Methods for detecting circulating cancer stem cells (CCSCs) as a novel approach for diagnosis of colon cancer relapse/metastasis

Carla Kantara, Malaney Ravae O'Connell, Gurinder Luthra, Aakash Gajjar, Shubhashish Sarkar, Robert Leo Ullrich, Pomila Singh

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Cancer stem cells (CSCs) are believed to be resistant to currently available therapies and may be responsible for relapse of cancer in patients. Measuring circulating tumor cells (CTCs) in the blood of patients has emerged as a non-invasive diagnostic procedure for screening patients who may be at high risk for developing metastatic cancers or relapse of the cancer disease. However, accurate detection of CTCs has remained a problem, as epithelial-cell markers used to date are not always reliable for detecting CTCs, especially during epithelial-mesenchymal transition. As CSCs are required to initiate metastatic tumors, our goal was to optimize and standardize a method for identifying circulating CSCs (CCSCs) in patients, using established CSC markers. Here, we report for the first time the detection of CCSCs in the blood of athymic nude mice, bearing metastatic tumors, and in the blood of patients positive for colonic adenocarcinomas. Using a simple and non-expensive method, we isolated a relatively pure population of CSCs (CD45-/CK19+), free of red blood cells and largely free of contaminating CD45+ white blood cells. Enriched CCSCs from patients with colon adenocarcinomas had a malignant phenotype and co-expressed CSC markers (DCLK1/LGR5) with CD44/Annexin A2. CSCs were not found in the blood of non-cancer patients, free of colonic growths. Enriched CCSCs from colon cancer patients grew primary spheroids, suggesting the presence of tumor-initiating cells in the blood of these patients. In conclusion, we have developed a novel diagnostic assay for detecting CSCs in circulation, which may more accurately predict the risk of relapse or metastatic disease in patients. As CSCs can potentially initiate metastatic growths, patients positive for CCSCs can be treated with inhibitory agents that selectively target CSCs, besides conventional treatments, to reduce the risk of relapse/metastatic disease for improving clinical outcomes.

Original languageEnglish (US)
Pages (from-to)100-112
Number of pages13
JournalLaboratory Investigation
Volume95
Issue number1
DOIs
StatePublished - Jan 31 2015

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Neoplastic Stem Cells
Colonic Neoplasms
Neoplasm Metastasis
Recurrence
Circulating Neoplastic Cells
Neoplasms
Nude Mice
Adenocarcinoma
Annexin A2
Epithelial-Mesenchymal Transition
Growth
Colon

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Cell Biology
  • Molecular Biology

Cite this

Methods for detecting circulating cancer stem cells (CCSCs) as a novel approach for diagnosis of colon cancer relapse/metastasis. / Kantara, Carla; O'Connell, Malaney Ravae; Luthra, Gurinder; Gajjar, Aakash; Sarkar, Shubhashish; Ullrich, Robert Leo; Singh, Pomila.

In: Laboratory Investigation, Vol. 95, No. 1, 31.01.2015, p. 100-112.

Research output: Contribution to journalArticle

Kantara, Carla ; O'Connell, Malaney Ravae ; Luthra, Gurinder ; Gajjar, Aakash ; Sarkar, Shubhashish ; Ullrich, Robert Leo ; Singh, Pomila. / Methods for detecting circulating cancer stem cells (CCSCs) as a novel approach for diagnosis of colon cancer relapse/metastasis. In: Laboratory Investigation. 2015 ; Vol. 95, No. 1. pp. 100-112.
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