PRADA: Portable Reusable Accurate Diagnostics with nanoAntennas for Multiplexed Biomarker Screening of Preterm Labor

PRADA：便携式可重复使用的纳米天线精确诊断，用于早产的多重生物标志物筛查

• 批准号: 10229351
• 负责人: Rizia Bardhan
• 金额: $ 22万
• 依托单位: IOWA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10229351
• 关键词: 37 weeks gestation; Address; Age; Age-Years; Bar Codes; Benchmarking; Biological Assay; Biological Markers; C-reactive protein; Cervical; Cessation of life; Childhood; Clinical; Clinical Trials; Cohort Studies; Complex; Corticotropin-Releasing Hormone; Detection; Diagnosis; Diagnostic; Enzyme-Linked Immunosorbent Assay; Functional disorder; Gestational Age; Gold; Granulocyte-Macrophage Colony-Stimulating Factor; Heterogeneity; High Risk Woman; Infant Mortality; Interleukin-10; Interleukin-6; Intervention; Label; Length; Linear Regressions; Liquid substance; Magnetism; Maternal Age; Measurement; Metadata; Microfluidic Microchips; Microfluidics; Microspheres; Modeling; Neonatal Mortality; Patient Recruitments; Patients; Peptides; Phenotype; Pilot Projects; Predictive Value; Pregnancy; Pregnancy Tests; Pregnant Women; Premature Birth; Premature Labor; Raman Spectrum Analysis; Recording of previous events; Reproducibility; Research Personnel; Risk; Risk Factors; Sampling; Sensitivity and Specificity; Serum; Specificity; Surface; Vagina; Validation; Variant; Woman; Work; accurate diagnosis; accurate diagnostics; age group; aggressive therapy; alpha-Fetoproteins; base; cervicovaginal; clinical Diagnosis; clinical practice; clinically relevant; clinically translatable; comorbidity; cost; detection limit; fetal fibronectin; high risk population; improved; infant death; innovation; magnetic beads; maternal risk; multiplex assay; multiplex detection; nanoGold; nanoparticle; neonatal morbidity; polyclonal antibody; portability; predictive marker; pregnant; recruit; response; risk prediction; risk stratification; screening; sensor

Project Summary/Abstract: Spontaneous preterm labor (PTL, <37 weeks gestation) is the leading cause of neonatal and infant mortality globally, representing ~60% of all infant deaths. The physiopathology of PTL remains poorly understood due to the heterogeneities associated with the risk factors including maternal age, multiple gestation, and co-morbidities among others. Current clinical standard to identify women at high risk of PTL include cervical length assessment, and screening for fetal fibronectin in the cervicovaginal fluid. However, these approaches are inadequate and often fail to identify women who may deliver preterm. Recent patient metadata studies have identified multiple biomarkers correlated to PTL but risk scoring based on single biomarkers has shown low positive predictive values and have been ineffective in PTL prediction. The scientific premise of the proposed study is that there is an urgent clinical need for accurate diagnostics with multiplexed biomarker detection capabilities that are simultaneously affordable to facilitate routine bedside screening for all patients. Further by benchmarking the biodiagnostic against commercial assays using the same sample type from the same patients will minimize inter- and intra-assay variation, which remains a challenge in current bedside diagnostics. The objective of this study is to address this unmet need with an innovative biodiagnostic, PRADA, which combines high sensitivity, specificity, multiplexing, use of low sample volumes, and low cost. PRADA, portable reusable accurate diagnostics with nanoantennas, consists of magnetic microbeads capture probes functionalized with polyclonal antibodies, and near-infrared resonant gold nanostar “antennas” barcoded with Raman tags and labeled with peptides to detect the biomarkers via surface-enhanced Raman spectroscopy (SERS). PRADA has successfully demonstrated multiplexed detection of 3 PTL biomarkers including alpha- fetoprotein (AFP), corticotrophin releasing hormone (CRH) and granulocyte macrophage colony-stimulating factor (GM-CSF) in pregnant patient serum. PRADA achieved ultrasensitive limit of detection (LOD, 1.7 pg/ml GM-CSF) ideal for risk stratification, and when compared to commercial Luminex and ELISA, PRADA showed far lower LODs. Further, PRADA is also reusable (due to the use of magnetic beads), lowering the overall cost by allowing >15 uses of a single microfluidic device. This work will leverage innovative PRADA to quantitatively detect 6 serum biomarkers which are the most predictive of PTL including C-reactive protein (CRP), AFP, IL-10, IL-6, CRH, and GM-CSF benchmarked against Luminex and ELISA (Aim 1). Further, we will validate multiplexing with PRADA in serum of patients recruited for this study in the 18 – 35 years age group with singleton pregnancy and gestation age 19 – 36 weeks (Aim 2). A maternal risk score will be established categorizing patients into high and low risk groups, and the scoring will be compared to their existing diagnosis to determine the accuracy of PRADA. Our central hypothesis is PRADA will enable a robust bedside diagnosis that accurately predicts the onset of PTL, identifies patients who need immediate treatment, and minimizes unnecessary intervention for those at low risk. PRADA is clinically translatable as gold nanoparticles are already in clinical trials (NCT00436410, NCT00356980) and integrated in commercial assays (First response pregnancy tests), and SERS has demonstrated utility in commercial sensors (Oxonica, Renishaw Diagnostics etc.). This impactful project will leverage the expertise of the PI (Bardhan) in nanoparticle diagnostics and SERS, with the complementary expertise of co-investigators Dr. Leon Bellan (microfluidics), Dr. Jeff Reese (mechanisms of PTL), and Dr. J. Michael Newton (PTL patient treatment).

项目摘要/摘要： 自发性早产（PTL，妊娠<37周）是新生儿和婴儿流产的主要原因 全球婴儿死亡人数约占所有婴儿死亡人数的 60%。 PTL 的病理生理学仍然很差 由于与危险因素相关的异质性，包括母亲年龄、多重 妊娠和合并症等。当前识别 PTL 高风险女性的临床标准 包括宫颈长度评估和宫颈阴道液中胎儿纤连蛋白的筛查。然而， 这些方法是不够的，而且往往无法识别可能早产的妇女。最近的病人 元数据研究已经确定了与 PTL 相关的多个生物标志物，但风险评分基于单个生物标志物 生物标志物显示出较低的阳性预测值并且在 PTL 预测中无效。这 该研究的科学前提是临床迫切需要准确的诊断 多重生物标志物检测功能，同时经济实惠，可促进常规床旁治疗 对所有患者进行筛查。进一步通过使用商业化验对生物诊断进行基准测试 来自相同患者的相同样本类型将最大限度地减少检测间和检测内的变异，这仍然是一个 当前床边诊断面临的挑战。 这项研究的目的是通过创新的生物诊断 PRADA 来解决这一未满足的需求， 它结合了高灵敏度、特异性、多重性、低样本量和低成本。普拉达， 便携式可重复使用的纳米天线精确诊断，由磁性微珠捕获探针组成 用多克隆抗体进行功能化，以及带有条形码的近红外共振金纳米星“天线” 拉曼标签并用肽标记，通过表面增强拉曼光谱检测生物标志物 （表面增强拉曼光谱）。 PRADA 已成功演示了 3 种 PTL 生物标志物的多重检测，包括 α- 甲胎蛋白 (AFP)、促肾上腺皮质激素释放激素 (CRH) 和粒细胞巨噬细胞集落刺激 妊娠患者血清中的因子（GM-CSF）。 PRADA 达到超灵敏检测限（LOD，1.7 pg/ml） GM-CSF）是风险分层的理想选择，与商业 Luminex 和 ELISA 相比，PRADA 表明 LOD 低得多。此外，PRADA还可以重复使用（由于使用了磁珠），降低了总体成本 允许使用单个微流体装置超过 15 次。这项工作将利用创新的 PRADA 定量检测最能预测 PTL 的 6 种血清生物标志物，包括 C 反应蛋白 (CRP)、AFP、IL-10、IL-6、CRH 和 GM-CSF 以 Luminex 和 ELISA 为基准（目标 1）。此外，我们 将在本研究招募的 18 – 35 岁患者血清中验证 PRADA 多重分析 单胎妊娠且胎龄 19 – 36 周的组（目标 2）。孕产妇风险评分将是 将患者分为高风险组和低风险组，并将评分与他们的评分进行比较 确定PRADA现有诊断的准确性。我们的中心假设是 PRADA 将使 强大的床边诊断可准确预测 PTL 的发作，识别需要立即治疗的患者 治疗，并最大限度地减少对低风险人群的不必要干预。 PRADA 具有临床可转化性，因为金纳米颗粒已进入临床试验（NCT00436410， NCT00356980）并集成到商业化验（第一反应妊娠测试）中，并且 SERS 已 已在商业传感器（Oxonica、Renishaw Diagnostics 等）中展示了实用性。这个影响深远的项目将 利用 PI (Bardhan) 在纳米颗粒诊断和 SERS 方面的专业知识，以及补充 共同研究者 Leon Bellan 博士（微流体）、Jeff Reese 博士（PTL 机制）和 J. Michael Newton（PTL 患者治疗）。

项目成果

First Trimester Prediction of Preterm Birth in Patient Plasma with Machine-Learning-Guided Raman Spectroscopy and Metabolomics.

• DOI: 10.1021/acsami.3c04260
• 发表时间: 2023-08
• 期刊: ACS applied materials & interfaces
• 影响因子: 9.5
• 作者: Lilly Synan;S. Ghazvini;Saji Uthaman;Gabriel Cutshaw;Che-Yu Lee;Joshua R. Waite;Xiaona Wen;S. Sarkar;Eugene C Lin;M. Santillan;D. Santillan;Rizia Bardhan