Automated and rapid detection of cancer in suspicious axillary lymph nodes in patients with breast cancer

Abstract

Preoperative staging of suspicious axillary lymph nodes (ALNs) allows patients to be triaged to ALN dissection or to sentinel lymph node biopsy (SLNB). Ultrasound-guided fine needle aspiration (FNA) and cytology of ALN is moderately sensitive but its clinical utility relies heavily on the cytologist’s experience. We proposed that the 5-h automated GeneXpert system-based prototype breast cancer detection assay (BCDA) that quantitatively measures DNA methylation in ten tumor-specific gene markers could provide a facile, accurate test for detecting cancer in FNA of enlarged lymph nodes. We validated the assay in ALN-FNA samples from a prospective study of patients (N = 230) undergoing SLNB. In a blinded analysis of 218 evaluable LN-FNAs from 108 malignant and 110 benign LNs by histology, BCDA displayed a sensitivity of 90.7% and specificity of 99.1%, achieving an area under the ROC curve, AUC of 0.958 (95% CI: 0.928–0.989; P < 0.0001). Next, we conducted a study of archival FNAs of ipsilateral palpable LNs (malignant, N = 72, benign, N = 53 by cytology) collected in the outpatient setting prior to neoadjuvant chemotherapy (NAC). Using the ROC-threshold determined in the prospective study, compared to cytology, BCDA achieved a sensitivity of 94.4% and a specificity of 92.5% with a ROC-AUC = 0.977 (95% CI: 0.953–1.000; P < 0.0001). Our study shows that the automated assay detects cancer in suspicious lymph nodes with a high level of accuracy within 5 h. This cancer detection assay, scalable for analysis to scores of LN FNAs, could assist in determining eligibility of patients to different treatment regimens.

High performance methylated DNA markers for detection of colon adenocarcinoma

“…The ultimate goal of our studies is to develop an automated method for detecting gene methylation in both tissue and blood, similar to the assays we are developing for breast cancer [ 28 , 29 ]. Therefore, as our first attempt in liquid biopsy of CRC, we tested plasma (300 μl) of stage IV CRC patients (carcinoma N = 20 and normal N = 20) using the cell-free circulating methylated DNA (cMethDNA) assay with the 8-gene subset TMEFF2 , COL6A2 , ZNF671, ARHGEF7 , TM6SF1 , MAL , GPX7 , and AKR1B1 .…”

“…We intend these studies to form the foundation for the development of a self-contained automated colon carcinoma assay. We have already demonstrated feasibility of this type of automated cancer detection approach in breast carcinoma [ 28 , 29 ].…”

“…In a pilot study, we tested these markers in cell-free plasma DNA of patients with stage IV CRC with our highly sensitive laboratory assay, cMethDNA. Our aim is for this gene marker panel to form the basis of development of a novel self-contained automated CC detection assay, similar to our prototype Breast Cancer Detection Assay (Research Use Only) [ 28 , 29 ]. The Breast Cancer Detection assay prototype is run on a GeneXpert ® system (available throughout Africa and India), and accurately and rapidly distinguishes between cancerous and benign growths both in fine needle aspirates of the breast lesion and enlarged axillary lymph nodes [ 28 , 29 ].…”

“…Our aim is for this gene marker panel to form the basis of development of a novel self-contained automated CC detection assay, similar to our prototype Breast Cancer Detection Assay (Research Use Only) [ 28 , 29 ]. The Breast Cancer Detection assay prototype is run on a GeneXpert ® system (available throughout Africa and India), and accurately and rapidly distinguishes between cancerous and benign growths both in fine needle aspirates of the breast lesion and enlarged axillary lymph nodes [ 28 , 29 ]. This novel automated CC screening technique, if applied to DNA from plasma or stool samples, has the potential to hasten cancer detection throughout the world by increasing screening participation rates due to its simplicity and through optimal risk stratification of patients who require follow-up with more invasive endoscopic techniques.…”

Associations between the Levels of Estradiol-, Progesterone-, and Testosterone-Sensitive MiRNAs and Main Clinicopathologic Features of Breast Cancer

“…Axillary lymph node (ALN) status is another important factor in the diagnosis of BC that predicts disease-free survival and overall survival. Conventional methods for diagnosing lymph node metastases (LNM) at the preoperative stage, such as ultrasound, mammography, and magnetic resonance imaging (MRI), have relatively low accuracy and sensitivity [2]. In recent years, sentinel lymph node biopsy (SLNB) has been used to diagnose lymph node status.…”