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

项目摘要 肿瘤的异质性正在成为癌症研究的一个主要焦点。许多器官的肿瘤，包括 肺，表现出肿瘤内的异质性，并且肿瘤的不同区域对 治疗。在治疗前更好地了解肿瘤的异质性可以用于更个性化的治疗， 知情治疗使患者的预后更好。福尔马林固定、石蜡包埋(FFPE) 对癌症标本进行切片，然后进行细胞染色，是病理诊断的标准做法。不像 较长的RNAs、microRNAs(MiRNAs)可以承受FFPE过程，并已知在许多情况下调节失调 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记物 和预后，目前还没有一种技术可以对miRNA进行定量、多路检测 来自肿瘤组织切片的生物标记物，同时保留了肿瘤活检的空间信息 劳动密集型。这个项目的目标是开发工具来空间和定量地解析多个miRNA 病理组织切片中的生物标记物。拟议的目标将通过以下具体措施实现 目的：1)建立凝胶后微孔平台，用于组织小区域的局部miRNA测量 部分，2)扩大传感阵列，推进样品处理和数据分析，3) 展示了集成系统在患者来源的有机物和小鼠异种移植样本上的应用， 以及基因工程小鼠模型。我们已经召集了一支工程师和病理学家团队来实现 这些目标。这些技术将以这样的方式开发，即它们将融入并补充 目前病理学家的工作流程，从而提供了一条被临床医生广泛采用的途径。该项目是 创新，因为目前还没有技术来执行空间分辨的miRNA表达谱 临床相关的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 个人化、知情的治疗为患者带来更好的预后。长期目标是包括 这项检测在标准护理中以组织生物标记物为基础，对每一位癌症患者进行检查。

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.