Supplement to Support the Development of a New Multiplexed Imaging Tool using Raman Spectroscopy for Breast Cancer

项目摘要 乳腺癌，特别是，是非常异质性与许多形态和分子特征， 不同的患者。未能完全理解分子表达和肿瘤异质性， 患者的肿瘤可导致无效治疗的施用，从而增加患者发病率和医疗保健 成本基因组学时代使我们有可能鉴定出几种与乳腺癌有关的新分子标记物 发展、存活、侵袭甚至预测治疗反应。我们目前缺乏一个简单的方法来获得 高含量的分子信息，同时提供跨患者肿瘤的高分辨率空间分布。这 该提案旨在为医生提供一种全新的多重分子成像策略， 以在单个组织学图像中提供高含量的分子表达和空间分布。拉曼 与表面增强拉曼散射（Sers）纳米颗粒结合的光谱是光学成像 技术，可以提供无与伦比的灵敏度（在fM的数量级）和多路复用能力的领域， 组织学成像具有在微观水平上提供丰富分子细节的潜力。临床医生将 能够在为组织学制备的相同组织切片上利用成像策略。将其纳入 病理学工作流程可以使医生更好地了解患者的肿瘤类型，并对患者进行分层， 接受最有效的治疗方案。这种独特的组织学成像策略也具有 有可能在患者的组织样本中发现新的分子趋势，这些分子趋势可用于预测 他们的肿瘤是什么或者病人对给定疗法的反应如何。这种创新的体外诊断 该策略具有很高的临床转化可能性;为多个组织提供快速的全组织切片成像， 生物标志物同时我们的方法首先开发了一套新的敏感的Sers纳米粒子（NP） 批次，每个批次都设计有独特的光谱条形码，以实现对整个样品的同时分子询问。 在单个图像内的组织样本。在我们新开发的多重Sers的制造和表征之后， 纳米粒子，我们将测试他们的多重成像能力和肿瘤靶向效率在各种乳腺癌 包括细胞培养和去识别组织切片的模型。我们的纳米粒子将主动靶向多个乳房 癌症受体通过化学结合的靶向配体。我们将评估的肿瘤靶向效率 我们新开发的纳米颗粒与显微拉曼成像工具，并与金标准进行比较 免疫组织化学（IHC）染色。这些结果将是这一新的临床转化的重要一步。 多路复用拉曼成像方法;以提供给定肿瘤的快速空间分子谱分析，同时使 为患者提供更有效的个性化治疗。

Project Summary Breast cancer, in particular, is very heterogeneous with many morphological and molecular features that present differently across patients. Failure to fully understand the molecular expression and tumor heterogeneity across a patient's tumor can lead to administration of ineffective therapies that increase patient morbidity and healthcare costs. The -omics era has made it possible to identify several new molecular markers involved in breast cancer development, survival, invasion and even predicting treatment response. We currently lack an easy way to obtain high content molecular information while providing high resolution spatial profiling across a patient's tumor. This proposal aims to provide physicians with an entirely new multiplexed molecular imaging strategy that has the potential to offer both high content molecular expression and spatial profiling in a single histology image. Raman spectroscopy in conjunction with surface enhanced Raman scattering (SERS) nanoparticles is an optical imaging technique that can offer unsurpassed sensitivity (on the order of fM) and multiplexing capabilities to the field of histology imaging with the potential to provide rich molecular details on the microscopic level. Clinicians will be able to utilize the imaging strategy on the same tissue sections prepared for histology. Incorporating it into the pathology workfiow could enable physicians to better understand the patient's tumor type and stratify patients to receive the most effective therapeutic regimen possible. This unique histology imaging strategy also has the potential to identify new molecular trends in patient's tissue samples that could be used to predict how aggressive their tumor is or how well the patient is likely to respond to given therapies. This innovative ex-vivo diagnostic strategy has a high likelihood for clinical translation; offering rapid whole tissue section imaging for multiple biomarkers simultaneously. Our approach begins by developing a new set of sensitive SERS nanoparticle (NP) batches, each designed with a unique spectral barcode to enable simultaneous molecular interrogation of an entire tissue sample within a single image. After fabrication and characterization of our newly developed multiplexed SERS nano particles, we will test their multiplexed imaging capabilities and tumor targeting efficiency in various breast cancer models including cell culture and on de-identified tissue sections. Our nanoparticles will actively target multiple breast cancer receptors through chemically conjugated targeting ligands. We will assess the tumor targeting efficiency of our newly developed nanoparticles with microscopic Raman imaging tools and compare with gold standard immunohistochemistry {IHC) staining. These results will be an important step in the clinical translation of this new multiplexed Raman imaging approach; to provide rapid spatial molecular profiling of a given tumor while enabling a more effective personalized therapy to the patient.