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).

项目概要/摘要： 自发性早产（PTL，<37孕周）是新生儿和婴儿的主要原因， 全球婴儿死亡率约占所有婴儿死亡的60%。PTL的病理生理学仍然很差 由于与风险因素相关的异质性，包括母亲年龄，多重 妊娠和合并症等。确定PTL高风险女性的现行临床标准 包括宫颈长度评估和宫颈阴道液中胎儿纤连蛋白的筛查。但是，在这方面， 这些方法是不充分的，往往不能识别可能早产的妇女。新近患者 元数据研究已经确定了与PTL相关的多种生物标志物，但基于单个 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。的 拟议研究的科学前提是，临床迫切需要准确的诊断， 多重生物标志物检测功能，同时负担得起，以促进常规床边 筛查所有患者。此外，通过将生物诊断与商业测定进行基准测试， 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变异，这仍然是 当前床旁诊断的挑战。 本研究的目的是通过一种创新的生物诊断产品PRADA， 其结合了高灵敏度、特异性、多路复用、使用低样品体积和低成本。普拉达， 便携式可重复使用的精确诊断与纳米天线，包括磁性微珠捕获探针 用多克隆抗体功能化，和近红外共振金纳米星"天线"条形码与 拉曼标签和肽标记，以通过表面增强拉曼光谱检测生物标志物 （SERS）。PRADA已成功证明可多重检测3种PTL生物标志物，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素 妊娠患者血清中的GM-CSF因子。PRADA达到了超灵敏的检测限（LOD，1.7 pg/ml GM-CSF）是风险分层的理想选择，与市售Luminex和ELISA相比，PRADA显示 更低的LODs。此外，PRADA还可重复使用（由于使用磁珠），降低了整体成本 通过允许单个微流体装置使用> 15次。这项工作将利用创新的普拉达， 定量检测包括C-反应蛋白在内的6种最能预测PTL的血清生物标志物 (CRP)、AFP、IL-10、IL-6、CRH和GM-CSF以Luminex和ELISA为基准（Aim 1）。我们还 将在本研究招募的18 - 35岁患者的血清中验证与PRADA的多重检测 单胎妊娠组，孕龄19~36周（Aim 2）。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 现有的诊断来确定PRADA的准确性。我们的核心假设是， 可靠的床旁诊断，可准确预测PTL的发作，识别需要立即治疗的患者， 治疗，并尽量减少对低风险患者的不必要干预。 PRADA在临床上是可转化的，因为金纳米颗粒已经在临床试验中（NCT 00436410， NCT 00356980）并整合到商业测定（第一反应妊娠试验）中，并且SERS具有 在商业传感器（Oxonica、Renishaw Diagnostics等）中证明了实用性。这个有影响力的项目将 利用PI（Bardhan）在纳米颗粒诊断和SERS方面的专业知识， 共同研究者Leon Bellan博士（微流体学）、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