Smartphone-based mobile detection platform for lung cancer detection in China

基于智能手机的中国肺癌移动检测平台

• 批准号: 10018939
• 负责人: Jinzhao Song
• 金额: $ 10.56万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-16 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10018939
• 关键词: Acute; Address; Advisory Committees; Air Pollution; Alleles; Area; Biological Assay; Biopsy; Blood; Blood specimen; Body Fluids; Cancer Detection; Cancer Diagnostics; Cancer Etiology; Cancer Patient; Cellular Phone; Cessation of life; China; Cleaved cell; Closure by clamp; Collaborations; Companions; Custom; DNA; Data; Detection; Devices; Diagnosis; Diagnostic; Drug Targeting; Drug resistance; Engineering; Enzymes; Epidemiologist; Epidermal Growth Factor Receptor; Evolution; Family; Funding Mechanisms; Future; Genetic; Genotype; Goals; Health Personnel; International; KRAS2 gene; Life; Liquid substance; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Medical; Medical Device Designs; Mentorship; Methods; Microbiology; Microfluidics; Monitor; Mutation; Oncologist; Patients; Peptide Nucleic Acids; Pharmaceutical Preparations; Polymerase; Process; Program Sustainability; Public Health; Records; Reporter; Reporting; Research; Research Personnel; Resources; Rural; Saliva; Sampling; Science; Signal Transduction; System; Test Result; Thermus thermophilus; Time; Tobacco use; Training; Treatment outcome; Urine; Whole Blood; Work; base; cancer cell; cancer diagnosis; cancer epidemiology; cancer genetics; cancer therapy; cancer type; career; cost; cost effective; design; effective therapy; epidemiology study; experience; global health; improved; individualized medicine; insertion/deletion mutation; instrument; interest; liquid biopsy; lung cancer screening; member; mobile computing; molecular diagnostics; multidisciplinary; multiplex detection; mutant; novel; point of care; point-of-care diagnostics; precision oncology; programs; skills; smartphone Application; spatial epidemiology; spatiotemporal; success; tool

ABSTRACT Lung cancer is the leading incident cancer and cause of cancer death in China. In the next five years, attributed to air pollution and high tobacco consumption, China will likely see an increase in the occurrence of lung cancer with an estimated of over 800,000 new diagnoses and 700,000 deaths per year. Although affordable cancer genotype-specific drugs are increasingly available in China, effective and affordable diagnostics of cancer genotypes is lacking. To address this critical need, I propose a novel inexpensive, point-of-care smartphone-based system for detecting rare mutant alleles in body fluids to enable liquid biopsy of lung cancer in rural China. The proposed system builds on my prior work with minimally instrumented and un-instrumented molecular diagnostics. My system will accept a raw sample, such as whole blood, process the sample, and provide rapid test results. Since the presence of a large abundance of wildtype (WT)-allele challenges detection of rare mutant alleles, my system will include a sample enrichment step that utilizes DNA guided cleaving enzymes of the Argonaute family to digest and deplete WT alleles while sparing the mutant alleles of interest. This will be followed with a loop mediated isothermal amplification (LAMP) that utilizes peptide nucleic acid (PNA) clamp to selectively amplify mutant alleles, but not WT-alleles. For multiplex detection of mutant alleles, the proposed system will firstly enrich the sample with the programmable cleaving enzyme and then subject the enriched sample to a novel two stage, multiplexed isothermal amplification process (Penn-RAMP), in the case of point/deletion/insertion mutations, with PNA clamps to discourage amplification of WT alleles in Penn-RAMP's second stage. Amplicons will be detected with bioluminescent reporters and a smartphone camera. A custom smartphone application will analyze the recorded signal; report test results; and, in the future, transmit these results to the patient's doctor and records and, in de-identified form, to the cloud for spatiotemporal surveillance, allowing public health officials identify hotspots. Our preliminary data indicates that our approach has high likelihood of success. I have developed a training plan in the following three areas: cancer epidemiology, cancer diagnostics and therapy, and medical device design and fabrication. In each of these areas, I have identified coursework and mentorship support from members of the K01 Advisory Committee. The Advisory Committee composed of internationally recognized experts in engineering, cancer epidemiology, lung cancer precision medicine, liquid biopsy, and microbiology has been assembled and will meet periodically to assess my progress and provide guidance. This research has the potential to greatly improve companion diagnosis and screening of lung cancer in China and enable effective therapies. The program will enable me to obtain new skills through field work, research, coursework, and collaborations, all of which will promote and accelerate my transition to an independent investigator for global health.

摘要 肺癌是我国第一大恶性肿瘤，也是我国癌症死亡的主要原因.在接下来的五年里，由于空气 污染和高烟草消费，中国可能会看到肺癌的发病率增加， 估计每年有超过80万例新诊断和70万例死亡。虽然负担得起的癌症基因型特异性 尽管中国越来越多的药物可用，但缺乏有效且负担得起的癌症基因型诊断。 为了满足这一关键需求，我提出了一种新颖的廉价的，基于即时智能手机的系统，用于检测罕见的 体液中突变等位基因，使中国农村肺癌液体活检成为可能。拟议的系统建立在我的 先前的工作与最低限度的仪器化和非仪器化的分子诊断。我的系统可以接受原始样本， 例如全血，处理样品，并提供快速测试结果。由于存在大量的 野生型（WT）等位基因挑战稀有突变等位基因的检测，我的系统将包括样品富集步骤， 利用Argonaute家族的DNA引导的切割酶消化和消耗WT等位基因，同时保留突变体 相关等位基因。随后进行环介导等温扩增（LAMP）， 核酸（PNA）钳以选择性扩增突变等位基因，但不扩增WT等位基因。用于突变体的多重检测 对于等位基因，所提出的系统将首先用可编程切割酶富集样品，然后使样品经受可编程切割酶。 富集的样品到一个新的两阶段，多重等温扩增过程（Penn-RAMP），在这种情况下， 点/缺失/插入突变，用PNA夹阻止Penn-RAMP第二次试验中WT等位基因的扩增。 阶段扩增子将用生物发光报告子和智能手机摄像头检测。定制智能手机 应用程序将分析记录的信号;报告测试结果;并在将来将这些结果传输到患者的 医生和记录，并在去识别的形式，到云的时空监测，使公共卫生官员 确定热点。我们的初步数据表明，我们的方法成功的可能性很高。 我已经制定了以下三个领域的培训计划：癌症流行病学，癌症诊断和治疗， 医疗器械设计和制造。在每一个领域，我都确定了课程和指导支持， K 01咨询委员会成员。咨询委员会由国际公认的专家组成， 工程学、癌症流行病学、肺癌精确医学、液体活检和微生物学已经组装 并将定期开会评估我的进展并提供指导。 这项研究有可能大大改善中国肺癌的伴随诊断和筛查， 有效的治疗。该计划将使我能够通过实地工作，研究，课程学习获得新技能， 所有这些都将促进和加速我向全球健康独立调查员的转变。