Abstract
Purpose
Increased vascular endothelial growth factor (VEGF) receptor expression has been found at the sites of angiogenesis, particularly in tumor growth areas, as compared with quiescent vasculature. An increase in VEGF receptor-2 is associated with colon cancer progression. The in vivo detection of VEGF receptor is of interest for the purposes of studying basic mechanisms of carcinogenesis, making clinical diagnoses, and monitoring the efficacy of chemopreventive and therapeutic agents. In this study, a novel single chain (sc)VEGF-based molecular probe is utilized in the azoxymethane (AOM)-treated mouse model of colorectal cancer to study delivery route and specificity for disease.
Procedures
The probe was constructed by site-specific conjugation of a near-infrared fluorescent dye, Cy5.5, to scVEGF and detected in vivo with a dual-modality optical coherence tomography/laser-induced fluorescence (OCT/LIF) endoscopic system. A probe inactivated via excessive biotinylation was utilized as a control for nonreceptor-mediated binding. The LIF excitation source was a 633-nm He:Ne laser, and red/near-infrared fluorescence was detected with a spectrometer. OCT was used to obtain two-dimensional longitudinal tomograms at eight rotations in the distal colon. Fluorescence emission levels were correlated with OCT-detected disease in vivo. OCT-detected disease was verified with hematoxylin and eosin stained histology slides ex vivo.
Results
High fluorescence emission intensity from the targeted probe was correlated with tumor presence as detected using OCT in vivo and VEGFR-2 immunostaining on histological sections ex vivo. The inactivated probe accumulated preferentially on the surface of tumor lesions and in lymphoid aggregate tissue and was less selective for VEGFR-2.
Conclusion
The scVEGF/Cy probe delivered via colonic lavage reaches tumor vasculature and selectively accumulates in VEGFR-2-positive areas, resulting in high sensitivity and specificity for tumor detection. The combination of OCT and LIF imaging modalities may allow the simultaneous study of tumor morphology and protein expression for the development of diagnostic and therapeutic methods for colorectal cancer.
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References
Cai W, Chen X (2007) Multimodality imaging of vascular endothelial growth factor and vascular endothelial growth factor receptor expression. Front Biosci 12:4267–4279
Folkman J (1971) Tumor angiogenesis: therapeutic implications. N Engl J Med 285:1182–1186
Jain RK (2008) Taming vessels to treat cancer. Sci Am 298:56–63
Galizia G, Lieto E, Ferreraccio F, Orditura M, De Vita F, Castellano P, Imperatore V, Romano C, Ciardiello F, Agostini B, Pignatelli C (2004) Determination of molecular marker expression can predict clinical outcome in colon carcinomas. Clin Cancer Res 10:3490–3499
Khorana AA, Ryan CK, Cox C, Eberly S, Sahasrabudhe DM (2003) Vascular endothelial growth factor, CD68, and epidermal growth factor receptor expression and survival in patients with stage II and stage III colon carcinoma: a role for the host response in prognosis. Cancer 97:960–968
Solban N, Selbo PK, Sinha AK, Chang SK, Hasan T (2006) Mechanistic investigation and implications of photodynamic therapy induction of vascular endothelial growth factor in prostate cancer. Cancer Res 66:5633–5640
Camphausen K, Moses MA, Beecken WD, Khan MK, Folkman J, O’Reilly MS (2001) Radiation therapy to a primary tumor accelerates metastatic growth in mice. Cancer Res 61:2207–2211
Chang SK, Rizvi I, Solban N, Hasan T (2008) In vivo optical molecular imaging of vascular endothelial growth factor for monitoring cancer treatment. Clin Cancer Res 14:4146–4153
Massoud TF, Gambhir SS (2007) Integrating noninvasive molecular imaging into molecular medicine: an evolving paradigm. Trends Mol Med 13:183–191
Weber WA, Czernin J, Phelps ME, Herschman HR (2008) Technology insight: novel imaging of molecular targets is an emerging area crucial to the development of targeted drugs. Nat Clin Pract Oncol 5:44–54
Blankenberg FG, Levashova Z, Sarkar SK, Pizzonia J, Backer MV, Backer JM (2010) Noninvasive assessment of tumor VEGF receptors in response to treatment with pazopanib: a molecular imaging study. Transl Oncol 3:56–64
Levashova Z, Backer M, Backer JM, Blankenberg FG (2009) Imaging vascular endothelial growth factor (VEGF) receptors in turpentine-induced sterile thigh abscesses with radiolabeled single-chain VEGF. J Nucl Med 50:2058–2063
Backer MV, Levashova Z, Patel V, Jehning BT, Claffey K, Blankenberg FG, Backer JM (2007) Molecular imaging of VEGF receptors in angiogenic vasculature with single-chain VEGF-based probes. Nat Med 13:504–509
Arevalo JF (2009) Retinal angiography and optical coherence tomography. Springer, New York
Evans JA, Poneros JM, Bouma BE, Bressner J, Halpern EF, Shishkov M, Lauwers GY, Mino-Kenudson M, Nishioka NS, Tearney GJ (2006) Optical coherence tomography to identify intramucosal carcinoma and high-grade dysplasia in Barrett’s esophagus. Clin Gastroenterol Hepatol 4:38–43
Hariri LP, Bonnema GT, Schmidt K, Winkler AM, Korde V, Hatch KD, Davis JR, Brewer MA, Barton JK (2009) Laparoscopic optical coherence tomography imaging of human ovarian cancer. Gynecol Oncol 114:188–194
Hariri LP, Qui Z, Tumlinson AR, Besselsen DG, Gerner EW, Ignatenko NA, Povazay B, Hermann B, Sattmann H, McNally J, Unterhuber A, Drexler W, Barton JK (2007) Serial endoscopy in azoxymethane treated mice using ultra-high resolution optical coherence tomography. Cancer Biol Ther 6:1753–1762
McNally JB, Kirkpatrick ND, Hariri LP, Tumlinson AR, Besselsen DG, Gerner EW, Utzinger U, Barton JK (2006) Task-based imaging of colon cancer in the Apc(Min/+) mouse model. Appl Opt 45:3049–3062
Saban M, Backer JM, Backer MV, Maier J, Fowler B, Davis CA, Simpson C, Wu XR, Birder L, Freeman MR, Soker S, Hurst RE, Saban R (2008) VEGF receptors and neuropilins are expressed in the urothelial and neuronal cells in normal mouse urinary bladder and are up-regulated in inflammation. Am J Physiol Renal Physiol 295:F60–F72
Saban R, Saban MR, Maier J, Fowler B, Tengowski M, Davis CA, Wu XR, Culkin DJ, Hauser P, Backer J, Hurst RE (2008) Urothelial expression of neuropilins and VEGF receptors in control and interstitial cystitis patients. Am J Physiol Renal Physiol 295:F1613–F1623
Tedesco MM, Terashima M, Blankenberg FG, Levashova Z, Spin JM, Backer M, Backer J, Sho M, Sho E, McConnell MV, Dalman RL (2009) Expression during experimental aortic aneurysm progression analysis of in situ and ex vivo vascular endothelial growth factor receptor. Arterioscler Thromb Vasc Biol 29:1452–1457
Takayama T, Katsuki S, Takahashi Y, Ohi M, Nojiri S, Sakamaki S, Kato J, Kogawa K, Miyake H, Niitsu Y (1998) Aberrant crypt foci of the colon as precursors of adenoma and cancer. N Engl J Med 339:1277–1284
Bird RP (1987) Observation and quantification of aberrant crypts in the murine colon treated with a colon carcinogen: preliminary findings. Cancer Lett 37:147–151
Papanikolaou A, Wang QS, Papanikolaou D, Whiteley HE, Rosenberg DW (2000) Sequential and morphological analyses of aberrant crypt foci formation in mice of differing susceptibility to azoxymethane-induced colon carcinogenesis. Carcinogenesis 21:1567–1572
Hariri LP, Tumlinson AR, Wade NH, Besselsen DG, Utzinger U, Gerner EW, Barton JK (2007) Ex vivo optical coherence tomography and laser-induced fluorescence spectroscopy imaging of murine gastrointestinal tract. Comp Med 57:175–185
Tumlinson AR, Hariri LP, Utzinger U, Barton JK (2004) Miniature endoscope for simultaneous optical coherence tomography and laser-induced fluorescence measurement. Appl Opt 43:113–121
Winkler AM, Rice PFS, Weichsel J, Backer MV, Backer JM, Barton JK (2009) In vivo imaging using a VEGF-based near-infrared fluorescence probe for early diagnosis in the AOM-treated mouse model. Proc SPIE 7190:71900M1–71900M9
Barton JK, Guzman F, Tumlinson AR (2004) Dual modality instrument for simultaneous optical coherence tomography imaging and fluorescence spectroscopy. J Biomed Opt 9:618–623
Wang TD, Friedland S, Sahbaie P, Soetikno R, Hsiung PL, Liu JT, Crawford JM, Contag CH (2007) Functional imaging of the colonic mucosa with a fibered confocal microscope for real time in vivo pathology. Clin Gastroenterol Hepatol 5:1300–1305
Acknowledgments
This work was supported in part by grants from the National Institutes of Health, R01CA109835 and R43CA132528. Additional funding was provided by Achievement Rewards for College Scientists, Technology Research Initiative Funding, and the Philanthropic Educational Organization. The authors also thank Erica Liebmann, Yue Zong, and Amber Luttmann who contributed to this study.
Conflict of interest statement
Dr. Joseph Backer has equity in SibTech, Inc., maker of scVEGF/Cy and inVEGF/Cy. The other authors have no conflict of interest.
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Winkler, A.M., Rice, P.F.S., Weichsel, J. et al. In Vivo, Dual-Modality OCT/LIF Imaging Using a Novel VEGF Receptor-Targeted NIR Fluorescent Probe in the AOM-Treated Mouse Model. Mol Imaging Biol 13, 1173–1182 (2011). https://doi.org/10.1007/s11307-010-0450-6
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DOI: https://doi.org/10.1007/s11307-010-0450-6