Development and characterization of a human single-chain antibody fragment against claudin-3: a novel therapeutic target in ovarian and uterine carcinomas

Romani, Chiara; Comper, Fabrizio; Bandiera, Elisabetta; Ravaggi, Antonella; Bignotti, Eliana; Tassi, Renata; Pecorelli, Sërgio; Santin, Alessandro D.

doi:10.1016/j.ajog.2009.02.010

Cited by 21 publications

(15 citation statements)

References 26 publications

(34 reference statements)

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“…We, therefore, included proper controls and confirmed the data with independent methods and systems. Nevertheless, MST can successfully be applied to Clds, as demonstrated with anti-Cld1 antibodies exhibiting a typical K d of several nM for such antibody-antigen interactions (42). Thus, K d values found for peptides and recombinant ECL *100 nM are also considered as exhibiting a high affinity.…”

Section: Discussionmentioning

confidence: 99%

Redox-Sensitive Structure and Function of the First Extracellular Loop of the Cell–Cell Contact Protein Claudin-1: Lessons from Molecular Structure to Animals

Dąbrowski

Staat

Zwanziger

et al. 2015

Antioxidants & Redox Signaling

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The paracellular cleft within epithelia/endothelia is sealed by tight junction (TJ) proteins. Their extracellular loops (ECLs) are assumed to control paracellular permeability and are targets of pathogenes. We demonstrated that claudin-1 is crucial for paracellular tightening. Its ECL1 is essential for the sealing and contains two cysteines conserved throughout all claudins. Aims: We prove the hypothesis that this cysteine motif forms a redox-sensitive intramolecular disulfide bridge and, hence, the claudin-1-ECL1 constitutes a functional structure which is associated to ECLs of this and other TJ proteins. Results: The structure and function of claudin-1-ECL1 was elucidated by investigating sequences of this ECL as synthetic peptides, C1C2, and as recombinant proteins, and exhibited a b-sheet binding surface flanked by an a-helix. These sequences bound to different claudins, their ECL1, and peptides with nanomolar binding constants. C-terminally truncated C1C2 (-4aaC) opened cellular barriers and the perineurium. Recombinant ECL1 formed oligomers, and bound to claudin-1 expressing cells. Oligomerization and claudin association were abolished by reducing agents, indicating intraloop disulfide bridging and redox sensitivity. Innovation: The structural and functional model based on our in vitro and in vivo investigations suggested that claudin-1-ECL1 constitutes a functional and ECL-binding bsheet, stabilized by a shielded and redox-sensitive disulfide bond. Conclusion: Since the b-sheet represents a consensus sequence of claudins and further junctional proteins, a general structural feature is implied. Therefore, our model is of general relevance for the TJ assembly in normal and pathological conditions. C1C2-4aaC is a new drug enhancer that is used to improve pharmacological treatment through tissue barriers.

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Section: Discussionmentioning

confidence: 99%

Redox-Sensitive Structure and Function of the First Extracellular Loop of the Cell–Cell Contact Protein Claudin-1: Lessons from Molecular Structure to Animals

Dąbrowski

Staat

Zwanziger

et al. 2015

Antioxidants & Redox Signaling

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“…However, their hydrophobicity and low immunogenicity has made it difficult to raise antibodies targeting claudins. Antibodies that specifically recognize the extracellular loops of human claudin-3 [148] or claudin-4 [149] have been successfully produced, and the anti-claudin-4 antibody has shown therapeutic antitumor activity in vitro and in vivo [149]. Furthermore, a dual-targeting monoclonal antibody against claudin-3 and claudin-4 was recently prepared and shown to possess antitumor effects in vitro and in vivo [150], supporting the potential role of claudins as targets for therapeutic antibodies.…”

Section: Claudins As Biomarkers and Therapeutic Targetsmentioning

confidence: 99%

Emerging Roles of Claudins in Human Cancer

Kwon

2013

IJMS

183

177

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Claudins are major integral membrane proteins of tight junctions. Altered expression of several claudin proteins, in particular claudin-1, -3, -4 and -7, has been linked to the development of various cancers. Although their dysregulation in cancer suggests that claudins play a role in tumorigenesis, the exact underlying mechanism remains unclear. The involvement of claudins in tumor progression was suggested by their important role in the migration, invasion and metastasis of cancer cells in a tissue-dependent manner. Recent studies have shown that they play a role in epithelial to mesenchymal transition (EMT), the formation of cancer stem cells or tumor-initiating cells (CSCs/TICs), and chemoresistance, suggesting that claudins are promising targets for the treatment of chemoresistant and recurrent tumors. A recently identified claudin-low breast cancer subtype that is characterized by the enrichment of EMT and stem cell-like features is significantly associated with disease recurrence, underscoring the importance of claudins as predictors of tumor recurrence. The critical role of epigenetic mechanisms in the regulation of claudin expression indicates the possible application of epigenetic therapy to target claudins. A better understanding of the emerging role of claudins in CSC/TICs and chemoresistance may help to develop therapies against recurrent cancers.

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“…Until recently, an antibody against the extracellular loop domain of claudin had never been successfully prepared, and C-CPE was the only known claudin binder (Sonoda et al, 1999). Recently, Romani et al (2009) prepared a single-chain antibody fragment against claudin-3 by using phage display technology. They found that the antibody fragment binds to ovarian and uterine carcinoma cells in vitro.…”

Section: Downloaded Frommentioning

confidence: 99%

A Claudin-Targeting Molecule as an Inhibitor of Tumor Metastasis

Saeki¹,

Kondoh²,

Kakutani³

et al. 2010

J Pharmacol Exp Ther

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Tumor metastasis of epithelium-derived tumors is the major cause of death from malignant tumors. Overexpression of claudin is observed frequently in malignant tumors. However, claudin-targeting antimetastasis therapy has never been investigated. We previously prepared a claudin-4-targeting antitumor molecule that consisted of the C-terminal fragment of Clostridium perfringens enterotoxin (C-CPE) fused to protein synthesis inhibitory factor (PSIF) derived from Pseudomonas exotoxin. In the present study, we investigated whether claudin CPE receptors can be a target for tumor metastasis by using the C-CPE-fused PSIF as a claudintargeting agent. One of the most popular murine metastasis models is the lung metastasis of intravenously injected B16 cells. Therefore, we first investigated the effects of the C-CPE-fused PSIF on lung metastasis of claudin-4-expressing B16 (CL4-B16) cells. Intravenous administration of the C-CPE-fused PSIF suppressed lung metastasis of CL4-B16 cells but not B16 cells. Injection of C-CPE-fused PSIF also inhibited tumor growth and spontaneous lung metastasis of murine breast cancer 4T1 cells inoculated into the subcutis. Treatment with C-CPE-fused PSIF did not show apparent side effects in mice. These findings indicate that claudin targeting may be a novel strategy for inhibiting some tumor metastases.

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Development and characterization of a human single-chain antibody fragment against claudin-3: a novel therapeutic target in ovarian and uterine carcinomas

Cited by 21 publications

References 26 publications

Redox-Sensitive Structure and Function of the First Extracellular Loop of the Cell–Cell Contact Protein Claudin-1: Lessons from Molecular Structure to Animals

Redox-Sensitive Structure and Function of the First Extracellular Loop of the Cell–Cell Contact Protein Claudin-1: Lessons from Molecular Structure to Animals

Emerging Roles of Claudins in Human Cancer

A Claudin-Targeting Molecule as an Inhibitor of Tumor Metastasis

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