上皮炎性衰老通过NLRC4/TLR5调控伴糖尿病牙周炎发生发展的研究

Cell inflammaging brings about a persistent proinflammatory state in human body. The changes in local microenvironment are what importantly account for diabetic mellitus and its complications. Recently, NLRC4 (NLR family CARD domain-containing protein 4) is reported to be a key protein in activating inflammaging process. Our previous study discovered that SASP (senescence-associated secretory phenotype) was elevated to affect diabetic periodontitis under stimulation of inflammaging environment. Meanwhile, NLRC4 could possibly interact with TLR5(Toll like receptor 5) in a LRR (Leucine-rich repeat) dependent manner according to our data. However, it still remains elusive whether the modulation of SASPs by NLRC4/TLR5 pathway is of biological prevalence and how the regulation in gingival epithelia works. In this study, experimental models both in vivo and in vitro are planned to be adopted, with CRISPR/Cas9 system, high-flux biological technology RNA sequencing, ChIP technique, and adeno-associated virus transfection utilizing as main methods. We aim to investigate the biological functions of NLRC4/TLR4 pathway on gingival epithelial inflammaging during the onset and development of diabetic periodontitis. An appropriate explanation will also be given to define the possible mechanisms of SASP and its potent role in gingival epithelial inflammaging. The implementation of this proposal will not only reveal the common pattern of inflammaging in periodontitis with diabetes, but also provide new strategies for the prevention and treatment of the disease.

细胞炎性衰老导致机体局部促炎微环境变化，是糖尿病及其并发症受关注的新机制。NOD样受体家族成员NLRC4是激活炎性衰老的关键节点。申请人前期发现衰老相关微环境通过分泌表型SASP影响伴糖尿病牙周炎，并提示NLRC4通过LRR结构域与TLR5分子作用调控上皮细胞炎性衰老。但在伴糖尿病牙周炎中，NLRC4/TLR5介导的SASP改变是否是一种普遍生物学现象，其如何调控牙龈上皮细胞炎性衰老都有待进一步阐明。本课题拟采用体内外模型，CRISPR/Cas9、ChIP、RNAseq、AAV病毒转染等技术，进一步研究上皮炎性衰老通过NLRC4/TLR5信号通路在伴糖尿病牙周炎发生发展时空进程中的生物学功能，及衰老相关分泌表型SASP变化调控上皮炎性衰老的潜在分子机制，明确NLRC4作为治疗靶标的潜在价值。本项目的实施不仅将揭示炎性衰老在伴糖尿病牙周炎发生发展中的共性规律，也为该疾病的防治提供新思路。

伴糖尿病牙周炎是牙周疾病研究中的重要领域也是难点之一。本项目聚焦伴糖尿病牙周炎发生发展过程中的生物特性、病理机理、分子机制和临床转化四个维度，首次揭示了炎性衰老在伴糖尿病牙周炎发生发展中的生物学特征和共性规律，解析了NLRC4/TLR5 信号通路在上皮炎性衰老调节糖尿病牙龈上皮防御的分子机制，为该疾病的防治提供新思路和新的分子靶标。课题组从组织层面发现了牙龈组织炎性衰老与牙周炎症程度成正相关，是造成伴糖尿病牙周炎症程度增加的原因；从分子层面对NLRC4炎性小体在高糖诱导上皮细胞炎症相关衰老过程中的机制作用深入研究，揭示高血糖激活NLRC4介导巨噬细胞炎性衰老，发现上皮衰老细胞SASP分泌增多可维持并加重局部炎性衰老；临床研究中，课题分析了不同类型糖尿病患者口腔基本特征及牙周破坏程度，并筛选糖尿病相关衰老分泌表型SASP因子，构建基于SASP网络的糖尿病口腔疾病精准评估系统，发现高糖相关SASP网络作为判断糖尿病患者牙周炎体液早期改变的依据，为探索早期检测并管理伴糖尿病口腔炎性疾病提供新途径。此外，本课题针对糖尿病患者的炎症相关衰老特征，发明了基于SASP网络的口腔炎症性疾病预测模型，应对糖尿病牙周炎高危人群及时采取恰当的措施医疗措施；发明了一种用于糖尿病牙周炎可防可控，快速恢复牙周缺损的血糖感应三维响应种植材料体系，有良好的推广应用的前景。研究成果发表论文27篇，其中SCI文章18篇 (包括1篇Nature Index 论文，最高影响因子24分)，中文核心文章5篇，会议论文4篇，国内特邀报告1次，学术交流报告2篇，参与编写专著2本，获得国家发明专利1项，申请国家发明专利1项。