Department of Oncology and Pathology, Cancer Center Karolinska, Karolinska Institutet and University Hospital, Stockholm, Sweden.
Department of Microbiology, Tumor and Cell Biology (MTC), Karolinska Institutet, Stockholm, Sweden.
Mol Oncol. 2019 Feb;13(2):376-391. doi: 10.1002/1878-0261.12410. Epub 2019 Jan 7.
There are increasing demands for informative cancer biomarkers, accessible via minimally invasive procedures, both for initial diagnostics and for follow-up of personalized cancer therapy, including immunotherapy. Fine-needle aspiration (FNA) biopsy provides ready access to relevant tissue samples; however, the minute amounts of sample require sensitive multiplex molecular analysis to be of clinical biomarker utility. We have applied proximity extension assays (PEA) to analyze 167 proteins in FNA samples from patients with breast cancer (BC; n = 25) and benign lesions (n = 32). We demonstrate that the FNA BC samples could be divided into two main clusters, characterized by differences in expression levels of the estrogen receptor (ER) and the proliferation marker Ki67. This clustering corresponded to some extent to established BC subtypes. Our analysis also revealed several proteins whose expression levels differed between BC and benign lesions (e.g., CA9, GZMB, IL-6, VEGFA, CXCL11, PDL1, and PCD1), as well as several chemokines correlating with ER and Ki67 status (e.g., CCL4, CCL8, CCL20, CXCL8, CXCL9, and CXCL17). Finally, we also identified three signatures that could predict Ki67 status, ER status, and tumor grade, respectively, based on a small subset of proteins, which was dominated by chemokines. To our knowledge, expression profiles of CCL13 in benign lesions and BC have not previously been described but were shown herein to correlate with proliferation (P = 0.00095), suggesting a role in advanced BC. Given the broad functional range of the proteins analyzed, immune-related proteins were overrepresented among the observed alterations. Our pilot study supports the emerging role of chemokines in BC progression. Due to the minimally traumatic sampling and clinically important molecular information for therapeutic decisions, this methodology is promising for future immunoscoring and monitoring of treatment efficacy in BC.
人们越来越需要能够通过微创程序获得的有信息价值的癌症生物标志物，无论是用于初始诊断还是用于个性化癌症治疗（包括免疫疗法）的后续监测。细针穿刺（FNA）活检可方便地获取相关组织样本；然而，由于样本量非常少，因此需要进行敏感的多重分子分析，才能将其用作有临床意义的生物标志物。我们应用邻近延伸分析（PEA）分析了 25 例乳腺癌（BC）患者和 32 例良性病变患者的 FNA 样本中的 167 种蛋白质。我们证明，FNA BC 样本可以分为两个主要簇，其特征是雌激素受体（ER）和增殖标志物 Ki67 的表达水平存在差异。这种聚类在一定程度上与已确立的 BC 亚型相对应。我们的分析还揭示了一些在 BC 和良性病变之间表达水平不同的蛋白质（例如，CA9、GZMB、IL-6、VEGFA、CXCL11、PDL1 和 PCD1），以及一些与 ER 和 Ki67 状态相关的趋化因子（例如，CCL4、CCL8、CCL20、CXCL8、CXCL9 和 CXCL17）。最后，我们还基于一小部分蛋白质（主要是趋化因子）分别鉴定出了三个可分别预测 Ki67 状态、ER 状态和肿瘤分级的特征。据我们所知，以前尚未描述过良性病变和 BC 中 CCL13 的表达谱，但本研究表明其与增殖相关（P=0.00095），提示其在晚期 BC 中发挥作用。鉴于分析的蛋白质具有广泛的功能范围，观察到的改变中免疫相关蛋白的含量过高。我们的初步研究支持趋化因子在 BC 进展中的新兴作用。由于采样创伤小，且为治疗决策提供了重要的临床分子信息，因此这种方法有望用于未来的免疫评分和 BC 治疗效果监测。
