A Laboratory-Friendly CTC Identification: Comparable Double-Immunocytochemistry with Triple-Immunofluorescence.
Raed SulaimanPradip DeJennifer Carlson AskeXiaoqian LinAdam DaleEthan VaselaarNischal KoiralaCheryl AgetonKris GasterJoshua PlordeBenjamin SolomonBradley ThaemertPaul MeyerLuis Rojas EspaillatDavid StarksNandini DeyPublished in: Cancers (2022)
The source of circulating tumor cells (CTC) in the peripheral blood of patients with solid tumors are from primary cancer, metastatic sites, and a disseminated tumor cell pool. As 90% of cancer-related deaths are caused by metastatic progression and/or resistance-associated treatment failure, the above fact justifies the undeniable predictive and prognostic value of identifying CTC in the bloodstream at stages of the disease progression and resistance to treatment. Yet enumeration of CTC remains far from a standard routine procedure either for post-surgery follow-ups or ongoing adjuvant therapy. The most compelling explanation for this paradox is the absence of a convenient, laboratory-friendly, and cost-effective method to determine CTC. We presented a specific and sensitive laboratory-friendly parallel double-detection format method for the simultaneous isolation and identification of CTC from peripheral blood of 91 consented and enrolled patients with various malignant solid tumors of the lung, endometrium, ovary, esophagus, prostate, and liver. Using a pressure-guided method, we used the size-based isolation to capture CTC on a commercially available microfilter. CTC identification was carried out by two expression marker-based independent staining methods, double-immunocytochemistry parallel to standard triple-immunofluorescence. The choice of markers included specific markers for epithelial cells, EpCAM and CK8,18,19, and exclusion markers for WBC, CD45. We tested the method's specificity based on the validation of the staining method, which included positive and negative spiked samples, blood from the healthy age-matched donor, healthy age-matched leucopaks, and blood from metastatic patients. Our user-friendly cost-effective CTC detection technique may facilitate the regular use of CTC detection even in community-based cancer centers for prognosis, before and after surgery.
Keyphrases
- circulating tumor cells
- circulating tumor
- peripheral blood
- squamous cell carcinoma
- prostate cancer
- papillary thyroid
- minimally invasive
- end stage renal disease
- poor prognosis
- chronic kidney disease
- newly diagnosed
- stem cells
- single cell
- escherichia coli
- ejection fraction
- cell therapy
- low cost
- real time pcr
- clinical practice
- coronary artery disease
- coronary artery bypass
- acute coronary syndrome
- bioinformatics analysis
- combination therapy
- replacement therapy
- benign prostatic hyperplasia
- decision making