Highly Integrated Nucleic-Acid Analysis Using Graphene Bioelectronics

使用石墨烯生物电子学进行高度集成的核酸分析

• 批准号: 10372664
• 负责人: Jinglei Ping
• 金额: $ 20.86万
• 依托单位: UNIVERSITY OF MASSACHUSETTS AMHERST
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10372664
• 关键词: Affinity; Animal Model; Biological Assay; Biological Markers; Biopsy; Biopsy Specimen; Blinded; Buffers; Clinical; Clinical Trials; Complex; Consumption; Core Facility; Custom; DNA; DNA Probes; Development; Devices; Diagnosis; Diagnostic; Disease; Disease Progression; Electrodes; Generations; Genomics; Goals; Hospitals; Hour; Immobilization; Immunoassay; Intervention; Investigation; Measurable; Measures; MicroRNAs; Monitor; Mus; Noninfiltrating Intraductal Carcinoma; Nucleic Acids; Oligonucleotides; Operative Surgical Procedures; Outcome; Pathway interactions; Patients; Performance; Plasma; Population; Prognosis; Progressive Disease; Property; Public Health; Reaction; Recurrence; Research; Risk Assessment; Sampling; Signal Pathway; Signal Transduction; Technology; Testing; Therapeutic; Time; Transducers; Transistors; Translating; Translational Research; Validation; Visit; Xenograft Model; base; bioelectronics; breast cancer progression; circulating microRNA; clinical care; cohort; cost; detection limit; diagnostic technologies; disease diagnosis; early screening; graphene; human subject; improved; individualized medicine; liquid biopsy; malignant breast neoplasm; micro-total analysis system; mouse model; new therapeutic target; next generation; outcome prediction; overtreatment; pH gradient; point of care; point-of-care diagnosis; pre-clinical; prevent; prognostic; programs; response; screening; sensor; targeted treatment; treatment response; treatment strategy; user-friendly; voltage

PROJECT SUMMARY The circulating population of microRNAs in biofluids are ideal biomarkers for various diseases. Point-of-care profiling of circulating microRNAs is in insatiable demand, but typical approaches, e.g., immunoassays and microRNA assays are lab-based/centralized, expensive ($400–1,000/test), and time-consuming (>6 hours). We will develop a highly integrated, all-nanobioelectronic platform technology for multiplex, high-accuracy circulating-microRNA analysis that is capable of profiling circulating microRNAs in a 50-μL plasma with ultra- high sensitivity (sub-fM) and efficiencies in time (<40 minutes) and cost (<$10/test), thereby enabling high- performance circulating-microRNA analysis at the point of test. The novelty of the program is to harness graphene-based bioelectronics to integrate circulating microRNA isolation, concentration, amplification, and quantification into a self-contained device. In order to proof the concept of this technology, the program will include the development and validation of two generations of graphene-based analytical platforms, GAP1 and GAP2. Three specific aims with measurable milestones will be pursued. (1) We will demonstrate that multiple microRNA analytes can be amplified via hybridization chain reaction on a probe-functionalized graphene sensor array and the analyte concentrations can be readily interrogated by the graphene sensor array and translated into electrical signals. We will develop GAP1 to selectively quantify eight pre-selected target microRNAs (MDCIS8) spiked in 5-μL buffer. The detection limit of the specific microRNAs is expected to be at fM level. (2) We will demonstrate that target circulating microRNAs can be isolated from plasma by immobilizing them on a DNA-functionalized graphene electrode and releasing them into a small-volume simple cargo solution upon the generation of pH gradient by applying voltage bias between the graphene-DNA electrode with a bare graphene electrode. We will develop a graphene-based circulating-microRNA isolation module, combine the module with GAP1 to form GAP2, and use GAP2 to profile circulating MDCIS8 in lysed samples of 50-μL plasma from NSG mice. The GAP2 is expected to concentrate the microRNAs by >5× and deliver sub-fM level sensitivity. (3) We will demonstrate the feasibility of using this platform technology for diagnostic applications. We will use GAP2 to quantify circulating MDCIS8, whose expression levels are indicative to pre-invasive breast cancer, in 50-μL plasma samples from a user blinded cohort of the MIND murine model. The profiling result will be analyzed to predict the progression of pre-invasive breast cancer whose rapid, inexpensive diagnosis remains a challenge. The GAP2 prediction outcome will be combined with that based on surgical biopsy to establish the accuracy of the technology for progression prediction. The expected prediction accuracy is >96%. If successful, the technology will offer a new pathway to next-generation point-of-care genomic diagnostic/prognostic micro total analysis systems that would be cheap enough and user friendly enough to be used in various clinical settings.

项目总结