Validation of circulating tumor DNA assays for detection of metastatic melanoma

循环肿瘤 DNA 检测转移性黑色素瘤检测的验证

基本信息

批准号：
10015828
负责人：
DAVID POLSKY
金额：
$ 3.39万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-07 至 2021-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10015828
关键词：
Adjuvant Adjuvant Therapy BRAF gene Biological Assay Biological Markers Cancer Patient Cells Characteristics Clinic Clinical Clinical Data Clinical Trials Collaborations Consensus DNA Data Data Set Detection Development Diagnostic Diagnostic radiologic examination Disease Disease Progression Excision Gene Mutation Genes Goals Hot Spot Laboratories Lactate Dehydrogenase Lymphatic MAP Kinase Gene Malignant Neoplasms Measures Metastatic Melanoma Molecular Monitor Mutation Mutation Detection Neoplasm Metastasis Nivolumab Operative Surgical Procedures Outcome Pathologic Pathway interactions Patients Performance Phase Pilot Projects Plasma Plasma Cells Procedures Process Promoter Regions Quality Control RNA-Directed DNA Polymerase Reagent Recurrence Relapse Reproducibility Research Resected Running Sampling Scanning Sensitivity and Specificity Serum Specificity Specimen Staging System Surgical Management Systemic disease Technology Telomerase Testing Time Tumor Burden United States Unresectable Validation Visceral base blood-based biomarker burden of illness circulating DNA design digital disorder later incidence prevention effective therapy efficacy study follow-up genetic testing immune checkpoint blockade immune checkpoint blockers improved melanoma mutant predictive modeling prospective reagent standardization response routine imaging serological marker specific biomarkers targeted treatment treatment comparison trial comparing tumor tumor DNA

项目摘要

Project Summary Leveraging the high sensitivity, specificity and quantitative capability of the droplet digital PCR (ddPCR) platform, this project seeks to analytically and clinically validate ddPCR assays for 7 melanoma-associated hot- spot mutations for use as blood-based biomarkers of disease recurrence in patients with resected metastatic melanoma. In the United States the only commonly used serologic marker to monitor disease recurrence in patients with resected metastatic melanoma is serum lactate dehydrogenase (LDH), which suffers from low sensitivity and specificity. With new, effective therapies for systemic disease being applied in the adjuvant setting, a sensitive and specific biomarker that can identify early disease recurrence could prompt clinicians to change patient management while the patient has a low tumor burden. Recently, highly sensitive and specific technologies, such as ddPCR, have revealed that cancer patients have abnormally high levels of cell-free, tumor-associated DNA (ctDNA) circulating in their plasma that generally correlate positively with metastatic tumor burden. We conducted a pilot study utilizing ddPCR to detect changes in mutant BRAF and NRAS ctDNA levels in patients with unresectable stage IV melanoma who were undergoing treatment with BRAF- targeted therapies or immune checkpoint blockade. Importantly for this application, we found that at the time of treatment initiation, ctDNA was a much more sensitive marker of low disease burden than LDH. ctDNA levels were elevated in 71% of patients with RECIST scores < 5cm at the start of systemic treatment compared to LDH, which was only elevated in 8% of patients. While these encouraging results were obtained with assays that detect 1 of the 5 of the most common melanoma-associated BRAF and NRAS mutations, collectively these 5 mutations are present in only 55% - 65% of patients. Recently, 2 functional, hotspot mutations in the promoter region of the telomerase reverse-transcriptase gene (TERT) were identified in many cancers including melanoma. Our preliminary studies have also found these TERT mutations frequently in melanoma and when evaluated with BRAF and NRAS mutations, 86% of patients have 1 or more of the 7 BRAF, NRAS or TERT mutations in their melanomas. In this application, we will analytically validate ddPCR assays for the 7 common mutations in BRAF, NRAS and TERT, and test the hypothesis that a rise in ctDNA levels, as measured by 1 of these 7 ddPCR assays will predict radiographic recurrence in resected metastatic patients prior to routine imaging scans. Development of plasma biomarkers that can accurately detect disease progression with high sensitivity can augment or replace the routine radiographic scans obtained every 3 months with scans ordered in response to a rise in the plasma ctDNA level. Also it could lay the groundwork for a clinical trial comparing the utility of adjuvant therapy for all resected patients versus selecting patients for treatment when evidence of micrometastatic disease emerges based on a rise in ctDNA levels.

项目概要利用液滴数字 PCR (ddPCR) 的高灵敏度、特异性和定量能力平台，该项目旨在分析和临床验证 7 种黑色素瘤相关热点的 ddPCR 检测点突变用作切除转移患者疾病复发的血液生物标志物黑色素瘤。在美国，唯一常用的监测疾病复发的血清学标志物切除的转移性黑色素瘤患者的血清乳酸脱氢酶（LDH）水平较低敏感性和特异性。随着新的、有效的全身性疾病疗法在佐剂中的应用在这种情况下，一种可以识别早期疾病复发的敏感且特异的生物标志物可能会促使临床医生在患者肿瘤负荷较低的情况下改变患者管理。最近，高度敏感和具体 ddPCR 等技术显示癌症患者体内的游离细胞水平异常高，血浆中循环的肿瘤相关 DNA (ctDNA) 通常与转移呈正相关肿瘤负荷。我们利用 ddPCR 进行了一项初步研究，以检测突变 BRAF 和 NRAS 的变化接受 BRAF 治疗的不可切除 IV 期黑色素瘤患者的 ctDNA 水平靶向治疗或免疫检查点阻断。对于这个应用程序来说重要的是，我们发现当时在治疗开始时，ctDNA 是比 LDH 更敏感的低疾病负担标志物。 ctDNA 与全身治疗开始时 RECIST 评分 < 5cm 的患者相比，71% 的患者水平升高 LDH，仅在 8% 的患者中升高。虽然这些令人鼓舞的结果是通过检测 5 种最常见的黑色素瘤相关 BRAF 和 NRAS 突变中的 1 种的检测方法，总的来说，这 5 种突变仅存在于 55% - 65% 的患者中。最近，2个功能性、热点端粒酶逆转录酶基因（TERT）启动子区域的突变在许多癌症，包括黑色素瘤。我们的初步研究也发现这些TERT突变频繁发生在黑色素瘤，当用 BRAF 和 NRAS 突变进行评估时，86% 的患者具有 7 种突变中的一种或多种黑色素瘤中的 BRAF、NRAS 或 TERT 突变。在此应用中，我们将分析验证 ddPCR 检测 BRAF、NRAS 和 TERT 中 7 种常见突变，并检验 ctDNA 增加的假设通过这 7 种 ddPCR 测定中的一种测定的水平将预测切除的转移性肿瘤的放射照相复发患者在常规影像扫描之前。开发可以准确检测疾病的血浆生物标志物高灵敏度的进展可以增强或取代每 3 次获得的常规放射线扫描几个月后，为了应对血浆 ctDNA 水平的升高而进行了扫描。也可以打下基础一项临床试验，比较辅助治疗对所有切除患者与选择患者进行治疗的效用当 ctDNA 水平升高出现微转移性疾病证据时进行治疗。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Cell-Free DNA in Dermatology Research.

皮肤病学研究中的无细胞 DNA。

DOI：
发表时间：
2022-06
期刊：
影响因子：
0
作者：
Wiggins, Jennifer M;Ali, Saim;Polsky, David
通讯作者：
Polsky, David

Validation of Circulating Tumor DNA Assays for Detection of Metastatic Melanoma.

循环肿瘤 DNA 检测转移性黑色素瘤检测的验证。

DOI：
发表时间：
2020
期刊：
影响因子：
0
作者：
Syeda, Mahrukh M;Wiggins, Jennifer M;Corless, Broderick;Spittle, Cindy;Karlin;Polsky, David
通讯作者：
Polsky, David

Circulating tumour DNA in patients with advanced melanoma treated with dabrafenib or dabrafenib plus trametinib: a clinical validation study.

使用达拉非尼或达拉非尼加曲美替尼治疗的晚期黑色素瘤患者的循环肿瘤 DNA：一项临床验证研究。

DOI：
发表时间：
2021-03
期刊：
影响因子：
0
作者：
Syeda, Mahrukh M;Wiggins, Jennifer M;Corless, Broderick C;Long, Georgina V;Flaherty, Keith T;Schadendorf, Dirk;Nathan, Paul D;Robert, Caroline;Ribas, Antoni;Davies, Michael A;Grob, Jean Jacques;Gasal, Eduard;Squires, Matthew;Marker, Mahtab;G
通讯作者：
G