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Human breast cancer and lymph node metastases express Gb3 and can be targeted by STxB-vectorized chemotherapeutic compounds.

Stimmer L, Dehay S, Nemati F, Massonnet G, Richon S, Decaudin D, Klijanienko J, Johannes L - BMC Cancer (2014)

Bottom Line: Gb3 positivity correlated with estrogen receptor expression (p≤0.01), whereas absence of Gb3 expression in primary tumors was correlated with the presence of lymph node metastases (p≤0.03). 65% of lymph node metastases were Gb3 positive and in 40% of tested patients, we observed a statistically significant increase of metastatic Gb3 expression (p≤0.04).Intravenous injections of fluorescent STxB into HBC xenografted mice showed preferential STxB accumulation in epithelial cells and cells with endothelial morphology of the tumor.Gb3 expressing HBCx can be used as a model for preclinical studies with STxB conjugates.

View Article: PubMed Central - PubMed

Affiliation: Endocytic Trafficking and Therapeutic Delivery Group, UMR3666 CNRS - U1143 INSERM, Institut Curie-Centre de Recherche, 26 rue d'Ulm, 75248 Paris Cedex 05, France. ludger.johannes@curie.fr.

ABSTRACT

Background: The B-subunit of Shiga toxin (STxB) specifically binds to the glycosphingolipid Gb3 that is highly expressed on a number of human tumors and has been shown to target tumor cells in mouse models and ex vivo on primary colon carcinoma specimen.

Methods: Using a novel ex vivo STxB labeling (ESL) method we studied Gb3 expression in cytological specimens of primary human breast tumors from 107 patients, and in synchronous lymph node metastases from 20 patients. Fluorescent STxB was incubated with fine-needle aspiration (FNA) specimens, and Gb3 expression was evaluated by fluorescence microscopy. Furthermore, 11 patient-derived human breast cancer xenografts (HBCx) were evaluated for expression of Gb3 by ESL and FACS. In addition, the biodistribution of fluorescent STxB conjugate was studied after intravenous injection in a Gb3 positive HBCx model.

Results: Gb3 expression was detected in 62 of 107 patients (57.9%), mainly in epithelial tumor cells. Gb3 positivity correlated with estrogen receptor expression (p≤0.01), whereas absence of Gb3 expression in primary tumors was correlated with the presence of lymph node metastases (p≤0.03). 65% of lymph node metastases were Gb3 positive and in 40% of tested patients, we observed a statistically significant increase of metastatic Gb3 expression (p≤0.04). Using concordant ESL and flow cytometry analysis, 6 out of 11 HBCx samples were scored positive. Intravenous injections of fluorescent STxB into HBC xenografted mice showed preferential STxB accumulation in epithelial cells and cells with endothelial morphology of the tumor.

Conclusion: The enhanced expression of Gb3 in primary breast carcinomas and its lymph node metastases indicate that the development of STxB-based therapeutic strategies is of interest in this pathology. Gb3 expressing HBCx can be used as a model for preclinical studies with STxB conjugates. Finally, the ESL technique on FNA represents a rapid and cost effective method for the stratification of patients in future clinical trials.

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Gb3 expression in normal breast tissue, adenofibroma, breast carcinoma and normal human kidney specimens.
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Fig1: Gb3 expression in normal breast tissue, adenofibroma, breast carcinoma and normal human kidney specimens.

Mentions: In order to evaluate global quantity of Gb3 in normal and tumor breast tissue, Gb3 was extracted and quantified using a previously published STxB overlay method [22] Normal breast tissue expressed Gb3 in the range from 13.0 to 53.0 ng per mg of tissue (mean 24.52 ng/mg), whereas breast carcinoma showed 1.5 times higher Gb3 expression (mean 37.9 ng/mg) with a higher range from 8.0 ng/mg to 121.0 ng/mg. However, this difference was not significant (p > 0.5). In contrast to breast carcinoma, adenofibroma showed comparable Gb3 levels to normal tissue (mean 28.51 ng/mg, min. 1.8 ng/mg, max. 45.0 ng/mg). In addition, normal kidney tissue, known for high Gb3 levels, was extracted as a positive control. This tissue showed mean values of 85.72 ng of Gb3 per mg of tissue (min. 31.5 ng/mg, max. 124.7 ng/mg), which was 2.3 times higher than the mean of Gb3 in breast carcinoma. These data are summarized in Figure 1.Figure 1


Human breast cancer and lymph node metastases express Gb3 and can be targeted by STxB-vectorized chemotherapeutic compounds.

Stimmer L, Dehay S, Nemati F, Massonnet G, Richon S, Decaudin D, Klijanienko J, Johannes L - BMC Cancer (2014)

Gb3 expression in normal breast tissue, adenofibroma, breast carcinoma and normal human kidney specimens.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4289340&req=5

Fig1: Gb3 expression in normal breast tissue, adenofibroma, breast carcinoma and normal human kidney specimens.
Mentions: In order to evaluate global quantity of Gb3 in normal and tumor breast tissue, Gb3 was extracted and quantified using a previously published STxB overlay method [22] Normal breast tissue expressed Gb3 in the range from 13.0 to 53.0 ng per mg of tissue (mean 24.52 ng/mg), whereas breast carcinoma showed 1.5 times higher Gb3 expression (mean 37.9 ng/mg) with a higher range from 8.0 ng/mg to 121.0 ng/mg. However, this difference was not significant (p > 0.5). In contrast to breast carcinoma, adenofibroma showed comparable Gb3 levels to normal tissue (mean 28.51 ng/mg, min. 1.8 ng/mg, max. 45.0 ng/mg). In addition, normal kidney tissue, known for high Gb3 levels, was extracted as a positive control. This tissue showed mean values of 85.72 ng of Gb3 per mg of tissue (min. 31.5 ng/mg, max. 124.7 ng/mg), which was 2.3 times higher than the mean of Gb3 in breast carcinoma. These data are summarized in Figure 1.Figure 1

Bottom Line: Gb3 positivity correlated with estrogen receptor expression (p≤0.01), whereas absence of Gb3 expression in primary tumors was correlated with the presence of lymph node metastases (p≤0.03). 65% of lymph node metastases were Gb3 positive and in 40% of tested patients, we observed a statistically significant increase of metastatic Gb3 expression (p≤0.04).Intravenous injections of fluorescent STxB into HBC xenografted mice showed preferential STxB accumulation in epithelial cells and cells with endothelial morphology of the tumor.Gb3 expressing HBCx can be used as a model for preclinical studies with STxB conjugates.

View Article: PubMed Central - PubMed

Affiliation: Endocytic Trafficking and Therapeutic Delivery Group, UMR3666 CNRS - U1143 INSERM, Institut Curie-Centre de Recherche, 26 rue d'Ulm, 75248 Paris Cedex 05, France. ludger.johannes@curie.fr.

ABSTRACT

Background: The B-subunit of Shiga toxin (STxB) specifically binds to the glycosphingolipid Gb3 that is highly expressed on a number of human tumors and has been shown to target tumor cells in mouse models and ex vivo on primary colon carcinoma specimen.

Methods: Using a novel ex vivo STxB labeling (ESL) method we studied Gb3 expression in cytological specimens of primary human breast tumors from 107 patients, and in synchronous lymph node metastases from 20 patients. Fluorescent STxB was incubated with fine-needle aspiration (FNA) specimens, and Gb3 expression was evaluated by fluorescence microscopy. Furthermore, 11 patient-derived human breast cancer xenografts (HBCx) were evaluated for expression of Gb3 by ESL and FACS. In addition, the biodistribution of fluorescent STxB conjugate was studied after intravenous injection in a Gb3 positive HBCx model.

Results: Gb3 expression was detected in 62 of 107 patients (57.9%), mainly in epithelial tumor cells. Gb3 positivity correlated with estrogen receptor expression (p≤0.01), whereas absence of Gb3 expression in primary tumors was correlated with the presence of lymph node metastases (p≤0.03). 65% of lymph node metastases were Gb3 positive and in 40% of tested patients, we observed a statistically significant increase of metastatic Gb3 expression (p≤0.04). Using concordant ESL and flow cytometry analysis, 6 out of 11 HBCx samples were scored positive. Intravenous injections of fluorescent STxB into HBC xenografted mice showed preferential STxB accumulation in epithelial cells and cells with endothelial morphology of the tumor.

Conclusion: The enhanced expression of Gb3 in primary breast carcinomas and its lymph node metastases indicate that the development of STxB-based therapeutic strategies is of interest in this pathology. Gb3 expressing HBCx can be used as a model for preclinical studies with STxB conjugates. Finally, the ESL technique on FNA represents a rapid and cost effective method for the stratification of patients in future clinical trials.

Show MeSH
Related in: MedlinePlus