REGULATION OF P120-CATENIN TUMOR SUPRESSOR ACTIVITIES
Douglas Stairs, Research Associate
University Of Pennsylvania, 3451 Walnut Street, Philadelphia, Pa 19104
Grant 1K99CA138498-01A1 from National Cancer Institute
Abstract: Esophageal cancer represents the 5th most frequent cancer in males worldwide. Given the poor survival rate, advanced stage of the disease at diagnosis and the increasing frequency of the disease it is increasingly important to understand the molecular mechanisms of initiation of these tumors as well as the genes involved in their metastasis. My research will focus on p120- catenin (p120ctn) and its ability to modulate tumorigenesis as well as cell migration and invasion in vitro and in vivo. P120ctn defines a subfamily of catenin proteins related to beta-catenin that also bind to E-cadherin and stabilizes E-cadherin at adherens junctions. As a result, expression of E-cadherin and p120ctn appear to be coordinately regulated in many cell lines and it is speculated that this may be another mechanism by which E-cadherin expression may be lost and lead to EMT. Functionally, we have demonstrated that successful genetic knockdown of p120ctn results in loss of the integrity of the adherens junctions with augmentation of tumor cell migration and invasion. This has been pursued as well in a genetically engineered mouse model through tissue specific ablation of p120ctn, resulting in inflammation and cancer in the esophagus. Therefore, we hypothesize that p120ctn regulates tumor cell migration and invasion by its ability to modulate effectors such as cdc42/rho/rac, and that p120ctn has a parallel and distinct role in tumorigenesis from that of other oncogenes and tumor suppressors. This hypothesis will be pursued by the following interrelated Specific Aims Aim 1 Identify the pathways regulated by p120ctn involved in cellular transformation Aim 1a Understand the functional role(s) of p120ctn in tumor initiation using immortalized esophageal epithelial cells (keratinocytes). Aim 1b Determine the functional consequences of p120ctn loss in primary esophageal keratinocytes. Aim2 Determine the functional interaction between p120ctn and EGFR overexpression in esophageal tumor initiation. Aim 3 Determine the functional role(s) of p120ctn in tumor progression in mouse models of esophageal cancer. Over 14,000 new cases of esophageal cancer were diagnosed in the United States last year and more than 90% of those diagnosed will die of their disease, primarily from metastatic lesions. The proposed studies will provide novel insights into the biological roles of p120ctn in the development and progression of esophageal cancer. Ultimately, these studies may translate into new diagnostic and therapeutic modalities for this deadly disease
Keywords: ARF; Ablation; Adenocarcinoma of the Esophagus; Adherens Junction; Adhering Junction; Adhesive Junction; Anchoring Junction; Anti-Oncogenes; Antioncogenes; Binding; Binding (Molecular Function); Biological; Biological Models; Body Tissues; CAM 120/80; CCND1 Protein; CDK4I; CDKN2; CDKN2A; CDKN2A gene; CMM2; CTNNB1; CUL-2; Cadherin-1; Cadherin-Associated Protein, Beta; Cadherin-Associated Protein, Beta 1 (88kD); Cancer Biology; Cancer Genes; Cancer of the Esophagus; Cancer-Promoting Gene; Cancers; Catenin, Beta-1; Cell Culture Techniques; Cell Line; Cell Lines, Strains; Cell Locomotion; Cell Migration; Cell Movement; CellLine; Cells; Cellular Migration; Cellular Transformation; Crossmatching, Tissue; Cyclin D1; Cyclin-Dependent Kinase Inhibitor 2A Gene; Cytoplasm; Development; Diagnosis; Disease; Disorder; E-Cadherin; EGFR; ERBB Protein; ERBB1; Emerogenes; Epidermal Growth Factor Receptor; Epidermal Growth Factor Receptor Kinase; Epidermal Growth Factor Receptor Protein-Tyrosine Kinase; Epidermoid Cell Cancer; Epithelial Calcium-Dependent Adhesion Protein; Epithelial Cells; Epithelial-Cadherin; Esophageal; Esophageal Adenocarcinoma; Esophageal Cancer; Esophageal Neoplasms; Esophageal Neoplasms, Benign and Malignant; Esophageal Squamous Cell; Esophageal Tumor; Esophagus; Esophagus Cancer; Esophagus Neoplasm; Forecast of outcome; Frequencies (time pattern); Frequency; G1/S-Specific Cyclin D1; Gastrointestinal Tract, Esophagus; Genes; Genes, CDKN2; Genes, Cancer Suppressor; Genes, Onco-Suppressor; Genes, p16; Genes, p16INK4A; Genes, p53; Genetic; Genetic Alteration; Genetic Change; Genetic defect; Genetically Engineered Mouse; HER1; Histocompatibility Testing; Human; Human, General; INFLM; INK4; INK4A; Immune; In Vitro; Inflammation; Intestinal; Intestines; Lead; Limited Stage; MLM; MTS1; MTS1 Genes; Malignant Epidermoid Cell Neoplasm; Malignant Epidermoid Cell Tumor; Malignant Esophageal Neoplasm; Malignant Esophageal Tumor; Malignant Neoplasms; Malignant Squamous Cell Neoplasm; Malignant Squamous Cell Tumor; Malignant Tumor; Malignant Tumor of the Esophagus; Malignant neoplasm of esophagus; Man (Taxonomy); Man, Modern; Mentors; Metaplastic; Metastasis; Metastasize; Metastatic Lesion; Metastatic Neoplasm; Metastatic Tumor; Modality; Model System; Models, Biologic; Molecular; Molecular Interaction; Motility; Motility, Cellular; Mutation; Neoplasm Metastasis; Oncogenes; Oncogenes, Recessive; Oncogenes-Tumor Suppressors; Oncogenesis; P53; PRAD1 Protein; PRO2286; Pathogenesis; Pathway interactions; Pb element; Phenotype; Prognosis; Proteins; Proto-Oncogene Proteins c-bcl-1; Receptor, EGF; Receptor, TGF-alpha; Receptor, Urogastrone; Receptors, Epidermal Growth Factor-Urogastrone; Regulation; Research; Research Proposals; Role; Secondary Neoplasm; Secondary Tumor; Squamous Cell Cancers; Squamous Epithelium; Staging; Survival Rate; TP16; TP53; TP53 gene; TRP53; TSG9A; Therapeutic; Therapeutic Intervention; Tissue Crossmatchings; Tissue Typing; Tissues; Transforming Genes; Transforming Growth Factor alpha Receptor; Translating; Translatings; Tumor Cell Migration; Tumor Protein p53 Gene; Tumor Suppressing Genes; Tumor Suppressor Genes; Tumor of the Esophagus; United States; Uvomorulin; base; bcl-1 Proto-Oncogene Products; bcl-1 Proto-Oncogene Proteins; bcl1 Proto-Oncogene Proteins; beta catenin; bowel; c-bcl-1 Proteins; c-erbB-1; c-erbB-1 Protein; cancer metastasis; cancer progression; catenin p120(ctn) protein; catenin p120ctn protein; cell motility; combinatorial; cultured cell line; cyclin D; disease/disorder; erbB-1; erbB-1 Proto-Oncogene Protein; erbBl; gene product; genome mutation; heavy metal Pb; heavy metal lead; histocompatibility typing; in vivo; insight; intervention therapy; keratinocyte; language translation; male; malignancy; metaplastic cell transformation; mouse model; neoplasm progression; neoplasm/cancer; neoplastic progression; new diagnostics; next generation diagnostics; novel; novel diagnostics; oncosuppressor gene; outcome forecast; overexpression; p120(ctn); p120ctn; p14ARF; p16INK4 Genes; p16INK4a; pathway; proto-oncogene protein c-erbB-1; public health relevance; rho; social role; therapeutic target; transdifferentiation; tumor; tumor initiation; tumor progression; tumorigenesis
Relevance: Over 14,000 new cases of esophageal cancer were diagnosed in the United States last year and more than 90% of those diagnosed will die of their disease, primarily from metastatic lesions. The proposed studies will provide novel insights into the biological roles of p120ctn in the development and progression of esophageal cancer. Ultimately, these studies may translate into new diagnostic and therapeutic modalities for this deadly disease
Project start date: 2009-09-29
Project end date: 2011-08-31
Budget start date: 29-SEP-2009
Budget end date: 31-AUG-2010
PFA/PA: PA-09-036
1K99CA138498-01A1 (2009): $137376
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to Douglas Stairs
REGULATION OF P120-CATENIN TUMOR SUPRESSOR ACTIVITIES
Douglas Stairs
University Of Pennsylvania, 3451 Walnut Street, Philadelphia, Pa 19104
Grant 5K99CA138498-02 from National Cancer Institute
Abstract: Esophageal cancer represents the 5th most frequent cancer in males worldwide. Given the poor survival rate, advanced stage of the disease at diagnosis and the increasing frequency of the disease it is increasingly important to understand the molecular mechanisms of initiation of these tumors as well as the genes involved in their metastasis. My research will focus on p120- catenin (p120ctn) and its ability to modulate tumorigenesis as well as cell migration and invasion in vitro and in vivo. P120ctn defines a subfamily of catenin proteins related to beta-catenin that also bind to E-cadherin and stabilizes E-cadherin at adherens junctions. As a result, expression of E-cadherin and p120ctn appear to be coordinately regulated in many cell lines and it is speculated that this may be another mechanism by which E-cadherin expression may be lost and lead to EMT. Functionally, we have demonstrated that successful genetic knockdown of p120ctn results in loss of the integrity of the adherens junctions with augmentation of tumor cell migration and invasion. This has been pursued as well in a genetically engineered mouse model through tissue specific ablation of p120ctn, resulting in inflammation and cancer in the esophagus. Therefore, we hypothesize that p120ctn regulates tumor cell migration and invasion by its ability to modulate effectors such as cdc42/rho/rac, and that p120ctn has a parallel and distinct role in tumorigenesis from that of other oncogenes and tumor suppressors. This hypothesis will be pursued by the following interrelated Specific Aims Aim 1 Identify the pathways regulated by p120ctn involved in cellular transformation Aim 1a Understand the functional role(s) of p120ctn in tumor initiation using immortalized esophageal epithelial cells (keratinocytes). Aim 1b Determine the functional consequences of p120ctn loss in primary esophageal keratinocytes. Aim2 Determine the functional interaction between p120ctn and EGFR overexpression in esophageal tumor initiation. Aim 3 Determine the functional role(s) of p120ctn in tumor progression in mouse models of esophageal cancer. Over 14,000 new cases of esophageal cancer were diagnosed in the United States last year and more than 90% of those diagnosed will die of their disease, primarily from metastatic lesions. The proposed studies will provide novel insights into the biological roles of p120ctn in the development and progression of esophageal cancer. Ultimately, these studies may translate into new diagnostic and therapeutic modalities for this deadly disease
Keywords: ARF; Ablation; Adenocarcinoma of the Esophagus; Adherens Junction; Adhering Junction; Adhesive Junction; Anchoring Junction; Anti-Oncogenes; Antioncogenes; Binding; Binding (Molecular Function); Biological; Biological Models; Body Tissues; CAM 120/80; CCND1 Protein; CDK4I; CDKN2; CDKN2A; CDKN2A gene; CMM2; CTNNB1; CUL-2; Cadherin-1; Cadherin-Associated Protein, Beta; Cadherin-Associated Protein, Beta 1 (88kD); Cancer Biology; Cancer Genes; Cancer of the Esophagus; Cancer-Promoting Gene; Cancers; Catenin, Beta-1; Cell Culture Techniques; Cell Line; Cell Lines, Strains; Cell Locomotion; Cell Migration; Cell Movement; CellLine; Cells; Cellular Migration; Cellular Transformation; Crossmatching, Tissue; Cyclin D1; Cyclin-Dependent Kinase Inhibitor 2A Gene; Cytoplasm; Development; Diagnosis; Disease; Disorder; E-Cadherin; EGFR; ERBB Protein; ERBB1; Emerogenes; Epidermal Growth Factor Receptor; Epidermal Growth Factor Receptor Kinase; Epidermal Growth Factor Receptor Protein-Tyrosine Kinase; Epidermoid Cell Cancer; Epithelial Calcium-Dependent Adhesion Protein; Epithelial Cells; Epithelial-Cadherin; Esophageal; Esophageal Adenocarcinoma; Esophageal Cancer; Esophageal Neoplasms; Esophageal Neoplasms, Benign and Malignant; Esophageal Squamous Cell; Esophageal Tumor; Esophagus; Esophagus Cancer; Esophagus Neoplasm; Forecast of outcome; Frequencies (time pattern); Frequency; G1/S-Specific Cyclin D1; Gastrointestinal Tract, Esophagus; Genes; Genes, CDKN2; Genes, Cancer Suppressor; Genes, Onco-Suppressor; Genes, p16; Genes, p16INK4A; Genes, p53; Genetic; Genetic Alteration; Genetic Change; Genetic defect; Genetically Engineered Mouse; HER1; Histocompatibility Testing; Human; Human, General; INFLM; INK4; INK4A; Immune; In Vitro; Inflammation; Intestinal; Intestines; Lead; Limited Stage; MLM; MTS1; MTS1 Genes; Malignant Epidermoid Cell Neoplasm; Malignant Epidermoid Cell Tumor; Malignant Esophageal Neoplasm; Malignant Esophageal Tumor; Malignant Neoplasms; Malignant Squamous Cell Neoplasm; Malignant Squamous Cell Tumor; Malignant Tumor; Malignant Tumor of the Esophagus; Malignant neoplasm of esophagus; Man (Taxonomy); Man, Modern; Mentors; Metaplastic; Metastasis; Metastasize; Metastatic Lesion; Metastatic Neoplasm; Metastatic Tumor; Modality; Model System; Models, Biologic; Molecular; Molecular Interaction; Motility; Motility, Cellular; Mutation; Neoplasm Metastasis; Oncogenes; Oncogenes, Recessive; Oncogenes-Tumor Suppressors; Oncogenesis; P53; PRAD1 Protein; PRO2286; Pathogenesis; Pathway interactions; Pb element; Phenotype; Prognosis; Proteins; Proto-Oncogene Proteins c-bcl-1; Receptor, EGF; Receptor, TGF-alpha; Receptor, Urogastrone; Receptors, Epidermal Growth Factor-Urogastrone; Regulation; Research; Research Proposals; Role; Secondary Neoplasm; Secondary Tumor; Squamous Cell Cancers; Squamous Epithelium; Staging; Survival Rate; TP16; TP53; TP53 gene; TRP53; TSG9A; Therapeutic; Therapeutic Intervention; Tissue Crossmatchings; Tissue Typing; Tissues; Transforming Genes; Transforming Growth Factor alpha Receptor; Translating; Translatings; Tumor Cell Migration; Tumor Protein p53 Gene; Tumor Suppressing Genes; Tumor Suppressor Genes; Tumor of the Esophagus; United States; Uvomorulin; base; bcl-1 Proto-Oncogene Products; bcl-1 Proto-Oncogene Proteins; bcl1 Proto-Oncogene Proteins; beta catenin; bowel; c-bcl-1 Proteins; c-erbB-1; c-erbB-1 Protein; cancer metastasis; cancer progression; catenin p120(ctn) protein; catenin p120ctn protein; cell motility; combinatorial; cultured cell line; cyclin D; disease/disorder; erbB-1; erbB-1 Proto-Oncogene Protein; erbBl; gene product; genome mutation; heavy metal Pb; heavy metal lead; histocompatibility typing; in vivo; insight; intervention therapy; keratinocyte; language translation; male; malignancy; metaplastic cell transformation; mouse model; neoplasm progression; neoplasm/cancer; neoplastic progression; new diagnostics; next generation diagnostics; novel; novel diagnostics; oncosuppressor gene; outcome forecast; overexpression; p120(ctn); p120ctn; p14ARF; p16INK4 Genes; p16INK4a; pathway; proto-oncogene protein c-erbB-1; public health relevance; rho; social role; therapeutic target; transdifferentiation; tumor; tumor initiation; tumor progression; tumorigenesis
Relevance: Over 14,000 new cases of esophageal cancer were diagnosed in the United States last year and more than 90% of those diagnosed will die of their disease, primarily from metastatic lesions. The proposed studies will provide novel insights into the biological roles of p120ctn in the development and progression of esophageal cancer. Ultimately, these studies may translate into new diagnostic and therapeutic modalities for this deadly disease
Project start date: 2009-09-29
Project end date: 2011-08-31
Budget start date: 1-SEP-2010
Budget end date: 31-AUG-2011
PFA/PA: PA-09-036
5K99CA138498-02 (2010): $140525
ROLE OF P120CTN IN ESOPHAGEAL CANCER
Douglas Stairs, Research Associate
University Of Pennsylvania, 3451 Walnut Street, Philadelphia, Pa 19104
Grant 5F32CA130851-03 from National Cancer Institute
Abstract: Esophageal cancer represents the 5th most frequent cancer in males worldwide. Given the poor survival rate, advanced stage of the disease at diagnosis and the increasing frequency of the disease it is increasingly important to understand the molecular mechanisms of initiation of these tumors as well as the genes involved in their metastasis. My research will focus on the catenin family member p120ctn and its ability to modulate tumorigenesis as well as cell migration and invasion in vitro and in vivo. p120ctn defines a family of catenin proteins related to beta-catenin that also bind to E-cadherin and stabilizes E-cadherin at adherens junctions. As a result, expression of E-cadherin and p120ctn appear to be coordinately regulated in many cell lines and it is speculated that this may be another mechanism by which E-cadherin expression may be lost and lead to EMT. Interestingly, p120ctn contains 16 different phosphorylation sites, 8 tyrosine and 8 serine/threonine. What kinases directly phosphorylate these sites and under what stimuli are largely unknown. However, it is clear that EGFR activation can induce phosphorylation of p120ctn at least at Y228. Additionally, many splice forms for p120ctn exist. We hypothesize that the different isoforms and phosphorylation sites of p120ctn may regulate its ability to interact with its binding partners and therefore alter its ability to promote tumorigenesis and metastasis. This hypothesis will be pursued by the following interrelated specific aims. Aim 1 To assess the effects different isoforms and phosphorylation mutants have on motility and invasiveness. Expression of p120ctn will be knocked-down in several esophageal cell lines. Various isoforms and phosphorylation-deficient mutants will be introduced and motility and invasiveness of the mutants will be determined through monolayer assays as well as three-dimensional Matrigel and organotypic culture models. Aim2 To understand the role of p120ctn in tumor initiation and progression. We will overexpress EGFR and knockdown p120ctn expressin in vivo and in vitro. We will monitor these EGFR-overexpressing, p120ctn-deleted mice and cell lines for esophageal tumorigenesis and metastasis. Over 14,000 new cases of esophageal cancer were diagnosed in the United States last year and more than 90% of those diagnosed will die of their disease, primarily from metastatic lesions. The proposed studies will provide novel insights into the biological roles of p120ctn in the development and progression of esophageal cancer. Ultimately, these studies may translate into new diagnostic and therapeutic modalities for this deadly disease
Keywords: Adherens Junction; Adhering Junction; Adhesive Junction; Agar; American Cancer Society; Amino Acids; Anchoring Junction; Assay; Binding; Binding (Molecular Function); Bioassay; Biochemical; Biologic Assays; Biological; Biological Assay; Burkitt Herpesvirus; Burkitt Lymphoma Virus; CAM 120/80; CTNNB1; CUL-2; Cadherin-1; Cadherin-Associated Protein, Beta; Cadherin-Associated Protein, Beta 1 (88kD); Cancer of the Esophagus; Cancers; Catenin, Beta-1; Cell Communication and Signaling; Cell Line; Cell Line, Tumor; Cell Lines, Strains; Cell Locomotion; Cell Migration; Cell Movement; Cell Signaling; CellLine; Cells; Cellular Migration; Cessation of life; Death; Development; Diagnosis; Disease; Disorder; E-B Virus; E-Cadherin; EB virus; EBV; EC 2.7; EGFR; EGFR Protein Overexpression; ERBB Protein; ERBB1; Epidermal Growth Factor Receptor; Epidermal Growth Factor Receptor Kinase; Epidermal Growth Factor Receptor Overexpression; Epidermal Growth Factor Receptor Protein-Tyrosine Kinase; Epithelial Calcium-Dependent Adhesion Protein; Epithelial-Cadherin; Epstein Barr Virus; Epstein-Barr Virus; Esophageal; Esophageal Cancer; Esophageal Epidermoid Carcinoma; Esophageal Neoplasms; Esophageal Neoplasms, Benign and Malignant; Esophageal SCC; Esophageal Squamous Cell Carcinoma; Esophageal Tumor; Esophagus; Esophagus Cancer; Esophagus Neoplasm; Event; Family; Family member; Frequencies (time pattern); Frequency; Gastrointestinal Tract, Esophagus; Generalized Growth; Genes; Genetic Alteration; Genetic Change; Genetic defect; Growth; HER1; HHV-4; Herpesvirus 4 (gamma), Human; Herpesvirus 4, Human; Human; Human Herpesvirus 4; Human, General; In Vitro; Infectious Mononucleosis Virus; Intracellular Communication and Signaling; Isoforms; Kinases; L-Serine; L-Threonine; L-Tyrosine; Lead; Lesion; Malignant Cell; Malignant Esophageal Neoplasm; Malignant Esophageal Tumor; Malignant Neoplasms; Malignant Tumor; Malignant Tumor of the Esophagus; Malignant neoplasm of esophagus; Mammals, Mice; Man (Taxonomy); Man, Modern; Metastasis; Metastasize; Metastatic Lesion; Metastatic Neoplasm; Metastatic Tumor; Mice; Modality; Modeling; Molecular; Molecular Interaction; Monitor; Motility; Motility, Cellular; Mouse Cell Line; Murine; Mus; Mutation; Neoplasm Metastasis; Oncogenesis; PRO2286; Pb element; Phosphorylation; Phosphorylation Site; Phosphotransferases; Promoter; Promoters (Genetics); Promotor; Promotor (Genetics); Protein Isoforms; Protein Phosphorylation; Proteins; RNA Splicing; Receptor Activation; Receptor, EGF; Receptor, TGF-alpha; Receptor, Urogastrone; Receptors, Epidermal Growth Factor-Urogastrone; Research; Role; SCC of the Esophagus; Secondary Neoplasm; Secondary Tumor; Serine; Signal Transduction; Signal Transduction Systems; Signaling; Site; Splicing; Squamous Cell Carcinoma of the Esophagus; Staging; Stimulus; Survival Rate; TYR; Therapeutic; Threonine; Tissue Growth; Transforming Growth Factor alpha Receptor; Translating; Translatings; Transphosphorylases; Tumor Cell Line; Tumor Cell Migration; Tumor of the Esophagus; Tyrosine; Tyrosine, L-isomer; United States; Uvomorulin; aminoacid; beta catenin; biological signal transduction; c-erbB-1; c-erbB-1 Protein; cancer cell; cancer metastasis; catenin p120(ctn) protein; catenin p120ctn protein; cell motility; cultured cell line; disease/disorder; epithelial to mesenchymal transition; erbB-1; erbB-1 Proto-Oncogene Protein; erbBl; experiment; experimental research; experimental study; gene product; genome mutation; heavy metal Pb; heavy metal lead; human herpesvirus 4 group; in vivo; insight; knock-down; language translation; male; malignancy; matrigel; migration; monolayer; mutant; neoplasm/cancer; new diagnostics; next generation diagnostics; novel; novel diagnostics; ontogeny; overexpression; p120(ctn); p120ctn; para-Tyrosine; proto-oncogene protein c-erbB-1; research study; shRNA; short hairpin RNA; small hairpin RNA; social role; tissue culture; tumor; tumor initiation; tumorigenesis; tumorigenic
Project start date: 2007-07-01
Project end date: 2010-06-30
Budget start date: 1-JUL-2009
Budget end date: 30-JUN-2010
PFA/PA: PA-06-373
5F32CA130851-03 (2009): $51710
5F32CA130851-02 (2008): $49646
1F32CA130851-01 (2007): $46826