MSH2 deficiency abolishes the anticancer and pro-aging activity of short telomeres
Paula Martinez
Telomeres and Telomerase Group, Molecular Oncology Program, Spanish National Cancer Centre, 28029 Madrid, Spain
Search for more papers by this authorIrene Siegl-Cachedenier
Telomeres and Telomerase Group, Molecular Oncology Program, Spanish National Cancer Centre, 28029 Madrid, Spain
Search for more papers by this authorJuana M. Flores
Animal Surgery and Medicine Department, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040 Madrid, Spain
Search for more papers by this authorMaria A. Blasco
Telomeres and Telomerase Group, Molecular Oncology Program, Spanish National Cancer Centre, 28029 Madrid, Spain
Search for more papers by this authorPaula Martinez
Telomeres and Telomerase Group, Molecular Oncology Program, Spanish National Cancer Centre, 28029 Madrid, Spain
Search for more papers by this authorIrene Siegl-Cachedenier
Telomeres and Telomerase Group, Molecular Oncology Program, Spanish National Cancer Centre, 28029 Madrid, Spain
Search for more papers by this authorJuana M. Flores
Animal Surgery and Medicine Department, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040 Madrid, Spain
Search for more papers by this authorMaria A. Blasco
Telomeres and Telomerase Group, Molecular Oncology Program, Spanish National Cancer Centre, 28029 Madrid, Spain
Search for more papers by this authorPaula Martinez and Irene Siegl-Cachedenier contributed equally to this work.
Summary
Mutations in the mismatch repair (MMR) pathway occur in human colorectal cancers with microsatellite instability. Mounting evidence suggests that cell-cycle arrest in response to a number of cellular stresses, including telomere shortening, is a potent anticancer barrier. The telomerase-deficient mouse model illustrates the anticancer effect of cell-cycle arrest provoked by short telomeres. Here, we describe a role for the MMR protein, MSH2, in signaling cell-cycle arrest in a p21/p53-dependent manner in response to short telomeres in the context of telomerase-deficient mice. In particular, progressively shorter telomeres at successive generations of MSH2−/–Terc−/–- mice did not suppress cancer in these mice, indicating that MSH2 deficiency abolishes the tumor suppressor activity of short telomeres. Interestingly, MSH2 deficiency prevented degenerative pathologies in the gastrointestinal tract of MSH2−/–Terc−/– mice concomitant with a rescue of proliferative defects. The abolishment of the anticancer and pro-aging effects of short telomeres provoked by MSH2 abrogation was independent of changes in telomere length. These results highlight a role for MSH2 in the organismal response to dysfunctional telomeres, which in turn may be important in the pathobiology of human cancers bearing mutations in the MMR pathway.
Supporting Information
Fig. S1 Kaplan–Meier survival curves of the different mouse cohorts. (A) MSH2+/+/Terc+/+, G1MSH2+/+/Terc−/–, G2MSH2+/+/Terc−/– and G3MSH2+/+/Terc−/–; (B) MSH2−/–/Terc+/+, G1MSH2−/–/Terc−/–, G2MSH2−/–/Terc−/–- and G3MSH2−/–/Terc−/–. n, number of mice of each genotype included in the analysis. Statistical comparisons among genotypes using the log rank test are shown. For statistical comparisons between generations, see Supporting Table S1.
Fig. S2 Histopathological analysis of mice of the different genotypes at their time of death. Percentage of mice of the indicated genotype showing degenerative pathologies in the indicated organs at time of death. Intestinal lesions include atrophy, hyperplasia, cysts, hemorrhage, edema and lymphoid infiltrations (see also Fig. 2). Kidney lesions include atrophy, tubular degeneration, amyloidosis, glomerulonephritis, hydronephrosis, intersticial nephritis, cysts and lymphoid infiltrations. Spleen lesions include atrophy, amyloidosis, lymphoid and myeloid hyperplasia. Liver lesions include vacuolar degeneration, necrosis, cyst and lymphoid infiltration. Testis, seminal vesicle, ovary and uterus compose the reproductive tract group. The reproductive tract lesions include atrophy, cyst, hyperplasia of Leydig cells, inflammation of seminal vesicles and cystic endometrial hyperplasia. Heart lesions include congestion, ventricular dilatation and myocardiopathy. Lung lesions include congestion, hemorrhage, alveolar hyperplasia and lymphoid infiltration. Statistical comparisons between genotypes using the chi-squared test are shown.
Fig. S3 Representative combined images of telomere fluorescence and DAPI staining on small intestine sections of mice of the indicated genotype as determined by Q-FISH.
Fig. S4 Telomere length as determined by TRF in MEF of the indicated genotypes. No significant differences in TRF fragment size were detected between single Terc−/– and double Terc−/– MSH2−/– mice of the indicated mouse generations.
Fig. S5 (a) Percentage of ATR-positive cells in small intestine sections of mice of the indicated genotypes. n, number of mice analyzed per genotype. The total number of cells scored is indicated. Bars represent standard error. Statistical comparisons using the chi-squared test are shown. Gray bars, MSH2+/+ mice; black bars, MSH2−/– mice. For statistical comparisons between generations, see Supporting Table S2. (b) Representative images of ATR-positive cells (upper panel) and of the DAPI combined-images (lower panel) in small intestine sections of G3Terc−/– MSH2+/+ and G3Terc−/– MSH2−/– mice.
Table S1 Statistical significance in median survival and mean lifespan analysis by log rank and Student’s t-test, respectively
Table S2 Comparisons between generations. Statistical significance in immunohistochemistry and in situ hybridization determinations by chi-squared test
Table S3 Statistical significance in Q-FISH analysis in small intestine sections
Table S4 Statistical significance in Q-FISH analysis in MEF metaphases
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