ADDING ADDITIONAL FUNDS UNDER OPTION PERIOD 1, TASK AREAS 5 AND 6, FOR CORE YEAR 5.

Broad将为TOPMed提供代谢组学和蛋白质组学服务。 代谢组学。 代谢组学技术提供了人血浆和组织样本的系统生化表征。我们已经建立了一个高通量代谢组学平台， 基于真实标准品测量了600多种已知代谢物，以及我们正在有条不紊地表征的数千个LC-MS峰。我们的方法整合了 靶向和非靶向LC-MS方法。我们严格处理了试验内和试验间的变异性，以“群体”水平规模采集数据。在FHS、糖尿病预防研究、妇女健康倡议和其他队列的先前分析中，我们已经确定了心脏代谢疾病的新生物标志物，并在模型系统中整合了遗传信息和研究，以确定新的生物学途径。为了响应2017年发布的关于2000个梅萨样本的试点代谢组学研究和2018年发布的关于约8000个样本的CORE第3年代谢物分析的任务订单请求，我们提供了完整及时的高质量靶向和非靶向数据，在两种情况下都满足了最后期限。我们目前正在与梅萨研究人员合作，为CORE第4年的数据生成生成约8000份血浆样本的代谢组学数据。自试点研究以来，我们一直积极响应TOPM研究人员的代谢组学数据工作，并支持和合作进行数据分析。 基于这些经验，我们寻求提供代谢组学能力，以响应任务订单请求（TORFP）#75N92020R0034;集中组学资源（CORE）;任务领域5：使用我们的高通量，多学科方法的代谢物谱。 Proteomics 新兴的蛋白质组学技术开始允许对人血浆样本进行系统的、无偏倚的表征，尽管在大型的、基于人群的队列研究中存在很少的数据。为了解决先前研究的局限性，我们在实验室建立了一个高通量蛋白质组学平台，使用广泛的收集来测量1，536种蛋白质 核苷酸标记抗体（Olink）。该测定利用了一种称为邻近延伸测定（PEA）的新方法，该方法可以通过下一代测序进行定量。 （NGS）。我们严格解决了分析内和分析间的变异性（均<13%），获得的数据比基于质谱的方法快20倍以上，具有一定的特异性 这似乎优于现有的使用DNA适体的基于高通量亲和力的方法。在杰克逊心脏研究和其他队列的新工作中，我们证实了已知生物标志物和心血管特征之间的关系，并发现了许多新的关联。 基于这些经验，我们寻求提供蛋白质组学能力，以响应任务订单请求提案（TORFP）#75N92020R0034;集中组学资源（CORE）;任务领域6：使用我们的高通量，多学科方法的蛋白质组学。我们的提案反映了我们实验室和布罗德研究所之间的长期合作努力。本申请还将使用质谱法将发现与新的“正交”验证方案结合。我们平台上生成的所有数据都将提供给TOPM研究人员，并将通过dbGap向科学界广泛提供真实的时间，就像我们以前所做的那样。我们坚信这一经验。 支持我们的计划，分析约8,000个样本，并提供蛋白质组学概况作为NHLBI TOPMed计划的一部分，在未来一年。

The Broad will provide both metabolomics and proteomics service for the TOPMed. Metabolomics. Metabolomic technologies provide the systematic biochemical characterization of human plasma and tissue samples. We have established a high-throughput metabolomics platform that measures over 600 known metabolites based on authentic standards, as well as thousands of LC-MS peaks that we are methodically working to characterize. Our methods integrate both targeted and non-targeted LC-MS approaches. We have rigorously addressed intra- and interassay variability, acquiring data at “population” level scale. In prior analyses in the Framingham Heart Study (FHS), the Diabetes Prevention Study, the Women’s Health Initiative and other cohorts, we have identified novel biomarkers of cardiometabolic disease and integrated genetic information and studies in model systems to identify novel biologic pathways. In response to Task Order Request for Proposals issued in 2017 for pilot metabolomics studies on 2000 MESA samples and in 2018 for CORE Year 3 metabolite profiling of approximately 8000 samples, we provided high quality targeted and nontargeted data delivered in a complete and timely manner, meeting deadlines in both cases. We are presently collaborating with MESA investigators to generate metabolomics data on approximately 8000 plasma samples for Data generation for CORE Year 4. Since the pilot studies, we have been responsive to TOPMed investigators working with metabolomics data and have supported and collaborated on data analysis. Based on these experiences, we seek to provide metabolomic capabilities in response to Task Order Request for Proposal (TORFP) #75N92020R0034; Centralized Omics REsource (CORE); Task Area 5: Metabolite Profiles using our high throughput, multidisciplinary approach. Proteomics Emerging proteomic technologies are beginning to permit the systematic, unbiased characterization of human plasma samples, though few data exist in large, population-based cohort studies. To address limitations of prior studies, we have established a high-throughput proteomic platform in our laboratory that measures 1,536 proteins using an extensive collection of nucleotide-labeled antibodies (Olink). The assay leverages a novel method referred to as a proximity extension assay (PEA) which can be quantified by Next Generation Sequencing (NGS). We have rigorously addressed intra- and inter-assay variability (both <13%) and acquired data >20-fold faster than mass spectrometry based methods with a level of specificity that appears to be better than existing high throughput affinity-based methods that use DNA aptamers. In new work in the Jackson Heart Study and other cohorts, we confirmed established relationships between known biomarkers and cardiovascular traits and discovered many new associations. Based on these experiences, we seek to provide proteomic capabilities in response to Task Order Request for Proposal (TORFP) #75N92020R0034; Centralized Omics REsource (CORE); Task Area 6: Proteomics using our high throughput, multidisciplinary approach. Our proposal reflects a longstanding collaborative effort between our laboratory and the Broad Institute. The present application will also couple discovery with novel “orthogonal” validation protocols using mass spectrometry. All of the data generated on our platform will be available for TOPMed investigators and will be made broadly available in real time to the scientific community via dbGap, as we have done previously. We believe this body of experience strongly. supports our plan to analyze ~8,000 samples and provide proteomic profiles as part of the NHLBI TOPMed program in the coming year.