Development of a Universal Assay for Minimal Residual Disease in Acute Myeloid Leukemia using Duplex Sequencing

急性髓系白血病(AML)是一种病态疾病，每年新增病例超过2万例，死亡1万人 在美国，虽然大多数患者都能得到缓解，但大多数患者的残留病数量都很少 (MRD)这最终会导致复发。检测MRD的能力很重要，因为它的存在与 随着复发和死亡风险的增加，人们对调整治疗强度很感兴趣 根据有无MRD。不幸的是，目前的MRD检测方法存在变量 敏感性，不同实验室的表现不一致，缺乏对所有患者的广泛适用性。的确有 对更好的MRD检测方法的迫切而未得到满足的需求，这将使患者以及 其他利益相关者。下一代测序(NGS)允许检测基因异常 亚克隆水平。由于几乎所有的AML都含有突变，NGS可能成为“通用的”MRD检测的平台。 与其他检测方法不同，NGS将揭示特定的突变，并可能建议有针对性的治疗。 到目前为止，人们对NGS检测MRD的热情一直受到这一方法相对较差的敏感性的影响 方法。双链测序是最准确的NGS技术，可以改变MRD检测的范式。 我们团队的成员开创了这项专有技术，并在主体证明研究中证明了 它可以在极低的水平上准确地检测到白血病克隆。在此Fast Track应用程序的第一阶段，我们 将完善我们测序程序中的步骤，以促进我们的MRD检测的工业规模部署，并 验证分析性能。在第二阶段，我们基于银行AML来评估我们的分析的性能 样本。在目标1中，我们将重点放在我们的检测是否可以预测疾病复发。在目标2中，我们比较了我们的 目前检测MRD的金标准是流式细胞术。在目标3中，我们比较了 我们对配对的骨髓和外周血样本的检测，以确定我们是否可以 结果侵入性更小。最终产品将是一个强大、经济高效且可实施的实验室 已开发的测试(LDT)可用于商业部署。这款产品将对患者有广泛的用途， 肿瘤学家和付款人都通过帮助最有可能受益的患者进行尖端治疗， 同时避免其他人不必要的医疗和经济上的毒害。它将允许研究人员和制药公司 公司迅速评估新疗法，允许未来的临床试验规模更小、成本更低。 急性髓细胞白血病每年夺走数千名患者的生命。患者既死于这种疾病，也死于 积极的治疗。有一种超准确的“通用”检测方法来检测MRD将会有更好的效果 并将为更有效的个性化治疗铺平道路。我们从根本上说 我相信这样的测试是必要的，也是我们可以做到的，我们的团队比任何人都处于更好的位置 世界上其他国家将这一进步带给患者。

Acute myeloid leukemia (AML) is a morbid condition, with over 20,000 new cases and 10,000 deaths annually in the U.S. While the majority of patients achieve remission, most harbor minimal amounts of residual disease (MRD) that will ultimately lead to relapse. The ability to detect MRD is important as its presence is associated with increased risk of relapse and death, and there is considerable interest in modulating treatment intensity based on the presence or absence of MRD. Unfortunately, current MRD detection methods suffer from variable sensitivity, non-uniform performance across laboratories, and lack of broad applicability to all patients. There is an urgent, unmet need for a better MRD detection method, which could substantially benefit patients as well as other stakeholders. Next generation sequencing (NGS) permits detection of genetic aberrations on the subclonal level. As virtually all AML harbors mutations, NGS could be a platform for a “universal” MRD assay. Unlike other detection methods, NGS would reveal specific mutations and could suggest targeted therapies. Enthusiasm for NGS for MRD detection has, until now, been tempered by the relatively poor sensitivity of this method. Duplex Sequencing, the most accurate NGS technology, can change the paradigm for MRD detection. Members of our team pioneered this proprietary technology and demonstrated in proof-of-principal studies that it can accurately detect leukemic clones at extremely low levels. In Phase I of this Fast Track application, we will refine steps in our sequencing procedures to facilitate industrial-scale deployment of our MRD assay and validate analytical performance. In Phase II, we assess our assay's performance based on banked AML samples. In Aim 1, we focus on whether our assay is prognostic of disease relapse. In Aim 2, we compare our assay to flow cytometry, the current gold standard for MRD detection. In Aim 3, we compare performance of our assay on paired bone marrow and peripheral blood samples to determine if we can achieve comparable results less invasively. The final product will be a robust, cost-effective, and implementable Laboratory Developed Test (LDT) ready for commercial deployment. This product will be widely useful for patients, oncologists, and payers alike by helping direct cutting-edge therapies to the patients most likely to benefit, while sparing others unnecessary medical and financial toxicities. It will allow researchers and pharmaceutical companies to rapidly evaluate novel therapies, permitting future clinical trials to be smaller and less costly. AML takes the lives of thousands of patients every year. Patients die both from the disease and from the aggressive treatment. Having an ultra-accurate “universal” test to detect MRD would allow for improved prognostication and would pave the way for more efficacious personalized treatment. We fundamentally believe that such a test is necessary and well within our reach, and that our team is positioned better than any other in the world to bring this advance to patients.