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

项目总结 基本上所有上皮性癌症的肿瘤微环境(TME)与显著的生化相关 以及细胞外基质(ECM)的结构变化。许多肿瘤，包括乳房、胰腺等 和卵巢的特点是胶原构筑发生了深刻的变化。ECM变化(~微米级) 低于传统成像模式的分辨率，但对这种结构的分析对于 了解癌症的发生和转移。我们已经使用了胶原蛋白特有的二次谐波 基于变化的第二代(SHG)光学显微镜区分癌组织和正常组织 在超分子结构、原纤维结构和纤维形态方面，我们重点关注高级浆液性 卵巢癌(HGSOC)。然而，SHG无法识别特定的分子变化，这可能会提供 关于疾病病因、预后和治疗反应的重要信息。现在我们将开发一种新的方法 它结合了空间配准的倍频和表面增强的中红外光谱成像(SE-MIRSI) 相关形态计量学和化学计量学信息以阐明促进肿瘤的细胞外基质改变。后者 从振动光谱中空间探测特定的分子特征，并提供更高的灵敏度 使用纳米光子衬底，允许对整个组织切片进行快速和大面积的化学成像。 具体来说，SE-MIRSI可以定量地识别翻译后亚型分布的具体变化 胶原纤维的修饰和改变的交联性。SHG和SE-MIRSI的空间配准将 提供全面、超灵敏、无标记、无损、高分辨率的结构和生化 研究ECM改变在促进肿瘤发生和转移中的作用的成像平台。 在这里，我们将开发一个多变量数据处理工作流来识别胶原蛋白的特定签名 和来自这两个模式的其他ECM组件，为准确的分类器奠定了基础。我们会 验证了HSGOC组织样本的多模式特征。在这个项目结束时，我们将拥有 开发了一个多模式成像平台，将唯一地识别胶原和其他ECM生化 TME的变化。我们将根据成像速度和吞吐量建立性能衡量标准， 敏感度和分类准确性。这些结构和生化分析将提供新的洞察力 几种癌症的致癌和疾病进展。我们提出以下目标： 目的1.通过联合使用确定体外细胞外基质模型的特定结构和生化特征 SHG和SE-MIRSI。 目的2.验证空间配准SHG/SE-MIRSI方法在高级别浆液性卵巢癌中的诊断价值 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.