Functional and molecular characterization of cancer stem-like cells in bladder cancer: a potential signature for muscle-invasive tumors

Abstract

Striking evidence associates cancer stem cells (CSCs) to the high recurrence rates and poor survival of patients with muscle-invasive bladder cancer (BC). However, the prognostic implication of those cells in risk stratification is not firmly established, mainly due to the functional and phenotypic heterogeneity of CSCs populations, as well as, to the conflicting data regarding their identification based on a single specific marker. This emphasizes the need to exploit putative CSC-related molecular markers with potential prognostic significance in BC patients.This study aimed to isolate and characterize bladder CSCs making use of different functional and molecular approaches. The data obtained provide strong evidence that muscle-invasive BC is enriched with a heterogeneous stem-like population characterized by enhanced chemoresistance and tumor initiating properties, able to recapitulate the heterogeneity of the original tumor. Additionally, a logistic regression analysis identified a 2-gene stem-like signature (SOX2 and ALDH2) that allows a 93% accurate discrimination between non-muscle-invasive and invasive tumors.Our findings suggest that a stemness-related gene signature, combined with a cluster of markers to more narrowly refine the CSC phenotype, could better identify BC patients that would benefit from a more aggressive therapeutic intervention targeting CSCs population.

β-Arrestins Regulate Stem Cell-Like Phenotype and Response to Chemotherapy in Bladder Cancer

“…We observed that expression of ARRB2 inversely correlated with CSC properties such as expression of CSC markers and selfrenewal potential in bladder cancer cell lines. A recent study demonstrated that ALDH2 and SOX2 provide a 2-gene stem-like signature that discriminates muscle-invasive from nonmuscle invasive tumors with an accuracy of 93% (43). Interestingly, we did not observe any significant effect on expression of SOX2 upon overexpression of ARRB2 in HT1376 and 5637 cells (Supplementary Fig.…”

Natural killer cell-based adoptive immunotherapy eradicates and drives differentiation of chemoresistant bladder cancer stem-like cells

“…We tested whether NK cells could induce CSCs towards a more differentiated phenotype rendering them susceptible to chemotherapy. Therefore, spheres were incubated with the supernatants of activated-NK cells for 4 h, followed by analysis of stemness related markers previously identified [5]. The ALDH activity, considered a functional readout of stemness, decreased by 60 % in spheres after 4 h incubation with NK supernatants (Fig.…”

“…Cultured cells were then stained with fluorochromeconjugated monoclonal antibodies against human blood surface antigens: CD3 PerCP/Cy5. 5 After a 4-h incubation period, the supernatants were harvested and counted for released radioactivity in a gamma counter (CRC-55tW Capintec), within a 51 Cr sensitivity energy window (300-400 keV). The specific lysis of target cells was calculated as follows: Percentage of specific lysis = (experimental release -spontaneous release)/(maximum release -spontaneous release) × 100.…”

“…The subcutaneous tumor model was induced by subcutaneous injection into the lower flank of 1 × 10 6 of Luc + HT-1376 cells suspended in 100 μL of a 1:1 PBS/Matrigel mixture. The orthotopic model that more closely resembles the clinical and histopathological features of primary MIBC was developed by intravesical instillation of Luc + HT-1376 cells as previously described [5]. Bioluminescent images were taken 24 h post-implantation and every 3 days to monitor engraftment and growth of tumor cells using an IVIS Lumina XR (Caliper LifeSciences, Hopkinton, MA, USA) after intraperitoneal injection with D-luciferin (150 mg/kg, Synchem, BHg, Germany) with the animals under anesthesia (100 mg/kg ketamine and 2.5 % of chlorpromazine solution).…”

PD‐1 blockade enhances the antitumor efficacy of GM‐CSF surface‐modified bladder cancer stem cells vaccine

“…Bladder cancer (BCa) is one of the most common malignancies, with high recurrence rate . As we all know, cancer stem cells (CSCs) may play an important role in tumor growth . Eliminating CSCs is a key issue in eradicating tumor.…”