East African Cichlids: A Natural Model for Dental Regeneration

东非慈鲷：牙齿再生的自然模型

• 批准号: 8657807
• 负责人: Ryan Fredric Bloomquist
• 金额: $ 3.14万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8657807
• 关键词: Address; Adult; African; Agonist; Animal Model; Animals; Antibodies; Biocompatible; Biological; Biological Assay; Biology; Biomaterials Research; Biomedical Engineering; Bromodeoxyuridine; Cell Therapy; Cell model; Cells; Characteristics; Chemicals; Cichlids; Congenital Abnormality; DNA; Data; Defect; Dental; Dental Models; Dentistry; Dentition; Development; Developmental Biology; Engineering; Epithelial; Evolution; Exhibits; Family; Fibroblast Growth Factor; Fishes; Funding; Generations; Genes; Growth; Human; In Situ Hybridization; Label; Learning; Length; Life; Location; Maintenance; Malawi; Mesenchymal; Mesenchymal Stem Cells; Messenger RNA; MicroRNAs; Modality; Modeling; Molecular; Molecular Profiling; Morbidity - disease rate; Mus; National Institute of Dental and Craniofacial Research; Natural regeneration; Nature; Nucleosides; Odontogenesis; Oral; Organ; Organism; Pathology; Pathway interactions; Phenotype; Physiologic pulse; Play; Population; Primary Dentition; Protocols documentation; Regenerative Medicine; Research Support; Role; Signal Transduction; Stem cells; Strategic Planning; System; Taste Buds; Testing; Therapeutic; Time; Tissues; Tooth Germ; Tooth structure; United States National Institutes of Health; Vertebrates; Work; adult stem cell; base; craniofacial; global health; improved; inhibitor/antagonist; insight; interest; notch protein; oral tissue; population based; public health relevance; reconstruction; regenerative; repaired; research study; small molecule; stem; stem cell biology; stem cell niche; stem cell population; therapy design; treatment effect

DESCRIPTION (provided by applicant): The most promising therapeutics for addressing morbidities that arise from congenital, traumatic, or infectious pathology lie in the field of regenerative medicine. On the forefront of these therapeutics is the field of regenerative dentistry. As we turn to cell-based therapies for repair of missing teeth and damaged oral tissues, we must look to developmental biology to understand how nature grew these tissues in the first place. What we know about tooth development or odontogenesis has been almost exclusively derived from the mouse. While we have learned a great deal from this system, in contrast to humans and most other vertebrates, the mouse lacks a successional or replacement set of teeth. We know relatively little about how to replace the dentition. To discover how nature perfected the growth of a tooth in an adult organism, we turn to the de-novo continuously replacing dentition of Lake Malawi cichlid fishes. In specific aim 1, I will identify dental stem cll populations responsible for natural de-novo tooth replacement in a perpetually replacing vertebrate dentition. I will utilize protocols I have developed to identify expression profiles of factors implicated in the stem cell biology of other vertebrate models. In supporting research, I noted that cichlid teeth and taste buds share a common epithelial ribbon, as well as gene co-expression, during development and regeneration. I will employ in-situ hybridization (ISH) for mRNAs and miRNAs to uncover factors expressed in both tooth and/or taste bud development which may be involved in their continuous turn-over. I will then employ pulse-chase analysis to verify the location and character of putative stem cells in support of my ISH experiments. In aim 2, I will manipulate molecular signals that putatively control the regeneration of teeth and taste buds. In this aim, I will treat fishes with small molecule chemical agonists and antagonists of major developmental pathways expressed in putative stem cell niches. I will use antagomirs to disrupt the function of miRNAs analyzed in aim one that are expressed in the putative stem niches. I will subsequently assay changes in mRNA, miRNA, and pulse-chase data in those treatments resulting in disrupted tooth replacement phenotypes. These experiments will demonstrate the importance of key factors in dental regeneration. As we move into an era where the replacement of missing or damaged tissues has become not only a promising therapy, but a viable possibility, it is imperative that we appreciate how those tissues were formed in the first place. With a more global health impact, we seek to identify the epithelial and mesenchymal stem cells responsible for the natural replacement of an adult organ. Part of the NIDCR strategic plan over the next years is to "facilitate reconstruction and regeneration of diseased or damaged oral and craniofacial tissues and organs through biological, bioengineering and biomaterials research approaches". These findings will advance our understanding of dental stem cells so that we may one day design therapies based on endogenous mechanisms of repair.

描述（由申请人提供）：用于解决由先天性、创伤性或感染性病理学引起的发病率的最有希望的治疗方法在于再生医学领域。在这些疗法的最前沿是再生牙科领域。当我们转向以细胞为基础的治疗方法来修复缺失的牙齿和受损的口腔组织时，我们必须从发育生物学的角度来理解自然界首先是如何生长这些组织的。我们对牙齿发育或牙齿发生的了解几乎完全来自小鼠。虽然我们已经从这个系统中学到了很多，但与人类和大多数其他脊椎动物相比，老鼠缺乏连续或替换的牙齿。我们对如何更换牙列知之甚少。为了发现大自然是如何完善成年生物牙齿的生长的，我们转向马拉维湖慈鲷鱼不断更换的牙齿。在具体目标1中，我将确定在永久替换的脊椎动物牙列中负责自然从头牙齿替换的牙干细胞群。我将利用我已经开发的协议来确定其他脊椎动物模型的干细胞生物学中涉及的因子的表达谱。在支持研究中，我注意到慈鲷的牙齿和味蕾在发育和再生过程中共享一个共同的上皮带，以及基因共表达。我将采用mRNA和miRNA的原位杂交（ISH）来揭示牙齿和/或味蕾发育中表达的因子，这些因子可能参与了它们的连续翻转。然后，我将采用脉冲追踪分析来验证假定的干细胞的位置和特征，以支持我的ISH实验。在目标2中，我将操纵分子信号来控制牙齿和味蕾的再生。在这个目标中，我将治疗鱼类与小分子化学激动剂和拮抗剂的主要发育途径中表达的假定干细胞壁龛。我将使用miRNAs干扰目标1中分析的miRNAs的功能，这些miRNAs在假定的茎小生境中表达。随后，我将分析导致牙齿替换表型破坏的那些治疗中mRNA、miRNA和脉冲追踪数据的变化。这些实验将证明关键因素在牙齿再生中的重要性。随着我们进入一个时代，缺失或受损组织的替代不仅成为一种有前途的疗法，而且成为一种可行的可能性，我们必须首先了解这些组织是如何形成的。随着全球健康的影响，我们寻求确定上皮和 间充质干细胞负责成人器官的自然替代。NIDCR未来几年战略计划的一部分是“通过生物、生物工程和生物材料研究方法促进患病或受损口腔和颅面组织和器官的重建和再生”。这些发现将促进我们对牙齿干细胞的理解，以便我们有一天可以设计基于内源性修复机制的治疗方法。