Point-of-Care Diagnosis of Esophageal Cancer in LMICs

中低收入国家食管癌的即时诊断

• 批准号: 10649166
• 负责人: Stephen J Meltzer
• 金额: $ 62.05万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10649166
• 关键词: Area; Benign; Biological Assay; Biological Markers; Biopsy; Cancer Detection; Cells; Cellular Phone; Cessation of life; Clinical; Cold Chains; Collection; Coupled; Cytolysis; DNA; DNA Integration; DNA Methylation; DNA purification; Data; Defect; Deglutition; Detection; Development; Devices; Diagnosis; Diagnostic; Disadvantaged; Disease; Dryness; Electricity; Endoscopy; Energy Supply; Engineering; Esophageal Squamous Cell Carcinoma; Esophageal Tissue; Esophagus; Evaluation; Feedback; Fluorescence; Freeze Drying; Healthcare; Histology; Histopathology; Hour; Human Resources; Hypermethylation; Impairment; Light; Magnetism; Malignant Neoplasms; Malignant neoplasm of esophagus; Measurement; Measures; Medical; Methylation; Microfluidic Microchips; Microfluidics; Modeling; Molecular; Mucous Membrane; Patients; Persons; Pilot Projects; Porifera; Reagent; Sampling; Sensitivity and Specificity; Services; Site; Specimen; Specimen Handling; Speed; Techniques; Temperature; Testing; Therapeutic; Time; Training; Uganda; Universities; University Hospitals; Waxes; bisulfite; candidate marker; capsule; design; diagnostic strategy; disorder control; follow-up; high risk; instrument; invention; low and middle-income countries; meetings; methylation biomarker; methylation testing; microchip; minimally invasive; molecular marker; mortality; point of care; point-of-care diagnosis; portability; rapid diagnosis; remote location; rural setting; sample collection; screening; sealant; tumor

SUMMARY: Despite substantial progress in clinical approaches to squamous cancer of the esophagus (ESCC), which causes most esophageal cancers (EC) in the world, this deadly tumor usually occurs at late disease stages, with very poor survival. Restricted availability of endoscopy (EGD), along with rarity and delays in histology, impairs detection of ESCC in LMICs, adversely impacting our ability to treat this disease effectively. Thus, in LMICs, inexpensive, safe, locally performable strategies for detecting ESCC are necessary to identify high-risk patients and refer them quickly to suitable diagnostic and therapeutic options. Therefore, a diagnostic approach featuring a retrievable swallowed sponge-on-a-string to gather esophageal specimens for molecular testing, combined with a point-of-care (POC) magnetofluidic chip for sample processing and DNA methylation detection, is proposed. The string-sponge is less expensive, more noninvasive, more convenient, and more rapid than EGD with biopsy. The magnetofluidic chip streamlines DNA purification, DNA bisulphite treatment, and PCR detection of methylation markers into a single POC apparatus. This approach does not necessitate EGD, can be performed in remote areas with portable energy supplies and does not require extensive medical training, and is thereby amenable to implementation in LMICs. Our Specific Aims are: 1: Using a sponge-capsule swallowed/tethered collection device, to construct a methylation marker-based strategy to detect ESCC. In 100 ESCC and 100 benign control patients, we propose (1) building a multivariate model containing biomarker candidates; (2) carrying out feedback-feasibility meetings with health care and endoscopy personnel at Makerere University Hospitals to fine-tune eventual POC usage; 2: In order to achieve a sample-to-answer assay, to implement DNA extraction, bisulfite treatment, and methylation-specific PCR into a magnetofluidic chip with dried reagents. We’ll use magnetofluidic techniques to streamline cell lysis, DNA extraction, bisulphite treatment, and methylation-specific PCR into a compact chip built from cheap thermoplastic materials. In addition, we’ll lyophilize reagents and use heat- deployed wax sealant plugs to permit storage at room temperature. In this fashion, we will fashion a sample-to- answer assay that is easy to use, inexpensive, and free of cold-chain steps; 3: In order to achieve fully automatic high-speed biomarker assaying, to design a small, light apparatus. We’ll engineer an instrument containing programmable magnetic actuation, temperature control, and detection of fluorescence to execute the test in a chip with very little user input. We’ll also design the apparatus to be small, light, easy to operate, portable electricity-powered, and mobile phone-controlled to ease integration with LMIC-based clinical tasking; and 4: Using our POC approach to carry out a diagnostic pilot study of ESCC in Uganda. While applying the magnetofluidic chip and apparatus used in Aim 2 and Aim 3, we’ll carry out a trial to measure specificity and sensitivity in 120 EGD-confirmed cases of ESCC and 360 benign disease control patients in Kampala, Uganda.

摘要：尽管食管鳞状癌 (ESCC) 的临床治疗方法取得了实质性进展， 它导致世界上大多数食道癌 (EC)，这种致命的肿瘤通常发生在疾病晚期 阶段，内窥镜检查 (EGD) 的可用性非常有限，并且稀有且延迟。 组织学，损害中低收入国家 ESCC 的检测，对我们有效治疗这种疾病的能力产生不利影响。 因此，在中低收入国家，需要廉价、安全、本地可实施的 ESCC 检测策略来识别 高风险患者，并迅速将他们转介给合适的诊断和治疗方案。 因此，一种诊断方法采用可回收吞咽的绳状海绵来收集食管 用于分子测试的样本，与用于样本处理的即时护理 (POC) 磁流体芯片相结合 和DNA甲基化检测，提出了绳状海绵更便宜、更无创、更有效。 磁流体芯片简化了 DNA 纯化、DNA 检测，比 EGD 更方便、更快速。 将亚硫酸氢盐处理和甲基化标记物的 PCR 检测放入单个 POC 装置中。 不需要 EGD，可以在具有便携式能源供应的偏远地区进行，并且不需要 广泛的医疗培训，因此适合在中低收入国家实施。 我们的具体目标是： 1：使用海绵胶囊吞咽/系留收集装置，构建一个 我们在 100 名食管癌患者和 100 名良性对照患者中采用基于甲基化标记的策略进行检测。 (1) 建立包含候选生物标志物的多变量模型 (2) 进行反馈可行性； 与麦克雷雷大学医院的医疗保健和内窥镜人员举行会议，以调整最终的 POC 用法；2：为了实现样品到答案的测定，实施DNA提取、亚硫酸氢盐处理， 和甲基化特异性 PCR 到磁流体芯片中，我们将使用磁流体。 技术将细胞裂解、DNA 提取、亚硫酸氢盐处理和甲基化特异性 PCR 简化为 由廉价的热塑性材料制成的紧凑芯片此外，我们将冻干试剂并使用热- 部署蜡密封剂塞以允许在室温下储存以这种方式，我们将制作样品。 答案测定易于使用、价格低廉且无需冷链步骤 3：为了完全实现； 自动高速生物标志物测定，设计一种小型、轻型设备我们将设计一种仪器。 包含可编程磁驱动、温度控制和荧光检测来执行 只需很少的用户输入即可在芯片中进行测试，我们还将将该设备设计得小巧、轻便、易于操作、便携。 电力驱动、手机控制，可轻松与基于 LMIC 的临床任务集成；4： 使用我们的 POC 方法在乌干达开展 ESCC 诊断试点研究。 Aim 2和Aim 3中使用的磁流控芯片和设备，我们将进行试验来测量特异性和 乌干达坎帕拉 120 例 EGD 确诊的 ESCC 病例和 360 例良性疾病控制患者的敏感性。