DENTIN SPECIFIC PHOSPHOPROTEINS AND MINERALIZATION
Mary B Macdougall, James R. Rosen Chair Of Dental Research
University Of Texas Hlth Sci Ctr San Ant San Antonio, Tx 78229
Grant 5R01DE011658-05 from National Institute Of Dental And Craniofacial Research IRG: ZDE1
Abstract: The goal of this project is to study the biological function of these dentin-specific phosphoproteins in the process of dentin biomineralization, thereby advancing our overall understanding of the process of odontoblast differentiation and dentinogenesis. This goal is based on an emerging body of literature showing that acidic proteins generally play a key role in direct regulation of crystal formation. Our hypothesis is that the dentin-specific phosphoproteins, DMP 1 and DPP(s), are markers for odontoblast differentiation which are important in the nucleation and regulation of hydroxyapatite formation during dentin extracellular matrix mineralization. Our first objective is to isolate and characterize the full-length cDNAs for DMP-1 and DPP(s). This will be accomplished by construction and screening of a mouse molar cDNA library. Once identified, the DMP-1 and DPP(s) cDNAs will be heterologously expressed in procaryotic and eukaryotic hosts, and the purified protein used to produce specific antibodies. These antibodies will be used to map the sequential expression patterns of DMP-1 and DPP(s) during odontoblast differentiation related to dentin extracellular matrix mineralization using immunohistochemistry. The expression of these dentin-specific phosphoproteins at a transcriptional level will also be examined using RT-PCR amplification and in situ hybridization techniques. Finally, the biological function of DMP-1 and DPP(s) will be investigated using antisense "knockout" and antibody perturbation strategies monitoring the resulting mineral phase and matrix formation using electron microscopy, X-ray diffraction and electron microprobe analyses. These experiments will take advantage of a unique monolayer odontoblast cell culture system established in our laboratory to determine the functions of the dentin-specific phosphoproteins. This culture system has previously been shown to be conducive for the cytodifferentiation of odontoblasts from dental papilla mesenchyme cells, maintenance of the odontoblast phenotype, sequential expression and secretion of dentin extracellular matrix proteins, and dentin biomineralization. In addition, we have recently established immortalized mouse odontoblast and dental papilla mesenchyme cell lines at key stages of the odontoblast differentiation which will be used to determine the effect of modulation of dentin-specific phosphoprotein expression on DECM production and subsequent mineralization. These two key research tools will be used to investigate the biological function of dentin-specific phosphoproteins during odontoblast differentiation and dentin mineralization in this application.
Keywords: dentin, dentinogenesis, normal ossification, phosphoprotein, cell differentiation, complementary DNA, extracellular matrix, extracellular matrix protein, hydroxyapatite, nucleic acid sequence, odontoblast, X ray crystallography, antibody formation, electron microscopy, immunocytochemistry, in situ hybridization, laboratory mouse, polymerase chain reaction, tissue /cell culture
Project start date: 1996-03-15
Project end date: 2002-03-14
5R01DE011658-05 (2000): $178092
Sponsored Links Excellgen http://Excellgen.com
DENTIN SPECIFIC PHOSPHOPROTEINS AND MINERALIZATION
Mary B Macdougall, James R. Rosen Chair Of Dental Research
University Of Texas Hlth Sci Ctr San Ant San Antonio, Tx 78229
Grant 5R01DE011658-04 from National Institute Of Dental And Craniofacial Research IRG: ZDE1
Abstract: The goal of this project is to study the biological function of these dentin-specific phosphoproteins in the process of dentin biomineralization, thereby advancing our overall understanding of the process of odontoblast differentiation and dentinogenesis. This goal is based on an emerging body of literature showing that acidic proteins generally play a key role in direct regulation of crystal formation. Our hypothesis is that the dentin-specific phosphoproteins, DMP 1 and DPP(s), are markers for odontoblast differentiation which are important in the nucleation and regulation of hydroxyapatite formation during dentin extracellular matrix mineralization. Our first objective is to isolate and characterize the full-length cDNAs for DMP-1 and DPP(s). This will be accomplished by construction and screening of a mouse molar cDNA library. Once identified, the DMP-1 and DPP(s) cDNAs will be heterologously expressed in procaryotic and eukaryotic hosts, and the purified protein used to produce specific antibodies. These antibodies will be used to map the sequential expression patterns of DMP-1 and DPP(s) during odontoblast differentiation related to dentin extracellular matrix mineralization using immunohistochemistry. The expression of these dentin-specific phosphoproteins at a transcriptional level will also be examined using RT-PCR amplification and in situ hybridization techniques. Finally, the biological function of DMP-1 and DPP(s) will be investigated using antisense "knockout" and antibody perturbation strategies monitoring the resulting mineral phase and matrix formation using electron microscopy, X-ray diffraction and electron microprobe analyses. These experiments will take advantage of a unique monolayer odontoblast cell culture system established in our laboratory to determine the functions of the dentin-specific phosphoproteins. This culture system has previously been shown to be conducive for the cytodifferentiation of odontoblasts from dental papilla mesenchyme cells, maintenance of the odontoblast phenotype, sequential expression and secretion of dentin extracellular matrix proteins, and dentin biomineralization. In addition, we have recently established immortalized mouse odontoblast and dental papilla mesenchyme cell lines at key stages of the odontoblast differentiation which will be used to determine the effect of modulation of dentin-specific phosphoprotein expression on DECM production and subsequent mineralization. These two key research tools will be used to investigate the biological function of dentin-specific phosphoproteins during odontoblast differentiation and dentin mineralization in this application.
Keywords: dentin, dentinogenesis, normal ossification, phosphoprotein, cell differentiation, complementary DNA, extracellular matrix, extracellular matrix protein, hydroxyapatite, nucleic acid sequence, odontoblast, X ray crystallography, antibody formation, electron microscopy, immunocytochemistry, in situ hybridization, laboratory mouse, polymerase chain reaction, tissue /cell culture
Project start date: 1996-03-15
Project end date: 2001-03-14
5R01DE011658-04 (1999): $171242
5R01DE011658-03 (1998): $164656
5R01DE011658-02 (1997): $190624
Grants awarded to Mary B Macdougall
Mary B Macdougall, James R. Rosen Chair Of Dental Research
University Of Southern California Department Of Contracts And Grants Los Angeles, Ca 90033
Grant 5R29DE009875-03 from National Institute Of Dental And Craniofacial Research IRG: OBM
Abstract: Understanding the molecular basis of human genetic diseases described to date has rapidly advanced in recent years due to the advent of recombinant DNA technology. Traditionally, defective gene products associated with a genetic disorder would be isolated and characterized based on a previous knowledge of the normal gene products. This process, known as "forward genetics," has limitations in that the protein associated with the disease must first be identified. This major drawback has recently been overcome with the establishment of "reverse genetics," which relies on the use of genetic linkage studies to define the chromosomal localization of the disease followed by cloning of chromosomal structural abnormalities at defined chromosomal regions. Although several diseases, which affect the human dentition, have been clinically defined and their patterns of inheritance determined, no conclusive protein defects have been established for any of these diseases. Defects of the dentin have been classified into two major subgroups dentin dysplasia and dentinogenesis imperfecta (DGI). Of these two groups, dentinogenesis imperfecta is the most common with an incidence of 18000. Dentinogenesis imperfecta is an autosomal dominant disorder which affects both dentitions. Linkage analysis has established this disease to be located on the long arm (q) of human chromosome 4 in the region ql3-q2l. The goal of this proposal is to delineate the primary genetic defects which are responsible for dentinogenesis imperfecta. Our hypothesis is that mutations within a single gene localized within human chromosome 4 q 13-q21, whose gene product, is solely expressed by odontoblast cells during dentinogenesis, is responsible for DGI. We plan to use DNA, isolated from a patient with a hemizygous interstitial deletion of the chromosome 4 ql3-q2l, to construct a regional genomic library using the phenol-enhanced differential reassociation (PERT) technique. This regional chromosome 4 library will be screened for sequences which are expressed during active formation of the dentin extracellular matrix. These sequences will be characterized by their temporal-spatial expression pattcrns using in situ hybridization thus demonstrating their relevance to dentin formation and DGI. The mutations which occur in available kindreds with DGI types II and III will be mapped once the putative DGI gene has be identified. Lastly, the "DGI" protein will be isolated from dentin matrix and characterized by production of synthetic polypeptide antibodies. These studies will provide a better understanding of genetic diseases associated to mineralizing tissues.
Keywords: autosomal dominant trait, chromosome deletion, dental disorder, dentinogenesis, genetic disorder, linkage mapping, dentin, extracellular matrix protein, genetic library, messenger RNA, molecular biology, mutation, normal ossification, phosphoprotein, polymerase chain reaction, protein sequence, synthetic peptide, SDS polyacrylamide gel electrophoresis, human subject, immunologic assay /test, laboratory mouse, tissue /cell culture, volunteer
Project start date: 1991-04-01
Project end date: 1993-09-30
5R29DE009875-03 (1993): $120659
UAB SOD´S INSTITUTE OF ORAL HEALTH RESEARCH ENHANCEMENT VIA FACULTY RECRUITMENT
Mary B Macdougall
University Of Alabama At Birmingham, 1530 3rd Avenue South, Birmingham, Al 35294
Grant 5P30DE020744-02 from National Institute Of Dental & Craniofacial Research
Abstract: This School of Dentistry (SOD) at the University of Alabama at Birmingham (UAB) application is in response to the NIDCR´s RFA-OD-09-005 "Recovery Act Limited Competition Supporting New Faculty Recruitment to Enhance Research Resources through Biomedical Research Core Centers (P-30)". This RFA is specifically designed to provide institutional awards through the Core Center Grants mechanism to provide "funding to hire, provide appropriate start-up packages, and develop pilot research projects for newly independent investigators, with the goal of augmenting and expanding the institution´s community of multidisciplinary researchers" focusing on areas relevant to NIDCR´s strategic mission " The UAB SOD is seeking support for the enhancement of our research intense faculty through our established "Biomedical Core Center"-The Institute of Oral Health Research (lOHR). This mechanism will allow the lOHR to hire, provide start-up packages, and develop pilot research projects for newly-independent investigators, with the goal of expanding multidisciplinary researchers within the SOD lOHR to establishing a UAB-wide supported pilot center "Center for Discoveries, Genomics and Therapeutics of Craniofacial and Dental Diseases (CDT-CADD). In particular, we will be seeking to enlarge our cadre of faculty within the area of dental and craniofacial developmental biology and genomic as relevant to NIDCR´s strategic plan. The lOHR provides support and shared research resources for SOD and UAB faculty from diverse disciplines in order to establish a joint multidisciplinary approach to the diagnosis, discovery of the underlying molecular causes and development of novel therapeutics for craniofacial diseases and syndromes. In order to achieve our goal, we will hire three new scientists/clinician-scientists with emerging expertise in developmental biology, genetics, proteomics, and/or gene therapy providing them a nurturing and supportive environment ensuring their successful career development. Research start-up resources for the new faculty will include salary support, needed specialized equipment, animal housing costs and materials and supplies for initiating their research. This RFA application will strengthen the UAB SOD´s Institute of Oral Health Research through the expansion of our scientific capacity within the focus area of craniofacial developmental biology and genomics of dental diseases through increasing our faculty workforce
Project start date: 2009-09-17
Project end date: 2011-08-31
Budget start date: 1-SEP-2010
Budget end date: 31-AUG-2011
PFA/PA: RFA-OD-09-005
5P30DE020744-02 (2010): $687974
1P30DE020744-01 (2009): $642866
Institutional Research Training Grant
Mary B Macdougall, James R. Rosen Chair Of Dental Research
University Of Texas Hlth Sci Ctr San Ant San Antonio, Tx 78229
Grant 1T32DE014318-01A1 from National Institute Of Dental And Craniofacial Research IRG: ZDE1
Abstract: This is a revised application in response to the NIDCR National Research Service Award Institutional Research Training Grant Public Announcement (PAR-00-l16). This research training program is focused on the area of Craniofacial Oral-Biology and is named "Craniofacial Oral-Biology Student Training in Academic Research (COSTAR)". It is based on the suggestions of the 1999 NDCR Blue Ribbon Panel on Research Training and Career Development. The grant will provide a broadened interdisciplinary research experience for 24 exceptionally motivated trainees (8 training positions, plus 2 DSTP position) consisting of predoctoral dental, dental scientist, graduate, postdoctoral students, and faculty members in a research-intensive academic Health Science Center environment. The program will emphasize two research themes Craniofacial Biomimetics and Craniofacial Pain and Response to Injury. The program will offer five complementary training tracks 1) short-term summer research experience for dental students; 2) a DDS/Ph.D. program; 3) a predoctoral Ph.D. program; 4) Postdoctoral fellowships; and 5) faculty development training. The purpose of the COSTAR program is to train a cadre of highly skilled, interactive scientists who can successfully address the expanding opportunities in dental, oral, and craniofacial research. A key element of COSTAR is providing trainees with positive, productive, successful faculty role models; many with DDS/M.S. and DDS/Ph.D. degrees obtained through NIDCR supported training programs. The specific aims of this program are 1) to identify and select qualified, motivated candidates who have the interest and potential to pursue training opportunities in dental, craniofacial, oral research to address the scientific opportunities emerging in these fields in the 21st century; 2) to provide a mechanism for integrating trainees into UTHSCSA s collaborative, research-intensive clinical and basic science research environment; 3) to develop trainees with broad, integrated research training in dental, oral and craniofacial health, thereby enhancing their competencies and research skills; 4) to provide flexibility for cross-disciplinary interaction through diverse entry points and opportunities for sustained learning; and 5) to assess the short and long-term impact of the program on the trainee, the institution, and our nation s need for dental academicians.
Project start date: 2002-07-08
Project end date: 2007-06-30
1T32DE014318-01A1 (2002): $423990
DENTAL ACADEMIC RESEARCH TRAINING - DART
Mary B Macdougall, Assoc Dean Research/ Director
University Of Alabama At Birmingham, 1530 3rd Avenue South, Birmingham, Al 35294
Grant 5T32DE017607-04 from National Institute Of Dental & Craniofacial Research
Abstract: This application is a NIDCR National Research Service Award Institutional Research Training Grant application from the University of Alabama at Birmingham (UAB) School of Dentistry (SOD). This comprehensive research-training program is focused on development of an innovative, integrated, multi-disciplinary approach to produce a cadre of well-trained, skilled, collaborative scientists and clinician scientists that are capable of addressing critical dental, oral and craniofacial research issues. We have named our training program Dental Academic Research Training - DART. This program is a culmination of our successful NIDCR T32 Research Training programs (Post-doctoral Training in Oral Health T32-DE14300 & Post-Doctoral Training in Caries Research T32-DE007026) and a Short-Term Research Training for Health Professional Students grant (T35DK07545) funded for over thirty years. The experience gained through these various training activities has lead to an innovative program designed to enhance the interest of dentists, graduate students and dental students in basic and clinical research, and introduce students to dental academic careers. The DART Program offers four complementary training tracks Track I- Short-term dental student; Track II- DMD/PhD; Track III- Pre-doctoral PhD; and Track IV- Post-doctoral training with two emphasis lines 1) a traditional post-doctoral experience; and 2) a Masters of Science degree in Public Health in Clinical Research program. Research training will be provided in one of the SOD´s programmatic research theme foci biomaterials and tissue engineering; microbiology and immunology; craniofacial developmental biology and genetics; and public health and clinical research. The primary objective of this new NIDCR National Research Service Award Institutional Research Training Grant application is to provide a broadened interdisciplinary research experience in Year 1 for 18 exceptionally motivated trainees in a research-intensive academic university environment 8 short-term dental students, 3 Dental Scientist Training Program (DSTP) students, 3 Predoctoral PhD students, and 4 postdoctoral trainees. The UAB SOD is uniquely position to provide research training since we are the only dental school that has obtained NIDCR U24 Research Infrastructure (DE016449), U01 Practice-based Research Network (DE016747) and R25 Research Curriculum (DE015614) grants
Keywords: Dental; Research Training
Project start date: 2007-07-01
Project end date: 2012-06-30
Budget start date: 1-JUL-2010
Budget end date: 30-JUN-2011
PFA/PA: PAR-05-101
5T32DE017607-04 (2010): $770522
5T32DE017607-03 (2009): $766940
1T32DE017607-01A1 (2007): $507468
CANDIDATE GENES FOR DGI-II/III
Mary B Macdougall, James R. Rosen Chair Of Dental Research
Pediatric Dentistryuniversity Of Texas Hlth Sci Ctr San Ant
san Antonio, Tx 78229
Grant 5R01DE009875-09 from National Institute Of Dental & Craniofacial Research IRG: OBM
Abstract: The goal of this research proposal is delineation of primary genetic defects which are responsible for dentinogenesis imperfecta type II (DGI- III) and type III (DGI-III) and the autosomal dominate local hypoplastic form of amelogenesis imperfecta (ADAI). Dentinogenesis imperfecta (DGI) and amelogenesis imperfecta (AI) are inherited disorders of both the primary and secondary dentitions which affect dentin and enamel formation, respectively. The genetic linkage of DGI- II in a number of multi- generation families has been studied and place this gene locus on human chromosome 4 in the region 4q21-q23. Furthermore, a linkage study on a DGI-III kindred with juvenile periodontitis (JP) has also suggested that this disease may be linked to the long arm (q) of human chromosome 4. Furthermore, the autosomal dominate local hypoplastic form of AI has also been recently linked to a 15.2 cM region of human chromosome 4 in the region 4q11-4q13. The proposal is based on the following hypotheses 1, that mutations within one of the human dentin matrix proteins, dentin matrix 1 (DMP1), dentin sialoprotein (DSP) or dentin phosphoprotein (DPP), genes localized to 4q21-q23, cause DGI types II and/or III; and 2, that mutations within the human ameloblastin (AMBN) gene localized to 4q21 causes the local hypoplastic form of ADAI. To test these hypotheses, a human third molar cDNA library will be constructed in order to isolate the human cDNAs for DMP1, DSP and DPP and Ambn. These human cDNAs will be used to facilitate screening of human P1 or PAC genomic libraries for DMP1, DSP, DPP and AMBN genomic clones. In parallel, studies will be initiated to regionally localize DSP and DPP genes on human chromosome 4 using fluorescence in situ hybridization. Refined linkage studies will be performed to further establish linkage of DGI-III to human chromosome 4 q21-q23 markers. Once the human sequences for DMP-1, DSP, DPP and AMBN have been determined, mutational analysis of these genes will be initiated using available DNA samples of both DGI-II and III (DMP1, DSP, DPP) and ADAI (AMBN) kindreds
Keywords: ameloblast, dental disorder, dentinogenesis, extracellular matrix protein, gene mutation, genetic disorder, linkage mapping autosomal dominant trait, bone sialoprotein, chromosome deletion, family genetics, genetic marker, genome, phosphoprotein clinical research, complementary DNA, fluorescent in situ hybridization, genetic library, human genetic material tag, human subject, northern blotting, polymerase chain reaction, single strand conformation polymorphism, southern blotting, tissue /cell culture
Project start date: 1991-04-01
Project end date: 2000-01-31
5R01DE009875-09 (1999): $259978
2R01DE009875-07 (1997): $318723
CORE--CELL IMMORTALIZATION AND TRANSFECTION FACILITY
Mary B Macdougall, James R. Rosen Chair Of Dental Research
University Of Texas Hlth Sci Ctr San Ant San Antonio, Tx 78229
Grant 5P01DE013221-059001 from National Institute Of Dental And Craniofacial Research IRG: ZDE1
Abstract: The GERO Cell Immortalization and Transfection Facility, or CIT, will be supervised by Dr. MacDougall as project leader. The Service Core is designed to serve the 4 scientific projects of the Research Core, and each project will require tooth specific and stably transfected cell lines. The Service Core will produce immortalized cell lines by gene transfer to several dental cell types in monolayer culture, through use of retroviral oncogene constructs. Stable cell lines will be isolated and gene expression profile and nature of extracellular matrix will be analyzed. These cell lines will serve as a resource for in vitro promoter studies in the Research Core projects. The Service Core will also perform stable cell transfections with various promoter-reporter constructs provided by the individual projects.
Keywords: biomedical facility, cell line, cell transformation, dental development, transfection
Sponsored Links Excellgen http://Excellgen.com
GENE EXPRESSION REGULATION DURING ODONTAGENESIS
Mary B Macdougall, James R. Rosen Chair Of Dental Research
University Of Texas Hlth Sci Ctr San Ant San Antonio, Tx 78229
Grant 5P01DE013221-05 from National Institute Of Dental And Craniofacial Research IRG: ZDE1
Abstract: The Gene Expression and Regulation during Odontogenesis (GERO) Program Project grant is a collaborative effort between investigators from the Department of Pediatric Dentistry at The University of Texas Health Science Center at San Antonio, The University of Missouri at Kansas City, and nineteen fellow investigators from eight different domestic and four different foreign institutions. The overall objective of this interdisciplinary, multi-disciplinary research team of scientists is to understand the fundamental multi-disciplinary research team of scientists is to understand the fundamental regulatory mechanisms involved in tooth formation, tooth-specific gene expression, and matrix protein function. The program is composed of three Cores, designed to integrate the four research projects into a coordinated, goal-oriented effort, which is efficiently coordinated for maximum productivity and oversee research activities while providing both internal and external reviews. The Research Core consists of four individual but related research projects to investigate 1) the role of BMP4 in tooth formation and potential regulation of this cytokine by the transcription factor Msx1; 2) the regulation and function of enamelin, and enamel matrix protein; 3) the regulation and function of dentin sialophosphoprotein (DSPP) during odontogenesis; and 4) the regulation and function of a newly isolated enamel proteinase, enamel matrix serine proteinase 1 during amelogenesis. These research projects will be supported by a Service Core, entitled the "Cell Immortalization and Transfection facility," (CIT) which will provide unique dental cell lines and stable cell transfections for the GERO investigators. While the individual projects all have the potential to produce significant advances related to their research goals, the integrated format of this Program Project grant will further facilitate the productivity, resources, and potential of significant contributions by this highly motivated team of research investigators.
Keywords: dental development, developmental genetics, gene expression
Project start date: 2000-04-01
Project end date: 2006-03-31
5P01DE013221-05 (2004): $969760
5P01DE013221-04 (2003): $950826
5P01DE013221-03 (2002): $926254
5P01DE013221-02 (2001): $911381
1P01DE013221-01A1 (2000): $925881
CANDIDATE GENES FOR DG1 AND ADAI
Mary B Macdougall, James R. Rosen Chair Of Dental Research
Pediatric Dentistryuniversity Of Texas Hlth Sci Ctr San Ant
san Antonio, Tx 78229
Grant 5R01DE009875-11 from National Institute Of Dental & Craniofacial Research IRG: OBM
Abstract: Relatively little in known about the molecular etiology of the human genetic diseases of enamel and dentin formation amelogenesis imperfecta (AI), dentinogenesis imperfecta (DGI), and dentin dysplasia (DD). Linkage studies for autosomal dominant (AD) forms of AI, approximately 85% of all cases, have mapped only one type, local hypoplastic AI (AIH2), to human chromosome 4q21 within a 4 Mb region containing the genes for ameloblastin and enamelin. This same general region of 4q21 has been shown to contain the overlapping critical gene loci for three dentin diseases, DGI type II (DGI-II), DGI type III (DGI-III), and DD type II (DD-II). This overlapping region contains a dentin/bone "gene cluster", including osteopontin, bone sialoprotein, dentin matrix protein I, and dentin sialophosphoprotein. Mutational analysis studies have yet to establish the genes causing these diseases. The goal of this R01 continuation renewal is the delineation of the primary genetic defects that are responsible for the diseases DGI-II, DGI-III, DD- II, and the AD forms of AI. We will expand our efforts to include DD type II and other AD forms of AI through newly ascertained informative families. This study is based on the hypothesis that specific mutations within single dentin and enamel matrix proteins/proteinases have a causative role in the pathogenesis of DGI, DD and the AD forms of AI. To test this hypothesis we will ascertain informative families with these dental genetic diseases, establish or exclude linkage to the DGI/DD or AIH2 Critical disease loci on 4q21, and perform mutational analysis of potential candidate genes. Refined linkage studies will be performed on these disease loci through more extensive marker genotyping of additional families. ADAI families excluded from 4q2i, will be tested for linkage to other candidate enamel gene loci on chromosomes 1, 11, and 19, and the critical disease loci mapped. If ADAI families are excluded from linkage to all potential gene loci, a genome-wide search will be implemented. Finally, through positional cloning approaches identify potential new candidate tooth genes on 4q21 using constructed DGI/DD and AIH2 contigs. This information will contribute to our understanding of the molecular bases of these diseases, provide better diagnosis tools, increase our knowledge related to the function(s) of tooth proteins, and assisting in the design of treatment therapies
Keywords: ameloblast, dental disorder, dentinogenesis, extracellular matrix protein, gene mutation, genetic disorder, linkage mapping autosomal dominant trait, bone sialoprotein, dentin, family genetics, genetic marker, genotype, molecular pathology, phosphoprotein artificial chromosome, clinical research, fluorescent in situ hybridization, genetic library, human genetic material tag, human subject, northern blotting, polymerase chain reaction, single strand conformation polymorphism, southern blotting
Project start date: 1991-04-01
Project end date: 2004-03-31
5R01DE009875-11 (2001): $214944
2R01DE009875-10 (2000): $236194
Sponsored Links Excellgen http://Excellgen.com
DENTIN SIALOPHOSPHOPROTEIN (DSPP) DURING ODONTOGENESIS
Mary B Macdougall, James R. Rosen Chair Of Dental Research
University Of Texas Hlth Sci Ctr San Ant San Antonio, Tx 78229
Grant 5P01DE013221-050003 from National Institute Of Dental And Craniofacial Research IRG: ZDE1
Abstract: Adapted from the investigator s ) Currently very little is known regarding the tissue-specific gene regulation of tooth extracellular matrix proteins during odontogenesis. The purpose of this proposal is to study the regulatory mechanisms that control the process of dentine Sialophosphoprotein (DSPP) gene expression during tooth formation, while gaining information to its biological function. DSPP is a large processor protein that is specifically cleaved giving rise to two dentine proteins dentine sialoprotein (DSP) and dentine phosphoprotein also known as phosphophoryn (DPP). In situ hybridization studies show the restricted pattern of DSPP. The application plan to elucidate the molecular mechanism that control the expression of DSPP in the odontoblasts (enhancers) and down-regulate expression (silencers) in ameloblasts. The hypothesis is that tissue and developmental regulation of DSPP is regulated both up and down through discrete interactions of transcription factors with their cognate cis-elements in specific region of the promoter and that this protein has an important role in tooth mineralization. To test this hypothesis, four specific aims are proposed 1) to determine the DNA sequences of the 5 upstream promoter region of previous identified mouse and human genomic clones; 2) to determine the region of the DSPP promoter that confers the potential tissue specific expression of DSPP gene using novel immortalized dental cell lines and in vivo using a transgenic approach; 3) to generate transgenic Knock-in DSPP LacZ mouse model to investigate the in vivo endogenous patterns of expression of native DSPP promoter, while allowing the functional role to be explored in homozygous null DSPP animals; and 4) to determine the effects of DSPP over-expression using trasgenic mouse model. These experiments will provide needed preliminary data and background for the future potential use of the DSPP promoter in human gene therapy strategy for the numerous dentine and enamel genetic diseases. Furthermore, once fully characterized, the DSPP promoter could be used in other therapeutic applications to derive tooth specific expression of beneficial target genes related to pulp capping modalities.
Keywords: dental development, dentin, dentinogenesis, developmental genetics, extracellular matrix protein, phosphoprotein, sialate, nucleic acid sequence, odontoblast, transcription factor, cell line, genetically modified animal, in situ hybridization, laboratory mouse
Transdentinal Induction Of Tertiary Dentin
Mary B Macdougall, James R. Rosen Chair Of Dental Research
University Of Texas Hlth Sci Ctr San Ant San Antonio, Tx 78229
Grant 2P01DE011688-060003 from National Institute Of Dental And Craniofacial Research IRG: ZDE1
Abstract: A major goal of restorative dentistry is the replacement of carious tooth structure with materials that closely resemble that natural mineralized tissue, while preserving the vitality of the pulpo-dentinal complex. Pulp tissue exhibits an intrinsic defense mechanism against insult or injury culminating in the differentiation of new odontoblasts and the deposition of tertiary dentin. While the formation of reactionary or reparative dentin by the pulp tissue is an inherent response to injury or insult, it is not routinely evoked in modern clinical treatments. Our goal is to develop a new dental material that can be applied to the floor of a cavity to evoke this biologically favorable response of self-healing. Studies suggest that cytokines of the TGF-beta family, in particular bone morphogenetic proteins, as well as other, recently identified mineralized matrix molecules (e.g. bone sialoprotein, amelogenin, dentin sialophosphoprotein, dentin matrix protein 1) are capable of inducing proliferative and/or phenotypic changes in dental pulp cells that are comparable to those occurring during the reparative process. Our hypothesis is that dental material containing dentin-inducing molecules when placed on the floor of a cavity preparation will induce tertiary dentin formation by the underlying dental pulp tissue in a predictable manner. To test this hypothesis, we propose the following four specific aims Specific Aim 1 will use newly developed human dental cell lines to screen potential dentin-inductive molecules in a novel DSPP gene activation assay. Specific Aim 2 will determine the mechanism of action of identified inductive molecules. Specific Aim 3 will characterize the optimal concentration and time-course of promising dentin inductive molecules on the phenotypic changes in human dental pulp and odontoblast cells in vitro. Specific Aim 4 will determine the parameters of transdentinal movement of the dentin inductive molecules in combination with our cyclodextrin carrier system in vitro. While our goal and hypothesis have not changed in this continuation proposal, we have broadened our scope to a more general survey of suitable dentin-inductive molecules to be used in combination with our novel restorative system. The ideal dentin-inductive molecule should be readily available, should have prolonged biological activity either alone or in complex with carrier molecules like cyclodextrins, and have excellent biocompatibility. We remain committed to see transdentinal tertiary dentin induction becoming an integral part of the restorative dental armentarium.
Keywords: biomaterial development /preparation, cytokine receptor, dental material, dentin, dentinogenesis, restorative dentistry, bone morphogenetic protein, cell differentiation, cyclodextrin, cytokine, drug delivery system, extracellular matrix, odontoblast, pulp, regeneration, cell line, human tissue
Project start date: 1997-06-01
Project end date: 2007-05-31
Institutional Research Training Grant
Mary B Macdougall, James R. Rosen Chair Of Dental Research
University Of Texas Hlth Sci Ctr San Ant San Antonio, Tx 78229
Grant 3T32DE014318-02S1 from National Institute Of Dental And Craniofacial Research IRG: ZDE1
Project start date: 2002-07-08
Project end date: 2007-06-30
3T32DE014318-02S1 (2004): $3456
3T32DE014318-03S1 (2004): $61059
Sponsored Links Excellgen http://Excellgen.com
Transdentinal Induction Of Tertiary Dentin
Mary B Macdougall, James R. Rosen Chair Of Dental Research
University Of Texas Hlth Sci Ctr San Ant San Antonio, Tx 78229
Grant 5P01DE011688-080003 from National Institute Of Dental And Craniofacial Research IRG: ZDE1
Project start date: 2004-06-01
Project end date: 2007-05-31
SHORT-TERM TRAINING: STUDENTS IN HEALTH PROFESSIONAL SCH
Mary B Macdougall, James R. Rosen Chair Of Dental Research
Pediatric Dentistryuniversity Of Texas Hlth Sci Ctr San Ant
san Antonio, Tx 78229
Grant 5T35DE007160-15 from National Institute Of Dental & Craniofacial Research IRG: DSR
Project start date: 1985-06-01
Project end date: 2002-05-31
5T35DE007160-15 (2001): $59953
5T35DE007160-14 (2000): $55287
5T35DE007160-13 (1999): $52449
2T35DE007160-11 (1997): $28071
DENTIN SPECIFIC PHOSPHOPROTEINS AND MINERALIZATION
Mary B Macdougall, James R. Rosen Chair Of Dental Research
Pediatric Dentistryuniversity Of Texas Hlth Sci Ctr San Ant
san Antonio, Tx 78229
Grant 1R01DE011658-01 from National Institute Of Dental & Craniofacial Research IRG: ZDE1
Project start date: 1996-03-15
Project end date: 2001-03-14
1R01DE011658-01 (1996): $159082
ESTABLISHMENT OF HUMAN DENTAL PULP MESENCHYME CELL LINES
Mary B Macdougall, James R. Rosen Chair Of Dental Research
Pediatric Dentistryuniversity Of Texas Hlth Sci Ctr San Ant
san Antonio, Tx 78229
Grant 5R03DE011493-02 from National Institute Of Dental & Craniofacial Research IRG: ZDE1
Project start date: 1995-09-30
Project end date: 1997-08-31
5R03DE011493-02 (1996): $33105