A unique approach combining avatar mice and targeted mass spectrometry to identify blood biomarkers for early detection of breast cancer

项目总结/摘要 数以千万计的妇女通过乳房X光检查进行基于人口的乳腺癌筛查。去- 尽管早期发现具有挽救生命的潜力，但乳房X光检查的筛查指南仍存在争议， 继续发展。争论的焦点是乳房X光检查区分良性与癌性乳腺癌的特异性。 乳腺X线摄影在某些患者人群（例如，乳腺致密的女性）中的敏感性，以及 cerns关于过度诊断（例如，一些导管原位癌）。我们的最终临床目标是开发一种 血液检查用于早期发现乳腺癌，可与乳房X光检查结合使用，以改善 敏感性和特异性，因此通过检测可治疗的癌症具有巨大的临床影响， 通过乳腺X线摄影（降低死亡率）和避免不必要的侵入性手术， nign或非危及生命的疾病。 尽管付出了相当大的努力，但试图鉴定基于血液的筛选生物标志物以用于早期检测。 由于技术和方法的限制，乳腺癌的治疗失败了。显然，新方法 有正当理由我们提出了一个全新的策略，基于候选双- "阿凡达小鼠"（携带早期传代人乳腺癌异种移植物）血浆中的肿瘤标志物， 通过生物标志物分类和使用基于质量的靶向形式的新型生物标志物管道的验证， 质谱（MS）、多反应监测（MRM）。如果成功的话，这项研究可以提供一个路线图， 将这种通用方法应用于乳腺癌以外的其他癌症部位。 简言之，将在血浆中验证在avatar小鼠血浆中鉴定的候选循环生物标志物 并优先进行进一步的测试，基于整合的蛋白质基因组分析， 由NCI基因组和蛋白质组联合体（TCGA和CPTAC）生成的大型乳腺癌数据集。十一月- el，将开发并分析验证基于多重MRM的测定（根据已建立的适用于 目的指南）来量化多达50 - 100种优先化的生物标志物候选物。候选生物标志物将是 在现有的、强无偏的血浆样本收集中进行评价，其中血浆是在 活检，符合PRoBE研究设计标准，根据SOP，来自未确诊乳腺癌的女性， 图形损伤。候选生物标志物的性能最初将在训练集中进行评估（100个案例- es，100个对照），以验证哪些候选人显示病例与对照之间血浆水平的平均差异， trols，并测试建立多标记预测模型的可能性。随后，个别candi- 日期生物标志物（以及潜在的多标志物预测模型）将在一个独立的 验证集（150例病例，150例对照），以估计标志物/组的灵敏度和特异性。

Project Summary/Abstract Tens of millions of women undergo population-based screening for breast cancer by mammography. De- spite the life-saving potential of early detection, screening guidelines for mammography are controversial and continue to evolve. At issue are the specificity of mammography for distinguishing benign vs cancerous le- sions, the sensitivity of mammography in some patient populations (e.g. women with dense breasts), and con- cerns regarding over-diagnosis (e.g. some ductal carcinoma in situ). Our ultimate clinical aim is to develop a blood test for early detection of breast cancer that can be used in conjunction with mammography to improve sensitivity and specificity during screening, and thus have enormous clinical impact by detecting treatable can- cers missed by mammography (reducing mortality) and by avoiding unnecessary invasive procedures for be- nign or non-life-threatening disease. Despite considerable effort, attempts to identify blood-based screening biomarkers for early detection of breast cancer have failed, due to technological and methodological limitations. Clearly new approaches are warranted. We are proposing a completely novel strategy, based on the initial discovery of candidate bi- omarkers in the plasma of “avatar mice” (harboring early passage human breast cancer xenografts), and fol- lowed by biomarker triage and validation using a novel biomarker pipeline based on a targeted form of mass spectrometry (MS), multiple reaction monitoring (MRM). If successful, this study could provide a road map for applying this general approach to other cancer sites, beyond breast cancer. Briefly, candidate circulating biomarkers identified in the avatar mouse plasma will be verified in the plasma from the human patients and prioritized for further testing based on integrative proteo-genomic analyses using large breast cancer datasets generated by NCI genomic and proteomic consortia (TCGA and CPTAC). A nov- el, multiplex MRM-based assay will be developed and analytically validated (according to established fit-for- purpose guidelines) to quantify up to 50-100 prioritized biomarker candidates. Candidate biomarkers will be evaluated in an existing, strongly unbiased collection of plasma samples in which plasmas were collected prior to biopsy, compliant with PRoBE study design criteria, under an SOP from women with undiagnosed mammo- graphic lesions. Performance of the candidate biomarkers will initially be assessed in a training set (100 cas- es, 100 controls) to verify which candidates show a mean difference in plasma levels between cases vs con- trols, and to test the possibility of building a multiple marker prediction model. Subsequently, individual candi- date biomarkers (as well as a potential multiple marker prediction model) will be assessed in an independent validation set (150 cases, 150 controls) to estimate the sensitivity and specificity of the markers/panel.