Spychalski Griffin B, Lin Andrew A, Yang Stephanie J, Shen Hanfei, Rosario Jean, Tien Kyle, French Kate, Ghali Miriyam, Yee Stephanie, Yin Melinda, Feldman Michael D, Conant Emily F, Weinstein Susan P, Carpenter Erica L, Issadore David, Nayak Anupma
Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
Department of Bioengineering, School of Engineering and Applied Science, University of Pennsylvania, Philadelphia, PA, USA.
Breast Cancer Res. 2025 May 22;27(1):90. doi: 10.1186/s13058-025-02029-2.
Mammography screening has improved early breast cancer detection, leading to reduced mortality and lower rates of advanced breast cancer. However, mammography has a high false positive rate that results in over a million invasive breast biopsies of benign lesions in the US each year. Therefore, there is a need for noninvasive, blood-based diagnostics that can accurately assess risk of malignancy for women with indeterminate lesions identified by mammography, such as BI-RADS category 4 breast lesions. The aim of this study is to identify biomarkers from multiplexed extracellular vesicle liquid biopsy that can accurately classify mammographically detected BI-RADS 4 lesions.
We analyzed plasma from 113 prospectively enrolled subjects with BI-RADS 4 breast lesions, including 86 women with benign lesions and 27 women with malignant lesions (including 12 with stage I invasive carcinoma and 14 with ductal carcinoma in situ). None of the invasive carcinomas were metastatic. From each plasma sample, we used track etched magnetic nanopore technology to separately isolate HER2 and CD24 expressing extracellular vesicles (EVs) and measured their miRNA cargo using next-generation sequencing. We evaluated the performance of EV-miRNA biomarkers for classifying malignancy and applied LASSO classification to identify a panel of four complementary EV miRNA biomarkers that we validated by qPCR.
We identified 19 differentially enriched miRNA from HER2+ EVs and 11 differentially enriched miRNA from CD24+ EVs of women with malignant lesions compared to benign lesions. We observed individual miRNA with an AUC of up to 0.87 for miR-340-5p from HER2+ EVs and 0.75 for miR-223-3p from CD24+ EVs. LASSO classification selected a panel of four complementary EV miRNA for classifying breast cancer: miR-340-5p (HER2+ EVs), miR-598-3p (CD24+), miR-15b-5p (HER2+), and miR-126-3p (CD24+).
HER2+ and CD24+ EV subpopulations contain complementary biomarkers suitable for validation in larger studies that can accurately detect early-stage breast cancer among women with BI-RADS category 4 breast lesions.
乳腺钼靶筛查改善了早期乳腺癌的检测，降低了死亡率和晚期乳腺癌的发病率。然而，乳腺钼靶检查的假阳性率很高，导致美国每年有超过100万例对良性病变进行的侵入性乳腺活检。因此，需要一种非侵入性的、基于血液的诊断方法，能够准确评估乳腺钼靶检查发现的不确定病变女性（如BI-RADS 4类乳腺病变）的恶性风险。本研究的目的是从多重细胞外囊泡液体活检中鉴定生物标志物，以准确分类乳腺钼靶检测到的BI-RADS 4类病变。
我们分析了113名前瞻性纳入的患有BI-RADS 4类乳腺病变受试者的血浆，其中包括86名良性病变女性和27名恶性病变女性（包括12名I期浸润性癌患者和14名原位导管癌患者）。所有浸润性癌均无转移。从每个血浆样本中，我们使用径迹蚀刻磁性纳米孔技术分别分离表达HER2和CD24的细胞外囊泡（EV），并使用下一代测序测量其miRNA含量。我们评估了EV-miRNA生物标志物对恶性肿瘤分类的性能，并应用套索分类法鉴定了一组四个互补的EV miRNA生物标志物，并通过qPCR进行了验证。
与良性病变女性相比，我们在恶性病变女性的HER2+ EV中鉴定出19种差异富集的miRNA，在CD24+ EV中鉴定出11种差异富集的miRNA。我们观察到，HER2+ EV中的miR-340-5p和CD24+ EV中的miR-223-3p的单个miRNA的AUC分别高达0.87和0.75。套索分类法选择了一组四个互补的EV miRNA用于乳腺癌分类：miR-340-5p（HER2+ EV）、miR-598-3p（CD24+）、miR-15b-5p（HER2+）和miR-126-3p（CD24+）。
HER2+和CD24+ EV亚群包含适合在更大规模研究中验证的互补生物标志物，可准确检测BI-RADS 4类乳腺病变女性中的早期乳腺癌。
