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）：K08 指导临床科学家研究职业发展奖的拟议申请由旧金山加利福尼亚大学牙医科学家 Orapin Horst 博士提交。该奖项将为 Horst 博士提供必要的支持和培训，使她能够接受指导，发展成为一名独立研究者，从而实现她的长期目标，即创建基于细胞的疗法和组织工程方法来再生牙本质牙髓复合物以重现正常发展的方式。她将有机会获得 UCSF 的众多机会，帮助她实现以下与职业发展相关的目标： 1) 学习发育生物学和 G 蛋白偶联受体 (GPCR) 信号传导学科； 2）与具有基础到临床转化工作经验的导师一起体验跨学科研究； 3）培养学术技能，成为优秀的研究导师和教员； 4）培养资助和获得资金的技能； 5) 为未来的 R01 应用收集试点数据。为了继续实现这些目标，霍斯特博士提出了一项研究计划，该计划将阐明牙上皮和间充质祖细胞之间的信号传导事件，以诱导成牙本质细胞分化并控制牙本质-牙髓复合物的形成。她将通过 GPCR 的功能研究这些信号事件，GPCR 有可能引发或修改任何信号通路。其他细胞和组织类型（特别是骨骼）中也有相关发现，但人们对牙齿中 GPCR 的功能知之甚少。本 K08 将测试的中心假设是 GPCR 调节的信号分子调节成牙本质细胞分化和牙本质基质形成。该假设通过以下具体目标得到解决：1) 确定牙间充质细胞（包括成牙本质细胞和前成牙本质细胞）中的 Gs 偶联 GPCR 信号如何控制牙本质形成，以及 2) 确定来自牙上皮细胞的 Gs 偶联 GPCR 信号如何调节成牙本质细胞分化。这些目标将使用新开发的矿化组织形成中 G 蛋白信号转导的转基因小鼠模型来实现。具体目标1将使用由I型胶原启动子驱动的成牙本质细胞和前成牙本质细胞中Gs偶联GPCR信号增加的转基因小鼠，而由角蛋白5启动子驱动的牙上皮细胞中Gs偶联GPCR信号增加的小鼠将被使用具体目标 2。这些研究将确定 GPCR 信号通路与其他调节成牙本质细胞发育和牙本质基质形成的机制之间的新相互作用。最终，这些数据将为跨学科研究计划和 R01 拨款申请奠定基础，以设计支持成牙本质细胞和牙髓的管状牙本质生理复合体，以治愈龋齿。这项工作的知识还将提高我们对遗传性牙齿畸形和适当细胞-基质相互作用所需的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的研究密集型环境中进行。牙齿发育生物学领域的两位国际专家 Pamela DenBesten DDS, MS（主要导师）和 Ophir Klein MD, Ph.D（共同导师）将监督 Horst 博士的上皮-间质相互作用和矿化组织形成的研究重点。 GPCR 信号传导方面的两位国际专家 Bruce Conklin MD（共同导师，干细胞中的 GPCR 信号传导）和 Robert Nissenson 博士（共同导师，骨骼中的 GPCR 信号传导）将监督转基因小鼠模型的工作，并分享他们丰富的经验与其他干细胞类型和矿化组织合作，了解 GPCR 信号在牙干细胞中的作用以及牙本质-牙髓复合物的形成。这些导师都是加州大学旧金山分校新成立的 Eli 和 Edythe Broad 再生医学和干细胞研究中心的成员，该中心将作为培训机会（讲习班、研讨会、期刊俱乐部和静修会）的中心资源，结合发展、具有医学相关焦点的分子和细胞生物学，例如本项目中提出的。拟议的研究是协调职业发展计划的一部分，旨在通过研究、课程和辅导使候选人成为一名杰出的、独立的牙医科学家。跨学科导师团队将共同指导职业发展活动，并促进 Horst 博士从当前的研究经验自然发展到受指导的临床科学家发展阶段，然后走向独立。