Gastroenterology

Gastroenterology

Volume 121, Issue 3, September 2001, Pages 678-684
Gastroenterology

Liver, Pancreas, and Biliary Tract
Claudin-4: A new target for pancreatic cancer treatment using Clostridium perfringens enterotoxin,☆☆,

https://doi.org/10.1053/gast.2001.27124Get rights and content

Abstract

Background & Aims: Recently, several members of the claudin family have been identified as integral constituents of tight junctions. Using expression profiling, we previously found claudin-4 to be overexpressed in pancreatic cancer. Because claudin-4 has been described as a receptor for the cytotoxic Clostridium perfringens enterotoxin (CPE), we investigated the effect of CPE on pancreatic cancer cells. Methods: Expression of claudin-4 was analyzed by Northern blots. In vitro toxicity of CPE was determined by trypan blue exclusion and the 86Rb-release assay. The in vivo effect of CPE was studied in claudin-4–expressing nude mouse xenografts of the Panc-1 cell line. Results: Expression analyses showed that claudin-4 was overexpressed in most pancreatic cancer tissues and cell lines and several other gastrointestinal tumors. CPE led to an acute dose-dependent cytotoxic effect, restricted to claudin-4–expressing cells and dependent on claudin-4 expression levels. Furthermore, transforming growth factor β was identified as a negative modulator of both claudin-4 expression and susceptibility to CPE. In vivo, intratumoral injections of CPE in Panc-1 xenografts led to large areas of tumor cell necrosis and significant reduction of tumor growth. Conclusions: Our findings suggest that targeting claudin-4–expressing tumors with CPE represents a promising new treatment modality for pancreatic cancer and other solid tumors.

GASTROENTEROLOGY 2001;121:678-684

Section snippets

Materials and cell lines

Human tissue from patients with histologically verified ductal adenocarcinoma of the pancreas, tumor tissues of different origin, human pancreatic tissue from organ donors, and chronic pancreatitis tissues were provided by the Department of Surgery at the University of Ulm and by the Hungarian Academy of Sciences (Budapest, Hungary). All tissues were obtained after approval by local ethics committees.

The human pancreatic cancer cell lines were obtained from the following suppliers: PANC-1 and

Expression pattern of Claudin-4 in pancreatic cancer and normal tissues

Northern blot analyses showed that claudin-4 expression was very weak or absent in all normal pancreas (n = 9) and chronic pancreatitis (n = 9) tissues. In contrast, it was strongly overexpressed in 24 of 31 pancreatic cancer tissues (Figure 1).

. (A) Overexpression of claudin-4 in pancreatic cancer tissues and colon carcinomas, as determined by Northern blot analysis. P1–P5, pancreatic adenocarcinomas, representative of 31 tested pancreatic adenocarcinomas; C1–C5, adenocarcinomas of the colon,

Discussion

CPE is considered the virulence factor responsible for the symptoms of C. perfringens strain A food poisoning and some cases of non–food-borne human gastrointestinal diseases, e.g., antibiotic-associated diarrhea.23 CPE-induced injury of intestinal epithelial cells, morphologically characterized by bleb balloon formation, is initiated by binding to receptors such as CPE-R.14, 24 The C-terminal domain of CPE has been shown to be responsible for high-affinity binding to the CPE-R1; the N-terminal

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      Interestingly, claudin receptors 3 and 4 are highly expressed in some cancers including PC. Initial studies, using recombinant CPE showed selective and efficient therapeutic effects in many cancers including breast and pancreatic cancer [64,65]. Resistance to treatment was also low but unfortunately some off-target effects in normal cells expressing claudin receptors have been described in a preclinical animal model [64].

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    Supported by grants of the Deutsche Forschungsgemeinschaft (SFB 518/Project B1), the European Community (BMH4-CT98-3085), and the National Institutes of Health (AI19844-17).

    ☆☆

    Address requests for reprints to: Thomas M. Gress, M.D., Universität Ulm, Abteilung Innere Medizin I, Robert-Koch-Strasse 8, 89081 Ulm, Germany. e-mail: [email protected]; fax: (49) 731-500-24302.

    Patrick Michl and Malte Buchholz contributed equally to this work.

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