Microfluidic intact cell platform: A novel tool for oral cancer detection

微流控完整细胞平台：口腔癌检测的新工具

• 批准号: 10043470
• 负责人: Umut A. Gurkan
• 金额: $ 41.4万
• 依托单位: CASE WESTERN RESERVE 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/10043470
• 关键词: Address; Aliquot; Bedside Testings; Benign; Biological; Biological Assay; Biological Markers; Biopsy; Caliber; Cancer Detection; Cancerous; Carcinoma in Situ; Cations; Cell membrane; Cells; Cellular Assay; Cellular Phone; Clinical Research; Coin; Collaborations; Collecting Cell; Computational algorithm; Computer Analysis; Consumption; Contralateral; Country; Custom; Dental Clinics; Detection; Developing Countries; Development; Devices; Diagnosis; Early Diagnosis; Early Intervention; Engineering; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Event; Expression Profiling; Fluorescence; Fluorescent Antibody Technique; Future; Goals; Gold; Head and Neck Cancer; Health care facility; Histopathology; Hour; Human; Image; Immobilization; India; Indigenous; Individual; Label; Laboratories; Lesion; Measures; Medicine; Methods; Microfluidic Microchips; Microfluidics; Molecular; Monitor; Mouth Carcinoma; Myelogenous; Non-Invasive Lesion; Observational Study; Oral; Oral Stage; Oral cavity; Oral mucous membrane structure; Pain; Pathology; Patient-Focused Outcomes; Patients; Peptides; Phase; Phenotype; Primary Health Care; Procedures; Proteins; Reporting; Research Personnel; Resources; Sampling; Screening for Oral Cancer; Screening procedure; Site; Specificity; Swab; System; Techniques; Technology; Testing; Time; Tissues; University Hospitals; Work; antimicrobial; antimicrobial peptide; base; beta pleated sheet; beta-Defensins; beta-defensin 3; biomarker development; cellular imaging; chemokine; clinical care; cost; cost effective; deep learning algorithm; diagnosis standard; diagnostic accuracy; experience; fluorescence imaging; imaging Segmentation; imaging capabilities; imaging platform; immunoregulation; improved; improved outcome; indexing; innovation; malignant mouth neoplasm; metaplastic cell transformation; microfluidic technology; monitoring device; mouth squamous cell carcinoma; mucosal site; novel; novel strategies; oral lesion; overexpression; patient population; point of care; portability; portable monitoring; premalignant; primary care setting; prospective; prototype; recruit; scalpel; screening; socioeconomics; success; tertiary care; tool

Oral squamous cell carcinoma (OSCC) claims the lives of thousands in the U.S. and hundreds of thousands worldwide annually. A biopsy followed by histopathology, the gold standard for the diagnosis of OSCC, is painful, invasive, costly, not practical if longitudinal assessments of the same lesion are required and oftentimes is not available or imprecise in third world countries. A tool that quickly distinguishes cancerous from non- cancerous lesions and identifies progressive or transforming lesions could allow for early intervention, which would improve outcomes and negate the need for unnecessary biopsies in patients whose lesions remain benign or haven’t begun to degenerate. Therefore, there is an unmet need for a rapid, non-invasive, objective and cost-effective test for OSCC. We and others have reported that an altered expression profile of human beta defensin 3 (hBD-3), an epithelial cell derived antimicrobial peptide (AMP), and hBD-2, another epithelial cell AMP, is an early event in OSCC. Therefore, the ratio of hBD-3 and hBD-2 in the lesion, when compared to the contralateral site, could be exploited in distinguishing OSCC from other lesions of the oral cavity. We refer to this ratio as the beta defensin index (BDI). Our ongoing clinical study of 78 subjects with suspicious oral lesions demonstrated high sensitivity (100%) and specificity (74%) of the ELISA based BDI in distinguishing cancerous from noncancerous oral lesions (P<0.0001). With the high accuracy (98%) of our BDI based molecular assay, we now wish to advance our novel platform from the laborious, time consuming ELISA format into an imaging-based point-of-care (POC) device that utilizes microfluidic technology to quantify the BDI with an expected turnover time of half an hour. Our microfluidic intact cell assay (MICA) approach to developing a POC device for oral cancer detection is unique; it utilizes intact epithelial cells trapped in a microfluidic chip encompassing microfabricated pillar arrays with varying spaces to allow the capture of epithelial cells. Upon capture, the cells are permeabilized and labeled with fluorescent antibodies for hBD ratio analysis. We employ automated fluorescence imaging and computational algorithm to enable automated calculation of the BDI scores. We now hypothesize that the ELISA format that can effectively detect oral cancer, can be configured for point of care MICA, retaining its high accuracy and making it easier to use worldwide. To advance the discovery of this new approach for oral cancer detection, we propose the following aims: 1. Develop a working prototype of a MICA POC device for oral cancer testing equipped with cell imaging and BDI calculation capabilities. 2. Conduct a discovery phase study where MICA POC and ELISA, as independent assays, will be compared with pathology review in their ability to detect oral cancer. The MICA POC, while not intending to replace biopsy, could be deployed, in the future, to objectively and non- invasively determine who actually needs a biopsy, monitor oral premalignant lesions in real world practice and fulfill a major unmet need in low-socio economic countries where pathology review is lacking and/or unreliable.

口腔鳞状细胞癌（OSCC）在美国夺走了成千上万的生命和成千上万的生命 每年在全球。活检后进行了组织病理学，即OSCC诊断的黄金标准是 如果需要对同一病变进行纵向评估，并且通常情况 在第三世界国家不可用或不可用。一种迅速将取消与非 - 的工具 癌性病变并识别渐进式或转化病变可以允许早期干预，这 将改善预后，否定病变仍在的患者中不必要的活检的需求 良性或没有开始退化。因此，未满足的需要快速，无创的 OSCC的客观和成本效益测试。我们和其他人报告说， 人β防御素3（HBD-3），一种衍生的抗菌肽（AMP）和HBD-2的上皮细胞，另一种 上皮细胞放大器是OSCC的早期事件。因此，病变中HBD-3和HBD-2的比率 与对侧部位相比，可以在区分OSCC和其他口腔病变时被利用 腔。我们将此比率称为β防御素指数（BDI）。我们正在进行的对78位受试者的临床研究 可疑的口腔病变表现出高灵敏度（100％）和基于ELISA BDI的特异性（74％） 区分癌性与非癌性口腔病变（p <0.0001）。我们的BDI准确度（98％） 基于分子测定，我们现在希望从实验室中推进我们的新颖平台，耗时ELISA 格式化为基于成像的即将点（POC）设备，该设备利用微流体技术来量化BDI 预期的失误时间为半小时。我们的微流体完整细胞测定法（MICA）的发展方法 用于口腔癌检测的POC装置是独一无二的。它利用被困在微流体芯片中的完整上皮细胞 包含带有不同空间的微生物柱阵列，以捕获上皮细胞。之上 捕获，将细胞透化并用荧光抗体标记，以进行HBD比分析。我们雇用 自动荧光成像和计算算法以实现BDI的自动计算 分数。现在，我们假设可以有效检测口腔癌的ELISA格式可以是 配置为用于护理点的云母，保留其高精度并使其更易于在全球使用。到 提出了这种新的口腔癌检测方法的发现，我们提出了以下目的：1。 开发用于配备细胞成像的口腔癌测试的云母POC设备的工作原型 和BDI计算功能。 2。进行发现阶段研究，其中云母POC和ELISA为 独立测定将与病理评论进行比较，以检测口腔癌的能力。这 云母POC虽然不打算替换活检，但将来可以部署 侵入性地确定谁实际上需要活检，监测现实世界实践中的口腔前病变，并且 在缺乏和/或不可靠的病理审查的低企业经济国家中，满足了主要的未满足需求。

项目成果

Integrating pathomics with radiomics and genomics for cancer prognosis: A brief review.

• DOI: 10.21147/j.issn.1000-9604.2021.05.03
• 发表时间: 2021-10-31
• 期刊: Chinese journal of cancer research = Chung-kuo yen cheng yen chiu
• 作者: Lu C;Shiradkar R;Liu Z
• 通讯作者: Liu Z

HBD-2 binds SARS-CoV-2 RBD and blocks viral entry: Strategy to combat COVID-19.

• DOI: 10.1016/j.isci.2022.103856
• 发表时间: 2022-03-18
• 期刊: iScience
• 影响因子: 5.8
• 作者: Zhang L;Ghosh SK;Basavarajappa SC;Chen Y;Shrestha P;Penfield J;Brewer A;Ramakrishnan P;Buck M;Weinberg A
• 通讯作者: Weinberg A

Feature-driven local cell graph (FLocK): New computational pathology-based descriptors for prognosis of lung cancer and HPV status of oropharyngeal cancers.

• DOI: 10.1016/j.media.2020.101903
• 发表时间: 2021-03
• 期刊: Medical image analysis
• 影响因子: 10.9
• 作者: Lu C;Koyuncu C;Corredor G;Prasanna P;Leo P;Wang X;Janowczyk A;Bera K;Lewis J Jr;Velcheti V;Madabhushi A
• 通讯作者: Madabhushi A

An unsupervised method for histological image segmentation based on tissue cluster level graph cut.

一种基于组织簇级图割的无监督组织学图像分割方法。

• DOI: 10.1016/j.compmedimag.2021.101974
• 发表时间: 2021
• 期刊: Computerized medical imaging and graphics : the official journal of the Computerized Medical Imaging Society
• 作者: Xu,Hongming;Liu,Lina;Lei,Xiujuan;Mandal,Mrinal;Lu,Cheng
• 通讯作者: Lu,Cheng