Why is Fam83h critical for enamel formation?

为什么 Fam83h 对于牙釉质形成至关重要？

• 批准号: 8441387
• 负责人: JAN Ching Chun HU
• 金额: $ 34.53万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8441387
• 关键词: 8q24.3; Ameloblasts; Amelogenesis; Amelogenesis Imperfecta; Bacteria; Biochemical; Biological; Candidate Disease Gene; Cells; Characteristics; Chromosomes; Chromosomes, Human, Pair 8; Co-Immunoprecipitations; Code; Complex; Defect; Dental Enamel; Development; Disease; Dominant-Negative Mutation; Enamel Formation; Etiology; Exons; Extracellular Matrix Proteins; Family; Genes; Genetic; Goals; Histologic; Immunohistochemistry; In Situ Hybridization; In Vitro; Inferior; Inherited; Lead; Learning; Link; MMP-20; Mass Spectrum Analysis; Monoclonal Antibodies; Mutate; Mutation; Nonsense Mutation; Odontogenesis; Oral health; Pain; Patients; Pattern; Peptides; Phenotype; Phosphoserine; Phosphothreonine; Phosphotyrosine; Play; Post-Translational Protein Processing; Principal Investigator; Proteins; Quality of life; Recombinants; Research; Role; Scanning Electron Microscopy; Secondary to; Structural Protein; Structure; System; Testing; Time Study; Tissues; Tooth structure; Transgenes; Transgenic Mice; Translations; Western Blotting; amelogenin; base; enamelin; genome-wide; glycosylation; in vivo; light microscopy; malformation; member; mouse model; novel; polyclonal antibody; programs; self esteem; yeast two hybrid system

Inherited diseases of dental enamel formation are grouped under the designation of amelogenesis imperfecta (AI). About 25% of all amelogenesis imperfecta (AI) cases are caused by the genes encoding four enamel extracellular matrix proteins, AMELX, ENAM, MMP20, and KLK4. Recently we identified mutations in FAM83H that cause AI in six different families suffering from amelogenesis imperfecta. This demonstrates that FAM83H is necessary for proper enamel formation. Virtually nothing is known about the Fam83h protein. Our objective is to learn the roles played by Fam83h in normal and defective enamel formation. Hypotheses: Because the phenotype in people with FAM83H mutations is limited to developing teeth, we hypothesize that Fam83H is expressed during odontogenesis. Because all of the AI- causing FAM83H mutations are dominant nonsense mutations that terminate translation in the last coding exon (exon 5), we further hypothesize that the Fam83h interacts with other cellular proteins to form functional complexes. We propose four Specific Aims: SA 1: To characterize the temporal and spatial pattern of Fam83h expression during odontogenesis and to determine its subcellular localization. SA 2: To isolate Fam83H protein for structural and functional characterization. SA 3: To identify Fam83H interacting proteins. SA 4: To determine if the expression of truncated Fam83h interferes with amelogenesis. Approach: The temporal and spatial pattern of Fam83h expression during odontogenesis is determined by in situ hybridization and immunohistochemistry. Recombinant and native Fam83h are used in structural and functional studies. Fam83h interacting proteins are identified using the yeast two-hybrid system and validated using a mammalian two-hybrid system, co- immunoprecipitation, and Far Western analyses. Normal and mutated Fam83h are expressed in ameloblasts as transgenes and their effects on enamel formation characterized. Heath Relatedness: Patients with inherited enamel defects (AI) have painful, disfigured teeth, lower self-esteem, and perceive themselves as having an inferior quality of life. The discovery that FAM83H is critical for dental enamel formation is a significant advance in our understanding of the etiology of AI. The proposed research may lead to a breakthrough in our understanding of important activities within ameloblasts and the discovery of new candidate gene(s) for AI.

遗传性牙釉质形成疾病被归类为 成釉不全(AI)。在所有的成釉发育不全(AI)病例中，约25%是由 由编码四种釉质细胞外基质蛋白AMELX、ENAM、MMP20和 KLK4.最近，我们在6个不同的家系中发现了导致AI的FAM83H基因突变 患有釉质发育不全。这表明FAM83H是必需的 适当的釉质形成。对Fam83h蛋白几乎一无所知。我们的目标是 了解Fam83h在正常和缺陷牙釉质形成中的作用。 假设：因为FAM83H突变患者的表型仅限于发育阶段 我们假设Fam83H在牙齿发育过程中表达。因为所有的人工智能- 引起FAM83H突变的是终止翻译的显性无义突变 最后一个编码外显子(外显子5)，我们进一步假设Fam83h与其他细胞相互作用 蛋白质形成功能复合体。我们提出了四个具体目标： SA 1：研究Fam83h基因表达的时间和空间模式 并确定其亚细胞定位。 SA 2：分离Fam83H蛋白，进行结构和功能鉴定。 SA 3：鉴定Fam83H相互作用蛋白。 SA 4：确定截短的Fam83h的表达是否干扰成釉发生。 方法：Fam83h在牙齿发育过程中表达的时空模式是 用原位杂交和免疫组织化学方法检测。重组和原生的 Fam83h用于结构和功能研究。鉴定出与Fam83h相互作用的蛋白质 使用酵母双杂交系统并使用哺乳动物双杂交系统进行验证，co- 免疫沉淀和Far Western分析。正常和突变的Fam83h在 成釉细胞作为转基因细胞及其对釉质形成的影响。 健康相关：遗传性釉质缺陷(AI)患者有疼痛、毁容的牙齿， 降低自尊，认为自己的生活质量较低。这一发现 FAM83H对牙釉质的形成至关重要，这是我们对牙釉质形成认识的重大进步 人工智能的病因学。这项拟议的研究可能会使我们对 成釉细胞内的重要活动和AI新候选基因(S)的发现。

项目成果

Effects of Fam83h overexpression on enamel and dentine formation.

• DOI: 10.1016/j.archoralbio.2013.03.001
• 发表时间: 2013-09
• 期刊: ARCHIVES OF ORAL BIOLOGY
• 影响因子: 3
• 作者: Kueon, Young-Sun;Lee, Kyung-Eun;Ko, Jiyeon;Hu, Jan C. -C.;Simmer, James P.;Kim, Jung-Wook
• 通讯作者: Kim, Jung-Wook

Fam83h is associated with intracellular vesicles and ADHCAI.

• DOI: 10.1177/0022034509349454
• 发表时间: 2009-11
• 期刊: Journal of dental research
• 影响因子: 7.6
• 作者: Ding Y;Estrella MR;Hu YY;Chan HL;Zhang HD;Kim JW;Simmer JP;Hu JC
• 通讯作者: Hu JC