"Molecular Signatures of Lethal and Indolent Prostate Cancer"

描述（申请人提供）：前列腺癌（PCA）的预后和临床管理是当代肿瘤学中最复杂和最有争议的问题之一。我们的首要目标是发展基于组织的分子测试，以区分治疗前的惰性和侵袭性PCA。我们的提案将解决与PCA临床相关分子特征开发相关的几个关键问题：(1)我们使用PCA特异性死亡作为临床终点；(2)我们将从瑞典招募3个明确的患者群体，进行长期完整的临床随访；(3)由于PCA的细胞异质性，我们采用了肿瘤富集方法，如激光捕获显微解剖（LCM）。这些指导原则被组织为3个具体目标：在目标1中，我们使用10OK单核苷酸多态性（SNP）阵列来检测杂合性缺失（LOH）、纯合缺失和基因扩增事件，以9Kb的分辨率生成全基因组遗传图谱。结合这些分子特征数据和来自瑞典Orebro的96个连续冷冻样本的PCA活检样本的临床数据，这些男性被诊断为临床局限性PCA，其中40人死于PCA， 30人死于其他原因，我们将开发一种分子特征来区分侵袭性和惰性PCA。在Aim 2中，我们使用基于头部的纳米技术测试并验证了这一分子图谱，以评估DNA中的体细胞改变，这些DNA可以应用于瑞典全国前列腺切除术队列（n=600）和两个观察等待队列（n=620）的福尔马林固定石蜡包埋样本，预计分别有200和120例PCA特异性死亡。在Aim 3中，我们将使用来自瑞典前列腺切除术和Watchful Waiting队列的组织微阵列，并通过免疫组织化学、免疫荧光（AQUA）、原位杂交（ISH）或荧光原位杂交（FISH）的定量分析来评估生物标志物的组合。我们期望开发可转移到其他实验室进行临床验证的原位测试。在这一建议的结论，我们期望有完善和充分验证的分子预测PCA死亡，以及适合进一步临床发展的惰性PCA。

DESCRIPTION (provided by applicant): Prognostication and clinical management of prostate cancer (PCA) is one the most complex and controversial issues in contemporary oncology. Our overarching goal is to develop tissue based molecular tests to distinguish indolent from aggressive PCA prior to treatment. Our proposal will address several critical issues related to the development of a clinically relevant molecular signature of PCA: (1) We use PCA specific death as the clinical endpoint; (2) We will employ 3 well defined patient populations from Sweden with long term and complete clinical follow up; (3) We use tumor enriching methods such as laser capture microdissection (LCM) due to the cellular heterogeneity of PCA. These guiding principles have been organized into 3 Specific Aims: In Aim 1, we use 10OK single nucleotide polymorphism (SNP) arrays for the detection of Loss of Heterozygosity (LOH), homozygous deletions and gene amplification events to generate genome-wide genetic maps at a resolution of 9Kb. Combining this molecular signature data with clinical data on PCA biopsy samples from 96 consecutive frozen samples from men diagnosed in Orebro, Sweden with clinically localized PCA of whom 40 died of PCA and 30 of other causes, we will develop a molecular signature to distinguish aggressive from indolent PCA. In Aim 2, we test and validate this molecular profile using a bead-based nanotechnology to assess for somatic alterations in the DNA that can be applied on formalin-fixed paraffin embedded samples from the nationwide cohort of prostatectomies in Sweden (n=600) and two Watchful Waiting cohorts (n=620) with a projected 200 and 120 PCA specific deaths, respectively. In Aim 3, we will employ tissue microarrays from the Swedish Prostatectomy and the Watchful Waiting cohorts and evaluate combinations of biomarkers by quantitative analysis of immunohistochemistry, immunofluorescence (AQUA), in situ hybridization (ISH) or fluorescence in situ hybridization (FISH). We expect to develop in situ tests that will be transportable to other laboratories for clinical validation. At the conclusion of this proposal, we expect to have refined and fully validated molecular predictors of PCA death as well as indolent PCA that is appropriate for further clinical development.