TET2 controls chemoresistant slow-cycling cancer cell survival and tumor recurrence
Isabel Puig, Stephan P. Tenbaum, Irene Chicote, Oriol Arqués, Jordi Martínez-Quintanilla, Estefania Cuesta-Borrás, Lorena Ramírez, Pilar Gonzalo, Atenea Soto, Susana Aguilar, Cristina Eguizabal, Ginevra Caratù, Aleix Prat, Guillem Argilés, Stefania Landolfi, Oriol Casanovas, Violeta Serra, Alberto Villanueva, Alicia G. Arroyo, Luigi Terracciano, Paolo Nuciforo, Joan Seoane, Juan A. Recio, Ana Vivancos, Rodrigo Dienstmann, Josep Tabernero, Héctor G. Palmer
Isabel Puig, Stephan P. Tenbaum, Irene Chicote, Oriol Arqués, Jordi Martínez-Quintanilla, Estefania Cuesta-Borrás, Lorena Ramírez, Pilar Gonzalo, Atenea Soto, Susana Aguilar, Cristina Eguizabal, Ginevra Caratù, Aleix Prat, Guillem Argilés, Stefania Landolfi, Oriol Casanovas, Violeta Serra, Alberto Villanueva, Alicia G. Arroyo, Luigi Terracciano, Paolo Nuciforo, Joan Seoane, Juan A. Recio, Ana Vivancos, Rodrigo Dienstmann, Josep Tabernero, Héctor G. Palmer
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Research Article Oncology

TET2 controls chemoresistant slow-cycling cancer cell survival and tumor recurrence

Abstract

Dormant or slow-cycling tumor cells can form a residual chemoresistant reservoir responsible for relapse in patients, years after curative surgery and adjuvant therapy. We have adapted the pulse-chase expression of H2BeGFP for labeling and isolating slow-cycling cancer cells (SCCCs). SCCCs showed cancer initiation potential and enhanced chemoresistance. Cells at this slow-cycling status presented a distinctive nongenetic and cell-autonomous gene expression profile shared across different tumor types. We identified TET2 epigenetic enzyme as a key factor controlling SCCC numbers, survival, and tumor recurrence. 5-Hydroxymethylcytosine (5hmC), generated by TET2 enzymatic activity, labeled the SCCC genome in carcinomas and was a predictive biomarker of relapse and survival in cancer patients. We have shown the enhanced chemoresistance of SCCCs and revealed 5hmC as a biomarker for their clinical identification and TET2 as a potential drug target for SCCC elimination that could extend patients’ survival.

Authors

Isabel Puig, Stephan P. Tenbaum, Irene Chicote, Oriol Arqués, Jordi Martínez-Quintanilla, Estefania Cuesta-Borrás, Lorena Ramírez, Pilar Gonzalo, Atenea Soto, Susana Aguilar, Cristina Eguizabal, Ginevra Caratù, Aleix Prat, Guillem Argilés, Stefania Landolfi, Oriol Casanovas, Violeta Serra, Alberto Villanueva, Alicia G. Arroyo, Luigi Terracciano, Paolo Nuciforo, Joan Seoane, Juan A. Recio, Ana Vivancos, Rodrigo Dienstmann, Josep Tabernero, Héctor G. Palmer

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Figure 9

SCCC survival depends on TET2 enzymatic activity.

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SCCC survival depends on TET2 enzymatic activity. (A) Expression of H19 ...
(A) Expression of H19 gene measured by qPCR. (B) Percentage of 5hmC in total genomic DNA from mouse brain or the indicated cell lines. (C) Evaluation of apoptosis by flow cytometry in RCCCs and SCCCs after cell-permeable 2-hydroxyglutarate (TFMB-2HG) exposure. Apoptosis measurements: shCTRL/shTET2 RCCC VEH/TFMB-2HG vs. shCTRL SCCC TFMB-2HG (P ≤ 0.0001); shCTRL/shTET2 RCCC VEH/TFMB-2HG vs. shTET2 SCCC VEH/TFMB-2HG (P ≤ 0.0001); shCTRL SCCC VEH vs. shCTRL/shTET2 SCCC TFMB-2HG (P ≤ 0.0001); shCTRL SCCC VEH vs. shTET2 SCCC VEH (P ≤ 0.0001); shTET2 SCCC VEH vs. shTET2 SCCC TFMB-2HG (P ≤ 0.01). (D) Expression of exogenous TET2 (TET2-WT or -HxD FLAG-tagged) was evaluated by qPCR in the indicated cell lines. Specific primers for TET2 (forward) and FLAG (reverse) were used. (E and F) Analysis of apoptosis (E) and proportion of SCCCs (F) was performed by flow cytometry in the indicated cell lines. Apoptosis measurements: TET2-KO RCCC vs. TET2-KO SCCC (P ≤ 0.001); TET2-KO/-HxD RCCC vs. TET2-HxD SCCC (P ≤ 0.01); TET2-KO SCCC vs. TET2-WT/-HxD RCCC (P ≤ 0.001); TET2-WT RCCC vs. TET2-HxD SCCC (P ≤ 0.001). (C, E, and F) Blue bars, RCCCs; green bars, SCCCs. (A–F) Data are represented as mean ± SEM (B, C, E, and F) or ± SD (A and D) of triplicates from 3 independent experiments. (A and B) 2-tailed Student’s t test. (C, E, and F) 1-way ANOVA. (A–C, E, and F) *P ≤ 0.05; **P ≤ 0.01; ****P ≤ 0.0001. All analyses were performed in the indicated SW1222-H2BeGFP cell lines growing as MTs.