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Cripto-1 as a novel therapeutic target for triple negative breast cancer.

Castro NP, Fedorova-Abrams ND, Merchant AS, Rangel MC, Nagaoka T, Karasawa H, Klauzinska M, Hewitt SM, Biswas K, Sharan SK, Salomon DS - Oncotarget (2015)

Bottom Line: The use of laser-capture microdissection combined with Nanostring mRNA and microRNA analysis revealed overexpression of either epithelial and miRNA-200 family or mesenchymal markers in adenocarcinoma and mesenchymal regions, respectively.Cripto-1 knockout by the CRISPR-Cas9 system inhibited tumor growth and pulmonary metastasis.Our findings show characterization of a novel mouse model that mimics the TNBC and reveal Cripto-1 as a TNBC target hence may offer alternative treatment strategies for TNBC.

View Article: PubMed Central - PubMed

Affiliation: Tumor Growth Factor Section, Mouse Cancer Genetics Program, National Cancer Institute, Frederick, MD, USA.

ABSTRACT
Triple-negative breast cancer (TNBC) presents the poorest prognosis among the breast cancer subtypes and no current standard therapy. Here, we performed an in-depth molecular analysis of a mouse model that establishes spontaneous lung metastasis from JygMC(A) cells. These primary tumors resembled the triple-negative breast cancer (TNBC) both phenotypically and molecularly. Morphologically, primary tumors presented both epithelial and spindle-like cells but displayed only adenocarcinoma-like features in lung parenchyma. The use of laser-capture microdissection combined with Nanostring mRNA and microRNA analysis revealed overexpression of either epithelial and miRNA-200 family or mesenchymal markers in adenocarcinoma and mesenchymal regions, respectively. Cripto-1, an embryonic stem cell marker, was present in spindle-like areas and its promoter showed activity in primary tumors. Cripto-1 knockout by the CRISPR-Cas9 system inhibited tumor growth and pulmonary metastasis. Our findings show characterization of a novel mouse model that mimics the TNBC and reveal Cripto-1 as a TNBC target hence may offer alternative treatment strategies for TNBC.

No MeSH data available.


Related in: MedlinePlus

Gene expression profiles of laser capture microdissected tumor progression samples through unsupervised hierarchical clusteringPrimary tumor adenocarcinoma versus EMT-like areas; B. Primary tumor adenocarcinoma versus lung metastasis; C. Primary tumor EMT-like areas versus lung metastasis; D. Primary tumor, lung metastasis, normal mammary gland and normal lung parenchyma. Each row represents a single gene and each column a single sample. The red color indicates up-regulation, the green color indicates down-regulation, and the black color indicates no change in expression level compared with the reference sample. The gray color indicates that no intensity was detected. EMT: epithelial-to-mesenchymal transition.
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Figure 3: Gene expression profiles of laser capture microdissected tumor progression samples through unsupervised hierarchical clusteringPrimary tumor adenocarcinoma versus EMT-like areas; B. Primary tumor adenocarcinoma versus lung metastasis; C. Primary tumor EMT-like areas versus lung metastasis; D. Primary tumor, lung metastasis, normal mammary gland and normal lung parenchyma. Each row represents a single gene and each column a single sample. The red color indicates up-regulation, the green color indicates down-regulation, and the black color indicates no change in expression level compared with the reference sample. The gray color indicates that no intensity was detected. EMT: epithelial-to-mesenchymal transition.

Mentions: To examine the gene expression profile of these mixed cell populations in the primary tumors and lung metastases, we integrated the use of laser capture microssection (LCM) and NanoString Technologies to identify expression levels of ESC and EMT-MET markers (Dataset S1A). Since ESCs are known for their ability to differentiate into somatic tissues derived from different germ layer cell types (ectoderm, mesoderm and endoderm) [7], we reasoned that the re-expression of ESC genes might also play a role during this metastatic process. Based on a collection of published literature, we built a customized NanoString nCounter® Gene Expression Codeset that contained significant ECS and EMT-MET markers, and targeted gene expression profiling on the following samples: primary tumor adenocarcinoma and EMT-like, lung metastases, normal mammary gland (NMG) and normal lung parenchyma. Figure 3 shows the most notably altered genes between these sample groups. An examination of gene expression patterns of the primary tumor that contained mixed areas revealed 31 genes differentially expressed (Figure 3A). Overexpression of ESC markers segregated distinctly in the epithelial cell-populated regions (Cd24 and Cd49f) and spindle-like cell areas (Klf4, Sox2 and Cripto-1). Moreover, overexpression of epithelial markers (Cadherin1, Epcam and Occludin) was observed in adenocarcinoma regions. For complete gene lists, see Dataset S1B.


Cripto-1 as a novel therapeutic target for triple negative breast cancer.

Castro NP, Fedorova-Abrams ND, Merchant AS, Rangel MC, Nagaoka T, Karasawa H, Klauzinska M, Hewitt SM, Biswas K, Sharan SK, Salomon DS - Oncotarget (2015)

Gene expression profiles of laser capture microdissected tumor progression samples through unsupervised hierarchical clusteringPrimary tumor adenocarcinoma versus EMT-like areas; B. Primary tumor adenocarcinoma versus lung metastasis; C. Primary tumor EMT-like areas versus lung metastasis; D. Primary tumor, lung metastasis, normal mammary gland and normal lung parenchyma. Each row represents a single gene and each column a single sample. The red color indicates up-regulation, the green color indicates down-regulation, and the black color indicates no change in expression level compared with the reference sample. The gray color indicates that no intensity was detected. EMT: epithelial-to-mesenchymal transition.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4494913&req=5

Figure 3: Gene expression profiles of laser capture microdissected tumor progression samples through unsupervised hierarchical clusteringPrimary tumor adenocarcinoma versus EMT-like areas; B. Primary tumor adenocarcinoma versus lung metastasis; C. Primary tumor EMT-like areas versus lung metastasis; D. Primary tumor, lung metastasis, normal mammary gland and normal lung parenchyma. Each row represents a single gene and each column a single sample. The red color indicates up-regulation, the green color indicates down-regulation, and the black color indicates no change in expression level compared with the reference sample. The gray color indicates that no intensity was detected. EMT: epithelial-to-mesenchymal transition.
Mentions: To examine the gene expression profile of these mixed cell populations in the primary tumors and lung metastases, we integrated the use of laser capture microssection (LCM) and NanoString Technologies to identify expression levels of ESC and EMT-MET markers (Dataset S1A). Since ESCs are known for their ability to differentiate into somatic tissues derived from different germ layer cell types (ectoderm, mesoderm and endoderm) [7], we reasoned that the re-expression of ESC genes might also play a role during this metastatic process. Based on a collection of published literature, we built a customized NanoString nCounter® Gene Expression Codeset that contained significant ECS and EMT-MET markers, and targeted gene expression profiling on the following samples: primary tumor adenocarcinoma and EMT-like, lung metastases, normal mammary gland (NMG) and normal lung parenchyma. Figure 3 shows the most notably altered genes between these sample groups. An examination of gene expression patterns of the primary tumor that contained mixed areas revealed 31 genes differentially expressed (Figure 3A). Overexpression of ESC markers segregated distinctly in the epithelial cell-populated regions (Cd24 and Cd49f) and spindle-like cell areas (Klf4, Sox2 and Cripto-1). Moreover, overexpression of epithelial markers (Cadherin1, Epcam and Occludin) was observed in adenocarcinoma regions. For complete gene lists, see Dataset S1B.

Bottom Line: The use of laser-capture microdissection combined with Nanostring mRNA and microRNA analysis revealed overexpression of either epithelial and miRNA-200 family or mesenchymal markers in adenocarcinoma and mesenchymal regions, respectively.Cripto-1 knockout by the CRISPR-Cas9 system inhibited tumor growth and pulmonary metastasis.Our findings show characterization of a novel mouse model that mimics the TNBC and reveal Cripto-1 as a TNBC target hence may offer alternative treatment strategies for TNBC.

View Article: PubMed Central - PubMed

Affiliation: Tumor Growth Factor Section, Mouse Cancer Genetics Program, National Cancer Institute, Frederick, MD, USA.

ABSTRACT
Triple-negative breast cancer (TNBC) presents the poorest prognosis among the breast cancer subtypes and no current standard therapy. Here, we performed an in-depth molecular analysis of a mouse model that establishes spontaneous lung metastasis from JygMC(A) cells. These primary tumors resembled the triple-negative breast cancer (TNBC) both phenotypically and molecularly. Morphologically, primary tumors presented both epithelial and spindle-like cells but displayed only adenocarcinoma-like features in lung parenchyma. The use of laser-capture microdissection combined with Nanostring mRNA and microRNA analysis revealed overexpression of either epithelial and miRNA-200 family or mesenchymal markers in adenocarcinoma and mesenchymal regions, respectively. Cripto-1, an embryonic stem cell marker, was present in spindle-like areas and its promoter showed activity in primary tumors. Cripto-1 knockout by the CRISPR-Cas9 system inhibited tumor growth and pulmonary metastasis. Our findings show characterization of a novel mouse model that mimics the TNBC and reveal Cripto-1 as a TNBC target hence may offer alternative treatment strategies for TNBC.

No MeSH data available.


Related in: MedlinePlus