In Vivo Imaging with Fluorescent Smart Probes to Assess Treatment Strategies for Acute Pancreatitis

Agarwal, Abhiruchi; Boettcher, Andreas; Kneuer, Rainer; Sari-Sarraf, Farid; Donovan, Adriana; Woelcke, Julian; Simic, Oliver; Brandl, Trixi; Krucker, Thomas
February 2013
PLoS ONE;Feb2013, Vol. 8 Issue 2, p1
Academic Journal
Background and Aims: Endoprotease activation is a key step in acute pancreatitis and early inhibition of these enzymes may protect from organ damage. In vivo models commonly used to evaluate protease inhibitors require animal sacrifice and therefore limit the assessment of dynamic processes. Here, we established a non-invasive fluorescence imaging-based biomarker assay to assess real-time protease inhibition and disease progression in a preclinical model of experimental pancreatitis. Methods: Edema development and trypsin activation were imaged in a rat caerulein-injection pancreatitis model. A fluorescent “smart” probe, selectively activated by trypsin, was synthesized by labeling with Cy5.5 of a pegylated poly-L-lysine copolymer. Following injection of the probe, trypsin activation was monitored in the presence or absence of inhibitors by in vivo and ex vivo imaging. Results: We established the trypsin-selectivity of the fluorescent probe in vitro using a panel of endopeptidases and specific inhibitor. In vivo, the probe accumulated in the liver and a region attributed to the pancreas by necropsy. A dose dependent decrease of total pancreatic fluorescence signal occurred upon administration of known trypsin inhibitors. The fluorescence-based method was a better predictor of trypsin inhibition than pancreatic to body weight ratio. Conclusions: We established a fluorescence imaging assay to access trypsin inhibition in real-time in vivo. This method is more sensitive and dynamic than classic tissue sample readouts and could be applied to preclinically optimize trypsin inhibitors towards intrapancreatic target inhibition.


Related Articles

  • HIGH MOBILITY GROUP A: A NOVEL BIOMARKER AND THERAPEUTIC ADENOCARCINOMA. Liau, S. S.; Whang, E. // Surgeon (Edinburgh University Press);Oct2009, Vol. 7 Issue 5, p297 

    High mobility group A1 (HMGA1) proteins are architectural transcriptional factors that are over-expressed in a wide range of human malignancies. Recently published evidence suggests HMGA1 is a promising candidate biomarker and therapeutic target in pancreatic cancer. This review summarises data...

  • Mutational analysis of the pancreatic secretory trypsin inhibitor gene in familial and juvenile pancreatitis in Japan. Kuwata, Kinuko; Hirota, Masahiko; Nishimori, Isao; Otsuki, Makoto; Ogawa, Michio // Journal of Gastroenterology;2003, Vol. 38 Issue 4, p365 

    Background. Mutations in the pancreatic secretory trypsin inhibitor (PSTI) gene have been reported in patients suffering from chronic pancreatitis. The aim of the present study was to investigate whether similar gene mutations are present in familial and juvenile pancreatitis patients in Japan....

  • Severity markers in patients with acute pancreatitis. Kisaoglu, Abdullah; Aydinli, Bulent; Ozturk, Gurkan; Atamanalp, Sabri; Ozogul, Bunyami; Yildirgan, Mehmet; Polat, Kamil // Central European Journal of Medicine;Aug2014, Vol. 9 Issue 4, p556 

    To evaluate the effectiveness of serum levels of resistin and CD14 expression in monocytes, and high-sensitivity C-reactive protein (hsCRP) in early stages of acute pancreatitis and correct prediction of the severity of acute pancreatitis (AP) using scoring systems. The study involved 10...

  • Do point mutations in the PSTI (SPINK1) gene truly contribute to the pathogenesis of chronic pancreatitis? Shimosegawa, Tooru // Journal of Gastroenterology;2001, Vol. 36 Issue 9, p645 

    Editorial. Examines the role of point mutations in the pancreatic secretory trypsin inhibitor (PSTI) gene in the pathogenesis of chronic pancreatitis. Characterization of chronic pancreatitis; Information on PSTI; Incidence of gene mutations in unrelated patients with idiopathic chronic...

  • Functional analysis of recombinant pancreatic secretory trypsin inhibitor protein with amino-acid substitution. Kuwata, Kinuko; Hirota, Masahiko; Shimizu, Hiroyuki; Nakae, Masanori; Nishihara, Shoji; Takimoto, Akio; Mitsushima, Kenji; Kikuchi, Norihisa; Endo, Kazuaki; Inoue, Masayasu; Ogawa, Michio // Journal of Gastroenterology;2002, Vol. 37 Issue 11, p928 

    Background. We hypothesized that mutation of the pancreatic secretory trypsin inhibitor (PSTI) gene may promote a predisposition to pancreatitis, possibly by reducing the inhibition of trypsin activity. Based on this hypothesis, we performed a biochemical analysis of recombinant PSTI protein....

  • Assisted Selection by Specific DNA Markers for Genetic Elimination of the Kunitz Trypsin Inhibitor and Lectin in Soybean Seeds. Alves de Moraes, Rita Maria; Bastos Soares, Tais; Colombo, Lucinete; Spegiorin Salla, Maria; Gomes de Almeida Barros, Josie; Deniz Piovesan, Newton; Gonçalves de Barros, Everaldo; Alves Moreira, Maurilio // Euphytica;Jun2006, Vol. 149 Issue 1/2, p221 

    The antinutritional factors found in soybean, lectin (SBL) and Kunitz trypsin inhibitor (SKTI), are usually inactivated by heat treatment. However, residual activity of these factors can be found in several types of soy-derived products. Heat treatment does not completely eliminate these...

  • Short-term variability of biomarkers of proteinase activity in patients with emphysema associated with type Z alpha-1-antitrypsin deficiency. Stolk, Jan; Veldhuisen, Barbara; Annovazzi, Laura; Zanone, Chiara; Versteeg, Elly M.; Van Kuppevelt, Toine H.; Nieuwenhuizen, Willem; Iadarola, Paolo; Luisetti, Maurizio // Respiratory Research;2005, Vol. 6, p47 

    Background: The burden of proteinases from inflammatory cells in the lung of subjects with type Pi ZZ of alpha-1-antitrypsin deficiency is higher than in those without the deficiency. Cross-sectional studies have shown increased levels of biomarkers of extracellular matrix degradation in vivo....

  • Effects of radiation therapy on tissue and serum concentrations of tumour associated trypsin inhibitor and their prognostic significance in rectal cancer patients.  // Radiation Oncology;2011, Vol. 6 Issue 1, p100 

    The article reports on a study investigating the effects of neoadjuvant radiotherapy (RT) on concentrations of tumor tissue-trypsin inhibitor (t-TATI) and serum-trypsin inhibitor (s-TATI) in patients with rectal cancer. 53 rectal cancer patients from whom serum, normal mucosa and tumour tissues...

  • Increased serum levels of tumour-associated trypsin inhibitor independently predict a poor prognosis in colorectal cancer patients. Gaber, Alexander; Nodin, Björn; Hotakainen, Kristina; Nilsson, Elise; Stenman, Ulf-Håkan; Bjartell, Anders; Birgisson, Helgi; Jirström, Karin // BMC Cancer;2010, Vol. 10, p498 

    Background: There is an insufficient number of reliable prognostic and response predictive biomarkers in colorectal cancer (CRC) management. In a previous study, we found that high tumour tissue expression of tumourassociated trypsin inhibitor (TATI) correlated with liver metastasis and an...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics