Optical Imaging of Spontaneous Breast Tumors Using Protease Sensing ‘Smart’ Optical Probes

Objective:The objective of this study was to determine if spontaneous breast cancer lesions can be detected by fluorescence reflectance imaging (FRI) and fluorescence mediated tomography (FMT) using protease-sensing optical probes. Materials and Methods:Transgenic (FVB/N-TgN (WapHRAS)69Lin YSJL)) mice, which spontaneously develop breast cancer, were injected intravenously with a cathepsin-sensing fluorescent imaging probe. FRI and FMT were performed 24 hours after probe injection and region of interest (ROI) analysis was performed. Magnetic resonance images were acquired for anatomic coregistration with the FMT data. Moreover, correlative immunohistochemistry and fluorescence microscopy were performed. Results:All tumor nodules were clearly delineated by FRI showing an average signal intensity of 380 ± 106 AU. Similarly, tumors were clearly detected by FMT imaging. Immunohistochemistry confirmed cathepsin-B expression of primary tumors and fluorescence microscopy revealed a strong Cy 5.5 deposition in the tissue. Conclusions:FRI and FMT using “smart” protease sensing probes permits detection of experimental spontaneous breast cancers. Because the expression levels of various proteases correlate with patient outcome, this technique may not only help to detect, but also to differentiate breast cancers noninvasively.

[1]  T. Helbich,et al.  Characterization of Benign and Malignant Breast Lesions With Computed Tomography Laser Mammography (CTLM): Initial Experience , 2005, Investigative radiology.

[2]  J. Ripoll,et al.  Visualization of antitumor treatment by means of fluorescence molecular tomography with an annexin V-Cy5.5 conjugate. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[3]  Chun Li,et al.  Near-infrared optical imaging of epidermal growth factor receptor in breast cancer xenografts. , 2003, Cancer research.

[4]  J. Ripoll,et al.  Free-space propagation of diffuse light: theory and experiments. , 2003, Physical review letters.

[5]  Christoph Bremer,et al.  Near-infrared fluorescence imaging of lymph nodes using a new enzyme sensing activatable macromolecular optical probe , 2003, European Radiology.

[6]  Vasilis Ntziachristos,et al.  A submillimeter resolution fluorescence molecular imaging system for small animal imaging. , 2003, Medical physics.

[7]  Woo Kyung Moon,et al.  Enhanced tumor detection using a folate receptor-targeted near-infrared fluorochrome conjugate. , 2003, Bioconjugate chemistry.

[8]  Victor X D Yang,et al.  A multispectral fluorescence imaging system: Design and initial clinical tests in intra‐operative Photofrin‐photodynamic therapy of brain tumors , 2003, Lasers in surgery and medicine.

[9]  Ralph Weissleder,et al.  Pan and sentinel lymph node visualization using a near-infrared fluorescent probe. , 2003, Molecular imaging.

[10]  Woo Kyung Moon,et al.  A Receptor‐Targeted Near‐Infrared Fluorescence Probe for In Vivo Tumor Imaging , 2002, Chembiochem : a European journal of chemical biology.

[11]  R. Weissleder,et al.  Fluorescence molecular tomography resolves protease activity in vivo , 2002, Nature Medicine.

[12]  R. Weissleder,et al.  Imaging of differential protease expression in breast cancers for detection of aggressive tumor phenotypes. , 2002, Radiology.

[13]  Ralph Weissleder,et al.  Detection of dysplastic intestinal adenomas using enzyme-sensing molecular beacons in mice. , 2002, Gastroenterology.

[14]  Ralph Weissleder,et al.  Cellular activation of the self-quenched fluorescent reporter probe in tumor microenvironment. , 2002, Neoplasia.

[15]  V. Ntziachristos,et al.  MRI-guided diffuse optical spectroscopy of malignant and benign breast lesions. , 2002, Neoplasia.

[16]  R. Weissleder,et al.  Fluorescence imaging with near-infrared light: new technological advances that enable in vivo molecular imaging , 2002, European Radiology.

[17]  Ralph Weissleder,et al.  In vivo molecular target assessment of matrix metalloproteinase inhibition , 2001, Nature Medicine.

[18]  W. Semmler,et al.  Receptor-targeted optical imaging of tumors with near-infrared fluorescent ligands , 2001, Nature Biotechnology.

[19]  R Weissleder,et al.  In vivo imaging of proteolytic enzyme activity using a novel molecular reporter. , 2000, Cancer research.

[20]  S. Achilefu,et al.  Novel receptor-targeted fluorescent contrast agents for in vivo tumor imaging. , 2000, Investigative radiology.

[21]  V. Ntziachristos,et al.  Concurrent MRI and diffuse optical tomography of breast after indocyanine green enhancement. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[22]  Bonnie F. Sloane,et al.  Imaging proteolysis by living human breast cancer cells. , 2000, Neoplasia.

[23]  R Weissleder,et al.  Near-infrared optical imaging of protease activity for tumor detection. , 1999, Radiology.

[24]  R. Weissleder.  Molecular imaging: exploring the next frontier. , 1999, Radiology.

[25]  R. Weissleder,et al.  In vivo imaging of tumors with protease-activated near-infrared fluorescent probes , 1999, Nature Biotechnology.

[26]  D. Edwards,et al.  Cancer: Proteases — invasion and more , 1998, Nature.

[27]  M. Duffy,et al.  High Levels of Cathepsin B Predict Poor Outcome in Patients with Breast Cancer , 1998, The International journal of biological markers.

[28]  R. Mason,et al.  Control of breast tumor cell growth using a targeted cysteine protease inhibitor. , 1998, Cancer research.

[29]  M. Schwartz.  Tissue cathepsins as tumor markers. , 1995, Clinica chimica acta; international journal of clinical chemistry.

[30]  J. Catino,et al.  In wap-ras transgenic mice, tumor phenotype but not cyclophosphamide-sensitivity is affected by genetic background. , 1995, Anticancer research.

[31]  L. Benítez-Bribiesca,et al.  Proteinase activity in invasive cancer of the breast. Correlation with tumor progression. , 1995, Archives of medical research.

[32]  G. Tanara,et al.  Evaluation of cathepsin D as prognostic predictor in breast cancer. , 1994, Oncology.

[33]  D. C. Schultz,et al.  Western blotting and enzymatic activity analysis of cathepsin D in breast tissue and sera of patients with breast cancer and benign breast disease and of normal controls. , 1994, Cancer research.

[34]  B. Wilson,et al.  Time resolved reflectance and transmittance for the non-invasive measurement of tissue optical properties. , 1989, Applied optics.

[35]  W. Totty,et al.  Light scanning versus mammography in breast cancer detection. , 1987, Radiology.

[36]  B. Monsees,et al.  Light scan evaluation of nonpalpable breast lesions. , 1987, Radiology.

[37]  B. Groner,et al.  Ha-ras oncogene expression directed by a milk protein gene promoter: tissue specificity, hormonal regulation, and tumor induction in transgenic mice. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[38]  J. Bartek,et al.  Cytochemical and biochemical evidence of cathepsin B in malignant, transformed and normal breast epithelial cells. , 1987, Journal of cell science.

[39]  B. Drexler,et al.  Diaphanography in the diagnosis of breast cancer. , 1985, Radiology.

[40]  G. E. Geslien,et al.  Transillumination in breast cancer detection: screening failures and potential. , 1985, AJR. American journal of roentgenology.

[41]  J. Abecassis,et al.  Proteinases and sialyltransferase in human breast tumors , 1984, International journal of cancer.

[42]  A. Recklies,et al.  Secretion of proteinases from malignant and nonmalignant human breast tissue. , 1980, Cancer research.

[43]  A. Poole,et al.  Differences in secretion of the proteinase cathepsin B at the edges of human breast carcinomas and fibroadenomas , 1978, Nature.