ABSTRACT
BACKGROUND: Personalized targeted treatment in metastatic breast cancer relies on accurate assessment of molecular aberrations, e.g. overexpression of Human Epidermal growth factor Receptor 2 (HER-2). Molecular interrogation of circulating tumor cells (CTCs) can provide an attractive alternative for real-time biomarker assessment. However, implementation of CellSearch-based HER-2 analysis has been limited. Immunofluorescent (IF) image interpretation is crucial, as different HER-2 categories have been described. Major questions in CTC research are how these IF categories reflect gene expression and amplification, and if we should consider 'medium' HER-2 expressing CTCs for patient selection. METHODS: Tumor cells from spiked cell lines (n = 8) and CTCs (n = 116 samples) of 85 metastatic breast cancer patients were enriched using CellSearch. Comparative analysis of HER-2 expression by IF imaging (ACCEPT, DEPArray, and visual scoring) with qRT-PCR and HER-2/neu FISH was performed. RESULTS: Automated IF HER-2-profiling by DEPArray and ACCEPT delivered comparable results. There was a 98% agreement between 17 trained observers (visual scoring) and ACCEPT considering HER-2neg and HER-2high expressing CTCs. However, 89% of HER-2med expressing CTCs by ACCEPT were scored negative by observers. HER-2high expressing tumor cells demonstrated HER-2/neu gene amplification, whereas HER-2neg and HER-2med expressing tumor cells and CTCs by ACCEPT were copy-number neutral. All patients with HER-2-positive archival tumors had ≥1 HER-2high expressing CTCs, while 80% of HER-2-negative patients did not. High relative gene expression of HER-2 measured on enriched CTC lysates correlated with having ≥1 HER-2high expressing CTCs. CONCLUSION: Automated images analysis has enormous potential for clinical implementation. HER-2 characterization and clinical trial design should be focused on HER-2high expressing CTCs.
Subject(s)
Biomarkers, Tumor , Breast Neoplasms/genetics , Breast Neoplasms/pathology , Neoplastic Cells, Circulating/metabolism , Neoplastic Cells, Circulating/pathology , Receptor, ErbB-2/genetics , Cell Line, Tumor , Female , Gene Amplification , Humans , In Situ Hybridization, Fluorescence , RNA, Messenger/genetics , WorkflowABSTRACT
OBJECTIVES: Renal cysts of the sinuses or parapelvic cysts constitute a rare form of renal cyst, usually associated with low back pain. Symptomatic forms may be treated by percutaneous aspiration, but the treatment of these cysts has changed considerably over recent years due to the proximity of the renal hilum. Percutaneous treatment is now contraindicated due to its high complication rate, and open surgery is an invasive technique. Laparoscopy appears to be the most appropriate and most effective solution. PATIENTS AND METHODS: The authors report five cases of parapelvic renal cysts all presenting with renal colic and diagnosed by ultrasound, and confirmed by CT in three cases. Treatment consisted of retroperitoneal laparoscopy using four trocars allowing cyst resection with no intraoperative complications or conversion. RESULTS: The mean operating time was 55 minutes (range: 40 to 70 minutes). The postoperative course was uneventful and all patients were discharged on postoperative D1 or D2. No recurrence was observed after a mean follow-up of 6 months (range: 3 to 15 months). CONCLUSION: Retroperitoneal laparoscopic treatment of parapelvic renal cysts is an effective approach, with a low morbidity and less postoperative pain, a short convalescence period and good cosmetic results.
