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Deciphering the cellular source of tumor relapse identifies CD44 as a major therapeutic target in pancreatic adenocarcinoma.

Molejon MI, Tellechea JI, Loncle C, Gayet O, Gilabert M, Duconseil P, Lopez-Millan MB, Moutardier V, Gasmi M, Garcia S, Turrini O, Ouaissi M, Poizat F, Dusetti N, Iovanna J - Oncotarget (2015)

Bottom Line: The origin and biological characteristics of residual tumor cells in PDAC still remain unclear.During PDAC relapse, proliferating CD44+ cells decrease expression of ZEB1, while overexpressing the MUC1 protein, and gain morphological and biological characteristics of differentiation.We confirmed the propagation of CD44+ cells in samples from cases of human relapse, following standard PDAC treatment.

View Article: PubMed Central - PubMed

Affiliation: Centre de Recherche en Cancérologie de Marseille (CRCM), INSERM U1068, CNRS UMR 7258, Aix-Marseille Université and Institut Paoli-Calmettes, Parc Scientifique et Technologique de Luminy, Marseille, France.

ABSTRACT
It has been commonly found that in patients presenting Pancreatic Ductal Adenocarcinoma (PDAC), after a period of satisfactory response to standard treatments, the tumor becomes non-responsive and patient death quickly follows. This phenomenon is mainly due to the rapid and uncontrolled development of the residual tumor. The origin and biological characteristics of residual tumor cells in PDAC still remain unclear. In this work, using PDACs from patients, preserved as xenografts in nude mice, we demonstrated that a residual PDAC tumor originated from a small number of CD44+ cells present in the tumor. During PDAC relapse, proliferating CD44+ cells decrease expression of ZEB1, while overexpressing the MUC1 protein, and gain morphological and biological characteristics of differentiation. Also, we report that CD44+ cells, in primary and residual PDAC tumors, are part of a heterogeneous population, which includes variable numbers of CD133+ and EpCAM+ cells. We confirmed the propagation of CD44+ cells in samples from cases of human relapse, following standard PDAC treatment. Finally, using systemic administration of anti-CD44 antibodies in vivo, we demonstrated that CD44 is an efficient therapeutic target for treating tumor relapse, but not primary PDAC tumors. We conclude that CD44+ cells generate the relapsing tumor and, as such, are themselves promising therapeutic targets for treating patients with recurrent PDAC.

No MeSH data available.


Related in: MedlinePlus

Expression of Cancer Stem Cell (CSC) markers in PDXsImmunofluorescent labeling of (A) CD44-FITC (green) and 4′,6-diamidino-2-phenylindole (DAPI), (blue); (B) EpCAM-FITC (green); DAPI (blue) and (C) CD133-FITC (green); DAPI (blue) in PDXs upon vehicle or gemcitabine treatment, show the distribution of each CSC marker in tumor tissues. (D) Quantification of the number of positive cells in each PDX. Scale bar represents 50 μm. Error bars ± SEM; n=3 per group. *P<0.05, **P<0.001 compared to vehicle treatment.
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Figure 2: Expression of Cancer Stem Cell (CSC) markers in PDXsImmunofluorescent labeling of (A) CD44-FITC (green) and 4′,6-diamidino-2-phenylindole (DAPI), (blue); (B) EpCAM-FITC (green); DAPI (blue) and (C) CD133-FITC (green); DAPI (blue) in PDXs upon vehicle or gemcitabine treatment, show the distribution of each CSC marker in tumor tissues. (D) Quantification of the number of positive cells in each PDX. Scale bar represents 50 μm. Error bars ± SEM; n=3 per group. *P<0.05, **P<0.001 compared to vehicle treatment.

Mentions: Considering that CSCs represent the only cell population with tumor-initiating potential, we hypothesized that these cells, which seem to play a crucial role in treatment resistance, are associated with the phenotypic transformation described above. These cells are capable of resisting toxicity by drugs, such as gemcitabine, that target highly proliferative cells, since they enter into the cell cycle less frequently and only divide in response to certain stimuli, which have yet to be fully identified. We sought to investigate the potential involvement of CSCs in driving residual tumor growth following gemcitabine treatment. We used immunohistochemistry to analyze the expression of some CSC-associated cell surface markers on these tumors. We selected PDXs which were either sensitive to, or moderately or highly resistant to gemcitabine therapy (X-IPC, C-NOR and AO-IPC, respectively). Those PDXs reaching a volume of 400 mm3 received either vehicle or 100 mg/kg gemcitabine, twice weekly. Immunofluorescence analysis revealed that the percentage of CD44+, EpCAM+ and CD133+ cells in gemcitabine-treated tumors increased systematically 3-4 folds when compared to vehicle-treated tumors. As shown in Figure 2A, CD44 expression was found in 22±2.8%, 25±2.3% and 30±4.4% of vehicle-treated tumors, while its expression increased to 59±7.0%, 65±7.2% and 73±7.9% in gemcitabine-treated X-IPC, C-NOR and AO-IPC, respectively. Similarly, EpCAM was expressed in 3.6±1.0%, 0.2±0.7% and 4.3±0.8% of vehicle-treated tumors and increased to 24±2.8%, 1.2±1.3% and 31±3.3% in gemcitabine-treated X-IPC, C-NOR and AO-IPC, respectively (Figure 2B). Finally, CD133 was expressed in 5.0±0.5%, 1.0±0.9% and 5.1±0.8% of vehicle-treated tumors, while in gemcitabine-treated tumors its expression was found in 18±2.7%, 3.2±1.2% and 5.3±1.2% of X-IPC, C-NOR and AO-IPC, respectively (Figure 2C). Quantification is shown in Figure 2D. Thus, these data strongly suggest that gemcitabine treatment may select a highly enriched population of cells expressing CSC-associated markers in vivo. Interestingly, CD44+ cells are the most abundant population, when compared with the expression of EpCAM and CD133. In addition, this selection is independent of the degree of gemcitabine chemosensitivity of each tumor.


Deciphering the cellular source of tumor relapse identifies CD44 as a major therapeutic target in pancreatic adenocarcinoma.

Molejon MI, Tellechea JI, Loncle C, Gayet O, Gilabert M, Duconseil P, Lopez-Millan MB, Moutardier V, Gasmi M, Garcia S, Turrini O, Ouaissi M, Poizat F, Dusetti N, Iovanna J - Oncotarget (2015)

Expression of Cancer Stem Cell (CSC) markers in PDXsImmunofluorescent labeling of (A) CD44-FITC (green) and 4′,6-diamidino-2-phenylindole (DAPI), (blue); (B) EpCAM-FITC (green); DAPI (blue) and (C) CD133-FITC (green); DAPI (blue) in PDXs upon vehicle or gemcitabine treatment, show the distribution of each CSC marker in tumor tissues. (D) Quantification of the number of positive cells in each PDX. Scale bar represents 50 μm. Error bars ± SEM; n=3 per group. *P<0.05, **P<0.001 compared to vehicle treatment.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4480689&req=5

Figure 2: Expression of Cancer Stem Cell (CSC) markers in PDXsImmunofluorescent labeling of (A) CD44-FITC (green) and 4′,6-diamidino-2-phenylindole (DAPI), (blue); (B) EpCAM-FITC (green); DAPI (blue) and (C) CD133-FITC (green); DAPI (blue) in PDXs upon vehicle or gemcitabine treatment, show the distribution of each CSC marker in tumor tissues. (D) Quantification of the number of positive cells in each PDX. Scale bar represents 50 μm. Error bars ± SEM; n=3 per group. *P<0.05, **P<0.001 compared to vehicle treatment.
Mentions: Considering that CSCs represent the only cell population with tumor-initiating potential, we hypothesized that these cells, which seem to play a crucial role in treatment resistance, are associated with the phenotypic transformation described above. These cells are capable of resisting toxicity by drugs, such as gemcitabine, that target highly proliferative cells, since they enter into the cell cycle less frequently and only divide in response to certain stimuli, which have yet to be fully identified. We sought to investigate the potential involvement of CSCs in driving residual tumor growth following gemcitabine treatment. We used immunohistochemistry to analyze the expression of some CSC-associated cell surface markers on these tumors. We selected PDXs which were either sensitive to, or moderately or highly resistant to gemcitabine therapy (X-IPC, C-NOR and AO-IPC, respectively). Those PDXs reaching a volume of 400 mm3 received either vehicle or 100 mg/kg gemcitabine, twice weekly. Immunofluorescence analysis revealed that the percentage of CD44+, EpCAM+ and CD133+ cells in gemcitabine-treated tumors increased systematically 3-4 folds when compared to vehicle-treated tumors. As shown in Figure 2A, CD44 expression was found in 22±2.8%, 25±2.3% and 30±4.4% of vehicle-treated tumors, while its expression increased to 59±7.0%, 65±7.2% and 73±7.9% in gemcitabine-treated X-IPC, C-NOR and AO-IPC, respectively. Similarly, EpCAM was expressed in 3.6±1.0%, 0.2±0.7% and 4.3±0.8% of vehicle-treated tumors and increased to 24±2.8%, 1.2±1.3% and 31±3.3% in gemcitabine-treated X-IPC, C-NOR and AO-IPC, respectively (Figure 2B). Finally, CD133 was expressed in 5.0±0.5%, 1.0±0.9% and 5.1±0.8% of vehicle-treated tumors, while in gemcitabine-treated tumors its expression was found in 18±2.7%, 3.2±1.2% and 5.3±1.2% of X-IPC, C-NOR and AO-IPC, respectively (Figure 2C). Quantification is shown in Figure 2D. Thus, these data strongly suggest that gemcitabine treatment may select a highly enriched population of cells expressing CSC-associated markers in vivo. Interestingly, CD44+ cells are the most abundant population, when compared with the expression of EpCAM and CD133. In addition, this selection is independent of the degree of gemcitabine chemosensitivity of each tumor.

Bottom Line: The origin and biological characteristics of residual tumor cells in PDAC still remain unclear.During PDAC relapse, proliferating CD44+ cells decrease expression of ZEB1, while overexpressing the MUC1 protein, and gain morphological and biological characteristics of differentiation.We confirmed the propagation of CD44+ cells in samples from cases of human relapse, following standard PDAC treatment.

View Article: PubMed Central - PubMed

Affiliation: Centre de Recherche en Cancérologie de Marseille (CRCM), INSERM U1068, CNRS UMR 7258, Aix-Marseille Université and Institut Paoli-Calmettes, Parc Scientifique et Technologique de Luminy, Marseille, France.

ABSTRACT
It has been commonly found that in patients presenting Pancreatic Ductal Adenocarcinoma (PDAC), after a period of satisfactory response to standard treatments, the tumor becomes non-responsive and patient death quickly follows. This phenomenon is mainly due to the rapid and uncontrolled development of the residual tumor. The origin and biological characteristics of residual tumor cells in PDAC still remain unclear. In this work, using PDACs from patients, preserved as xenografts in nude mice, we demonstrated that a residual PDAC tumor originated from a small number of CD44+ cells present in the tumor. During PDAC relapse, proliferating CD44+ cells decrease expression of ZEB1, while overexpressing the MUC1 protein, and gain morphological and biological characteristics of differentiation. Also, we report that CD44+ cells, in primary and residual PDAC tumors, are part of a heterogeneous population, which includes variable numbers of CD133+ and EpCAM+ cells. We confirmed the propagation of CD44+ cells in samples from cases of human relapse, following standard PDAC treatment. Finally, using systemic administration of anti-CD44 antibodies in vivo, we demonstrated that CD44 is an efficient therapeutic target for treating tumor relapse, but not primary PDAC tumors. We conclude that CD44+ cells generate the relapsing tumor and, as such, are themselves promising therapeutic targets for treating patients with recurrent PDAC.

No MeSH data available.


Related in: MedlinePlus