Discovery and verification of methylated circulating tumor DNA markers for the detection of colorectal cancer in subjects under 50 years of age

发现并验证用于检测 50 岁以下受试者结直肠癌的甲基化循环肿瘤 DNA 标记

• 批准号: 10658886
• 负责人: ROBERT William HAILE
• 金额: $ 65.88万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10658886
• 关键词: Address; Adherence; Adult; Age; Age Years; Age of Onset; Biological Assay; Biological Markers; Blood; Case/Control Studies; Characteristics; Colon; Colonoscopy; Colorectal; Colorectal Cancer; DNA Markers; DNA Methylation; DNA methylation profiling; Data; Data Set; Detection; Diagnosis; Diagnostic; Disease; Early Diagnosis; Eligibility Determination; Ensure; FDA approved; Feces; Formalin; Freezing; Friends; Human; Hypermethylation; Incidence; Leukocytes; Life; Malignant Neoplasms; Methods; Methylation; Mucous Membrane; Mutation; Neoplasms; Paraffin Embedding; Pathway interactions; Patients; Performance; Persons; Plasma; Population; Primary Neoplasm; Recommendation; Research Design; Resistance; Risk; SEER Program; Sampling; Screening for cancer; Series; Signal Transduction; Site; Specificity; Symptoms; Testing; The Cancer Genome Atlas; Time; Tissues; Triage; Tumor stage; United States Preventative Services Task Force; Validation; adenoma; advanced disease; age group; aged; bioinformatics pipeline; biomarker identification; biomarker panel; cancer type; colon cancer patients; colorectal cancer screening; colorectal cancer treatment; cost effective; diagnostic accuracy; early onset; early onset colorectal cancer; epigenomics; follow-up; head-to-head comparison; high risk; improved; metastatic colorectal; methylation biomarker; methylation testing; minimally invasive; mortality; novel; nuclease; population based; pyrosequencing; screening; screening guidelines; specific biomarkers; surveillance data; tumor; tumor DNA; uptake

Abstract In recent years, the incidence of colorectal cancer (CRC) under the age of 50 years has increased significantly and population-based screening is currently not offered to persons under 50 years. Consequently, persons with early-onset (<50 years) CRC (EOCRC) more frequently present with symptomatic disease at advanced stages (III/IV) resulting in greater loss of life in young cases. In 2020, the U.S. Preventive Services Task Force recommended the age for CRC screening by colonoscopy or fecal tests be reduced to 45 years. However, the uptake of screening by these methods in all screen-eligible populations is low, including in those <50 years who are genetically at high risk, so adherence in the asymptomatic population <50 years is also likely to be low. Also, over half of EOCRC occur in persons under <45 years of age, where no such screening would be offered. A minimally invasive, blood-based screening test for EOCRC would provide a cost-effective and patient-friendly option for triaging and identifying those warranting a follow-up colonoscopy, while increasing screening adherence in the younger population. Cancer-specific methylated DNA biomarkers are highly suited for population-based cancer screening via the detection of circulating tumor DNA (ctDNA) in blood plasma because they are more prevalent across patients with a given cancer type than tumor mutations and are more stable (nuclease-resistant) in plasma. The methylated SEPTIN9 plasma ctDNA test, “mSEPT9” (Epi proColon V2.0, Epigenomics), is FDA-approved for CRC screening in persons aged ≥50 years who decline colonoscopy and fecal tests. This test urgently needs to be assessed in persons <50 years to determine its suitability for the detection of EOCRC. Even so, a multi-marker test is likely to have superior sensitivity to a single-marker test. Thus, the objectives of our study are to identify and confirm a panel of methylated ctDNA markers for the plasma-based detection of EOCRC, and to compare the diagnostic performance of this panel to the mSEPT9 test in persons <50 years of age. In Aim 1, we will identify the most prevalent differentially-methylated (tumor vs. normal) DNA markers in CRC cases <50 years by performing deep Methyl-Seq across 4.2 million CpG sites in paired primary tumor vs. normal colon tissues from EOCRC cases, and leukocytes from healthy controls (to filter out non-specific markers). The identified markers will be validated in independent sample series. For the top-ranked markers, we will then develop methylation-specific real-time PCR assays with high analytical sensitivity and test these in pooled plasma from metastatic CRC cases (to select markers producing the strongest signals) and healthy controls (to eliminate any producing low-level non-specific signals) to finalize the marker panel. In Aim 2, we will evaluate the diagnostic performance characteristics of the mSEPT9 test vs. the multi-marker panel test in plasma from colonoscopy-verified CRC cases and controls <50 years. This study will yield a multi-marker methylated ctDNA panel with improved diagnostic performance over the single-marker mSEPT9 test for cost-effective, blood-based, CRC screening in asymptomatic persons <50 years of age.

摘要 近年来，50岁以下结直肠癌(CRC)的发病率显著上升 目前不向50岁以下的人提供基于人口的筛查。因此，人们 早发性(&lt；50岁)结直肠癌(EOCRC)更容易在晚期出现症状性疾病 阶段(III/IV)，在年轻病例中造成更大的生命损失。2020年，美国预防服务工作组 建议通过结肠镜或粪便检查进行结直肠癌筛查的年龄降至45岁。然而， 在所有符合筛查条件的人群中，通过这些方法进行筛查的接受率很低，包括在过去50年里 他们的遗传风险很高，所以在没有症状的人群中坚持50年也很可能是 很低。此外，超过一半的卵巢癌发生在45岁以下的人中，在这些人中不会进行这种筛查 出价了。一种基于血液的微创EOCRC筛查试验将提供一种成本效益高和 患者友好的选项，用于分诊和识别那些需要进行后续结肠镜检查的患者，同时增加 在年轻人群中进行依从性筛查。癌症特异性甲基化DNA生物标记物非常适合 通过检测血浆中循环肿瘤DNA(CtDNA)进行基于人群的癌症筛查 因为在特定癌症类型的患者中，它们比肿瘤突变更常见，而且更多 在血浆中稳定(抗核酸酶)。甲基化Septin9血浆ctDNA检测，“mSEPT9”(Epi proColon V2.0，表观基因组学)，被FDA批准用于拒绝结肠镜检查的50岁≥人群的结直肠癌筛查 还有粪便检测。这项测试迫切需要在50年内进行人员评估，以确定其适用于 EOCRC的检测。即便如此，多标记检测可能比单标记检测具有更高的灵敏度。 因此，我们研究的目标是确定和确认一组甲基化的ctDNA标记 基于血浆的EOCRC检测，并将该面板的诊断性能与mSEPT9进行比较 在50岁的人身上进行测试。在目标1中，我们将确定最普遍的差异甲基化(肿瘤 对420万cpG进行深层甲基化序列检测50年来结直肠癌患者的DNA标记 来自EOCRC患者的配对原发肿瘤与正常结肠组织以及来自健康人的白细胞中的位置 控件(以过滤掉非特定标记)。识别出的标记将在独立样本中进行验证 系列片。对于排名靠前的标记，我们将开发甲基化特异的实时荧光聚合酶链式反应分析 分析敏感性，并在转移性结直肠癌患者的混合血浆中进行检测(以选择产生 最强信号)和健康对照(消除任何产生低水平非特定信号)以完成 标记面板。在目标2中，我们将评估mSEPT9测试与MSEPT9测试的诊断性能特征。 结肠镜检查证实的结直肠癌患者和对照50年来的血浆多标记物小组试验。本研究 将产生多标记甲基化的ctDNA面板，其诊断性能比单标记更好 MSEPT9试验对50岁无症状的人进行成本效益高的血液结直肠癌筛查。