Translational Molecular and Cellular Imaging Technologies for Prostate Tumor Pathology

摘要 在美国，每年有近24万名男性被诊断患有前列腺癌（PCa），但没有 临床测试，可以有效地确定他们的肿瘤是否会发展到危及生命的疾病或仍然存在 懒惰因此，大多数患有低风险PCa的男性应该简单地接受主动监测（AS） 而是接受具有严重长期不良影响的昂贵治疗。一个多学科的调查小组 包括临床医生，生物学家和生物工程师，他们各自开发了关键技术，现在寻求 来联合收割机资源直接解决这个长期存在的临床问题。拟议项目的重点是 验证一种新的技术平台，该平台结合了无标记和量子点标记的光谱成像， 预测PCa进展。说明我们的技术的需要和实用性是我们所选择的具体测定方法， 在这里使用。马约团队已经证明，重排和/或拷贝数变异水平为5 福尔马林固定和石蜡包埋（FFPE）活检标本中肿瘤细胞中的基因组区域可能是有用的 来确定前列腺癌的侵袭性。然而，这些标记物不能发展成一种稳健的、临床上可接受的标记物。 由于目前技术的限制，检测：针吸活检标本通常只含有少量 即使是切成薄片，使用5探针分析也通常是不可能的 因为多重常规FISH探针的能力有限。此外，不可能 同时识别标记为荧光的切片中的细胞类型，因此不清楚染色的细胞是否 FISH探针阳性或阴性的细胞包括癌细胞或基质细胞。这是我们的普遍问题。 技术将解决缺乏多重分子定量和鉴定（非）应答细胞的问题。 伊利诺斯州的研究小组已经证明，使用红外（IR）光谱成像，可以描绘肿瘤微环境 并且可以获得新的预测信息。然而，这种方法需要在更大的试验中进行验证。 我们的项目通过结合（a）基于量子的FISH探针来解决技术和验证需求 点，用于识别特定的分子改变，（B）使用无标记红外线进行细胞/组织识别 光谱成像虽然这两种技术都是独立开发的， 尽管这些措施没有发挥有效作用，但并未以互补的方式加以整合，也未共同用于满足PCa的需求。 在这里，我们提出了一个测试和验证这种组合技术通过队列的PCa标本， 已经进行了基因分析该组合技术的验证还将测试其在以下方面的有效性： 为PCa预后提供了一个实用的测试与统计模型和措施，将比较， 目前的黄金标准。该提案的成功将使我们能够制定一个稳健的检测策略， 确定哪些男性是AS或更积极治疗的最佳候选人-这将是 前列腺保健的变革。这项技术的验证也为结合分子生物学铺平了道路。 以及所有组织和所有类型分子靶点的细胞分析，这可以极大地扩展我们的能力， 提供准确和特异性的诊断，可以指导各种病理的治疗。

ABSTRACT Nearly 240,000 men are diagnosed with prostate cancer (PCa) every year in the United States but there is no clinical test that can effectively determine whether their tumors will progress to life-threatening disease or remain indolent. Consequently, a majority of men with low-risk PCa who should simply undergo active surveillance (AS) instead receive costly treatments with major long-term adverse effects. A multidisciplinary team of investigators comprising clinicians, biologists, and bioengineers, who individually have developed key technologies, now seek to combine resources to directly address this long-standing clinical problem. The proposed project focuses on validating a novel technology platform that combines label-free and quantum dot-labeled spectral imaging to predict PCa progression. Illustrating the need and utility of our technology is the specific choice of assays we are utilizing here. The Mayo team has shown that rearrangements and/or copy number variant levels of five genomic regions in tumor cells in formalin fixed and paraffin embedded (FFPE) biopsy specimens can be useful in determining risk of PCa aggressiveness. However, these markers cannot be developed into a robust, clinical assay due to the current limitations of technology: Needle biopsy specimens often contain only a small amount of cancer, and even when cut into thin sections, the employment of a 5-probe assay is often simply not possible because of the limited capacity to multiplex conventional FISH probes. Furthermore, it is not possible to simultaneously identify cell types in sections labeled for fluorescence, so it is not clear whether cells that stain positive or negative for the FISH probes comprise cancer or stromal cells. This is the general problem our technology will address – lack of multiplexed molecular quantitation and identification of (non)responsive cells. The Illinois team has shown that using infrared (IR) spectral imaging, the tumor microenvironment can be profiled and new predictive information can be obtained. However, this approach needs to be validated in a larger trial. Our project addresses the technology and validation needs by combining (a) FISH-probes based on quantum dots to identify specific molecular alterations with (b) cell/tissue identification using label-free infrared spectroscopic imaging. While both technologies has been independently developed and demonstrated to be effective, they have not been integrated in a complementary manner nor together used to address PCa needs. Here, we propose a test and validation of this combination technology via a cohort of PCa specimens that have already been genomically profiled. The combined technology's validation will also test its effectiveness in providing a practical test for PCa prognosis with statistical models and measures that will be compared to the current gold standards. Success in this proposal will enable the production of a robust assay strategy to determine which men are best candidates for AS or for more aggressive treatment – which would be transformative for prostate healthcare. Validation of this technology also paves the way for combined molecular and cellular analysis in all tissues and for all types of molecular targets, which can vastly expand our capacity to provide accurate and specific diagnoses that can guide therapy in a wide range of pathologies.