Rapid Estimation of Gene Expression in Tissue Slices

描述(由申请人提供)： 外科医生、病理学家和其他内科医生在癌症治疗过程中做出的决定在很大程度上取决于如何对恶性肿瘤进行分类，这一过程历史上仅限于组织学标准。最近的数据表明，当肿瘤的基因表达谱已知时，分类过程可以显著改善。利用微阵列方法获得了最完整的图谱，它具有同时监测数万个基因表达的能力。这些分析已经确定了对分类各种肿瘤最重要的mRNA转录本。然而，微阵列技术并不容易适用于单个细胞。对整个组织的分析产生的数据包括反映未转化细胞和转化细胞中基因表达的值。此外，肿瘤可能包含不止一种转化的细胞类型，这一现象混淆了这种分析，并可能导致分类错误。我们希望绕过整个组织的微阵列分析固有的限制，这促使我们寻找替代策略来获得这一信息。我们建议建立一种仪器，能够同时监测冷冻切片中单个细胞中的几个mRNA转录本。它的设计利用了众所周知的荧光光谱特性，应该能够在手术期间获得这些数据。这将允许病理学家和外科医生在必须决定手术范围的时候更好地评估肿瘤的性质。它还将允许医生更好地定制可能涉及放射和化疗的后续治疗，这些治疗可能会产生令人虚弱的副作用。

DESCRIPTION (provided by applicant): Decisions made by surgeons, pathologists, and other physicians during cancer therapy depend largely on how the malignancy is classified, a process limited historically to histological criteria. Recent data suggest that the classification process can be improved significantly when the gene expression profile of the tumor is known. The most complete profiles have been obtained using microarray methods, which have the capabilities of monitoring the expression of tens of thousands of genes simultaneously. These analyses have identified mRNA transcripts that are most important for classifying a variety of tumors. Microarray techniques are not readily applicable to single cells, however. Data produced by analyses of whole tissues include values that reflect gene expression in non-transformed as well as transformed cells. Furthermore, tumors may contain more than one transformed cell type, a phenomenon that confounds this analysis and that may lead to errors in classification. Our desire to circumvent the limitations inherent in microarray analyses of whole tissues led us to seek alternate strategies for obtaining this information. We propose to build an instrument capable of monitoring simultaneously several mRNA transcripts in single cells of frozen sections. Its design makes use of well-known properties of fluorescence spectroscopy and should enable these data to be acquired during surgery. This will permit pathologists and surgeons to better assess the nature of the tumor at the time when decisions must made regarding the extent of the surgical procedure. It will also permit physicians to better tailor subsequent treatments that may involve radiation and chemotherapy, treatments that can have debilitating side effects.