单一液体活检分析物(LBA)已用于转移性乳腺癌(MBC)的治疗选择。我们进行了综合统计分析,以检查使用多个LBA的临床相关性:匹配的循环肿瘤细胞(CTC)mRNA,CTC基因组DNA(gDNA),细胞外囊泡(EV)mRNA,和无细胞DNA(cfDNA)。
从26名激素受体阳性患者中抽取血液,HER2阴性MBC患者。使用多标记qPCR分析CTCmRNA和EVmRNA。来自去除了CTC的血液的血浆用于cfDNA分离。从mRNA耗尽的CTC裂解物中分离来自CTC的gDNA。通过靶向测序分析CTCgDNA和cfDNA。在每个分析物中进行分层聚类,并将其结果合并为一个评分,称为转移性乳腺癌患者中多种液体活检分析物的评估全部来自一个血液样本(ELIMA。得分),计算每种分析物对总体生存预测的贡献。奇异值分解(SVD),互信息计算,k均值聚类,并进行了图论分析以阐明各个分析物之间的依赖性。
两种/三种/四种LBA的组合增加了具有可操作信号的患者的患病率。将各个LBA的分层聚类结果汇总到ELIMA中。评分与总生存率高度显着相关,从而为使用多个LBA的附加价值提供了证据。互信息的计算表明,没有一个LBA独立于其他LBA,但是单个LBA描述其他LBA的能力相当有限-只有CTCgDNA可以部分描述其他三个LBA。SVD显示最强的奇异向量来自所有四个LBA,但大多数起源于CTCgDNA。在基于所有四个LBA的参数对患者进行k均值聚类之后,图论分析显示CTCERBB2变异仅存在于属于一个特定集群的患者中.
在本试验研究中,客观地证明了使用所有四种LBA的额外益处。这也表明CTCgDNA相对于其他LBA的相对优势。因此,多参数液体活检方法去卷积基因组和转录组的复杂性,应在临床实践中加以考虑.
Single liquid biopsy analytes (LBAs) have been utilized for therapy selection in metastatic breast cancer (MBC). We performed integrative statistical analyses to examine the clinical relevance of using multiple LBAs: matched circulating tumor cell (CTC) mRNA, CTC genomic DNA (gDNA), extracellular vesicle (EV) mRNA, and cell-free DNA (cfDNA).
Blood was drawn from 26 hormone receptor-positive, HER2-negative MBC patients. CTC mRNA and EV mRNA were analyzed using a multi-marker qPCR. Plasma from CTC-depleted blood was utilized for cfDNA isolation. gDNA from CTCs was isolated from mRNA-depleted CTC lysates. CTC gDNA and cfDNA were analyzed by targeted sequencing. Hierarchical clustering was performed within each analyte, and its results were combined into a score termed Evaluation of multiple Liquid biopsy analytes In Metastatic breast cancer patients All from one blood sample (ELIMA.score), which calculates the contribution of each analyte to the overall survival prediction. Singular value decomposition (SVD), mutual information calculation, k-means clustering, and graph-theoretic analysis were conducted to elucidate the dependence between individual analytes.
A combination of two/three/four LBAs increased the prevalence of patients with actionable signals. Aggregating the results of hierarchical clustering of individual LBAs into the ELIMA.score resulted in a highly significant correlation with overall survival, thereby bolstering evidence for the additive value of using multiple LBAs. Computation of mutual information indicated that none of the LBAs is independent of the others, but the ability of a single LBA to describe the others is rather limited-only CTC gDNA could partially describe the other three LBAs. SVD revealed that the strongest singular vectors originate from all four LBAs, but a majority originated from CTC gDNA. After k-means clustering of patients based on parameters of all four LBAs, the graph-theoretic analysis revealed CTC ERBB2 variants only in patients belonging to one particular cluster.
The additional benefits of using all four LBAs were objectively demonstrated in this pilot study, which also indicated a relative dominance of CTC gDNA over the other LBAs. Consequently, a multi-parametric liquid biopsy approach deconvolutes the genomic and transcriptomic complexity and should be considered in clinical practice.