Molecular imaging of epidermal growth factor-receptor and survivin in vivo in porcine esophageal and gastric mucosae using probe-based confocal laser-induced endomicroscopy: proof of concept.

UNLABELLED Confocal laser-induced endomicroscopy (CLE) enables in vivo, real time visualization of the subsurface cells and tissue structures in gastrointestinal mucosa at a subcellular resolution of ≈1000x magnification. The aims of this pilot study were to establish a principle of molecular imaging and determine in vivo expression of epidermal growth factor receptor (EGF-R) and survivin in porcine esophageal and gastric mucosa using probe-based CLE (pCLE) and topically applied FITC-labeled antibodies. Studies were performed in anesthetized pigs. During endoscopy FITC-labeled antibodies against EGF-R and survivin were either sprayed onto esophageal and gastric mucosa in preselected areas or administered via submucosal injection. Thirty minutes later pCLE was performed using a through-the-scope probe (GastroFlex UHD, Cellvizio, Mauna Kea Technologies, Paris, France) to determine cellular and tissue localization of EGF-R and survivin. Then the pigs were euthanized and esophageal and gastric walls from the areas sprayed or injected with antibodies were collected for histologic examination under epifluorescence microscopy. RESULTS CLE enabled visualization of EGF-R and survivin in esophageal and gastric mucosa and this was confirmed by histology. In the esophagus both EGF-R and survivin were localized predominantly to the keratinocyte progenitor cells. In the stomach, EGF-R was localized to progenitor zone cells and some epithelial cells. Localization of survivin was similar, but involved more surface epithelial cells. This study demonstrated feasibility of using CLE and topical administration of FITC labeled antibodies for in vivo localization of EGF-R and survivin in esophageal and gastric mucosa.

[1]  A. Polglase,et al.  Confocal laser endoscopy for diagnosing intraepithelial neoplasias and colorectal cancer in vivo. , 2004, Gastroenterology.

[2]  D. Altieri,et al.  A novel anti-apoptosis gene, survivin, expressed in cancer and lymphoma , 1997, Nature Medicine.

[3]  S. Chiou,et al.  Survivin expression in the stomach: implications for mucosal integrity and protection. , 2003, Biochemical and biophysical research communications.

[4]  R. Pai,et al.  Induction of mitogen-activated protein kinase signal transduction pathway during gastric ulcer healing in rats. , 1998, Gastroenterology.

[5]  R. Kiesslich,et al.  Local barrier dysfunction identified by confocal laser endomicroscopy predicts relapse in inflammatory bowel disease , 2011, Gut.

[6]  S. Chiou,et al.  Survivin: a novel target for indomethacin-induced gastric injury. , 2005, Gastroenterology.

[7]  Michael K. Jones,et al.  Survivin is a key factor in the differential susceptibility of gastric endothelial and epithelial cells to alcohol‐induced injury , 2009, Journal of physiology and pharmacology : an official journal of the Polish Physiological Society.

[8]  Markus F Neurath,et al.  Molecular imaging of VEGF in gastrointestinal cancer in vivo using confocal laser endomicroscopy , 2010, Gut.

[9]  G. D. De Palma,et al.  Confocal laser endomicroscopy in the "in vivo" histological diagnosis of the gastrointestinal tract. , 2009, World journal of gastroenterology.

[10]  Markus F Neurath,et al.  In vivo subsurface morphological and functional cellular and subcellular imaging of the gastrointestinal tract with confocal mini-microscopy. , 2007, World journal of gastroenterology.

[11]  J. Galmiche,et al.  Colonic mucosal biopsies obtained during confocal endomicroscopy are pre-stained with fluorescein in vivo and are suitable for histologic evaluation , 2012, Endoscopy.

[12]  Vivek Kaul,et al.  Confocal laser endomicroscopy. , 2009, Gastrointestinal endoscopy.

[13]  A. Tarnawski,et al.  Prostaglandin E2 transactivates EGF receptor: A novel mechanism for promoting colon cancer growth and gastrointestinal hypertrophy , 2002, Nature Medicine.

[14]  J. Stachura,et al.  Increased expression of epidermal growth factor receptor during gastric ulcer healing in rats. , 1992, Gastroenterology.

[15]  J. Diebold,et al.  Developmentally regulated expression of the novel cancer anti-apoptosis gene survivin in human and mouse differentiation. , 1998, The American journal of pathology.

[16]  Michael Vieth,et al.  In vivo molecular imaging of colorectal cancer with confocal endomicroscopy by targeting epidermal growth factor receptor. , 2010, Gastroenterology.

[17]  S. Chiou,et al.  Survivin - an anti-apoptosis protein: its biological roles and implications for cancer and beyond. , 2003, Medical science monitor : international medical journal of experimental and clinical research.

[18]  Michael K. Jones,et al.  The anti‐apoptosis protein, survivin, mediates gastric epithelial cell cytoprotection against ethanol‐induced injury via activation of the p34cdc2 cyclin‐dependent kinase , 2008, Journal of cellular physiology.

[19]  Martin Goetz,et al.  Advances of endomicroscopy for gastrointestinal physiology and diseases. , 2010, American journal of physiology. Gastrointestinal and liver physiology.

[20]  Koji Takeuchi,et al.  Gastric mucosal defense and cytoprotection: bench to bedside. , 2008, Gastroenterology.

[21]  Martin Goetz,et al.  New imaging techniques and opportunities in endoscopy , 2011, Nature Reviews Gastroenterology &Hepatology.