DEVELOPMENT OF A SALIVARY ASSAY FOR MEASURING PERIODONTAL DISEASE ACTIVITY

开发用于测量牙周疾病活动的唾液测定法

• 批准号: 10600906
• 负责人: Robert Gellibolian
• 金额: $ 32.06万
• 依托单位: CELLECTGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-03 至 2024-06-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10600906
• 关键词: Acetates; Adult; Affect; Age Years; Antibodies; Biological; Biological Assay; Biological Markers; Centers for Disease Control and Prevention (U.S.); Classification; Clinic; Clinical; Clinical Data; Collagen; Collection; Communities; Coupled; Detection; Development; Devices; Diagnostic; Disease; Disease Management; Disease Progression; Disease remission; Enzyme-Linked Immunosorbent Assay; Enzymes; Event; FDA approved; Future; Gelatinase B; Goals; Guidelines; Human; Hybridomas; Individual; Interstitial Collagenase; Lateral; Legal patent; Lesion; Matrix Metalloproteinases; Measurable; Measurement; Measures; Methods; Molecular Target; Monoclonal Antibodies; Neutrophil Collagenase; Outcome; Patient Care; Patients; Pattern; Periodontal Diseases; Periodontitis; Phase; Phenotype; Positioning Attribute; Prediction of Response to Therapy; Prevention strategy; Process; Protein Engineering; Proxy; Recording of previous events; Records; Research; Risk; Saliva; Salivary; Sampling; Sensitivity and Specificity; Signs and Symptoms; Site; Specificity; Standardization; Technology; Testing; Time; Tissues; Tooth Loss; Validation; Vitelliform macular dystrophy; assay development; chronic inflammatory disease; clinical diagnostics; clinical phenotype; collagenase; cost estimate; cross reactivity; detection limit; experience; high risk; improved; innovation; insight; interest; novel strategies; point of care; point-of-care diagnostics; prototype; radiological imaging; saliva sample; salivary assay; screening; smartphone application; stromelysin 2; success; technological innovation; technology platform

Project Summary / Abstract Periodontal disease (PD) is globally the most common chronic inflammatory disease in humans, which according to the Centers for Disease Control and Prevention (CDC) affect 47% of U.S. adults 30 years of age or older and >60% of those over 65 years.1 Given that the course of PD is marked by a discontinuous pattern of disease activity and inactivity showing exacerbation and remission of tissue destruction,2-4 a critical challenge for clinicians is not detection of clinical features of periodontal disease, but rather identification of patients who have an elevated risk for expressing active/progressing disease. Unfortunately, current guidelines and classifications rely exclusively on clinical and radiographic measurements to identify existing disease. While these records are a good reflection of a patient’s past history,5, 6 they fail to provide information on the current status of active disease or identify individuals and sites at risk for future disease.4, 5 This is because active disease involves tissue destructive processes (biologic phenotype)6, 8, 9 that precede and subsequently trigger the resulting clinical signs and symptoms (clinical phenotype). A host of salivary biomarkers for PD have been identified7, 10, 11 and multiple commercial assays are currently available for use in the clinic, but most lack specificity for tissue destruction and/or are not amenable to point-of-care (POC) real-time testing. Matrix metalloproteinase-8 is amongst the most widely documented, abundant biomarkers in saliva, and uniquely suited to quantitatively assess the extent and degree of tissue destruction.8, 9, 12-14 As an enzyme, it is present in both active and latent (inactive) forms, the relative expression of which dictate the extent of tissue breakdown during active periods of disease.8, 9 Hence, we hypothesize that the ratio of active and total MMP-8 (MMP-8Active/MMP-8Total) will constitute a more meaningful biological measure of disease activity than total MMP-8. However, measures of disease activity to date have not been possible due to the lack of commercially available antibodies for “active” MMP-8. Building off our team’s success in development of Disease Activity Specific Monoclonal Antibody (DASMAB) against MMP-8, we propose here to leverage these unique assets to develop a targeted molecular assay for quantifying disease activity. In this Phase I proposal, we plan to develop and validate an optimized DASMAB- based ELISA assay through the following aims: (1) identify and select the best mAb pairs (capture and detection) for each MMP-8 form to produce ELISA kit prototypes, and (2) analytical and clinical validation of disease activity of ELISA prototypes. Successful development and validation of the first DASMAB-based disease activity assay will precede a clinical POC diagnostic product that will offer clinicians with not only the means to quantify the extent and estimate the rate of disease progression in real-time, but also assess response to therapy and predict the most likely outcome of future events. These technological innovations will allow clinicians to make more effective outcome-driven decisions and personalize preventive strategies to improve patient care.

项目摘要/摘要 牙周病(PD)是全球最常见的慢性炎症性疾病。 美国疾病控制和预防中心(CDC)影响47%的30岁或30岁以上的美国成年人 65岁以上人群中的60%。1鉴于帕金森病的病程以不连续的疾病模式为标志 活动和不活动显示组织破坏的加剧和缓解，2-4是对 临床医生不是检测牙周病的临床特征，而是识别哪些患者有 表现为活动性/进展性疾病的风险增加。不幸的是，目前的指南和分类 完全依靠临床和放射测量来识别现有疾病。虽然这些记录是 很好地反映了病人过去的病史，5，6他们不能提供有关当前活动状态的信息 4、5这是因为活动性疾病涉及 组织破坏性过程(生物表型)6、8、9，在导致临床症状之前和随后触发 体征和症状(临床表型)。已经确定了一系列PD的唾液生物标志物7、10、11和 目前有多种商业检测方法可用于临床，但大多数缺乏对组织的特异性。 销毁和/或不服从护理点(POC)实时测试。基质金属蛋白酶-8是 唾液中记录最广泛、含量最丰富的生物标记物之一，并且唯一适合于定量 评估组织破坏的程度和程度。8、9、12-14作为一种酶，它存在于活性和潜伏期 (非活动性)形式，其相对表达决定了活跃期组织破坏的程度 因此，我们假设活性与总的基质金属蛋白酶-8的比率(基质金属蛋白酶-8活性/基质金属蛋白酶-8总量)将构成 这是一种比总的基质金属蛋白酶-8更有意义的疾病活动性生物学指标。然而，疾病的衡量标准 到目前为止，由于缺乏“活性”的基质金属蛋白酶-8的商业化抗体，活性还不可能实现。 在我们团队成功开发疾病活性特异性单抗(DASMAB)的基础上 针对基质金属蛋白酶-8，我们建议利用这些独特的资产来开发一种靶向的分子检测方法 量化疾病活跃度。在此第一阶段提案中，我们计划开发和验证优化的DASMAB- 基于酶联免疫吸附试验通过以下目的：(1)确定和选择最佳单抗对(捕获和检测) 对于每种基质金属蛋白酶-8形式，以产生ELISA试剂盒原型，以及(2)疾病活动性的分析和临床验证 所有的酶联免疫吸附试验原型。首个基于DASMAB的疾病活动性分析的成功开发和验证 将领先于临床POC诊断产品，该产品将不仅为临床医生提供量化 实时评估疾病进展的程度和速度，还评估对治疗的反应和预测 未来事件最有可能的结果。这些技术创新将使临床医生能够做出更多 有效的以结果为导向的决策和个性化的预防策略，以改善患者护理。