Development of a multi-RNA signature in blood towards a rapid diagnostic test to robustly distinguish patients with acute myocardial infarction

摘要 胸痛是急性心肌梗死的主要症状，约占所有急诊的5%。 部门(ED)访问。在无心电图异常的情况下，急性心肌梗死的诊断金标准依赖于序列 肌钙蛋白(CTn)测量在20%-40%的患者中不确定，需要额外的测试和 在急诊室长期观察。如果没有适当的治疗，急性心肌梗死的漏诊将危及生命和 因此，排除诊断需要非常高的灵敏度。因此，快速护理点(POC)测试可用作 具有增强诊断性能的cTn附件将是风险分层的革命性产品，并将及时和 急诊室疑似心肌梗塞患者的安全分诊。 Inflammatx是一家分子诊断公司，专注于开发同类最好的产品并将其推向市场。 基于免疫反应、数据驱动的测试。我们开发了一种护理点式仪器，Myrna™，能够 直接从患者血液中定量，在不到30分钟的时间内(操作时间为2分钟)最多可检测到的mRNA， 装在一个完全一次性的墨盒里。我们专门使用最先进的多队列分析和机器学习 (Ml)识别和验证跨现实世界数据异质性、不同类型的强大生物标记 临床情况。以前的工作证明了血液基因表达作为心肌梗塞的生物标记物的潜力， 然而，基于血液中基因表达的免疫反应的临床测试还有待开发。我们申请了 我们对6个公开可用的数据集的分析框架，并在外周识别出一个多基因急性心肌梗死特征 使我们能够区分AUC~0.95的急性心肌梗死患者和临床相关的对照组的血液。 在这个项目中，我们通过研究和初步的研究，从初步的结果中提取了AMI签名 开发阶段，直到正式的临床诊断开发。我们将产生大量的 独立数据，利用Inflammatx ML功能进一步细化信使核糖核酸签名，并提供强大的 分类器准备好在前瞻性研究中进行验证。在具体目标1中，我们将生成、处理和分析 来自密切代表目标测试人群的回溯性队列的900份血液样本的RNA-SEQ数据。 在具体目标2中，我们将首先提炼、优化和验证mrna签名；然后开发一个原型。 ML分级机。具体地说，我们将1)整合来自所有队列的表达数据，同时将偏差降至最低；2)应用 300个新样本的最终基因选择的贝叶斯多队列框架；3)开发和评估 AMI型分类器样机的判别性能；4)在600上验证了AMI型分类器样机 看不见的样本。这些步骤将产生：i)一组有效的急性心肌梗死基因；ii)一个完整的数据集；以及iii) 分类器原型(AUC&GT；0.90)，准备通过第二阶段研究的前瞻性研究进行临床验证。这 当测试被开发为Inflammatx的POC仪器Myrna™上的试剂盒时，将有助于临床医生进行分类 疑似心肌梗死患者的决策，改善患者预后，并降低医疗成本。

ABSTRACT Chest pain, the main symptom of acute myocardial infarction (AMI), accounts for ~5% of all emergency department (ED) visits. In the absence of ECG abnormalities, diagnostic gold standard for AMI relies on serial troponin (cTn) measurements which are inconclusive in 20-40% of patients, requiring additional testing and prolonged observation in the ED. A missed diagnosis of AMI without proper treatment is life threatening and thus rule-out diagnosis requires very high sensitivity. Hence, a rapid point of care (POC) test utilized as an adjunct to cTn with enhanced diagnostic performance would be revolutionary for risk stratification and timely and safe triaging of patients with suspected MI in ED. Inflammatix is a molecular diagnostics company focused on developing and bringing to market best in class, immune response based, data-driven testing. We developed a point-of-care instrument, MyRNA™, capable of quantitating up to 64 mRNAs in under 30 minutes (with <2 minutes operator time), directly from patients’ blood, in a fully disposable cartridge. We specialize in use of state-of-art multi-cohort analysis and machine learning (ML) to identify and validate robust biomarkers that generalize across real-world data heterogeneity, in diverse clinical contexts. Previous work demonstrated the potential of blood gene expression as a biomarker for MI, however a clinical test based on immune response in blood gene expression is yet to be developed. We applied our analytical framework to 6 publicly available datasets and identified a multi-gene AMI signature in peripheral blood that allows us to differentiate patients with AMI from clinically relevant controls with AUC ~ 0.95. In this project, we propose to take the AMI signature from preliminary results through research and initial development stages, up to formal clinical diagnostic development. We will generate a significant amount of independent data, leverage Inflammatix ML capabilities to further refine the mRNA signature and deliver a robust classifier ready for validation in prospective studies. In Specific Aim 1, we will generate, process, and analyze RNA-seq data for 900 blood samples from retrospective cohorts closely representing the target test population. In Specific Aim 2, we will first refine, optimize, and validate the mRNA signature; and then develop a prototype ML classifier. Specifically, we will 1) integrate expression data from all cohorts while minimizing bias; 2) apply Bayesian multi-cohort framework for final gene set selection with 300 new samples; 3) develop and evaluate discriminatory performance of AMI classifier prototype; and 4) validate the AMI classifier prototype on 600 unseen samples. These steps will produce: i) a validated set of genes for AMI; ii) an integrated dataset; and iii) a classifier prototype (AUC > 0.90), ready for clinical validation via prospective studies in Phase 2 research. This test, when developed as a cartridge on Inflammatix’s POC instrument, MyRNA™, will facilitate clinician’s triaging decisions of patients with suspected MI, improve patient outcomes, and reduce healthcare costs.