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: 10306165
• 负责人: ROBERT William HAILE
• 金额: $ 68.55万
• 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/10306165
• 关键词: Address; Adherence; Adult; Advisory Committees; Age; Age of Onset; Age-Years; Biological Assay; Biological Markers; Blood; Case-Control Studies; Characteristics; Colon; Colonoscopy; Colorectal; Colorectal Cancer; DNA; DNA Markers; DNA methylation profiling; Data; Data Set; Detection; Diagnosis; Diagnostic; Disease; Early Diagnosis; Ensure; FDA approved; Feces; Formalin; Freezing; Gold; Human; Incidence; Leukocytes; Life; Malignant Neoplasms; Methods; Methylation; Mucous Membrane; Mutation; Neoplasms; Paraffin Embedding; Pathway interactions; Patients; Performance; Persons; Plasma; Population; Preventive service; Primary Neoplasm; 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; Validation; adenoma; advanced disease; age group; aged; base; bioinformatics pipeline; biomarker panel; cancer type; case control; 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; 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.

抽象的 近年来，50岁以下结直肠癌（CRC）发病率明显上升 目前不向 50 岁以下的人群提供基于人群的筛查。 早发性（<50 岁）CRC (EOCRC) 更常在晚期出现症状性疾病 美国预防服务工作组表示，2020 年，阶段（III/IV）导致年轻病例死亡人数增加。 建议通过结肠镜检查或粪便检测进行 CRC 筛查的年龄降低至 45 岁。 所有符合筛查资格的人群（包括 50 岁以下人群）对这些方法的筛查采用率较低 遗传上处于高风险的人，因此 50 岁以下的无症状人群的依从性也可能是 此外，超过一半的 EOCRC 发生在 45 岁以下的人群中，而无需进行此类筛查。 为 EOCRC 提供一种微创、基于血液的筛查测试将提供一种经济高效的方法。 对患者进行分类和识别那些需要进行后续结肠镜检查的患者友好的选择，同时增加 癌症特异性甲基化 DNA 生物标志物的筛查依从性非常适合。 通过检测血浆中的循环肿瘤 DNA (ctDNA) 进行基于人群的癌症筛查 因为它们在特定癌症类型的患者中比肿瘤突变更普遍，并且 甲基化 SEPTIN9 血浆 ctDNA 测试“mSEPT9”（Epi proColon）在血浆中稳定（抗核酸酶）。 V2.0（表观基因组学）经 FDA 批准用于年龄≥50 岁拒绝结肠镜检查的人群的 CRC 筛查 该测试迫切需要在 50 岁以下的人群中进行评估，以确定其是否适合。 即便如此，多标记物检测可能比单标记物检测具有更高的敏感性。 因此，我们研究的目的是鉴定并确认一组甲基化 ctDNA 标记 基于血浆的 EOCRC 检测，并将该面板与 mSEPT9 的诊断性能进行比较 在年龄 <50 岁的人群中进行测试 在目标 1 中，我们将确定最常见的差异甲基化（肿瘤）。 通过对 420 万个 CpG 进行深度甲基测序，在 50 岁以下的 CRC 病例中进行 DNA 标记 配对原发肿瘤与 EOCRC 病例正常结肠组织以及健康白细胞中的位点 对照（以过滤掉非特异性标记）。所识别的标记将在独立样本中进行验证。 对于排名靠前的标记物，我们将开发具有高甲基化特异性的实时 PCR 检测方法。 分析敏感性并在转移性 CRC 病例的混合血浆中进行测试（以选择产生的标记物） 最强的信号）和健康的对照（以消除任何产生低水平的非特异性信号）来最终确定 在目标 2 中，我们将评估 mSEPT9 测试与其他测试的诊断性能特征。 本研究对经结肠镜检查证实的 CRC 病例和对照的血浆进行多标记物检测。 将产生多标记甲基化 ctDNA 组合，其诊断性能优于单标记 mSEPT9 测试可对 50 岁以下的无症状人群进行经济有效的基于血液的 CRC 筛查。