Wei Dai
New York University School Of Medicine
Project start date: 2012-01-16
Project end date: 2016-10-31
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to Wei Dai
TIF AND ERYTHROLEUKEMIA CELL DIFFERENTIATION
Wei Dai, Professor
Institute For Cancer Prevention
1 Dana Rd
valhalla, Ny 105951549
Grant 7R01CA059985-06 from National Cancer Institute IRG: HEM
Abstract: This revised application describes studies to elucidate the functional role and molecular mechanism of action of tif, a new member of the axl family of receptors. Tif, which was cloned by RTPCR from a K562 library, is an extremely interesting receptor in that it bears extracellular fibronectin domains as well as Ig domains, suggesting a role in cell-matrix attachment and signalling. In addition, its pattern of RNA expression, gonad>brain>other tissues, suggests a potential role in primitive cell maintenance and self-renewal. In this application, proposes to 1) Study the transforming potential of tif by enforced over- expression; 2) Investigate the role of tif in hematopoietic determination and differentiation by analyzing the phenotype of both K562 and Ba/F3 cells constitutively expressing activated tif, and by differentiating ES cells with a targeted gene disruption of the tif locus; 3)Identify the immediate downstream components of the tif signaling pathway via immonocoprecipitation and via the yeast two-hybrid system; 4)Search for a functional interaction between tif and the Epo receptor by enforced expression of an EpoR/tif chimera, analyzing signal transduction and cellular phenotype in transfected Ba/F3 and K562 cells; and 5)Identify and clone the tif ligand via expression or affinity column chromatography. These studies are designed to provide the first important steps towards understanding the role of Tif in hematopoietic differentiation
Keywords: biological signal transduction, cell differentiation, erythroleukemia, growth factor receptor, neoplastic cell, protein tyrosine kinase cell cycle, colony stimulating factor, enzyme activity, erythroid stem cell, erythropoietin, gene expression, hematopoiesis, intermolecular interaction, phenotype, receptor expression affinity chromatography, immunoprecipitation, tissue /cell culture, yeast
Project start date: 1995-07-01
Project end date: 2001-04-30
7R01CA059985-06 (2000): $63188
5R01CA059985-05 (1999): $243103
5R01CA059985-04 (1998): $233752
5R01CA059985-03 (1997): $224763
PROTEIN SER/THR KINASE PRK AND CELL CYCLE PROGRESSION
Wei Dai, Professor
Institute For Cancer Prevention
1 Dana Rd
valhalla, Ny 105951549
Grant 5R01CA074229-05 from National Cancer Institute IRG: HEM
Abstract: This proposal focuses upon the role of a newly cloned serine-threonine kinase termed "prk". This kinase is homologous to the budding yeast cdc5 and Drosophila polo and moreover is strongly homologous with the previously described murine kinase fnk. Prk is most likely to be the human homolog of the murine fnk. The physiological role of this kinase has been investigated in several systems by these workers. Specifically, it has been found to enhance progesterone induced meiotic maturation of Xenopus oocytes whereas antisense prk transcripts inhibit their maturation. It is capable of rescuing a thermosensitive cdc5 mutant of Saccharomyces cerevisiae. Its cell cycle regulation has been investigated and found to peak in the late S and G2 phases. Most interestingly, its expression is activated by thrombopoietin in MO7e megakaryocyte cells and other megakaryocytic cell lines and correlated with megakaryocytic differentiation of Dami cells. Finally, it has been found to map to chromosomal locus 8p21, a region proposed to contain tumor suppressor genes. It has been found to be commonly down-regulated in lung tumors compared to normal tissues. Ectopic expression in fibroblasts reduces growth rate and there is some indication of interaction with pRB. The objectives of the proposed studies are 1) To examine whether prk plays a role in regulating endomitosis in megakaryocytes studied by ectopic expression and by inhibition using antisense and dominant negative mutants; 2) To investigate whether prk is mutated, deleted, and/or inactivated in spontaneous lung and breast cancers; 3) To identify proteins interacting with prk through affinity purification, the yeast two-hybrid system and in vitro phosphorylation screening of an expression library
Keywords: cell cycle, cell cycle protein, cell differentiation, cell growth regulation, cell proliferation, enzyme activity, protein kinase breast neoplasm, gene deletion mutation, gene expression, lung neoplasm, mutant, neoplasm /cancer genetics, phosphorylation, tumor suppressor gene affinity chromatography, antisense nucleic acid, athymic mouse, flow cytometry, nucleic acid sequence, polymerase chain reaction, protein purification, southern blotting, tissue /cell culture, yeast two hybrid system
Project start date: 1997-12-15
Project end date: 2001-09-30
5R01CA074229-05 (2001): $252060
7R01CA074229-04 (2000): $217748
5R01CA074229-03 (2000): $30745
5R01CA074229-02 (1999): $233050
1R01CA074229-01A1 (1998): $209204
BUBR1 In The Spindle Checkpoint And Tumor Suppression
Wei Dai, Professor
Institute For Cancer Prevention
1 Dana Rd
valhalla, Ny 105951549
Grant 1R01CA090658-01 from National Cancer Institute IRG: MEP
Abstract: The spindle checkpoint delays the progression of metaphase to anaphase until all chromosomes are properly attached to the mitotic spindle. A loss of the checkpoint function often results in genetic instability, which predisposes cells to malignant transformation. To understand the molecular basis of functional inactivation of this surveillance mechanism in human cancer development, the PI´s laboratory has focused on the roles of two protein kinases, hBUB1 and hBUBR1 (42;53;81), mutations of which are detected in colorectal cancer cells (4). Recent studies suggest that hBUBR1 is an important component of the spindle checkpoint signaling pathway, and that it targets the anaphase-promoting complex (APC) through interaction with p55CDC/hCdc2O. Specifically, whereas hBUBR1 is expressed as a 120-kDa entity in various cell lines during interphase, nocodazole (Noc)-arrested mitotic cells contain a distinct hBUBR1 species that exhibits a reduced mobility on SDS-polyacrylamide gels. Phosphatase treatment restored the mobility of the mitotic hBUBR1 species to that characteristic of the interphase state, indicating that the decrease in mobility of hBUBR1 is due to phosphorylation. Yeast two-hybrid, glutathione S-transferase pull-down, and immunoprecipitation analyses revealed that p55CDC interacts with hBUBR1. hBUBR1 also phosphorylated p55CDC in vitro. Furthermore, the APC components CDC16 and CDC27 present in lysates of Noc-treated cells interact with hBUBR1 affinity resins. Finally, we have shown that the stress-activated protein kinase p38 interacts with hBUBR1 and that Noc-induced phosphorylation of hBUBR1 in HeLa cells is blocked by pretreatment with a specific inhibitor of p38. On the basis of the known biochemical and biological activities of hBUBR1, we therefore hypothesize that activation of hBUBR1 by p38 induces its interaction with p55CDC and subsequent inhibition of the APC during spindle checkpoint activation, and that a loss of hBUBR1 function due to structural abnormalities may result in aneuploidy and cancer. To test this hypothesis, we will (i) investigate whether a dominant negative mutant of mouse BUBR1 promotes tumorigenesis in transgenic mice; (ii) determine whether p38 is an immediate upstream activator of hBUBR1 during spindle checkpoint activation; (iii) define the domain of p55CDC that interacts with hBUBR1; (iv) examine whether hBUBR1 phosphorylates p5SCDC in vivo and whether hBUBR1 and the CDK1-cyclin B complex phosphorylate p55CDC on different sites; and (v) evaluate whether hBUBR1 inhibits APC activity by phosphorylating CDC16 and CDC27. The long-term goal of this project is to determine the mechanism by which protein phosphorylation regulates the spindle checkpoint and how deregulation of it may result in cancer
Keywords: cell cycle, intestine neoplasm, mitotic spindle, neoplastic transformation, protein kinase aneuploidy, cell cycle protein, cyclin dependent kinase, enzyme activity, gene mutation, genetic promoter element, phosphorylation 3T3 cell, athymic mouse, immunoprecipitation, transfection, transgenic animal, yeast two hybrid system
Project start date: 2001-04-06
Project end date: 2001-09-30
1R01CA090658-01 (2001): $256835
Sponsored Links Excellgen http://Excellgen.com
A NOVEL MOUSE COLON CANCER MODEL AND CHEMOPREVENTION
Wei Dai
New York University School Of Medicine, New York, Ny 10016
Grant 5R01CA113349-05 from National Cancer Institute
Abstract: The spindle checkpoint delays the progression from metaphase to anaphase until all condensed chromosomes are properly attached to mitotic spindles. An impaired checkpoint function often results in genomic instability, which predisposes cells to malignant transformation. To understand the molecular basis of functional inactivation of this surveillance mechanism in human cancer development, the Pi´s laboratory has focused on the role of BubRl, a key spindle checkpoint kinase, in the maintenance of genomic stability and suppression of tumorigenesis. The Pi´s group has generated BubRl+/- mice as well as BubRl+/-ApcMin/+ compound mutant mice. BubRl+/- mice develop intestinal adenocarcinomas at an accelerated rate after azoxymethane treatment. Whereas ApcMin/+ mice develop many adenomatous polyps generally within the small intestine, BubRl+/-ApcMin/+ compound mutant mice develop significantly more spontaneous colonic tumors than ApcMin/+ mice. The colon tumors in BubRl+/-ApcMin/+ mice are clinically more advanced than those observed in ApcMin/+ mice. Moreover, chemopreventive compounds such as sulindac sulfide and S-allylmercaptocysteine are capable of inducing apoptosis in HT-29 and SW-480 colon tumor cells; BubRl - deficient cells are more resistant to apoptosis induced by these compounds. Given that both BubRl and Ape are involved in the regulation of genomic stability in normal cells, we hypothesize that BubRl+/-ApcMin+/- mice would be an excellent rodent model for evaluating the efficacy of anti-tumor activities of various chemopreventive compounds that target colon and understanding the in vivo role of spindle checkpoint components in the maintenance of genomic stability. To test this hypothesis, the Pi´s lab will (i) validate and test the chemopreventive effect of sulindac and S-allylmercaptocysteine, which (or the derivative of which) directly target microtubules or mitotic spindles, on suppression of spontaneous intestinal tumorigenesis in BubRl+/- ApcMin/+ compound mutant mice, and (ii) investigate the molecular basis of intestinal carcinogenesis in these mutant mice by studying (a) the genomic instability and the rate of spontaneous transformation of cells deficient in BubRl and/or Ape, (b) the dependence of anti-proliferative effect of sulindac and S-allylmercaptocysteine on the integrity of the spindle checkpoint, and (c) the mechanism by which ApcMin/+ mice shift in tumor burden from the small intestine to colon in the BubRl-deficient genetic background. The long-term goal of this project is to elucidate the mechanism by which cell cycle checkpoints and the Wnt signaling pathway regulate cell proliferation and differentiation as well as genomic stability
Keywords: (8S-cis)-10-[(3-Amino-2, 3, 6-trideoxy-alpha-L-lyxo-hexopyranosyl)oxy]-7, 8, 9, 10-tetrahydro-6, 8, 11-trihydroxy-8-(hydroacetyl)-1-methoxy-5, 12-naphthacenedione; (Z)-2-[4(1, 2-diphenyl-1-butenyl)-phenoxyl]-N, N-dimethylethanamine; (Z)-5-fluoro-2-methyl-1-[[4-(methylsulfinyl)phenyl]methylene]-1H-indene-3-acetic acid; 1-p-beta-dimethylamino-ethoxyphenyl-trans-1, 2-diphenylbut-1-ene; 14-Hydroxydaunomycin; 1H-Indene-3-acetic acid, 5-fluoro-2-methyl-1-(4-(methylthio)phenyl)methylene-, (Z)-; 5, 12-Naphthacenedione, 10-((3-amino-2, 3, 6-trideoxy-alpha-L-lyxo-hexopyranosyl)oxy)-7, 8, 9, 10-tetrahydro-6, 8, 11-trihydroxy-8-(hydroxyacetyl)-1-methoxy-, (8S-cis)-; APC - Adenomatous Polyposis Coli; Adenocarcinoma; Adenoma, Malignant; Adenomatous Polyposis Coli; Adenomatous Polyposis Coli, Familial; Adenomatous Polyposis of the Colon; Adenomatous Polyps; Adriamycine; Anaphase; Animal Model; Animal Models and Related Studies; Anti-Cancer Agents; Anti-Inflammatory Agents, Non-Steroidal; Anti-Tumor Agents; Anti-Tumor Drugs; Antiinflammatory Agents, Non Steroidal; Antiinflammatory Agents, Nonsteroidal; Antineoplastic Agents; Antineoplastic Drugs; Antineoplastics; Antiproliferative Agents; Antiproliferative Drugs; Anzatax; Apes; Apoptosis; Apoptosis Pathway; Asotax; Azoxymethane; Biochemical; Biological; Birth; Bristaxol; Cancer Causing Agents; Cancer Drug; Cancer Induction; Cancer Model; Cancer Susceptibility Gene; Cancer-Predisposing Gene; CancerModel; Cancers; Carcinogens; Cell Cycle Checkpoint; Cell Death, Programmed; Cell Function; Cell Growth in Number; Cell Multiplication; Cell Process; Cell Proliferation; Cell physiology; Cells; Cellular Function; Cellular Physiology; Cellular Process; Cellular Proliferation; Chemoprevention; Chemopreventive; Chemopreventive Agent; Chemotherapeutic Agents, Neoplastic Disease; Chromosomal Stability; Chromosomes; Cola; Colon; Colon Cancer; Colon Carcinoma; Colon Neoplasms; Colonic Cancer; Colonic Carcinoma; Colonic Mass; Colonic Neoplasms; Colonic Tumor; DNA; DOX; Defect; Deoxyribonucleic Acid; Dependence; Development; Diazene, dimethyl-, 1-oxide; Doxorubicin; Doxorubicina; Drugs; EC 2.7; Epithelial Cells; Ethanamine, 2-(4-(1, 2-diphenyl-1-butenyl)phenoxy)-N, N-dimethyl-, (Z)-; Familial Adenomatous Polyposis Syndrome; Gastrointestinal Tract, Small Intestine; Gene Targeting; Genetic; Genetic Alteration; Genetic Change; Genetic defect; Genome; Genome Instability; Genome Stability; Genomic Instability; Genus Cola; Goals; HT-29; HT-29 Cells; HT29 Cells; Hereditary Adenomatous Polyposis Coli; Human; Human, General; Hydroxyl Daunorubicin; Hydroxyldaunorubicin; Induction of Apoptosis; Interruption; Intestinal; Intestines; Intestines, Small; Investigators; Kinases; Laboratories; Link; Lung; Maintenance; Maintenances; Malignant; Malignant - descriptor; Malignant Colon Neoplasm; Malignant Colonic Neoplasm; Malignant Colonic Tumor; Malignant Neoplasms; Malignant Tumor; Malignant tumor of colon; Mammals, Mice; Man (Taxonomy); Man, Modern; Medication; Metaphase; Mice; Mice, Mutant Strains; Micro-tubule; Microtubules; Mitotic; Mitotic Anaphase; Mitotic Metaphase; Mitotic spindle; Modeling; Molecular; Murine; Mus; Mutant Strains Mice; Mutation; N-Debenzoyl-N-(tert-butoxycarbonyl)-10-deacetyltaxol; NSAIDs; Neoplasm of the Small Bowel; Non-Steroidal Anti-Inflammatory Agents; Nonsteroidal Anti-Inflammatory Agents; Nonsteroidal Antiinflammatory Drug; Normal Cell; Oncogenesis; Oncogens; Paclitaxel; Paclitaxel (Taxol); Parturition; Patients; Pharmaceutic Preparations; Pharmaceutical Preparations; Phosphotransferases; Play; Polyposis Coli; Polyposis Coli, Familial; Polyposis Syndrome, Familial; Polyps; Pongidae; Praxel; Predisposition gene; Programs (PT); Programs [Publication Type]; Property; Property, LOINC Axis 2; Regulation; Research Personnel; Researchers; Resistance; Respiratory System, Lung; Rodent Model; Role; Screening procedure; Signal Pathway; Small Intestinal Neoplasm; Small Intestine Neoplasm; Small Intestines; Stabilities, Chromosome; Stability, Genomic; Subcellular Process; Sulindac; Sulindac Sulfide; Susceptibility Gene; TAM; Tamoxifen; Targetings, Gene; Taxol; Taxol (Old NSC); Taxol A; Taxol Konzentrat; Taxotere; Testing; Transphosphorylases; Tumor Burden; Tumor Cell; Tumor Load; Tumor Suppression; Tumor Suppression, Molecular; Tumor of the Colon; Tumor of the Small Bowel; Tumor of the Small Intestine; Tumor-Specific Treatment Agents; anaphase-promoting complex; anticancer agent; anticancer drug; anticarcinogenic; base; bowel; carcinogenesis; cell transformation; chemotherapeutic agent; cis-5-fluoro-2-methyl-1-[p-(methylsulfinyl)benzylidene]indene-3-acetic acid; colon carcinogenesis; cyclosome; docetaxel; docetaxol; drug/agent; efficacy testing; familial adenomatous polyposis; familial polyposis; genome mutation; great ape; in vivo; loss of function mutation; malignancy; malignant colon tumor; model organism; mouse mutant; neoplasm/cancer; neoplastic cell; nonsteroidal anti-inflammatory drugs; novel; polypoid adenoma; pre-clinical; preclinical; predisposing gene; programs; pulmonary; resistant; screening; screenings; small bowel; social role; transformed cells; tumor; tumorigenesis
Project start date: 2006-08-24
Project end date: 2011-07-31
Budget start date: 1-AUG-2010
Budget end date: 31-JUL-2011
5R01CA113349-05 (2010): $295185
NOVEL MOUSE COLON CANCER MODELS AND CHEMOPREVENTION
Wei Dai
New York University School Of Medicine, New York, Ny 10016
Grant 5R01CA113349-04 from National Cancer Institute
Abstract: The spindle checkpoint delays the progression from metaphase to anaphase until all condensed chromosomes are properly attached to mitotic spindles. An impaired checkpoint function often results in genomic instability, which predisposes cells to malignant transformation. To understand the molecular basis of functional inactivation of this surveillance mechanism in human cancer development, the Pi´s laboratory has focused on the role of BubRl, a key spindle checkpoint kinase, in the maintenance of genomic stability and suppression of tumorigenesis. The Pi´s group has generated BubRl+/- mice as well as BubRl+/-ApcMin/+ compound mutant mice. BubRl+/- mice develop intestinal adenocarcinomas at an accelerated rate after azoxymethane treatment. Whereas ApcMin/+ mice develop many adenomatous polyps generally within the small intestine, BubRl+/-ApcMin/+ compound mutant mice develop significantly more spontaneous colonic tumors than ApcMin/+ mice. The colon tumors in BubRl+/-ApcMin/+ mice are clinically more advanced than those observed in ApcMin/+ mice. Moreover, chemopreventive compounds such as sulindac sulfide and S-allylmercaptocysteine are capable of inducing apoptosis in HT-29 and SW-480 colon tumor cells; BubRl - deficient cells are more resistant to apoptosis induced by these compounds. Given that both BubRl and Ape are involved in the regulation of genomic stability in normal cells, we hypothesize that BubRl+/-ApcMin+/- mice would be an excellent rodent model for evaluating the efficacy of anti-tumor activities of various chemopreventive compounds that target colon and understanding the in vivo role of spindle checkpoint components in the maintenance of genomic stability. To test this hypothesis, the Pi´s lab will (i) validate and test the chemopreventive effect of sulindac and S-allylmercaptocysteine, which (or the derivative of which) directly target microtubules or mitotic spindles, on suppression of spontaneous intestinal tumorigenesis in BubRl+/- ApcMin/+ compound mutant mice, and (ii) investigate the molecular basis of intestinal carcinogenesis in these mutant mice by studying (a) the genomic instability and the rate of spontaneous transformation of cells deficient in BubRl and/or Ape, (b) the dependence of anti-proliferative effect of sulindac and S-allylmercaptocysteine on the integrity of the spindle checkpoint, and (c) the mechanism by which ApcMin/+ mice shift in tumor burden from the small intestine to colon in the BubRl-deficient genetic background. The long-term goal of this project is to elucidate the mechanism by which cell cycle checkpoints and the Wnt signaling pathway regulate cell proliferation and differentiation as well as genomic stability
Keywords: (8S-cis)-10-[(3-Amino-2, 3, 6-trideoxy-alpha-L-lyxo-hexopyranosyl)oxy]-7, 8, 9, 10-tetrahydro-6, 8, 11-trihydroxy-8-(hydroacetyl)-1-methoxy-5, 12-naphthacenedione; (Z)-2-[4(1, 2-diphenyl-1-butenyl)-phenoxyl]-N, N-dimethylethanamine; (Z)-5-fluoro-2-methyl-1-[[4-(methylsulfinyl)phenyl]methylene]-1H-indene-3-acetic acid; 1-p-beta-dimethylamino-ethoxyphenyl-trans-1, 2-diphenylbut-1-ene; 14-Hydroxydaunomycin; 1H-Indene-3-acetic acid, 5-fluoro-2-methyl-1-(4-(methylthio)phenyl)methylene-, (Z)-; 5, 12-Naphthacenedione, 10-((3-amino-2, 3, 6-trideoxy-alpha-L-lyxo-hexopyranosyl)oxy)-7, 8, 9, 10-tetrahydro-6, 8, 11-trihydroxy-8-(hydroxyacetyl)-1-methoxy-, (8S-cis)-; APC - Adenomatous Polyposis Coli; Adenocarcinoma; Adenoma, Malignant; Adenomatous Polyposis Coli; Adenomatous Polyposis Coli, Familial; Adenomatous Polyposis of the Colon; Adenomatous Polyps; Adriamycine; Anaphase; Animal Model; Animal Models and Related Studies; Anti-Cancer Agents; Anti-Inflammatory Agents, Non-Steroidal; Anti-Tumor Agents; Anti-Tumor Drugs; Antiinflammatory Agents, Non Steroidal; Antiinflammatory Agents, Nonsteroidal; Antineoplastic Agents; Antineoplastic Drugs; Antineoplastics; Antiproliferative Agents; Antiproliferative Drugs; Anzatax; Apes; Apoptosis; Apoptosis Pathway; Asotax; Azoxymethane; Biochemical; Biological; Birth; Bristaxol; Cancer Causing Agents; Cancer Drug; Cancer Induction; Cancer Model; Cancer Susceptibility Gene; Cancer-Predisposing Gene; CancerModel; Cancers; Carcinogens; Cell Cycle Checkpoint; Cell Death, Programmed; Cell Function; Cell Growth in Number; Cell Multiplication; Cell Process; Cell Proliferation; Cell physiology; Cells; Cellular Function; Cellular Physiology; Cellular Process; Cellular Proliferation; Chemoprevention; Chemopreventive; Chemopreventive Agent; Chemotherapeutic Agents, Neoplastic Disease; Chromosomal Stability; Chromosomes; Cola; Colon; Colon Cancer; Colon Carcinoma; Colon Neoplasms; Colonic Cancer; Colonic Carcinoma; Colonic Mass; Colonic Neoplasms; Colonic Tumor; DNA; DOX; Defect; Deoxyribonucleic Acid; Dependence; Development; Diazene, dimethyl-, 1-oxide; Doxorubicin; Doxorubicina; Drugs; EC 2.7; Epithelial Cells; Ethanamine, 2-(4-(1, 2-diphenyl-1-butenyl)phenoxy)-N, N-dimethyl-, (Z)-; Familial Adenomatous Polyposis Syndrome; Gastrointestinal Tract, Small Intestine; Gene Targeting; Genetic; Genetic Alteration; Genetic Change; Genetic defect; Genome; Genome Instability; Genome Stability; Genomic Instability; Genus Cola; Goals; HT-29; HT-29 Cells; HT29 Cells; Hereditary Adenomatous Polyposis Coli; Human; Human, General; Hydroxyl Daunorubicin; Hydroxyldaunorubicin; Induction of Apoptosis; Interruption; Intestinal; Intestines; Intestines, Small; Investigators; Kinases; Laboratories; Link; Lung; Maintenance; Maintenances; Malignant; Malignant - descriptor; Malignant Colon Neoplasm; Malignant Colonic Neoplasm; Malignant Colonic Tumor; Malignant Neoplasms; Malignant Tumor; Malignant tumor of colon; Mammals, Mice; Man (Taxonomy); Man, Modern; Medication; Metaphase; Mice; Mice, Mutant Strains; Micro-tubule; Microtubules; Mitotic; Mitotic Anaphase; Mitotic Metaphase; Mitotic spindle; Modeling; Molecular; Murine; Mus; Mutant Strains Mice; Mutation; N-Debenzoyl-N-(tert-butoxycarbonyl)-10-deacetyltaxol; NSAIDs; Neoplasm of the Small Bowel; Non-Steroidal Anti-Inflammatory Agents; Nonsteroidal Anti-Inflammatory Agents; Nonsteroidal Antiinflammatory Drug; Normal Cell; Oncogenesis; Oncogens; Paclitaxel; Paclitaxel (Taxol); Parturition; Patients; Pharmaceutic Preparations; Pharmaceutical Preparations; Phosphotransferases; Play; Polyposis Coli; Polyposis Coli, Familial; Polyposis Syndrome, Familial; Polyps; Pongidae; Praxel; Predisposition gene; Programs (PT); Programs [Publication Type]; Property; Property, LOINC Axis 2; Regulation; Research Personnel; Researchers; Resistance; Respiratory System, Lung; Rodent Model; Role; Screening procedure; Signal Pathway; Small Intestinal Neoplasm; Small Intestine Neoplasm; Small Intestines; Stabilities, Chromosome; Stability, Genomic; Subcellular Process; Sulindac; Sulindac Sulfide; Susceptibility Gene; TAM; Tamoxifen; Targetings, Gene; Taxol; Taxol (Old NSC); Taxol A; Taxol Konzentrat; Taxotere; Testing; Transphosphorylases; Tumor Burden; Tumor Cell; Tumor Load; Tumor Suppression; Tumor Suppression, Molecular; Tumor of the Colon; Tumor of the Small Bowel; Tumor of the Small Intestine; Tumor-Specific Treatment Agents; anaphase-promoting complex; anticancer agent; anticancer drug; anticarcinogenic; base; bowel; carcinogenesis; cell transformation; chemotherapeutic agent; cis-5-fluoro-2-methyl-1-[p-(methylsulfinyl)benzylidene]indene-3-acetic acid; colon carcinogenesis; cyclosome; docetaxel; docetaxol; drug/agent; efficacy testing; familial adenomatous polyposis; familial polyposis; genome mutation; great ape; in vivo; loss of function mutation; malignancy; malignant colon tumor; model organism; mouse mutant; neoplasm/cancer; neoplastic cell; nonsteroidal anti-inflammatory drugs; novel; polypoid adenoma; pre-clinical; preclinical; predisposing gene; programs; pulmonary; resistant; screening; screenings; small bowel; social role; transformed cells; tumor; tumorigenesis
Project start date: 2006-08-24
Project end date: 2011-07-31
Budget start date: 1-AUG-2009
Budget end date: 31-JUL-2010
5R01CA113349-04 (2009): $295185
5R01CA113349-03 (2008): $295185
A Novel Mouse Colon Cancer Model And Chemoprevention
Wei Dai, Professor
New York University School Of Medicine New York, Ny 10016
Grant 5R01CA113349-02 from National Cancer Institute IRG: CDP
Abstract: The spindle checkpoint delays the progression from metaphase to anaphase until all condensed chromosomes are properly attached to mitotic spindles. An impaired checkpoint function often results in genomic instability, which predisposes cells to malignant transformation. To understand the molecular basis of functional inactivation of this surveillance mechanism in human cancer development, the Pi s laboratory has focused on the role of BubRl, a key spindle checkpoint kinase, in the maintenance of genomic stability and suppression of tumorigenesis. The Pi s group has generated BubRl+/- mice as well as BubRl+/-ApcMin/+ compound mutant mice. BubRl+/- mice develop intestinal adenocarcinomas at an accelerated rate after azoxymethane treatment. Whereas ApcMin/+ mice develop many adenomatous polyps generally within the small intestine, BubRl+/-ApcMin/+ compound mutant mice develop significantly more spontaneous colonic tumors than ApcMin/+ mice. The colon tumors in BubRl+/-ApcMin/+ mice are clinically more advanced than those observed in ApcMin/+ mice. Moreover, chemopreventive compounds such as sulindac sulfide and S-allylmercaptocysteine are capable of inducing apoptosis in HT-29 and SW-480 colon tumor cells; BubRl - deficient cells are more resistant to apoptosis induced by these compounds. Given that both BubRl and Ape are involved in the regulation of genomic stability in normal cells, we hypothesize that BubRl+/-ApcMin+/- mice would be an excellent rodent model for evaluating the efficacy of anti-tumor activities of various chemopreventive compounds that target colon and understanding the in vivo role of spindle checkpoint components in the maintenance of genomic stability. To test this hypothesis, the Pi s lab will (i) validate and test the chemopreventive effect of sulindac and S-allylmercaptocysteine, which (or the derivative of which) directly target microtubules or mitotic spindles, on suppression of spontaneous intestinal tumorigenesis in BubRl+/- ApcMin/+ compound mutant mice, and (ii) investigate the molecular basis of intestinal carcinogenesis in these mutant mice by studying (a) the genomic instability and the rate of spontaneous transformation of cells deficient in BubRl and/or Ape, (b) the dependence of anti-proliferative effect of sulindac and S-allylmercaptocysteine on the integrity of the spindle checkpoint, and (c) the mechanism by which ApcMin/+ mice shift in tumor burden from the small intestine to colon in the BubRl-deficient genetic background. The long-term goal of this project is to elucidate the mechanism by which cell cycle checkpoints and the Wnt signaling pathway regulate cell proliferation and differentiation as well as genomic stability.
Keywords: cell, chemoprevention, colon neoplasm, model, neoplasm /cancer, DNA, adenocarcinoma, adenomatous polyp, apoptosis, birth, carcinogen, carcinogenesis, cell cycle, cell proliferation, cell transformation, chromosome, colon, doxorubicin, gene, gene mutation, gene targeting, genetics, genome, human, lung, microtubule, mitotic spindle apparatus, mutant, paclitaxel, play, role, small intestine, sulfide, suppression, tamoxifen
Project start date: 2006-08-24
Project end date: 2011-07-31
5R01CA113349-02 (2007): $295185
1R01CA113349-01A2 (2006): $285331
BUBR1 In The Spindle Checkpoint And Tumor Suppression
Wei Dai, Professor
New York Medical College Administration Building Valhalla, Ny 10595
Grant 3R01CA090658-06S1 from National Cancer Institute IRG: MEP
Abstract: The spindle checkpoint delays the progression of metaphase to anaphase until all chromosomes are properly attached to the mitotic spindle. A loss of the checkpoint function often results in genetic instability, which predisposes cells to malignant transformation. To understand the molecular basis of functional inactivation of this surveillance mechanism in human cancer development, the PI s laboratory has focused on the roles of two protein kinases, hBUB1 and hBUBR1 (42;53;81), mutations of which are detected in colorectal cancer cells (4). Recent studies suggest that hBUBR1 is an important component of the spindle checkpoint signaling pathway, and that it targets the anaphase-promoting complex (APC) through interaction with p55CDC/hCdc2O. Specifically, whereas hBUBR1 is expressed as a 120-kDa entity in various cell lines during interphase, nocodazole (Noc)-arrested mitotic cells contain a distinct hBUBR1 species that exhibits a reduced mobility on SDS-polyacrylamide gels. Phosphatase treatment restored the mobility of the mitotic hBUBR1 species to that characteristic of the interphase state, indicating that the decrease in mobility of hBUBR1 is due to phosphorylation. Yeast two-hybrid, glutathione S-transferase pull-down, and immunoprecipitation analyses revealed that p55CDC interacts with hBUBR1. hBUBR1 also phosphorylated p55CDC in vitro. Furthermore, the APC components CDC16 and CDC27 present in lysates of Noc-treated cells interact with hBUBR1 affinity resins. Finally, we have shown that the stress-activated protein kinase p38 interacts with hBUBR1 and that Noc-induced phosphorylation of hBUBR1 in HeLa cells is blocked by pretreatment with a specific inhibitor of p38. On the basis of the known biochemical and biological activities of hBUBR1, we therefore hypothesize that activation of hBUBR1 by p38 induces its interaction with p55CDC and subsequent inhibition of the APC during spindle checkpoint activation, and that a loss of hBUBR1 function due to structural abnormalities may result in aneuploidy and cancer. To test this hypothesis, we will (i) investigate whether a dominant negative mutant of mouse BUBR1 promotes tumorigenesis in transgenic mice; (ii) determine whether p38 is an immediate upstream activator of hBUBR1 during spindle checkpoint activation; (iii) define the domain of p55CDC that interacts with hBUBR1; (iv) examine whether hBUBR1 phosphorylates p5SCDC in vivo and whether hBUBR1 and the CDK1-cyclin B complex phosphorylate p55CDC on different sites; and (v) evaluate whether hBUBR1 inhibits APC activity by phosphorylating CDC16 and CDC27. The long-term goal of this project is to determine the mechanism by which protein phosphorylation regulates the spindle checkpoint and how deregulation of it may result in cancer.
Keywords: cell cycle, intestine neoplasm, mitotic spindle apparatus, neoplastic transformation, protein kinase, aneuploidy, cell cycle protein, cyclin dependent kinase, enzyme activity, gene mutation, genetic promoter element, phosphorylation, 3T3 cell, athymic mouse, genetically modified animal, immunoprecipitation, transfection, yeast two hybrid system
Project start date: 2001-04-06
Project end date: 2007-03-31
3R01CA090658-06S1 (2006): $39500
5R01CA090658-06 (2005): $275471
5R01CA090658-05 (2004): $279048
5R01CA090658-04 (2003): $279353
Sponsored Links Excellgen http://Excellgen.com
MECHANISM OF TOXOPLASMA INFECTION IN AIDS PATIENTS
Wei Dai, Professor
University Of Cincinnati Sponsored Research Services Cincinnati, Oh 45221
Grant 5R29AI032483-05 from National Institute Of Allergy And Infectious Diseases IRG: ARRE
Abstract: The molecular mechanisms of opportunistic infection by Toxoplasma gondii in AIDS patients will be investigated. Central nervous system toxoplasmosis has caused disability and death in up to 30% of patients with AIDS. Current clinical treatment for toxoplasma infection is mainly chemotherapeutic, which is often associated with dose-related bone marrow suppression. Complete remission of the infection depends largely on the adequate restoration of the cell-mediated immune system of the patients and thus, it poses a continuous threat to AIDS patients. An alternative treatment using recombinant interferon-gamma (IFN-gamma) has been proposed since IFN-gamma, a CD4+ T cell product, blocks the growth of T. gondii in human epithelial and peripheral blood mononuclear cells, and macrophages. Treatment of mice with monoclonal anti-IFN-gamma antibody abrogates resistance to toxoplasma infection. The IFN-gamma-induced anti-toxoplasmic activity in human cells is strongly correlated with the degradation of the essential amino acid L-tryptophan in vitro. Destruction of L-tryptophan is due to increased activity of indoleamine 2,3-dioxygenase (IDO) which is in turn transcriptionally activated by IFN-gamma. It has been suggested that T. gondii infection is primarily controlled by IDO activity induced by IFN-gamma derived from CD4+ T cells. Therefore, we propose the following four specific aims to investigate the role of IDO in host defense against T. gondii infection. (1) To determine whether IDO activity alone, expressed via stable transfection of IDO cDNA controlled by a metallothionein inducible promoter, blocks the growth of T. gondii in cultured human fibroblast and macrophage cell lines; (2) To investigate whether IDO activity expressed in brain or hematopoietic system of mice via gene transfer approaches would confer resistance to T gondii infection in mice; (3) To examine the levels of IDO expression in mice either vaccinated with an avirulent strain of T. gondii or vaccinated but depleted of IFN-gamma through specific antibody injection; (4) Since the mechanisms of macrophage inhibition of parasite replication are unclear, to examine the role of IDO against T. gondii replication by asking if IDO activity shows an inverse correlation with the parasite growth in hosts depleted of CD4+ T cells and if macrophage activators (or deactivators), other than IFN-gamma, induce IDO and anti-toxoplasmic activity. The long term objective of this proposed research is to provide molecular information used for developing new and effective drugs for the prevention and treatment of toxoplasmosis in AIDS patients.
Keywords: AIDS, Toxoplasma gondii, enzyme activity, human immunodeficiency virus, opportunistic infection, toxoplasmosis, tryptophan 2, 3 dioxygenase, helper T lymphocyte, host organism interaction, interferon gamma, laboratory mouse, tissue /cell culture, transfection
Project start date: 1992-07-01
Project end date: 1997-04-30
5R29AI032483-05 (1995): $109915
5R29AI032483-03 (1994): $114523
5R29AI032483-02 (1993): $114257
1R29AI032483-01A1 (1992): $113290
Wei Dai
New York University School Of Medicine, New York, Ny 10016
Abstract: The Growth Control Program was created in 2001 as part of the reorganization of the NYU Cancer Institute. In the initial phase of organizing the Program, we found it useful to have co-leaders, Drs. Pagano and Skolnik. After this initial period, the synergetic exertion by two leaders was no longer necessary. Accordingly, since 2003 the Program has been led only by Dr. Pagano. Among the reasons for this change was the recommendation of the last site visitors who concluded that there was undesirable overlap in responsibilities with two co-leaders. The program started with 17 members. However, Dr. Pagano was committed to creating a critical mass around the central theme of the program by broadening its base and developing new initiatives in important areas (e.g., the integration of bench research with translational research and the promotion of productive interactions between basic scientists and clinicians, see sections 8.2.g and 8.2.h). Therefore, he selected a Steering Committee (see below) to solicit new members. The Steering Committee decided to focus on issues beyond the scientific stature of the candidate members, seeking investigators who would truly make a concrete contribution to the objectives of the Program and devote >50% of their time to the scientific themes of the Program. Following several strategic planning sessions, additional scientists whose expertise would advance the program´s objectives were enrolled. This growth was also fueled by the new faculty recruitment efforts of the NYUCI. Currently, the program includes 44 members of which 18 (41%) are junior faculty members. Importantly, 17 members of the program (39%) were recruited to NYU after 2002 (see details in the Table below). The extraordinary growth that the Program has experienced in the last four years represents an essential transition period that is expected to maximize programmatic cohesion and foster research interactions. The criteria used for selecting solicited and unsolicited applications for membership in the Growth Control Program include 1) Demonstrated commitment, expertise and funding in basic and translational cancer-relevant research; 2) Evidence of ongoing collaborative efforts with other investigators in these areas; 3) Commitment to the shared objectives of the Growth Control Program and the NYU Cancer Institute as a whole; 4) Funding from NCI or equivalent national organization for peer-reviewed, cancer-relevant and investigator-initiated research (junior investigators excluded). The membership of the Growth Control Program is reviewed annually by the Steering Committee. Members who fail to participate in the program activities during the previous year are presented with an opportunity to resign or increase their participation
Keywords: Animal Model; Animal Models and Related Studies; Anti-Oncogenes; Antioncogenes; Area; CCSG; Cancer Center Support Grant; Cancers; Clinical; Commit; Core Grant; Development; Emerogenes; Enrollment; Exertion; Faculty; Fostering; Funding; Gene Transcription; Generalized Growth; Genes, Cancer Suppressor; Genes, Onco-Suppressor; Genes, Suppressor; Genetic Alteration; Genetic Change; Genetic Transcription; Genetic defect; Goals; Grant; Growth; Human; Human, General; Institutes; Investigator-Initiated Research; Investigators; Knowledge; Malignant; Malignant - descriptor; Malignant Neoplasms; Malignant Tumor; Man (Taxonomy); Man, Modern; Molecular; Mutation; Neoplasms; Oncogenes, Recessive; Oncogenes-Tumor Suppressors; Organizations, Peer Review; P30 Grant; Pathway interactions; Pattern; Peer Review Organizations; Phase; Programs (PT); Programs [Publication Type]; Publications; RNA Expression; Recommendation; Recruitment Activity; Research; Research Personnel; Researchers; Resource Sharing; Sampling; Scientific Publication; Scientist; Second-Site Suppressor Genes; Signal Transduction Pathway; Site; Strategic Planning; Structure; Suppressor Genes; Therapeutic; Time; Tissue Growth; Transcription; Transcription, Genetic; Translational Research; Translational Research Enterprise; Translational Science; Tumor Cell; Tumor Suppressing Genes; Tumor Suppressor Genes; Tumors; Work; base; cohesion; design; designing; enroll; experience; genome mutation; instrument; interest; malignancy; member; model organism; neoplasia; neoplasm/cancer; neoplastic cell; neoplastic growth; novel; oncosuppressor gene; ontogeny; pathway; programs; recruit; translation research enterprise; tumor
Project start date: 2010-03-01
Project end date: 2013-02-28
Budget start date: 1-MAR-2010
Budget end date: 28-FEB-2011
5P30CA016087-30_0001 (2010): $24559
Mammal Cecropins As Antibiotics For Corneal Infections
Wei Dai, Professor
Cytopro, Inc. 455 N. Grafield Ave, #200 Monterey Park, Ca 917541201
Grant 1R41EY015354-01 from National Eye Institute IRG: ZRG1
Abstract: The emerging bacterial infections that are resistant to the current antibiotic arsenal have accelerated search for therapeutic agents with a novel mechanism of action. Recent studies suggest that the gene-encoded peptide antibiotics may hold great promise as a new generation of therapeutic agents. These agents have the potential to become medications used on a daily basis by ophthalmologists to treat or prevent ocular infections, especially those infections caused by bacteria resistant to other drugs. Cecropins, a family of structurally related small peptides originally isolated from invertebrates, are potent against a wide spectrum of gram-negative and gram-positive bacteria. Cecropins form an amphipathic helix, which anchors to and penetrate the surface of bacterial cell membrane, resulting in cell lysis. Whereas mammals have evolved an adaptive immune response they also retain many ancient genes encoding products with bactericidal activities, functioning as an innate immune response. Given that a porcine cecropin (cecropin P1) has been identified and characterized and that cecropins are highly specific for killing microorganisms including those commonly found in corneal infections we propose that humans have retained cecropin-homologous genes whose products can be explored as new antibiotics used in the ophthalmic clinics. The major goals of this project are to identify and characterize the human homologue of cecropin P1 and to explore cecropin P1 and its human counterpart for commercial applications as novel antibiotics in ophthalmology. It is anticipated that the studies proposed in this Phase I application will result in structural and functional characterization of human cecropin. Phase II will be aimed at testing the efficacy and potential side-effects of mammalian cecropins using animal model systems as well as at regulatory submission and clinical testing.
Keywords: antibiotic, cornea, drug discovery /isolation, eye infection, pore forming protein, cell differentiation, cell proliferation, complementary DNA, corneal epithelium, peptide chemical synthesis, genetic screening, molecular cloning, polymerase chain reaction, tissue /cell culture
Project start date: 2004-01-01
Project end date: 2005-12-31
1R41EY015354-01 (2004): $100000
PLK3 IN THE DNA DAMAGE CHECKPOINT AND TUMOR SUPPRESSION
Wei Dai
New York University School Of Medicine, 550 1st Ave, New York, Ny 10016
Abstract: The DNA damage checkpoint delays progress of the cell cycle until damage to the genome is repaired. To understand the molecular basis of this surveillance mechanism in human cells, the P.I. laboratory has been studying Plk3, a member of the Polo family of protein kinases. Recent studies suggest that Plk3 is an important component of the DNA damage checkpoint machinery and that it targets both the mitotic activator Cdc25C and the tumor suppressor p53. The kinase activity of Plk3 is rapidly increased by exposure of cells to genotoxic stresses. Ectopic expression of an active form of Plk3 (Plk3-A) suppresses cell proliferation and induces apoptosis. Plk3 phosphorylates p53 in vitro on two major sites that includes serine-20, and a kinase-defective mutant of Plk3 (Plk3K52R) inhibits phosphorylation of p53 on serine-20 in vivo. Moreover, Plk3 concentrates at unduplicated centrosomes during the G1 phase of the cell cycle, and enforced expression of Plk3K52R results in centrosome amplification and the formation of multiple microtubule organization centers. Furthermore, human PLK3 localizes to chromosome 1p32, a locus thought to contain cancer susceptibility genes. The expression of PLK3 is down-regulated in human lung and head-neck carcinomas as well as in carcinogen-induced colon tumors in rats. On the basis of these various observations, we hypothesize that PLK3 is a tumor-suppressor gene whose product integrates signals that control genomic instability and achieves its effects, at least in part, through regulation of p53 and centrosome function. To test this hypothesis, the P.I.´s laboratory will (i) map all the phosphorylation sites of p53 targeted by Plk3 and determine the functional consequence of such phosphorylation; (ii) determine whether Plk3 acts immediately downstream of the protein kinases Chkl and Chk2 in the signaling pathways that underlie the cellular response to DNA damage; (iii) investigate whether Plk3 plays a role in the arrest of centrosome duplication in response to activation of the DNA damage checkpoint; and (iv) examine whether mice with a targeted disruption of PLK3 show an increase susceptibility to the development of colon, lung, or other cancers. The long-term goal of this project is to define mechanisms by which Polo family kinases function as key regulatory enzymes in the DNA damage checkpoint and by which dysregulation of these enzymes may result in genomic instability and neoplastic transformation
Keywords: DNA damage; DNA replication; biological signal transduction; cell cycle proteins; cell growth regulation; centrosome; cytogenetics; embryonic stem cell; genetic regulation; genetic susceptibility; genetically modified animals; laboratory mouse; mitotic spindle apparatus; neoplastic transformation; p53 gene /protein; phosphorylation; serine threonine protein kinase; tumor suppressor genes
Project start date: 1997-12-15
Project end date: 2010-03-31
Budget start date: 7-MAY-2007
Budget end date: 31-MAR-2010
5R01CA074229-12 (2007): $0
5R01CA074229-10 (2006): $316855
5R01CA074229-09 (2005): $324480
Sponsored Links Excellgen http://Excellgen.com
5R01CA074229-08 (2004): $324220
2R01CA074229-07A2 (2003): $323960
Wei Dai
New York University School Of Medicine
Project start date: 2011-01-01
Project end date: 2015-12-31
A Novel Mouse Colon Cancer Model And Chemoprevention
Wei Dai, Professor
New York University School Of Medicine New York, Ny 10016
Grant 3R01CA113349-01A2S1 from National Cancer Institute IRG: CDP
Project start date: 2006-08-24
Project end date: 2011-07-31
3R01CA113349-01A2S1 (2007): $20000
PROTEIN SER/THR KINASE PRK AND CELL CYCLE PROGRESSION
Wei Dai, Professor
Internal Medicineuniversity Of Cincinnati
sponsored Research Services
cincinnati, Oh 45221
Grant 3R01CA074229-01A1S1 from National Cancer Institute IRG: HEM
Project start date: 1997-12-15
Project end date: 2001-11-30
3R01CA074229-01A1S1 (1998): $10775
TIF AND ERYTHROLEUKEMIA CELL DIFFERENTIATION
Wei Dai, Professor
University Of Cincinnati Sponsored Research Services Cincinnati, Oh 45221
Grant 5R01CA059985-02 from National Cancer Institute IRG: HEM
Project start date: 1995-07-01
Project end date: 2000-04-30
5R01CA059985-02 (1996): $215882
MECHANISM OF TOXOPLASMA INFECTION IN AIDS PATIENTS
Wei Dai, Professor
University Of Cincinnati Sponsored Research Services Cincinnati, Oh 45221
Grant 5R29AI032483-06 from National Institute Of Allergy And Infectious Diseases IRG: ARRE
Project start date: 1992-07-01
Project end date: 1997-04-30
5R29AI032483-06 (1996): $116075