Cancer Stem Cells Sensitivity Assay (STELLA) in Patients with Advanced Lung and Colorectal Cancer: A Feasibility Study

Manolo D'Arcangelo, Matilde Todaro, Jessica Salvini, Antonina Benfante, Maria Luisa Colorito, Armida D'Incecco, Lorenza Landi, Tiziana Apuzzo, Elisa Rossi, Spartaco Sani, Giorgio Stassi, Federico Cappuzzo, Manolo D'Arcangelo, Matilde Todaro, Jessica Salvini, Antonina Benfante, Maria Luisa Colorito, Armida D'Incecco, Lorenza Landi, Tiziana Apuzzo, Elisa Rossi, Spartaco Sani, Giorgio Stassi, Federico Cappuzzo

Abstract

Background: Cancer stem cells represent a population of immature tumor cells found in most solid tumors. Their peculiar features make them ideal models for studying drug resistance and sensitivity. In this study, we investigated whether cancer stem cells isolation and in vitro sensitivity assay are feasible in a clinical setting.

Methods: Cancer stem cells were isolated from effusions or fresh cancer tissue of 23 patients who progressed after standard therapy failure. Specific culture conditions selected for immature tumor cells that express markers of stemness. These cells were exposed in vitro to chemotherapeutic and targeted agents.

Results: Cancer stem cells were extracted from liver metastases in 6 cases (25%), lung nodules in 2 (8%), lymph node metastases in 3 (12.5%) and pleural/peritoneal/pericardial effusion in 13 (54%). Cancer stem cells were successfully isolated in 15 patients (63%), including 14 with lung cancer (93.3%). A sensitivity assay was successfully performed in 7 patients (30.4%), with a median of 15 drugs/combinations tested (range 5-28) and a median time required for results of 51 days (range 37-95).

Conclusion: The approach used for the STELLA trial allowed isolation of cancer stem cells in a consistent proportion of patients. The low percentage of cases completing the full procedure and the long median time for obtaining results highlights the need for a more efficient procedure.

Trial registration: ClinalTrials.gov NCT01483001.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exists.

Figures

Fig 1. Flow chart of the study.
Fig 1. Flow chart of the study.
Fig 2. Phenotypical characterization of immature tumor…
Fig 2. Phenotypical characterization of immature tumor cells isolated from tumor samples with immunofluorescence assay.
The following panel of images refers to the characterization of cells isolated from the sample of a patient enrolled in the study. Green labelling is typical of positive cells. All markers of stemness (CD133, CD166, Oct 3/4, ALDH1, CD90) are positive in this CSC sample.
Fig 3. Evaluation of cell mortality with…
Fig 3. Evaluation of cell mortality with acridine orange/ethidium bromide (AO/EB) staining after exposition to chemotherapeutic agents.
AO is a vital dye and stains both live and dead cells. EB penetrates into cells with disrupted cytoplasmic membrane, staining only dead cells. Therefore, viable cells appear uniformly green and dead cells are labelled in orange. a) Low sensitivity (cell mortality 5%). b) Average sensitivity (cell mortality 40%). c) High sensitivity (cell mortality 80%).

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