Potentials of Epigenetic Molecules in Attenuating the Phenotypes of Periodontitis

牙周病是成人中最常见的疾病之一，47%的人 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， 以三个主要病理特征为特征：加剧炎症反应，过度 破骨细胞性牙槽骨吸收，成骨细胞性骨形成减少。不幸的是，在那里 目前在诊所里只有姑息治疗。我们的实验室探索了各种策略来改善 疾病的严重程度包括骨髓基质细胞(BMSCs)的应用和诱导多能性 干细胞(IPSCs)与主要转录因子，如Runx2，OSX和SATB2，在基于细胞的基因治疗中， 但与此同时，也遇到了这些方法的缺陷和局限性。因此，我们正在积极 寻找一种阻断主要致病因素并刺激内生因素治疗的新疗法 再生因牙周炎而丢失的组织。表观遗传分子最近已成为一种强有力的 基因表达和多种人类疾病的治疗剂。我们第一次发现 新型合成BET蛋白抑制剂JQ1特异性抑制牙周炎症并显著 减少小鼠牙周炎模型的牙槽骨丢失。此外，我们还率先鉴定出miR-335-5P 并表征了其在促进骨形成和再生方面的作用。过表达miR-335-5p 改善实验性牙周炎的组织损伤，进一步提供了基础和强大的 这是当前提案的前提。在这份提案中，我们将使用一种精心设计和精确合成的 将携带JQ1和miR-335-5P并专门将它们释放到主要靶点的纳米颗粒(NP)系统 细胞在牙周炎中的作用，导致内源性再生因子的发病和激活受阻。 在目标1中，我们将在基因水平上阐明mir-335-5p在牙周炎中作用的分子机制。 利用我们新产生的基因操纵的啮齿动物，缺乏和过度表达条件。在AIM 2，我们将定义纳米颗粒介导的JQ1的精确度和特异性递送的表观遗传效应 和miR-335-5p在其相应的靶细胞上。在目标3中，我们将确定治疗效果 表观调节剂在小鼠牙周炎模型中的联合应用。我们会把EPI送到- 生物纳米产品同时、单独或交替地评估不同疾病的治疗效果 疾病分期。结果测量将包括无偏见的单细胞RNA测序，16S rDNA图谱， 牙周定性和定量分析及药代动力学。使用创新的表观遗传学方法 理解牙周病的病理生理学及其治疗潜力是一种范式的转变。 这项调查将由一个由PI和三名C-IS组成的跨学科小组进行，他们 在以下领域有深厚的知识和互补的技能：实验病理学和骨 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.