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 牙周疾病（PD）是人类最常见的慢性炎症性疾病，这是 疾病控制与预防中心（CDC）影响47％的美国成年人30岁或以上， > 65年以上的人中有60％。1鉴于PD的过程以不连续的疾病模式标记 活动和不活动表明组织破坏加剧和缓解，2-4是一个至关重要的挑战 临床医生不是发现牙周疾病的临床特征，而是鉴定患者 表达活跃/进步疾病的风险升高。不幸的是，当前的指南和分类 仅依靠临床和射线照相测量来鉴定现有疾病。这些记录是 很好地反映了患者过去的历史，5，6他们无法提供有关活动当前状态的信息 疾病或识别患有未来疾病风险的个人和部位。4，5这是因为主动疾病涉及 组织破坏性过程（生物表型）6、8、9之前并随后触发所得的临床 体征和症状（临床表型）。已经确定了许多PD的唾液生物标志物7、10、11和 目前可在诊所使用多种商业测定，但大多数对组织缺乏特殊性 破坏和/或不适合实时测试（POC）实时测试。基质金属蛋白酶8是 在唾液中最广泛，最丰富的生物标志物中，非常适合定量 评估组织破坏的程度和程度。8、9、12-14作为酶，它都存在于活性和潜在中 （非活动）形式，其相对表达决定了在活动期间组织分解程度 疾病8，9因此，我们假设活性和总MMP-8（MMP-8活动/MMP-8TOTAL）的比率将构成 与总MMP-8相比，对疾病活性的生物学测量更有意义。但是，疾病的措施 迄今为止，由于缺乏“活动” MMP-8的市售抗体，因此无法进行活动。 建立我们团队在疾病活动特定单克隆抗体（DASMAB）方面的成功发展 针对MMP-8，我们在这里提议利用这些独特的资产来开发针对性的分子测定 量化疾病活动。在此阶段I建议中，我们计划开发和验证优化的dasmab- 通过以下目的进行基于ELISA测定：（1）识别并选择最佳的mAb对（捕获和检测） 对于每种MMP-8形式，生产ELISA试剂盒原型，以及（2）疾病活性的分析和临床验证 Elisa原型。成功开发和验证第一个基于达斯马布的疾病活动测定法 将在临床POC诊断产品之前，该产品将为临床医生提供量化的手段 程度和估计实时疾病进展率，但也评估对治疗的反应并预测 未来事件的最可能结果。这些技术创新将使临床医生能够做更多 有效以结果为导向的决策并个性化改善患者护理的预防策略。