Daily Quantification of Cancer-Associated Exosomal miRNA in Patient Blood by Photonic Crystal-Enhanced Quantum Dot Emission

通过光子晶体增强量子点发射对患者血液中癌症相关外泌体 miRNA 进行每日定量

• 批准号: 10362538
• 负责人: Manish Kohli
• 金额: $ 30.83万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-03-19 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10362538
• 关键词: Acetates; Adoption; Affect; Aftercare; Award; Biological Assay; Biological Markers; Biometry; Blood; Blood specimen; Cancer Patient; Clinical; Clinical Data; Clinical Oncology; Clinical Trials; Closure by clamp; Collection; Coupling; Crystallization; Detection; Devices; Diagnosis; Disease; Drops; Effectiveness; Electronics; Engineering; Enrollment; Epidemiologist; Epidemiology; Equilibrium; Equipment; Genetic; Goals; Growth; Home; Human; Individual; Institutional Review Boards; Intervention; Knowledge; Label; Laboratories; Length; Light; Malignant Neoplasms; Measurement; Measures; Metastatic Prostate Cancer; MicroRNAs; Microscope; Molecular; Molecular Profiling; Monitor; Nucleic Acids; Oncologist; Optics; Outcome; Pathologist; Patient-Focused Outcomes; Patients; Peptide Nucleic Acids; Pharmacotherapy; Prednisone; Prognostic Marker; Prospective Studies; Quantum Dots; Regimen; Reproducibility; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Sampling; Screening for cancer; Series; Signal Transduction; Specificity; Specimen; Spottings; Surface; Technology; Temperature; Therapeutic; Time; Training; Translations; Treatment Efficacy; Treatment outcome; Variant; Work; abiraterone; advanced prostate cancer; assay development; base; biomarker discovery; cancer biomarkers; cancer survival; castration resistant prostate cancer; charge coupled device camera; chemotherapy; clinical application; clinical diagnostics; cost; density; design; detection limit; detection method; digital; docetaxel; drug efficacy; electric field; individual patient; insight; instrument; interest; molecular imaging; nanoprobe; new technology; novel; photonics; portability; predicting response; predictive marker; prospective; rapid technique; ratiometric; response; routine screening; single molecule; standard of care; success; translational barrier; tumor; tumor heterogeneity

ABSTRACT The primary goal of this proposal is to develop a new technology that can allow patients to repeatedly measure nucleic acid-based cancer biomarkers from a single drop of blood on a daily basis. This assay is being developed for specific clinical applications to determine drug treatment efficacies, prognosticate survival, and monitor post- treatment intervention by evaluating candidate nucleic acids shed from the tumor into blood of a cancer patient. We are focusing on detecting microRNA due to a very strong correlation with survival that our team has recently identified for patients with metastatic prostate cancer. We hypothesize that by measuring the concentrations of these markers in patients on a frequent basis during the course of therapy, we can precisely adjust therapeutic regimens for individual patients. However, accurately measuring microRNA in blood requires an extremely high limit of detection due to low concentrations, detection over a broad range of concentrations, and high sequence- specificity, attributes that are not currently possible for routine screening of fingerstick blood samples using standard methods of detection, such as PCR. Toward this end, our clinical needs have inspired a new form of assay to measure nucleic acids in blood through direct molecular counting in a microscope. This is now possible because we have developed novel ways to amplify the signals from individual molecules through a series of synergistic technologies, including light-emitting quantum dots, electric field-enhancing photonic crystals, and single-step sequence-specific enzymatic growth of microRNA. We now will combine these technologies to set the stage for measurement of microRNA using low-cost equipment that is already available in clinical diagnostic laboratories to minimize translational barriers to clinical adoption. To achieve these goals, our multi-investigator team has extensive expertise in probes for single-molecule imaging (Andrew Smith), optical detection in low- cost devices (Brian Cunningham), clinical oncology (Manish Kohli), biomarker discovery (Liang Wang), and epidemiology/biostatistics (Rebecca Smith). We will optimize our platform using a combination of synthetic and clinical blood specimens and thoroughly analyze the sequence selectivity of our assay, particularly focusing on microRNA variants, and closely compare our results with those from quantitative PCR assays. By the end of this award period, we expect to have developed the first direct-readout microRNA assay for use in human samples that is compatible with low-cost equipment, optimized the synergistic integration between quantum dots and photonic crystals, and measured, for the first time, the precise (digital) concentrations of microRNA in the blood of 100 subjects, prospectively enrolled and followed over 6 days each during the course of standard of care treatments for which no predictive or prognostic biomarkers currently exist in the treatment of metastatic prostate cancer. If successful, the outcome of this work will fill a major clinical gap in knowledge of how to match and finely tune treatments to individual molecular profiles.

抽象的 该提案的主要目标是开发一种新技术，可以让患者重复测量 每天从一滴血液中提取基于核酸的癌症生物标志物。该检测方法正在开发中 用于特定的临床应用，以确定药物治疗效果、预测生存率并监测术后情况 通过评估从肿瘤脱落到癌症患者血液中的候选核酸来进行治疗干预。 我们专注于检测 microRNA，因为我们的团队最近发现它与生存有很强的相关性 确定用于患有转移性前列腺癌的患者。我们假设通过测量 在治疗过程中，患者经常出现这些标志物，我们可以精确调整治疗方案 针对个别患者的治疗方案。然而，准确测量血液中的 microRNA 需要极高的 由于浓度低、检测浓度范围广和高序列而导致检测限 特异性、属性目前无法用于常规筛查指尖采血样本 标准检测方法，例如PCR。为此，我们的临床需求激发了一种新形式的 通过在显微镜中直接进行分子计数来测量血液中核酸的测定。现在这是可能的 因为我们已经开发出新的方法来通过一系列的方法来放大单个分子的信号 协同技术，包括发光量子点、电场增强光子晶体和 microRNA 的单步序列特异性酶促生长。我们现在将结合这些技术来设置 使用临床诊断中已有的低成本设备测量 microRNA 的阶段 实验室，以尽量减少临床采用的转化障碍。为了实现这些目标，我们的多位研究者 团队在单分子成像探针（Andrew Smith）、低浓度光学检测方面拥有丰富的专业知识 成本设备（Brian Cunningham）、临床肿瘤学（Manish Kohli）、生物标志物发现（Liang Wang）和 流行病学/生物统计学（丽贝卡·史密斯）。我们将结合合成和优化我们的平台 临床血液样本并彻底分析我们测定的序列选择性，特别关注 microRNA 变体，并将我们的结果与定量 PCR 检测的结果进行密切比较。到此结束时 颁奖期间，我们预计将开发出第一个用于人类样本的直接读出 microRNA 检测方法 兼容低成本设备，优化量子点与量子点之间的协同集成 光子晶体，并首次测量了血液中 microRNA 的精确（数字）浓度 前瞻性招募 100 名受试者，并在标准护理过程中对每名受试者进行超过 6 天的随访 目前在转移性前列腺治疗中尚无预测或预后生物标志物的治疗方法 癌症。如果成功，这项工作的成果将填补如何匹配和 根据个体分子特征微调治疗。

项目成果

Label-Free Digital Detection of Intact Virions by Enhanced Scattering Microscopy.

• DOI: 10.1021/jacs.1c09579
• 发表时间: 2022-02-02
• 期刊: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
• 影响因子: 15
• 作者: Li, Nantao;Wang, Xiaojing;Tibbs, Joseph;Che, Congnyu;Peinetti, Ana Sol;Zhao, Bin;Liu, Leyang;Barya, Priyash;Cooper, Laura;Rong, Lijun;Wang, Xing;Lu, Yi;Cunningham, Brian T.
• 通讯作者: Cunningham, Brian T.

Expanding the Dynamic Range of Fluorescence Assays through Single-Molecule Counting and Intensity Calibration.

• DOI: 10.1021/jacs.8b08879
• 发表时间: 2018-10-24
• 期刊: Journal of the American Chemical Society
• 影响因子: 15
• 作者: Smith L;Kohli M;Smith AM
• 通讯作者: Smith AM

Photonic crystal enhanced fluorescence emission and blinking suppression for single quantum dot digital resolution biosensing.

• DOI: 10.1038/s41467-022-32387-w
• 发表时间: 2022-08-08
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Xiong, Yanyu;Huang, Qinglan;Canady, Taylor D.;Barya, Priyash;Liu, Shengyan;Arogundade, Opeyemi H.;Race, Caitlin M.;Che, Congnyu;Wang, Xiaojing;Zhou, Lifeng;Wang, Xing;Kohli, Manish;Smith, Andrew M.;Cunningham, Brian T.
• 通讯作者: Cunningham, Brian T.

Digital and Absolute Assays for Low Abundance Molecular Biomarkers.

低丰度分子生物标志物的数字和绝对测定。

• DOI: 10.1021/acs.accounts.3c00030
• 发表时间: 2023
• 期刊: Accounts of chemical research
• 影响因子: 18.3
• 作者: Kuo,Chia-Wei;Smith,AndrewM
• 通讯作者: Smith,AndrewM