Modeling Dentin by G Protein Coupled Receptor Signaling

通过 G 蛋白偶联受体信号转导模拟牙本质

• 批准号: 8721747
• 负责人: Orapin V Horst
• 金额: $ 13.22万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-05 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8721747
• 关键词: Address; Affect; Annual Reports; Applications Grants; Award; BMP4; Bone Matrix; California; Cell Communication; Cell Density; Cell Therapy; Cell physiology; Cells; Cellular biology; Centers for Disease Control and Prevention (U.S.); Clinical; Collagen Type I; Complex; Coupled; Cyclic AMP; Cytokeratin-14 Staining Method; Data; Dental; Dental Pulp; Dental caries; Dental crowns; Dentin; Dentin Formation; Dentists; Development; Development Plans; Developmental Biology; Discipline; Doctor of Philosophy; Drug Design; Educational workshop; Engineering; Environment; Environmental Risk Factor; Epithelial; Epithelial Cells; Epithelium; Event; FGF9 gene; Faculty; Failure; Fostering; Foundations; Funding; Future; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; GTP-Binding Proteins; Goals; Growth Factor; Healed; Healthcare; Histocompatibility Testing; Hour; In Situ; Interdisciplinary Study; International; Journals; K-Series Research Career Programs; Knowledge; Lead; Learning; Ligands; Mediating; Medicine; Mentors; Mentorship; Mesenchymal; Mesenchymal Differentiation; Mesenchymal Stem Cells; Mesenchyme; Modeling; Molecular Biology; Morphology; Mus; National Institute of Dental and Craniofacial Research; Natural regeneration; Odontoblasts; Oral health; Phase; Physiological; Plant Roots; Production; Proteins; Pulp Chambers; Recurrence; Reporting; Research; Research Activity; Research Personnel; Research Training; Resources; Role; San Francisco; Schools; Scientist; Signal Pathway; Signal Transduction; Signaling Molecule; Specificity; Stem Cell Research; Stem cells; Testing; Tissue Engineering; Tissues; Tooth Cell; Tooth structure; Training; Training Support; Transgenic Mice; Tubular formation; Universities; Work; bone; career development; cell type; cost; cost effectiveness; dental genetics; experience; healing; improved; insight; keratin 5; malformation; material fatigue; member; mouse model; novel strategies; osteoblast differentiation; precursor cell; progenitor; programs; promoter; receptor; receptor function; repaired; research and development; restorative material; skills; transcription factor

DESCRIPTION (provided by applicant): This proposed application for a K08 Mentored Clinical Scientist Research Career Development Award is being submitted by Dr. Orapin Horst, a dentist-scientist at the University of California, San Francisco. This award will provide the support and training necessary for Dr. Horst to receive the mentoring to develop into an independent investigator, whereupon she will pursue her long-term goal of creating cell-based therapies and tissue engineering approaches to regenerate the dentin-pulp complex in a manner recapitulating normal development. She will have access to the many opportunities of UCSF to help her accomplish the following goals related to her career development: 1) to learn the disciplines of developmental biology and G protein coupled receptor (GPCR) signaling; 2) to experience interdisciplinary research with mentors experienced in basic to clinical translational work; 3) to build academic skills to become an outstanding research mentor and faculty member; 4) to develop skills in grantsmanship and obtaining funding; and 5) to collect pilot data for a future R01 application. To continue her progress towards these goals, Dr. Horst proposes a research plan that will elucidate signaling events between dental epithelial and mesenchymal progenitor cells to induce odontoblast differentiation and control the formation of the dentin-pulp complex. She will study these signaling events through the function of GPCRs, which have the potential to elicit or modify essentially any signaling pathway. Related findings have been reported in other cell and tissue types, in particular bone, but very little is known about GPCR function in teeth. The central hypothesis that will be tested in this K08 is that GPCR-modulated signaling molecules regulate odontoblast differentiation and dentin matrix formation. This hypothesis is addressed by the following specific aims: 1) To determine how Gs-coupled GPCR signals in dental mesenchymal cells including odontoblasts and preodontoblasts control dentin formation, and 2) To determine how Gs-coupled GPCR signals from dental epithelial cells modulate odontoblast differentiation. These aims will be pursued using a newly developed transgenic mouse model of G-protein signaling in mineralized tissue formation. Transgenic mice with increased Gs-coupled GPCR signals in odontoblasts and preodontoblasts driven by the type I collagen promoter will be used in Specific Aim 1, while mice with increased Gs-coupled GPCR signals in dental epithelial cells driven by the keratin 5 promoter will be used in Specific Aim 2. These studies will identify new interactions between GPCR signaling pathways and other mechanisms regulating odontoblast development and dentin matrix formation. Ultimately, these data will lay a foundation for an interdisciplinary research program and a R01 grant application to engineer a physiologic complex of tubular dentin supporting odontoblasts and pulp to heal carious teeth. The knowledge from this work will also improve our conceptual understanding of genetic dental malformations and environmental factors required for proper cell-matrix interactions to support normal development of dentin and other mineralized tissues. The proposed career development and research activities will take place in highly supportive, research- intensive environments. Two international experts in the developmental biology of teeth, Pamela DenBesten DDS, MS (primary mentor) and Ophir Klein MD, PhD (co-mentor), will supervise Dr. Horst's research focus on epithelial-mesenchymal interactions and formation of mineralized tissues. Two international experts in GPCR signaling, Bruce Conklin MD (co-mentor, GPCR signaling in stem cells) and Robert Nissenson PhD (co- mentor, GPCR signaling in bone), will supervise work with the transgenic mouse models, and relate their vast experience with other stem cell types and mineralized tissues, to understand the role of GPCR signaling in dental stem cells and the formation of the dentin-pulp complex. The mentors are all members of the new Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research of UCSF, which will serve as a central resource for training opportunities (workshops, seminars, journal clubs, and retreats) in combining expertise in developmental, molecular, and cellular biology with medically relevant foci such as that proposed in this project. The proposed research is part of a coordinated career development plan to prepare the candidate to be an outstanding, independent dentist-scientist through research, coursework, and tutorials. The interdisciplinary mentorship team will mentor the career development activities together, and facilitate Dr. Horst's natural progression from current research experiences to the mentored clinical scientist development phase and then to independence.

描述（由申请人提供）：该拟议的K08指导临床科学家研究职业发展奖的申请由加利福尼亚大学旧金山分校的牙医科学家Orapin Horst博士提交。该奖项将为霍斯特博士提供所必需的支持和培训，以接收指导，以发展为独立的研究者，于是她将追求长期的目标，即创建基于细胞的疗法和组织工程方法，以缩减正常发展的方式来再生牙本质 - 脉络膜复合体。她将获得UCSF的许多机会，以帮助她实现与职业发展有关的以下目标：1）学习发育生物学和G蛋白偶联受体（GPCR）信号的学科； 2）与在基础转化工作中经历的导师经历的跨学科研究； 3）建立学术技能，成为一名杰出的研究导师和教职员工； 4）发展授予技巧和获得资金的技能； 5）收集未来R01应用程序的试点数据。为了继续朝着这些目标迈进，霍斯特博士提出了一项研究计划，该计划将阐明牙皮上皮和间质祖细胞之间的信号传导事件，以诱导牙本质细胞分化并控制牙本质 - 脉络膜复合物的形成。她将通过GPCR的功能来研究这些信号事件，这些功能有可能引起或修改任何信号通路。已经报道了其他细胞和组织类型，尤其是骨骼，但对牙齿中的GPCR功能知之甚少。在此K08中将测试的中心假设是GPCR调节的信号分子调节牙糖细胞分化和牙本质基质形成。以下特定目的解决了这一假设：1）确定牙齿间充质细胞中的GS耦合GPCR信号（包括牙糖细胞和前牙本质细胞）如何控制牙本质的形成，以及2）如何确定GS耦合的GPCR信号如何调节牙皮上皮细胞调节牙型甲状腺细胞的分化。这些目的将使用新开发的G蛋白信号传导的转基因小鼠模型在矿化组织形成中追求。在特定目标1中，将使用具有I类胶原蛋白启动子驱动的ODONTOBLAST和前体细胞中GS耦合GPCR信号增加的GS耦合GPCR信号的转基因小鼠，而具有GS耦合的GPCR信号的小鼠在牙齿上皮细胞中的GS耦合GPCR信号在牙齿上上皮细胞中的其他指示剂将在特定的目标中使用。 Odontoblast发育和牙本质基质形成。最终，这些数据将为跨学科研究计划和R01赠款应用奠定基础，以设计肾小管牙本质的生理综合体，以支撑牙本质细胞和果肉来治愈龋齿。这项工作的知识还将提高我们对适当细胞玛底相互作用所需的遗传牙齿畸形和环境因素的概念理解，以支持牙本质和其他矿化组织的正常发育。拟议的职业发展和研究活动将在高度支持，研究密集的环境中进行。牙齿发育生物学的两位国际专家Pamela Denbesten DDS，MS（主要导师）和Ophir Klein MD博士（Co-Mentor）将监督Horst博士对上皮 - 间质相互作用的研究重点和矿化组织的形成。 Two international experts in GPCR signaling, Bruce Conklin MD (co-mentor, GPCR signaling in stem cells) and Robert Nissenson PhD (co- mentor, GPCR signaling in bone), will supervise work with the transgenic mouse models, and relate their vast experience with other stem cell types and mineralized tissues, to understand the role of GPCR signaling in dental stem cells and the formation of the dentin-pulp complex.这些导师都是UCSF的新Eli和Edythe Regeneration Medicine和Stem细胞研究中心的成员，该中心将成为培训机会的中心资源（研讨会，研讨会，期刊，期刊俱乐部和务虚会），以结合发育，分子和细胞生物学的专业知识，并在此项目中提供了医学上相关的焦点。拟议的研究是一项协调的职业发展计划的一部分，旨在通过研究，课程和教程使候选人成为一名杰出的独立牙医科学家。跨学科的指导团队将共同指导职业发展活动，并促进霍斯特博士从当前的研究经验到指导的临床科学家发展阶段，然后再到独立性的自然发展。