Epigenetic and non-epigenetic role of SIRT1 in fluoride-induced cell stress

SIRT1 在氟化物诱导的细胞应激中的表观遗传和非表观遗传作用

基本信息

批准号：
10408040
负责人：
Maiko Suzuki
金额：
$ 36.09万
依托单位：
AUGUSTA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-06-01 至 2023-03-23
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10408040
关键词：
Ablation Acquired Dental Fluorosis Ameloblasts Apoptosis Attenuated Autophagocytosis Biological CRISPR/Cas technology Cell Line Cell Survival Cells Cellular Stress ChIP-seq Child Chronic DNA Sequence Deacetylation Dental Enamel Dental caries Development Disease Epigenetic Process Exposure to Fluorides Functional disorder Future Gene Expression Gene Expression Alteration Genes Genetic Transcription Goals Histone Acetylation Histone Deacetylase Histone Deacetylation Histones Homologous Gene In Vitro Ingestion Knock-out Knockout Mice Longevity MAPK8 gene Mating Types Mediating Metabolic Modification Molecular Mus Oral health Oxidative Stress Pathway interactions Pharmacological Treatment Pharmacology Phenotype Play Population Prevalence Reactive Oxygen Species Regulation Reporting Role SIRT1 gene Signal Transduction Stream TP53 gene Testing Tooth structure Toxic effect Water fluoridation aged biological adaptation to stress conditional knockout endoplasmic reticulum stress epigenetic regulation fluorosis in vivo innovation malformation mitochondrial metabolism non-histone protein novel novel strategies novel therapeutic intervention overexpression oxidative damage prevent prophylactic response

项目摘要

The goal of this application is to characterize epigenetic and non-epigenetic functions of SIRT1 in adaptive responses during dental fluorosis. Fluoride is an effective caries prophylactic, however chronic over-exposure can result in dental fluorosis. More than 30% of children in the U.S. suffer from dental fluorosis. Our ultimate goal is to develop novel strategies that prevent or reduce dental fluorosis while keeping the benefit of prophylactic public water fluoridation to prevent caries. Recently we reported that fluoride activates SIRT1 and autophagy as an adaptive response to protect cells from cell stress. It is known that SIRT1 deacetylates histones to repress gene expression (epigenetic deacetylation). SIRT1 also deacetylates a number of non-histone proteins (non-epigenetic deacetylation) functioning in the regulation of autophagy, mitochondria metabolism, cell survival and organismal lifespan. However, the exact functions of SIRT1 and its downstream targets in dental fluorosis are unknown. Our goal in this proposal is to identify how SIRT1 functions in fluoride-induced stress responses and to find appropriate targets that could be modulated pharmacologically for the treatment of dental fluorosis. Our central hypothesis is that SIRT1 can play protective roles by intiating autophagy and regulating epigenetics in dental fluorosis. To test our hypothesis we propose three specific AIMs. AIM 1. Identify SIRT1 non-histone targets in ameloblast-derived cells treated with fluoride in vitro. AIM 2. Identify SIRT1 histone targets in ameloblast-derived cells treated with fluoride in vitro. AIM 3. Determine if SIRT1 over expression or conditional knockout effects enamel development and dental fluorosis in vivo. To attain these AIMs, we will use SIRT1 overexpressor and knockout ameloblast-like cells LS8 (LS8 Sirt1/over and LS8Sirt1/KO cells) established by the CRISPR/Cas9 technology in vitro. We will analyze the role of SIRT1 using SIRT1 over expressing mice (SIRT1super) and conditional knockout mice (SIRT1cKO) in vivo. Once epigenetic and non-epigenetic SIRT1 function in enamel pathophysiology is revealed, it may be possible to develop novel strategies to pharmacologically manipulate SIRT1 function to prevent dental fluorosis.

该应用的目标是表征表观遗传和非表观遗传功能 SIRT1 在氟斑牙期间适应性反应中的作用。氟化物是一种有效的防龋剂预防性的，但长期过度接触会导致氟斑牙。超过30% 美国的儿童患有氟斑牙。我们的最终目标是制定新颖的策略预防或减少氟斑牙，同时保持预防性公共水的益处氟化物可预防龋齿。最近我们报道氟化物激活 SIRT1 和自噬作为保护细胞免受细胞应激的适应性反应。众所周知，SIRT1 可以脱乙酰基组蛋白抑制基因表达（表观遗传脱乙酰化）。 SIRT1 还可以使在调节中起作用的非组蛋白（非表观遗传脱乙酰化）的数量自噬、线粒体代谢、细胞存活和有机体寿命。然而，确切的 SIRT1 及其下游靶点在氟斑牙中的功能尚不清楚。我们在这方面的目标该提案的目的是确定 SIRT1 在氟化物诱导的应激反应中如何发挥作用，并找到可以通过药理学调节来治疗牙科疾病的适当目标氟中毒。我们的中心假设是 SIRT1 可以通过启动自噬发挥保护作用以及调节氟斑牙的表观遗传学。为了检验我们的假设，我们提出了三个具体的目标。目的 1. 鉴定经氟化物处理的成釉细胞衍生细胞中的 SIRT1 非组蛋白靶标体外。目标 2. 鉴定经氟化物处理的成釉细胞衍生细胞中的 SIRT1 组蛋白靶点体外。目的 3. 确定 SIRT1 过度表达或条件敲除是否影响牙釉质体内发育和氟斑牙。为了实现这些目标，我们将使用 SIRT1 过表达和敲除成釉细胞样细胞 LS8（LS8 Sirt1/over 和 LS8Sirt1/KO 细胞）通过CRISPR/Cas9技术在体外建立。我们将使用以下方法分析 SIRT1 的作用体内 SIRT1 过表达小鼠 (SIRT1super) 和条件敲除小鼠 (SIRT1cKO)。一旦表观遗传和非表观遗传 SIRT1 在牙釉质病理生理学中的功能被揭示，或许可以开发出新的策略来药理学地操纵 SIRT1 功能预防氟斑牙。