Microengineered Technologies for Quantitative, Multiplexed and Spatially Resolved Measurement of miRNA in Tissue Sections

用于组织切片中 miRNA 的定量、多重和空间分辨测量的微工程技术

• 批准号: 10407515
• 负责人: Patrick S Doyle
• 金额: $ 34.76万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-10 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10407515
• 关键词: 3-Dimensional; Address; Area; Biological; Biological Assay; Biological Markers; Biopsy; Cancer Patient; Cancer Prognosis; Cells; Clinical; Complement; Cytotoxic agent; Data; Data Analyses; Diagnosis; Diagnostic; Disease; Drug resistance; Engineering; Exhibits; Formalin; Gel; Gene Expression; Genetically Engineered Mouse; Goals; Heterogeneity; Home; Human; Hydrogels; Image; Immunotherapy; Individual; Lung; Lung Neoplasms; Malignant Neoplasms; Malignant neoplasm of lung; Measurement; Measures; Messenger RNA; MicroRNAs; Mus; Mutation; Organ; Organoids; Paraffin Embedding; Pathologic; Pathologist; Pathology; Pathway interactions; Patients; Pattern; Performance; Pharmaceutical Preparations; Pharmacotherapy; Phenotype; Process; RNA; Relapse; Reproducibility; Research; Research Personnel; Resistance; Resolution; Sampling; Slice; Slide; Specimen; Stains; System; Techniques; Technology; Testing; Therapeutic; Time; Tissue Embedding; Tissue Sample; Tissues; Tumor Tissue; Xenograft procedure; anticancer research; base; cancer biomarkers; cancer diagnosis; clinically relevant; data acquisition; design; experience; innovation; large datasets; large scale production; lithography; miRNA expression profiling; microRNA biomarkers; microsystems; multiplex detection; neoplastic cell; novel; patient prognosis; personalized medicine; preservation; prognostic; provider adoption; scale up; software development; standard of care; targeted treatment; therapy resistant; tissue mapping; tool; treatment strategy; tumor; tumor heterogeneity

Project Summary Tumor heterogeneity is emerging as a major focus of cancer research. Tumors of many organs, including of the lung, exhibit intra-tumoral heterogeneity and different regions of a tumor can have varying resistance to therapies. A better understanding of tumor heterogeneity prior to treatment could be used for more personalized, informed treatments resulting in a better prognosis for patients. Formalin-fixed, paraffin-embedded (FFPE) sectioning of cancer specimens followed by cell staining is standard practice in pathological diagnosis. Unlike longer RNAs, microRNAs (miRNAs) can withstand the FFPE process and are known to be dysregulated in many diseases, especially cancer. Despite promise of miRNAs as sensitive and robust markers for cancer diagnosis and prognosis, there is no technology that can perform a quantitative, multiplexed detection of miRNA biomarkers from tumor tissue sections while preserving the spatial information of tumor biopsies and not being labor-intensive. The goal of this project is to develop tools to spatially and quantitatively resolve multiple miRNA biomarkers within pathological tissue sections. The proposed goals will be achieved via the following specific aims: 1) establishing a gel-post microwell platform for local miRNA measurement in small regions of tissue sections, 2) scaling up the sensing array and advancing the sample handling and data analysis, and 3) demonstrating the utility of the integrated system on patient-derived organoid and mouse xenograft samples, and genetically engineered mouse models. We have assembled a team of engineers and pathologists to achieve these goals. The technologies will be developed in a manner such that they integrate into and complement the current workflow of pathologists, thus providing a pathway to widespread adoption by clinicians. The project is innovative because no technology currently exists to perform spatially resolved miRNA expression profiles in clinically relevant FFPE tissue samples. The project is significant because it will provide researchers and pathologists a better understanding of tumor heterogeneity prior to treatment which could be used for more personalized, informed treatments resulting in a better prognosis for patients. The long-term goal is to include this assay in the standard of care tissue-based biomarker workup of every cancer patient.

项目摘要 肿瘤异质性是癌症研究的主要重点。许多器官的肿瘤，包括 肺，表现出肿瘤内异质性，肿瘤的不同区域可能具有不同的抵抗力 疗法。可以在治疗前更好地了解肿瘤异质性，以用于更个性化， 知情的治疗可为患者提供更好的预后。福尔马林固定，石蜡包裹（FFPE） 癌症标本的切片紧随细胞染色是病理诊断的标准实践。与众不同 较长的RNA，microRNA（miRNA）可以承受FFPE过程，并且已知在许多人中失调 疾病，尤其是癌症。尽管有保证miRNA作为癌症诊断的敏感和强大标记 和预后，没有技术可以执行miRNA的定量，多重检测 来自肿瘤组织切片的生物标志物，同时保留肿瘤活检的空间信息，而不是 劳动密集型。该项目的目的是开发工具以空间和定量解决多个miRNA 病理组织切片中的生物标志物。提出的目标将通过以下特定来实现 目的：1）建立一个小型组织中局部miRNA测量的凝胶 - 孔微孔平台 部分，2）扩展传感阵列并推进样品处理和数据分析，以及3） 证明集成系统在患者衍生的类器官和小鼠异种移植样品上的实用性， 和基因工程的鼠标模型。我们已经组建了一个工程师和病理学家团队来实现 这些目标。这些技术将以使其整合并补充的方式开发 病理学家的当前工作流程，因此为临床医生提供了广泛采用的途径。该项目是 创新性是因为目前尚无技术来执行空间解决的miRNA表达曲线 临床相关的FFPE组织样品。该项目很重要，因为它将为研究人员和 病理学家在治疗之前对肿瘤异质性有更好的了解，可以用于更多 个性化的知情治疗，导致患者的预后更好。长期目标是包括 该测定在每个癌症患者的基于护理组织的生物标志物检查中。