A claudin-4 modulator enhances the mucosal absorption of a biologically active peptide

Uchida, Hiroshi; Kondoh, Masuo; Hanada, Takeshi; Takahashi, Azusa; Hamakubo, Takao; Yagi, Kiyohito

doi:10.1016/j.bcp.2010.01.010

Cited by 69 publications

(58 citation statements)

References 40 publications

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“…This observation suggests that substantial gross or even histological damage may not be essential for CPE to gain access to the circulation. Many studies are investigating CPE as a tool for drug delivery, and those studies have shown that nontoxic CPE fragments are capable of increasing intestinal permeability simply by interacting with claudins in the tight junctions (11,25). Perhaps, a threshold lower concentration exists where native CPE can also initiate permeability alterations without causing severe epithelium damage and yet still allow for toxin absorption into the circulation.…”

Section: Discussionmentioning

confidence: 99%

Development and Application of a Mouse Intestinal Loop Model To Study the In Vivo Action of Clostridium perfringens Enterotoxin

et al. 2011

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Clostridium perfringens enterotoxin (CPE) is responsible for causing the gastrointestinal symptoms of C. perfringens type A food poisoning, the second most commonly identified bacterial food-borne illness in the United States. CPE is produced by sporulating C. perfringens cells in the small intestinal lumen, where it then causes epithelial cell damage and villous blunting that leads to diarrhea and cramping. Those effects are typically self-limiting; however, severe outbreaks of this food poisoning, particularly two occurring in psychiatric institutions, have involved deaths. Since animal models are currently limited for the study of the CPE action, a mouse ligated intestinal loop model was developed. With this model, significant lethality was observed after 2 h in loops receiving an inoculum of 100 or 200 g of CPE but not using a 50-g toxin inoculum. A correlation was noted between the overall intestinal histological damage and lethality in mice. Serum analysis revealed a dose-dependent increase in serum CPE and potassium levels. CPE binding to the liver and kidney was detected, along with elevated levels of potassium in the serum. These data suggest that CPE can be absorbed from the intestine into the circulation, followed by the binding of the toxin to internal organs to induce potassium leakage, which can cause death. Finally, CPE pore complexes similar to those formed in tissue culture cells were detected in the intestine and liver, suggesting that (i) CPE actions are similar in vivo and in vitro and (ii) CPE-induced potassium release into blood may result from CPE pore formation in internal organs such as the liver.

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Section: Discussionmentioning

confidence: 99%

Development and Application of a Mouse Intestinal Loop Model To Study the In Vivo Action of Clostridium perfringens Enterotoxin

et al. 2011

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“…The plasmids encoding C-CPE 194 and C-CPE m19 were kind gifts from the Laboratory of Bio-Functional Molecular Chemistry, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan, as part of our joint research (Uchida et al, 2010;Takahashi et al 2012). They were transfected into Escherichia coli BL-21 (DE 3) and protein expression was stimulated by the addition of isopropyl-D-thiogalactopyranoside.…”

Section: Preparation Of C-cpe Mutantsmentioning

confidence: 99%

“…Recently, it was found that a C-CPE mutant with 10 amino acids at the N-terminal of C-CPE had highly solubility in phosphatebuffered saline (PBS) and binding ability with claudin-4 (Uchida et al, 2010;Takahashi et al, 2011). Furthermore, a C-CPE mutant called C-CPE m19, which has binding ability not only with claudin-4 but also with claudin-1, was found after screening claudin binders from a C-CPE mutantdisplaying library by using claudin-displaying budded baculovirus (Takahashi et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

Claudin-binder C-CPE mutants enhance permeability of insulin across human nasal epithelial cells

Kojima

Kondoh

Keira³

et al. 2015

Drug Delivery

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Objective: Intranasal insulin administration has therapeutic potential for Alzheimer's disease and in intranasal administration across the nasal mucosa, the paracellular pathway regulated by tight junctions is important. The C-terminal fragment of Clostridium perfringens enterotoxin (C-CPE) binds the tight junction protein claudin and disrupts the tight junctional barrier without a cytotoxic effect. The C-CPE mutant called C-CPE 194 binds only to claudin-4, whereas the C-CPE 194 mutant called C-CPE m19 binds not only to claudin-4 but also to claudin-1. Methods: In the present study, to investigate the effects of C-CPE mutants on the tight junctional functions of human nasal epithelial cells (HNECs) and on the permeability of human recombinant insulin across the cells, HNECs were treated with C-CPE 194 and C-CPE m19. Results: C-CPE 194 and C-CPE m19 disrupted the barrier and fence functions without changes in expression of claudin-1, -4, -7, and occludin or cytotoxicity, whereas they transiently increased the activity of ERK1/2 phosphorylation. The disruption of the barrier function caused by C-CPE 194 and C-CPE m19 was prevented by pretreatment with the MAPKK inhibitor U0126. Furthermore, C-CPE 194 and C-CPE m19 significantly enhanced the permeability of human recombinant insulin across HNECs and the permeability was also inhibited by U0126. Conclusion: These findings suggest that C-CPE mutants 194 and m19 can regulate the permeability of insulin across HNECs via the MAPK pathway and may play a crucial role in therapy for the diseases such as Alzheimer's disease via the direct intranasal insulin administration.

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“…Human CLDN-4 recombinant proteins were purified by using a Sf-9 cell expression system as previously described (Uchida et al, 2010). To determine the binding kinetics of 5A5 to human CLDN-4 protein, we performed a surface plasmon resonance analysis by using a Biacore T200 instrument (GE Healthcare, Little Chalfont, UK) as previously described (Uchida et al, 2010). We used 10 mM HEPES, 150 mM NaCl, 3 mM EDTA, and 0.05% Tween 20 (pH 7.4) as the running buffer.…”

Section: Methodsmentioning

confidence: 99%

Development of an Anti–Claudin-3 and -4 Bispecific Monoclonal Antibody for Cancer Diagnosis and Therapy

Iida

Tada

et al. 2014

J Pharmacol Exp Ther

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Most malignant tumors are derived from epithelium, and claudin (CLDN)-3 and CLDN-4 are frequently overexpressed in such tumors. Although antibodies have potential in cancer diagnostics and therapy, development of antibodies against CLDNs has been difficult because the extracellular domains of CLDNs are too small and there is high homology among human, rat, and mouse sequences. Here, we created a monoclonal antibody that recognizes human CLDN-3 and CLDN-4 by immunizing rats with a plasmid vector encoding human CLDN-4. A hybridoma clone that produced a rat monoclonal antibody recognizing both CLDN-3 and -4 (clone 5A5) was obtained from a hybridoma screen by using CLDN-3-and -4-expressing cells; 5A5 did not bind to CLDN-1-, -2-, -5-, -6-, -7-, or -9-expressing cells. Fluorescenceconjugated 5A5 injected into xenograft mice bearing human cancer MKN74 or LoVo cells could visualize the tumor cells. The human-rat chimeric IgG1 monoclonal antibody (xi5A5) activated FcgRIIIa in the presence of CLDN-3-or -4-expressing cells, indicating that xi5A5 may exert antibody-dependent cellular cytotoxicity. Administration of xi5A5 attenuated tumor growth in xenograft mice bearing MKN74 or LoVo cells. These results suggest that 5A5 shows promise in the development of a diagnostic and therapeutic antibody for cancers.

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A claudin-4 modulator enhances the mucosal absorption of a biologically active peptide

Cited by 69 publications

References 40 publications

Development and Application of a Mouse Intestinal Loop Model To Study the In Vivo Action of Clostridium perfringens Enterotoxin

Development and Application of a Mouse Intestinal Loop Model To Study the In Vivo Action of Clostridium perfringens Enterotoxin

Claudin-binder C-CPE mutants enhance permeability of insulin across human nasal epithelial cells

Development of an Anti–Claudin-3 and -4 Bispecific Monoclonal Antibody for Cancer Diagnosis and Therapy

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