Journal: Molecular Imaging

Article Title: Ex Vivo and In Vivo Noninvasive Imaging of Epidermal Growth Factor Receptor Inhibition on Colon Tumorigenesis Using Activatable Near-Infrared Fluorescent Probes

doi: 10.1177/1536012117729044

Figure Lengend Snippet: Detection of decreased colonic tumor load in epidermal growth factor receptor (EGFR) inhibitor diet–fed azoxymethane (AOM)-treated KK-HIJ animals by 2-D fluorescence imaging and ProSense 680 activatable probe. A, The 2-D fluorescence reflectance imaging (FRI) images demonstrate reduced tumor load in EGFR inhibitor diet–fed animals. B, The 2-D quantification analysis suggests significant lowered FRI signals in the colon of EGFR inhibitor diet–fed mice versus chow diet–fed animals. C, Graph of distal colon 2-D FRI signal intensity (counts/energy) and tumor load in each animal indicates EGFR inhibitor diet–fed animals have smaller and less colonic tumors. Each dot represents individual animal (red dots represent animals from EGFR inhibitor diet–fed group, green dots represent animals from chow diet–fed group). D and E, Graph of 2-D FRI signal intensity (counts/energy) and lesion size for individual lesion from chow diet–fed animals (D, green plot graph) and EGFR inhibitor diet–fed animals (E, red plot graph).

Article Snippet: We further compared in vivo abdominal images from chow-fed animals with those from liquid diet–fed animals and found, in the latter animals, a significantly reduced whole abdomen signal intensity after giving cathepsin-activatable ProSense 680 probe injections, which demonstrated that 4 days of liquid diet feeding is sufficient to reduce the GI background during in vivo imaging.

Techniques: Fluorescence, Imaging

Journal: Molecular Imaging

Article Title: Ex Vivo and In Vivo Noninvasive Imaging of Epidermal Growth Factor Receptor Inhibition on Colon Tumorigenesis Using Activatable Near-Infrared Fluorescent Probes

doi: 10.1177/1536012117729044

Figure Lengend Snippet: The 2-D ex vivo imaging with ProSense 680 activatable probe can differentiate large nuclear β-catenin-positive tumors versus small lesions, but not small tumors and lymphoid follicle from normal epithelium. A, Representative 2-D fluorescence reflectance imaging (FRI) images of lesions in each category. B, β-Catenin immunostaining photomicrographs to illustrate lesions in each category (1-4) based on the grading scheme in . C, Scatter plot of each categorized lesion based on 2-D fluorescence signal intensity (counts/energy). * P < .05 versus categories 1, 2 and 3.

Article Snippet: We further compared in vivo abdominal images from chow-fed animals with those from liquid diet–fed animals and found, in the latter animals, a significantly reduced whole abdomen signal intensity after giving cathepsin-activatable ProSense 680 probe injections, which demonstrated that 4 days of liquid diet feeding is sufficient to reduce the GI background during in vivo imaging.

Techniques: Ex Vivo, Imaging, Fluorescence, Immunostaining

Journal: Molecular Imaging

Article Title: Ex Vivo and In Vivo Noninvasive Imaging of Epidermal Growth Factor Receptor Inhibition on Colon Tumorigenesis Using Activatable Near-Infrared Fluorescent Probes

doi: 10.1177/1536012117729044

Figure Lengend Snippet: Three-dimensional (3-D) in vivo fluorescence tomography imaging using ProSense 680 molecular probe is not effective to detect the reduction of colonic tumor load in epidermal growth factor receptor (EGFR)-inhibitor-diet-treated azoxymethane (AOM)-injected animals. A, Representative 3-D Fluorescence Molecular Tomography (FMT) images from chow diet– and EGFR inhibitor diet–fed animals. Region of interest (ROI) in purple is placed in abdominal area. B, Three-dimensional (3D) quantification by FMT in abdominal area (ROI volumes are the same across the animals) suggests no difference in fluorescence signal intensity between chow-diet and EGFR inhibitor diet–fed animals. C, Representative 3-D FMT images from chow-diet and EGFR inhibitor diet–fed animals. Region of interest in purple is placed in the colon area. D, Three-dimensional quantification by FMT in the colon area suggests no difference in fluorescence signal intensity between chow-diet and EGFR inhibitor diet–fed animals.

Article Snippet: We further compared in vivo abdominal images from chow-fed animals with those from liquid diet–fed animals and found, in the latter animals, a significantly reduced whole abdomen signal intensity after giving cathepsin-activatable ProSense 680 probe injections, which demonstrated that 4 days of liquid diet feeding is sufficient to reduce the GI background during in vivo imaging.

Techniques: In Vivo, Fluorescence, Tomography, Imaging, Injection

Journal: Molecular Imaging

Article Title: Ex Vivo and In Vivo Noninvasive Imaging of Epidermal Growth Factor Receptor Inhibition on Colon Tumorigenesis Using Activatable Near-Infrared Fluorescent Probes

doi: 10.1177/1536012117729044

Figure Lengend Snippet: In vivo 3-dimensional (3D) fluorescence imaging using ProSense 680 activatable probe can detect abdominal tumor load in Apc Min/+ mice. A, Representative 3-D in vivo fluorescence images of Apc Min/+ mice and nontumor control mice injected with ProSense 680 probe. Region of interest (ROI) in purple is placed in abdominal area. B, Three-dimensional quantification results in the abdominal area indicate significant higher fluorescence signals in Apc Min/+ mice than nontumor control animals. Data are expressed as mean ± standard error (SE). n = 4 to 5/group. * P < .05 versus nontumor control mice. C and D, Linear regression analysis on ex vivo 2-D whole intestine fluorescence signal intensity and in vivo 3-D abdominal fluorescence signal intensity suggests no significant correlation in control mice (C) or Apc Min/+ mice (D).

Article Snippet: We further compared in vivo abdominal images from chow-fed animals with those from liquid diet–fed animals and found, in the latter animals, a significantly reduced whole abdomen signal intensity after giving cathepsin-activatable ProSense 680 probe injections, which demonstrated that 4 days of liquid diet feeding is sufficient to reduce the GI background during in vivo imaging.

Techniques: In Vivo, Fluorescence, Imaging, Injection, Ex Vivo

Journal: Molecular Imaging

Article Title: Ex Vivo and In Vivo Noninvasive Imaging of Epidermal Growth Factor Receptor Inhibition on Colon Tumorigenesis Using Activatable Near-Infrared Fluorescent Probes

doi: 10.1177/1536012117729044

Figure Lengend Snippet: Dual-model microCT/FMT imaging on intestinal adenomas in Apc Min/+ mice. A, Tumor nodules were detected by contrast-enhanced microCT imaging (white arrows). B, Coregistration of microCT image with 3-dimensional (3D) fluorescence image activated by ProSense 680 probe. Three-dimensional image was taken at 24 hours after injection of ProSense 680 activatable probe and fused with microCT image for anatomical colocalization. C, Two-dimensional fluorescence reflectance imaging (FRI) images of small intestine. After coregistration of in vivo imaging, the small intestine was dissected and imaged ex vivo by FRI for validation.

Article Snippet: We further compared in vivo abdominal images from chow-fed animals with those from liquid diet–fed animals and found, in the latter animals, a significantly reduced whole abdomen signal intensity after giving cathepsin-activatable ProSense 680 probe injections, which demonstrated that 4 days of liquid diet feeding is sufficient to reduce the GI background during in vivo imaging.

Techniques: Imaging, Fluorescence, Injection, In Vivo Imaging, Ex Vivo

Journal: PLoS ONE

Article Title: Live Imaging of Cysteine-Cathepsin Activity Reveals Dynamics of Focal Inflammation, Angiogenesis, and Polyp Growth

doi: 10.1371/journal.pone.0002916

Figure Lengend Snippet: (Α) Histology of polyps from APC Δ468 mouse. (a–b) CEA stained paraffin sections counterstained with Gill's II Hematoxylin; (c&d) methylene blue staining; arrows point to (a) granulocytes, (b) mast cells, (c) plasma cells, (d) lymphocytes. (B) Immuno-fluorescence of polyps from APC Δ468 mice. Cryosections were stained with (a) DAPI, (b) CD11b-AlexaFluor 488, and (c) Gr1-AlexaFluor 594. Arrows point to polyp, and arrowheads to the adjacent healthy villus; note accumulation of CD11b + cells and/or Gr1 + cells in the polyps. (C) FACS analysis of leukocytes prepared from micro-dissected polyps, and from adjacent tissue (n = 4), and of intestinal tissue from age-matched healthy control mice (n = 3). Mean values and SEM are shown for frequencies of CD11b + and of CD11b + Gr1 + cells in 6 month-old mice.

Article Snippet: The frequencies of ProSense-680 + , CD11b + F4/80 + cells remain unchanged (mean 5.36% to 5.03% respectively).

Techniques: Staining, Clinical Proteomics, Fluorescence, Control

Journal: PLoS ONE

Article Title: Live Imaging of Cysteine-Cathepsin Activity Reveals Dynamics of Focal Inflammation, Angiogenesis, and Polyp Growth

doi: 10.1371/journal.pone.0002916

Figure Lengend Snippet: (A) FACS analyses of pro-inflammatory cells. (a) Exemplar FACS analyses of mononuclear cells prepared from the intestine of (a&c) a 5 month-old APC Δ468 mouse and (b&d) an age matched wt C57BL/6J mouse; note increases in the frequencies of (a) CD11b + Gr1 + and (c) CD11b + F4/80 + myeloid type cells in the polyposis intestine, compared to wt tissue (b&d respectively). (B) Summary of FACS analysis of MNCs from wt (black bars) or APC Δ468 (open bars) intestine, showing mean frequencies and absolute numbers of cells with SEM values; n = 6 for APC Δ468 , n = 3 for wt control mice, per age group.

Article Snippet: The frequencies of ProSense-680 + , CD11b + F4/80 + cells remain unchanged (mean 5.36% to 5.03% respectively).

Techniques: Control

Journal: PLoS ONE

Article Title: Live Imaging of Cysteine-Cathepsin Activity Reveals Dynamics of Focal Inflammation, Angiogenesis, and Polyp Growth

doi: 10.1371/journal.pone.0002916

Figure Lengend Snippet: Cryosections of ProSense-680 in vivo stained intestine from APC Δ468 mice were stained with antibodies to CD11b (AlexaFluor 488), Gr1 (AlexaFluor 594) and DAPI. The merged images of CD11b with DAPI (a, CD11b green, DAPI gray), Gr1 with DAPI (b, Gr1 red, DAPI gray), and ProSense-680 with DAPI (c, ProSense-680 blue, DAPI gray) were produced with the “RGB gray” plug-in of Image J. The “colocalization finder” plug-in produced images where the colocalized pixels appear white while the ProSense-680 was red (d&e), the CD11b was green (d, colocalization analysis of ProSense-680 and CD11b staining) and the Gr1 was green (e, colocalization analysis of ProSense-680 and Gr1 staining). ×400 magnification. Arrows mark a CD11b + Gr1 ProSense 680 + cell. Representative FACS dot-plots of MNCs prepared from polyposis intestine and ex vivo stained with ProSense-680 followed by CD11b and Gr1 staining. The live MNCs were gated for ProSence-680 + cells, which were analyzed for CD11b + Gr1 + (f, MDSCs) and CD11b + F4/80 + (g, macrophages) cells. Cumulative results of 6 FACS experiments showing % of CD11b + Gr1 + ProSense-680 + and CD11b + F4/80 + ProSense 680 + among total infiltrating MNCs. Note that among the ProSense-680 + cells (11±0.69% of total MNCs) over 75% were either CD11b + Gr1 + (3.4±0.6% of total MNCs) or CD11b + F4/80 (5.0±0.34% of total MNCs).

Article Snippet: The frequencies of ProSense-680 + , CD11b + F4/80 + cells remain unchanged (mean 5.36% to 5.03% respectively).

Techniques: In Vivo, Staining, Produced, Ex Vivo

Journal: PLoS ONE

Article Title: Live Imaging of Cysteine-Cathepsin Activity Reveals Dynamics of Focal Inflammation, Angiogenesis, and Polyp Growth

doi: 10.1371/journal.pone.0002916

Figure Lengend Snippet: (a) Non-linear regression analysis of polyp number and diameter, assuming Gaussian distribution; APC Δ468 Ctsb −/− (continued line, open squares), and APC Δ468 (dotted line, closed triangles). Note that Cathepsin B −/− mice had fewer and smaller polyps. (b) Frequencies of ProSense-680 active leukocytes amongst total MNCs prepared from the intestine of APC Δ468 (open bar, 6.7%±086%) or APC Δ468 Ctsb −/− (filled bar, 11%±0.69%, P = 0.0037; unpaired t test with Welsh correction). (c) Frequencies of ProSense-680 active CD11b + Gr1 + (mean 0.56%) or CD11b + F4/80 + (mean 4.46%) cells, from the intestines of APC Δ468 (open bar) and APC Δ468 Ctsb −/− mice (filled bars); P<0.001, n = 6, 2way ANOVA. Note that Cathepsin B deficiency predominantly impacted the abundance of CD11b + Gr1 + cells. (d) Attenuation of polyposis in anti-TNFα treated mice (solid line, open squares, n = 6), as compared to the APC Δ468 (dotted line & closed triangles). (e) Frequencies of CD11b + Gr1 + amongst total intestine live MNCs; APC Δ468 Ctsb −/− intestine (light gray bar, 0.56±0.15%, P<0.001), anti-TNFα treated (dark gray bar, 0.71±0.22%, P<0.001), untreated APC Δ468 (open bar, 4.2±0.093%), wt control intestine (black bar, 0.15±0.051%). (f) Frequencies of CD11b + F4/80 + in the APC Δ468 (5.03±0.78%), APC Δ468 Ctsb −/− intestine (5.36±0.92%), and anti-TNFα treated intestine (dark gray bar, 3.0±0.67%).

Article Snippet: The frequencies of ProSense-680 + , CD11b + F4/80 + cells remain unchanged (mean 5.36% to 5.03% respectively).

Techniques: Control