Department of Translational Molecular Medicine, John Wayne Cancer Institute, Saint John's Health Center, PHS, Santa Monica, CA.
Medical Data Research Center, Providence Saint Joseph Health, Portland, OR.
Clin Chem. 2020 Jan 1;66(1):169-177. doi: 10.1373/clinchem.2019.307140.
Blood molecular profiling of circulating tumor cells (CTCs) can enable monitoring of patients with metastatic melanoma during checkpoint inhibitor immunotherapy (CII) and in combination with targeted therapies. We developed a microfluidics-based CTC platform to explore CTC profiling utility in CII-treated patients with melanoma using a melanoma messenger RNA (mRNA)/DNA biomarker panel.
Blood samples (n = 213) were collected prospectively from 75 American Joint Committee on Cancer-staged III/IV melanoma patients during CII treatment and those enriched for CTCs. CTC profiling was performed using 5 known melanoma mRNA biomarkers and BRAF V600E DNA mutation. CTC biomarker status associations with clinical outcomes were assessed.
CTCs were detected in 88% of blood samples from patients with melanoma. CTC-derived biomarkers and clinical variables analyzed using classification and regression tree analysis revealed that a combination of lactate dehydrogenase, CTC-mRNA biomarkers, and tumor BRAF-mutation status was indicative of clinical outcomes for patients with stage IV melanoma (n = 52). The panel stratified low-risk and high-risk patients, whereby the latter had poor disease-free (P = 0.03) and overall survival (P = 0.02). Incorporation of a DNA biomarker with mRNA profiling increased overall CTC-detection capability by 57% compared to mRNA profiling only. RNA sequencing of isolated CTCs identified significant catenin beta 1 (CTNNB1) overexpression (P <0.01) compared to nondisease donor blood. CTC-CTNNB1 was associated with progressive disease/stable disease compared to complete-responder patient status (P = 0.02). Serial CTC profiling identified subclinical disease in patients who developed progressive disease during treatment/follow-up.
CTC-derived mRNA/DNA biomarkers have utility for monitoring CII, targeted, and combinatorial therapies in metastatic melanoma patients.
循环肿瘤细胞（CTC）的血液分子谱分析可实现对接受免疫检查点抑制剂治疗（CII）的转移性黑色素瘤患者以及接受靶向治疗联合 CII 治疗的患者进行监测。我们开发了一种基于微流控的 CTC 平台，使用黑色素瘤信使 RNA（mRNA）/DNA 生物标志物面板来探索 CTC 谱分析在接受 CII 治疗的黑色素瘤患者中的应用。
前瞻性地从 75 名接受 CII 治疗的美国癌症联合委员会分期 III/IV 期黑色素瘤患者的血液样本中收集（n=213），这些样本均富集了 CTC。使用 5 种已知的黑色素瘤 mRNA 生物标志物和 BRAF V600E DNA 突变对 CTC 进行了谱分析。评估了 CTC 生物标志物与临床结果的关联。
88%的黑色素瘤患者的血液样本中检测到 CTC。使用分类和回归树分析分析 CTC 衍生的生物标志物和临床变量，发现乳酸脱氢酶、CTC-mRNA 生物标志物和肿瘤 BRAF 突变状态的组合可预测 IV 期黑色素瘤患者（n=52）的临床结局。该面板将低风险和高风险患者分层，后者无疾病进展生存期（P=0.03）和总生存期（P=0.02）较差。与仅进行 mRNA 谱分析相比，将 DNA 生物标志物与 mRNA 谱分析相结合可将总体 CTC 检测能力提高 57%。对分离的 CTC 进行 RNA 测序发现，与非疾病供体血液相比，连环蛋白 beta 1（CTNNB1）明显过表达（P<0.01）。与完全缓解患者相比，CTC-CTNNB1 与进展性疾病/稳定疾病相关（P=0.02）。对接受治疗/随访过程中发生进展性疾病的患者进行连续 CTC 谱分析，发现亚临床疾病。
CTC 衍生的 mRNA/DNA 生物标志物可用于监测转移性黑色素瘤患者的 CII、靶向和联合治疗。
