Potentials of Epigenetic Molecules in Attenuating the Phenotypes of Periodontitis

牙周病是成年人中最常见的疾病之一， 30岁以上的美国人患有这种疾病。牙周炎是一种复杂的免疫和炎症性疾病， 其特征在于三个主要病理特征：炎症反应加剧，过度 骨细胞牙槽骨吸收和成骨细胞骨形成减少。可惜 目前在临床上只是治标不治本。我们的实验室已经探索了各种策略来改善 包括应用骨髓基质细胞（BMSCs）和诱导多能干细胞（MSCs） 干细胞（iPSC）与主要转录因子，如Runx 2，Osx和SATB 2在基于细胞的基因治疗， 但同时也经历了这些方法的缺点和限制。因此，我们积极 寻找一种新的治疗方法，阻断主要的致病因素，刺激内源性因素， 再生牙周炎中失去的组织。表观遗传分子最近已经成为一种有效的调节因子， 用于多种人类疾病的基因表达和治疗剂。我们第一次发现 新型合成BET蛋白抑制剂JQ 1特异性抑制牙周炎症， 减少小鼠牙周炎模型中的牙槽骨丢失。此外，我们是第一个发现miR-335- 5 p的人。 并表征其在促进骨形成和再生中的作用。过表达miR-335- 5 p 改善实验性牙周炎的组织损伤，进一步提供了基础和强有力的 当前提案的前提。在这个建议中，我们将使用一个优雅的设计和精确的合成， 一种纳米颗粒（NP）系统，其将携带JQ 1和miR-335- 5 p，并将它们特异性地释放到主要靶点中， 牙周炎中的细胞，导致发病机制和内源性再生因子的激活受阻。 在目的1中，我们将描述miR-335- 5 p在牙周炎中作用的分子机制， 缺乏和过度表达的条件下使用我们新产生的基因操纵的啮齿动物。在Aim中 2，我们将定义纳米颗粒介导的JQ 1的精确性和特异性递送的表观遗传效应 和miR-335- 5 p在其相应的靶细胞上的作用。在目标3中，我们将确定 在小鼠牙周炎模型中联合应用表调剂。我们会把肾上腺素- 生物纳米产品，同时，单独或交替评估在不同的治疗效果， 疾病阶段。结果测量将包括无偏倚的单细胞RNA测序，16 S rDNA分析， 牙周定性和定量分析以及药代动力学。使用创新的表观遗传方法 了解牙周病的病理生理学及其治疗潜力是一个范式的转变。 本研究将由PI和三名C-I组成的跨学科团队进行， 在以下领域拥有深厚的知识和互补技能：实验病理学和骨骼 生物学;细胞和分子生物学;炎症和免疫学，生物材料和药物输送。

Periodontal disease represents one of the most prevalent diseases in adult population as 47% of Americans over age 30 have the disease. Periodontitis is a complex immune and inflammatory disease, characterized by three major pathological features: exacerbating inflammatory response, excess osteoclastic alveolar bone resorption and decreased osteoblastic bone formation. Unfortunately, there are only palliative treatments in clinics currently. Our laboratory has explored a variety of strategies to ameliorate the severity of the disease including application of bone marrow stromal cells (BMSCs) and induced pluripotent stem cells (iPSCs) with major transcription factors, such as Runx2, Osx and SATB2 in cell-based gene-therapy, but at the meantime experienced drawbacks and limitations of these approaches. Therefore, we are actively searching for a new therapy that blocks the major pathogenetic elements and stimulates endogenous factors to regenerate the tissues lost in periodontitis. Epigenetic molecules have recently emerged as potent regulators of gene expression and therapeutic agents for a variety of human diseases. We for the first time found that the novel synthetic BET protein inhibitor, JQ1, specifically inhibits periodontal inflammation and significantly reduces alveolar bone loss in murine periodontitis model. Additionally, we are the first to identify miR-335-5p and characterized its role in promoting bone formation and regeneration. Overexpressing miR-335-5p ameliorates tissue damage in experimental periodontitis which further provides a foundation and a strong premise for the current proposal. In this proposal we will use an elegantly designed and precisely synthesized nanoparticle (NP) system that will carry both JQ1 and miR-335-5p and specifically release them into major target cells in periodontitis, resulting in a block of the pathogenesis and activation of endogenous regenerative factors. In Aim 1, we will delineate the molecular mechanisms of miR-335-5p effects in periodontitis under gene deficient and overexpressing conditions using our newly generated genetically manipulated rodents. In Aim 2, we will define the epigenetic effects of nanoparticle-mediated precision and specific delivery of JQ1 and miR-335-5p on their corresponding target cells. In Aim 3, we will determine the therapeutic efficacy of combined application of epi-modulators in a mouse model of periodontitis. We will deliver the epi- biological nano-products simultaneously, individually, or alternatingly to assess the therapeutic impact at different disease stages. Outcome measures will include unbiased single cell RNA-sequencing, 16S rDNA profiling, qualitative and quantitative periodontal analyses and pharmacokinetics. Using innovative epigenetic approaches to understand the pathophysiology of periodontal disease and its therapeutic potentials is a paradigm shift. This investigation will be conducted by an interdisciplinary team composed of the PI and three C-Is who have profound knowledge and complementary skills in the following areas: Experimental Pathology and Bone Biology; Cell and Molecular Biology; Inflammation and Immunology, and Biomaterials and Drug Delivery.