A novel multimodal ECM analysis platform for tumor characterization combining morphological and spectrochemical tissue imaging approaches.

项目摘要 基本上所有上皮癌中的肿瘤微环境（TME）与显著的生物化学相关。 和细胞外基质（ECM）的结构变化。许多肿瘤，包括乳腺癌，胰腺癌 和卵巢的特征在于胶原结构的深刻变化。ECM变化（~微米级） 低于传统成像模式的分辨率，但对该结构的分析对于 了解癌的发生和转移。我们已经使用了胶原蛋白特异性模态的二次谐波 第二代（SHG）光学显微镜，根据变化区分癌症标本和正常组织 在超分子结构、原纤结构和纤维形态方面，我们重点研究了高级浆液 卵巢癌（HGSOC）。然而，SHG不能识别特定的分子改变，这可以提供 关于疾病病因、预后和治疗反应的关键信息。现在我们将开发一种新的方法 结合了空间配准SHG和表面增强中红外光谱成像（SE-MIRSI） 关联形态计量学和化学计量学信息以阐明促肿瘤ECM改变。后者 空间探测来自振动光谱的特定分子特征， 使用纳米光子衬底，允许整个组织切片的快速和大面积化学成像。 具体而言，SE-MIRSI可以定量鉴定异构体分布、翻译后 胶原纤维的改性和改变的交联。然后，SHG和SE-MIRSI的空间配准将 提供一个全面的，超灵敏的，无标记的，非破坏性的，高分辨率的结构和生物化学 成像平台，以研究ECM改变在促进肿瘤发生和转移中的作用。 在这里，我们将开发一个多元数据处理工作流程，识别胶原蛋白的特定签名 以及来自两种模态的其他ECM成分，建立准确分类器的基础。我们将 验证HSGOC组织样本的多模态表征。在这个项目结束时，我们将有 开发了一个多模式成像平台，将独特地识别胶原蛋白和其他ECM生化 TME的变化。我们将根据成像速度和吞吐量建立性能指标， 灵敏度和分类精度。这些结构和生物化学分析将提供新的见解， 在几种癌中的致癌作用和疾病进展。我们提出这些目标： 目标1。通过组合使用，识别体外ECM模型的特定结构和生化特征 SHG和SE-MIRSI。 目标2.高级别浆液性卵巢癌的SHG/SE-MIRSI三维配准成像及鉴别诊断 特定的相关结构形态和生化特征。

PROJECT SUMMARY The tumor microenvironment (TME) in essentially all epithelial cancers is associated with significant biochemical and structural changes in the extracellular matrix (ECM). Many tumors including those of the breast, pancreas and ovary are characterized by profound changes in the collagen architecture. ECM changes (~micron scale) are below the resolution of conventional imaging modalities but analysis of this structure is critical for understanding carcinogenesis and metastasis. We have used the collagen-specific modality of Second Harmonic Generation (SHG) optical microscopy to discriminate cancer specimens from normal tissues based on changes in supramolecular structure, fibril structure, and fiber morphology, where we have focused on high grade serous ovarian cancer (HGSOC). However, SHG cannot identify the specific molecular alterations, which could provide critical information on disease etiology, prognosis, and response to therapy. Now we will develop a novel method that combines spatially registered SHG and surface enhanced mid-infrared spectral imaging (SE-MIRSI) correlating morphometric and chemometric information to elucidate tumor-promoting ECM alterations. The latter spatially probes specific molecular signatures from vibrational spectroscopy and provides increased sensitivity using nanophotonic substrates, allowing rapid and large-area chemical imaging of whole tissue sections. Specifically, SE-MIRSI can quantitatively identify specific changes in isoform distribution, posttranslational modifications and altered crosslinking of the collagen fibers. Spatial registration of SHG and SE-MIRSI then will provide a comprehensive, ultrasensitive, label free, non-destructive, high-resolution structural and biochemical imaging platform to investigate the role of ECM alterations in promoting tumor carcinogenesis and metastasis. Here, we will develop a multivariate data processing workflow that identifies the specific signatures of collagen and other ECM components from the two modalities establishing the basis of an accurate classifier. We will validate the multimodal characterizations on HSGOC tissue samples. At the end of this project, we will have developed a multimodal imaging platform that will uniquely identify collagen and other ECM biochemical alterations in the TME. We will establish performance measures based on imaging speed and throughput, sensitivity and classification accuracy. These structural and biochemical analyses will provide new insight into carcinogenesis and disease progression in several carcinomas. We propose these Aims: Aim 1. Identify specific structural and biochemical signatures of in vitro ECM models through the combined use of SHG and SE-MIRSI. Aim 2. Validate spatially registered SHG/SE-MIRSI method on high grade serous ovarian cancer and identify specific associated structural morphology and biochemical signatures.