Translational Molecular and Cellular Imaging Technologies for Prostate Tumor Pathology

摘要 在美国，每年有近24万名男性被诊断出患有前列腺癌(PCA)，但没有 可以有效地确定他们的肿瘤是进展为危及生命的疾病还是仍然存在的临床测试 懒惰。因此，大多数患有低风险前列腺癌的男性只需接受积极的监测(AS) 取而代之的是接受昂贵的治疗，但会产生重大的长期不良影响。由多个学科组成的调查小组 由临床医生、生物学家和生物工程师组成，他们各自开发了关键技术，现在寻求 整合资源，直接解决这一长期存在的临床问题。拟议的项目侧重于 验证一种结合了无标记和量子点标记光谱成像的新技术平台 预测前列腺癌的进展。说明我们的技术的需求和实用性是我们对分析的具体选择 正在这里利用。梅奥团队已经证明，重新编排和/或拷贝的数量变化程度为5 福尔马林固定石蜡包埋(FFPE)活检标本中肿瘤细胞的基因组区域可能是有用的 在确定前列腺癌侵袭性的风险方面。然而，这些标记物不能发展成强大的、临床的 由于目前技术的限制，化验结果：针吸活检样本通常只含有少量 对于癌症，即使切成薄片，使用5探针分析通常也是不可能的 因为多路传输常规鱼类探头的能力有限。此外，不可能 同时识别标记为荧光的切片中的细胞类型，因此尚不清楚细胞是否会染色 FISH探针阳性或阴性包括癌细胞或间质细胞。这是我们的普遍问题 技术将解决--缺乏对(无)反应细胞的多重分子定量和鉴定。 伊利诺伊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的轮廓 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得到验证。 我们的项目通过结合(A)基于量子的FISH探针来满足技术和验证需求 使用无标记红外识别细胞/组织的斑点以识别特定分子改变 光谱成像。虽然这两项技术都是独立开发的，并被证明是 尽管这两种方案都是有效的，但它们没有以互补的方式整合在一起，也没有一起用来满足PCA的需求。 在这里，我们建议通过一组具有以下特征的PCA样本来测试和验证这种组合技术 已经被基因分析过了。联合技术的验证还将测试其在 为PCA预测提供了一个实用的检验，其统计模型和度量将与 现行的黄金标准。这一提议的成功将使产生一种强有力的检测策略 确定哪些男性最适合AS或更积极的治疗-这将是 为前列腺医疗带来变革性的改变。这项技术的验证也为组合分子铺平了道路 以及所有组织和所有类型的分子靶标的细胞分析，这可以极大地扩展我们的能力 提供准确和具体的诊断，以指导广泛的病理治疗。

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.