004317:Doxorubicin / Q000008:administration & dosage; D004998:Ethiodized Oil / Q000008:administration & dosage; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D009593:Nitroimidazoles / Q000008:administration & dosage*; D010752:Phosphoramide Mustards / Q000008:administration & dosage*; D011817:Rabbits; D000072258:Tumor Hypoxia
chemical_list: D014408:Biomarkers, Tumor; D000074141:Circulating Tumor DNA; D009593:Nitroimidazoles; D010752:Phosphoramide Mustards; C552526:TH 302; D004998:Ethiodized Oil; D004317:Doxorubicin