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

项目摘要 肿瘤异质性正成为癌症研究的主要焦点。许多器官的肿瘤，包括 肺表现出肿瘤内异质性，且肿瘤不同区域可对 治疗在治疗前更好地了解肿瘤异质性可以用于更个性化， 知情治疗，为患者带来更好的预后。福尔马林固定，石蜡包埋（FFPE） 在病理学诊断中，对癌症样本进行切片，然后进行细胞染色是标准的做法。不像 更长的RNA，microRNA（miRNAs）可以承受FFPE过程，并且已知在许多细胞中失调。 疾病，尤其是癌症。尽管miRNAs有望成为癌症诊断的敏感和可靠的标志物， 和预后，没有技术可以进行定量，多重检测miRNA 本发明提供了一种用于从肿瘤组织切片中提取生物标志物的方法，同时保留肿瘤活检的空间信息， 劳动密集型。该项目的目标是开发空间和定量解析多种miRNA的工具， 病理组织切片中的生物标志物。将通过以下具体措施实现拟议目标 目的：1）建立一种用于小区域组织中局部miRNA测量的凝胶柱微孔平台 部分，2）按比例放大传感阵列并推进样品处理和数据分析，以及3） 证明了该集成系统对患者来源的类器官和小鼠异种移植物样品的效用， 和基因工程小鼠模型。我们已经组建了一个工程师和病理学家团队， 这些目标。这些技术的开发方式将使其融入并补充 病理学家的当前工作流程，从而为临床医生的广泛采用提供了途径。该项目 这是因为目前还没有技术可以在细胞中进行空间分辨的miRNA表达谱， 临床相关FFPE组织样本。该项目意义重大，因为它将为研究人员和 病理学家在治疗前更好地了解肿瘤的异质性，这可以用于更多 个性化的、知情的治疗，为患者带来更好的预后。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的标准中。

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.