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VEGFR2-Targeted Contrast-Enhanced Ultrasound to Distinguish between Two Anti-Angiogenic Treatments

      Abstract

      The aim of this study was to evaluate the capacity of BR55, an ultrasound contrast agent specifically targeting vascular endothelial growth factor receptor 2 (VEGFR2), to distinguish the specific anti-VEGFR2 therapy effect of sunitinib from other anti-angiogenic effects of a therapy (imatinib) that does not directly inhibit VEGFR2. Sunitinib, imatinib and placebo were administered daily for 11 d (264 h) to 45 BalbC mice bearing ectopic CT26 murine colorectal carcinomas. During the course of therapy, B-mode ultrasound, contrast-enhanced ultrasound and VEGFR2-targeted contrast-enhanced ultrasound were performed to assess tumor morphology, vascularization and VEGFR2 expression, respectively. The angiogenic effects on these three aspects were characterized using tumor volume, contrast-enhanced area and differential targeted enhancement. Necrosis, microvasculature and expression of VEGFR2 were also determined by histology and immunostaining. B-Mode imaging revealed that tumor growth was significantly decreased in sunitinib-treated mice at day 11 (p < 0.05), whereas imatinib did not affect growth. Functional evaluation revealed that the contrast-enhanced area decreased significantly (p < 0.02) and by similar amounts under both anti-angiogenic treatments by day 8 (192 h): −23% for imatinib and −21% for sunitinib. No significant decrease was observed in the placebo group. Targeted contrast-enhanced imaging revealed lower differential targeted enhancement, that is, lower levels of VEGFR2 expression, in sunitinib-treated mice relative to placebo-treated mice from 24 h (p < 0.05) and relative to both placebo- and imatinib-treated mice from 48 h (p < 0.05). Histologic assessment of tumors after the final imaging indicated that necrotic area was significantly higher for the sunitinib group (21%) than for the placebo (8%, p < 0.001) and imatinib (11%, p < 0.05) groups. VEGFR2-targeted ultrasound was able to sensitively differentiate the anti-VEGFR2 effect from the reduced area of tumor with functional flow produced by both anti-angiogenic agents. BR55 molecular imaging was, thus, able both to detect early therapeutic response to sunitinib in CT26 tumors as soon as 24 h after the beginning of the treatment and to provide early discrimination (48 h) between tumor response during anti-angiogenic therapy targeting VEGFR2 expression and response during anti-angiogenic therapy not directly acting on this receptor.

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      References

        • Agrawal G.
        • Su M.Y.
        • Nalcioglu O.
        • Feig S.A.
        • Chen J.H.
        Significance of breast lesion descriptors in the ACR BI-RADS MRI lexicon.
        Cancer. 2009; 115: 1363-1380
        • Balachandran V.P.
        • Cavnar M.J.
        • Zeng S.
        • Bamboat Z.M.
        • Ocuin L.M.
        • Obaid H.
        • Sorenson E.C.
        • Popow R.
        • Ariyan C.
        • Rossi F.
        • Besmer P.
        • Guo T.
        • Antonescu C.R.
        • Taguchi T.
        • Yuan J.
        • Wolchok J.D.
        • Allison J.P.
        • DeMatteo R.P.
        Imatinib potentiates antitumor T cell responses in gastrointestinal stromal tumor through the inhibition of Ido.
        Nat Med. 2011; 17: 1094-1100
        • BaronToaldo M.
        • Salvatore V.
        • Marinelli S.
        • Palamà C.
        • Milazzo M.
        • Croci L.
        • Venerandi L.
        • Cipone M.
        • Bolondi L.
        • Piscaglia F.
        Use of VEGFR-2 targeted ultrasound contrast agent for the early evaluation of response to sorafenib in a mouse model of hepatocellular carcinoma.
        Mol Imaging Biol. 2015; 17: 29-37
        • Capdeville R.
        • Buchdunger E.
        • Zimmermann J.
        • Matter A.
        Glivec (STI571, imatinib), a rationally developed, targeted anticancer drug.
        Nat Rev Drug Discov. 2002; 1: 493-502
        • Constantinides C.
        • Mean R.
        • Janssen B.J.
        Effects of isoflurane anesthesia on the cardiovascular function of the C57 BL/6 mouse.
        ILAR J. 2011; 52: e21-e31
        • Dizeux A.
        • Payen T.
        • Lechuga G.
        • Bridal S.L.
        Implementation of a controlled injection system for dynamic contrast-enhanced ultrasonography.
        Proc 2012 IEEE Int Ultrason Symp. 2012; 1: 1-4
        • Ewan L.C.
        • Jopling H.M.
        • Jia H.
        • Mittar S.
        • Bagherzadeh A.
        • Howell G.J.
        • Walker J.H.
        • Zachary I.C.
        • Ponnambalam S.
        Intrinsic tyrosine kinase activity is required for vascular endothelial growth factor receptor 2 ubiquitination, sorting and degradation in endothelial cells.
        Traffic. 2006; 7: 1270-1282
        • Faivre S.
        • Demetri G.
        • Sargent W.
        • Raymond E.
        Molecular basis for sunitinib efficacy and future clinical development.
        Nat Rev Drug Discov. 2007; 6: 734-745
        • Giatromanolaki A.
        • Koukourakis M.I.
        • Theodossiou D.
        • Barbatis K.
        • O'Byrne K.
        • Harris A.L.
        • Gatter K.C.
        Comparative evaluation of angiogenesis assessment with anti-factor-VIII and anti-CD31 immunostaining in non-small cell lung cancer.
        Clin Cancer Res. 1997; 3: 2485-2492
        • Gillies R.J.
        • Schornack P.A.
        • Secomb T.W.
        • Raghunand N.
        Causes and effects of heterogeneous perfusion in tumors.
        Neoplasia. 1999; 1: 197-207
        • Hicklin D.J.
        • Ellis L.
        Role of the vascular endothelial growth factor pathway in tumor growth and angiogenesis.
        J Clin Oncol. 2005; 23: 1011-1027
        • Hyvelin J.
        • Tardy I.
        • Arbogast C.
        • Costa M.
        • Emmel P.
        • Helbert A.
        • Theraulaz M.
        • Nunn A.
        • Tranquart F.
        Use of ultrasound contrast agent microbubbles in preclinical research.
        Invest Radiol. 2013; 48: 570-583
        • Jain R.K.
        Determinants of tumor blood flow: a review.
        Cancer Res. 1988; 48: 2641-2658
        • Junttila M.R.
        • de Sauvage F.J.
        Influence of tumour micro-environment heterogeneity on therapeutic response.
        Nature. 2013; 501: 346-354
        • Kano M.R.
        • Komuta Y.
        • Iwata C.
        • Oka M.
        • Shirai Y.
        • Morishita Y.
        • Ouchi Y.
        • Kataoka K.
        • Miyazono K.
        Comparison of the effects of the kinase inhibitors imatinib, sorafenib, and transforming growth factor-beta receptor inhibitor on extravasation of nanoparticles from neovasculature.
        Cancer Sci. 2009; 100: 173-180
        • Lamuraglia M.
        • Bridal S.L.
        • Santin M.
        • Izzi G.
        • Rixe O.
        • Paradiso A.
        • Lucidarme O.
        Clinical relevance of contrast-enhanced ultrasound in monitoring anti-angiogenic therapy of cancer: current status and perspectives.
        Crit Rev Oncol Hematol. 2010; 73: 202-212
        • Leach M.O.
        • Brindle K.M.
        • Evelhoch J.L.
        • Griffiths J.R.
        • Horsman M.R.
        • Jackson A.
        • Jayson G.C.
        • Judson I.R.
        • Knopp M.V.
        • Maxwell R.J.
        • McIntyre D.
        • Padhani A.R.
        • Price P.
        • Rathbone R.
        • Rustin G.J.
        • Tofts P.S.
        • Tozer G.M.
        • Vennart W.
        • Waterton J.C.
        • Williams S.R.
        • Workman P.
        The assessment of antiangiogenic and antivascular therapies in early-stage clinical trials using magnetic resonance imaging: issues and recommendations.
        Br J Cancer. 2005; 92: 1599-1610
        • Leguerney I.
        • Lassau N.
        • Koscielny S.
        • Rodrigues M.
        • Massard C.
        • Rouffiac V.
        • Benatsou B.
        • Thalmensi J.
        • Bawa O.
        • Opolon P.
        • Peronneau P.
        • Roche A.
        Combining functional imaging and interstitial pressure measurements to evaluate two anti-angiogenic treatments.
        Invest New Drugs. 2012; 30: 144-156
        • Levashova Z.
        • Backer M.
        • Hamby C.V.
        • Pizzonia J.
        • Backer J.M.
        • Blankenberg F.G.
        Molecular imaging of changes in the prevalence of vascular endothelial growth factor receptor in sunitinib-treated murine mammary tumors.
        J Nucl Med. 2010; 51: 959-966
        • Liu L.
        • Cao Y.
        • Chen C.
        • Zhang X.
        • McNabola A.
        • Wilkie D.
        • Wilhelm S.
        • Lynch M.
        • Carter C.
        Sorafenib blocks the RAF/MEK/ERK pathway, inhibits tumor angiogenesis, and induces tumor cell apoptosis in hepatocellular carcinoma model PLC/PRF/5.
        Cancer Res. 2006; 66: 11851-11858
        • Marinelli S.
        • Salvatore V.
        • Baron Toaldo M.
        • Milazzo M.
        • Croci L.
        • Venerandi L.
        • Pecorelli A.
        • Palamà C.
        • Diana A.
        • Bolondi L.
        • Piscaglia F.
        Evaluation of the impact of transient interruption of antiangiogenic treatment using ultrasound-based techniques in a murine model of hepatocellular carcinoma.
        BMC Cancer. 2014; 14: 403
        • McCarty M.F.
        • Somcio R.J.
        • Stoeltzing O.
        • Wey J.
        • Fan F.
        • Liu W.
        • Bucana C.
        • Ellis L.M.
        Overexpression of PDGF-BB decreases colorectal and pancreatic cancer growth by increasing tumor pericyte content.
        J Clin Invest. 2007; 117: 2114-2122
        • Mendel D.
        • Laird A.
        • Xin X.
        • Louie S.
        • Christensen J.
        • Li G.
        • Schreck R.
        • Abrams T.
        • Ngai T.
        • Lee L.
        • Murray L.
        • Carver J.
        • Chan E.
        • Moss K.
        • Haznedar J.
        • Sukbuntherng J.
        • Blake R.
        • Sun L.
        • Tang C.
        • Miller T.
        • Shirazian S.
        • McMahon G.
        • Cherrington J.
        In vivo antitumor activity of SU11248, a novel kinase inhibitor targeting vascular endothelial growth factor and platelet-derived growth factor receptors: Determination of a pharmacokinetic/pharmacodynamic relationship.
        Clin Cancer Res. 2003; 9: 327-337
        • Osusky K.L.
        • Hallahan D.E.
        • Fu A.
        • Ye F.
        • Shyr Y.
        • Geng L.
        The receptor tyrosine kinase inhibitor SU11248 impedes endothelial cell migration, tubule formation, and blood vessel formation in vivo, but has little effect on existing tumor vessels.
        Angiogenesis. 2004; 7: 225-233
        • Pastuskovas C.V.
        • Mundo E.E.
        • Williams S.P.
        • Nayak T.K.
        • Ho J.
        • Ulufatu S.
        • Clark S.
        • Ross S.
        • Cheng E.
        • Parsons-Reponte K.
        • Cain G.
        • Van Hoy M.
        • Majidy N.
        • Bheddah S.
        • dela Cruz Chuh J.
        • Kozak K.R.
        • Lewin-Koh N.
        • Nauka P.
        • Bumbaca D.
        • Sliwkowski M.
        • Tibbitts J.
        • Theil F.P.
        • Fielder P.J.
        • Khawli L.A.
        • Boswell C.A.
        Effects of anti-VEGF on pharmacokinetics, biodistribution, and tumor penetration of trastuzumab in a preclinical breast cancer model.
        Mol Cancer Ther. 2012; 11: 752-762
        • Payen T.
        • Coron A.
        • Lamuraglia M.
        • Le Guillou-Buffello D.
        • Gaud E.
        • Arditi M.
        • Lucidarme O.
        • Bridal S.L.
        Echo-power estimation from log-compressed video data in dynamic contrast-enhanced ultrasound imaging.
        Ultrasound Med Biol. 2013; 39: 1826-1837
        • Pillai R.
        • Marinelli E.R.
        • Fan H.
        • Nanjappan P.
        • Song B.
        • von Wronski M.A.
        • Cherkaoui S.
        • Tardy I.
        • Pochon S.
        • Schneider M.
        • Nunn A.D.
        • Swenson R.E.
        A phospholipid–PEG2000 conjugate of a vascular endothelial growth factor receptor 2 (VEGFR2)-targeting heterodimer peptide for contrast-enhanced ultrasound imaging of angiogenesis.
        Bioconjug Chem. 2010; 2: 556-562
        • Pochon S.
        • Tardy I.
        • Bussat P.
        • Bettinger T.
        • Brochot J.
        • von Wronski M.
        • Passantino L.
        • Schneider M.
        BR55: A lipopeptide-based VEGFR2-targeted ultrasound contrast agent for molecular imaging of angiogenesis.
        Invest Radiol. 2010; 45: 89-95
        • Potapova O.
        • Laird A.D.
        • Nannini M.A.
        • Barone A.
        • Li G.
        • Moss K.G.
        • Cherrington J.M.
        • Mendel D.B.
        Contribution of individual targets to the antitumor efficacy of the multitargeted receptor tyrosine kinase inhibitor SU11248.
        Mol Cancer Ther. 2006; 5: 1280-1289
        • Pysz M.A.
        • Foygel K.
        • Rosenberg J.
        • Gambhir S.S.
        • Schneider M.
        • Willmann J.K.
        Antiangiogenic cancer therapy: Monitoring with molecular US and a clinically translatable contrast agent (BR55).
        Radiology. 2010; 256: 519-527
        • Shrivastava A.
        • von Wronski M.A.
        • Sato A.K.
        • Dransfield D.T.
        • Sexton D.
        • Bogdan N.
        • Pillai R.
        • Nanjappan P.
        • Song B.
        • Marinelli E.
        • DeOliveira D.
        • Luneau C.
        • Devlin M.
        • Muruganandam A.
        • Abujoub A.
        • Connelly G.
        • Wu Q.L.
        • Conley G.
        • Chang Q.
        • Tweedle M.F.
        • Ladner R.C.
        • Swenson R.E.
        • Nunn A.D.
        A distinct strategy to generate high-affinity peptide binders to receptor tyrosine kinases.
        Protein Eng Des Sel. 2005; 18: 417-424
        • Sridharan A.
        • Eisenbrey J.R.
        • Liu J.-B.
        • Machado P.
        • Halldorsdottir V.G.
        • Dave J.K.
        • Zhao H.
        • He Y.
        • Park S.
        • Dianis S.
        • Wallace K.
        • Thomenius K.E.
        • Forsberg F.
        Perfusion estimation using contrast-enhanced 3-dimensional subharmonic ultrasound imaging: An in vivo study.
        Invest Radiol. 2013; 48: 654-660
        • Tardy I.
        • Pochon S.
        • Theraulaz M.
        • Emmel P.
        • Passantino L.
        • Tranquart F.
        • Schneider M.
        Ultrasound molecular imaging of VEGFR2 in a rat prostate tumor model using BR55.
        Invest Radiol. 2010; 45: 573-578
        • Tranquart F.
        • Mercier L.
        • Frinking P.
        • Gaud E.
        • Arditi M.
        Perfusion quantification in contrast-enhanced ultrasound (CEUS)—Ready for research projects and routine clinical use.
        Ultraschall Med. 2012; 33: S31-S38
        • Wei K.
        • Jayaweera A.R.
        • Firoozan S.
        • Linka A.
        • Skyba D.M.
        • Kaul S.
        Quantification of myocardial blood flow with ultrasound-induced destruction of microbubbles administered as a constant venous infusion.
        Circulation. 1998; 97: 473-483
        • Weis S.M.
        • Cheresh D.A.
        Tumor angiogenesis: Molecular pathways and therapeutic targets.
        Nat Med. 2011; 17: 1359-1370
        • Willmann J.K.
        • Paulmurugan R.
        • Chen K.
        • Gheysens O.
        • Rodriguez-Porcel M.
        • Lutz A.M.
        • Chen I.Y.
        • Chen X.
        • Gambhir S.S.
        US imaging of tumor angiogenesis with microbubbles targeted to vascular endothelial growth factor receptor type 2 in mice.
        Radiology. 2008; 246: 508-518
        • Wolff N.C.
        • Randle D.E.
        • Egorin M.J.
        • Minna J.D.
        • Ilaria R.L.
        Imatinib mesylate efficiently achieves therapeutic intratumor concentrations in vivo but has limited activity in a xenograft model of small cell lung cancer.
        Clin Cancer Res. 2004; 10: 3528-3534
        • Zubair A.C.
        • Malik S.
        • Paulsen A.
        • Ishikawa M.
        • McCoy C.
        • Adams P.X.
        • Amrani D.
        • Costa M.
        Evaluation of mobilized peripheral blood CD34(+) cells from patients with severe coronary artery disease as a source of endothelial progenitor cells.
        Cytotherapy. 2010; 12: 178-189