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1.
Proc Natl Acad Sci U S A ; 119(20): e2121499119, 2022 05 17.
Artículo en Inglés | MEDLINE | ID: mdl-35537048

RESUMEN

As the global elderly population grows, it is socioeconomically and medically critical to provide diverse and effective means of mitigating the impact of aging on human health. Previous studies showed that the adeno-associated virus (AAV) vector induced overexpression of certain proteins, which can suppress or reverse the effects of aging in animal models. In our study, we sought to determine whether the high-capacity cytomegalovirus vector (CMV) can be an effective and safe gene delivery method for two such protective factors: telomerase reverse transcriptase (TERT) and follistatin (FST). We found that the mouse cytomegalovirus (MCMV) carrying exogenous TERT or FST (MCMVTERT or MCMVFST) extended median lifespan by 41.4% and 32.5%, respectively. We report CMV being used successfully as both an intranasal and injectable gene therapy system to extend longevity. Specifically, this treatment significantly improved glucose tolerance, physical performance, as well as preventing body mass loss and alopecia. Further, telomere shortening associated with aging was ameliorated by TERT and mitochondrial structure deterioration was halted in both treatments. Intranasal and injectable preparations performed equally well in safely and efficiently delivering gene therapy to multiple organs, with long-lasting benefits and without carcinogenicity or unwanted side effects. Translating this research to humans could have significant benefits associated with quality of life and an increased health span.


Asunto(s)
Infecciones por Citomegalovirus , Terapia Genética , Esperanza de Vida , Telomerasa , Administración por Inhalación , Animales , Folistatina/genética , Terapia Genética/efectos adversos , Terapia Genética/métodos , Vectores Genéticos/genética , Inyecciones Intraperitoneales , Ratones , Modelos Animales , Neoplasias , Telomerasa/genética , Telomerasa/metabolismo
2.
Angiogenesis ; 22(1): 95-102, 2019 02.
Artículo en Inglés | MEDLINE | ID: mdl-30168024

RESUMEN

Vascular complications such as bleeding due to gastrointestinal telangiectatic anomalies, pulmonary arteriovenous malformations, hepatopulmonary syndrome, and retinal vessel abnormalities are being reported in patients with telomere biology disorders (TBDs) more frequently than previously described. The international clinical care consortium of telomere-associated ailments and family support group Dyskeratosis Congenita Outreach, Inc. held a workshop on vascular abnormalities in the TBDs at the National Cancer Institute in October 2017. Clinicians and basic scientists reviewed current data on vascular complications, hypotheses for the underlying biology and developed new collaborations to address the etiology and clinical management of vascular complications in TBDs.


Asunto(s)
Fístula Arteriovenosa , Arteria Pulmonar/anomalías , Venas Pulmonares/anomalías , Telangiectasia , Telómero , Animales , Fístula Arteriovenosa/genética , Fístula Arteriovenosa/metabolismo , Fístula Arteriovenosa/patología , Educación , Humanos , Arteria Pulmonar/metabolismo , Arteria Pulmonar/patología , Venas Pulmonares/metabolismo , Venas Pulmonares/patología , Telangiectasia/genética , Telangiectasia/metabolismo , Telangiectasia/patología , Telómero/genética , Telómero/metabolismo , Telómero/patología
3.
Proc Natl Acad Sci U S A ; 113(34): E5024-33, 2016 08 23.
Artículo en Inglés | MEDLINE | ID: mdl-27503890

RESUMEN

Oncogene-induced senescence (OIS) is a critical tumor-suppressing mechanism that restrains cancer progression at premalignant stages, in part by causing telomere dysfunction. Currently it is unknown whether this proliferative arrest presents a stable and therefore irreversible barrier to cancer progression. Here we demonstrate that cells frequently escape OIS induced by oncogenic H-Ras and B-Raf, after a prolonged period in the senescence arrested state. Cells that had escaped senescence displayed high oncogene expression levels, retained functional DNA damage responses, and acquired chromatin changes that promoted c-Myc-dependent expression of the human telomerase reverse transcriptase gene (hTERT). Telomerase was able to resolve existing telomeric DNA damage response foci and suppressed formation of new ones that were generated as a consequence of DNA replication stress and oncogenic signals. Inhibition of MAP kinase signaling, suppressing c-Myc expression, or inhibiting telomerase activity, caused telomere dysfunction and proliferative defects in cells that had escaped senescence, whereas ectopic expression of hTERT facilitated OIS escape. In human early neoplastic skin and breast tissue, hTERT expression was detected in cells that displayed features of senescence, suggesting that reactivation of telomerase expression in senescent cells is an early event during cancer progression in humans. Together, our data demonstrate that cells arrested in OIS retain the potential to escape senescence by mechanisms that involve derepression of hTERT expression.


Asunto(s)
Neoplasias de la Mama/genética , Regulación Neoplásica de la Expresión Génica , Proteína Oncogénica p21(ras)/genética , Proteínas Proto-Oncogénicas B-raf/genética , Neoplasias Cutáneas/genética , Telomerasa/genética , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Línea Celular , Línea Celular Tumoral , Supervivencia Celular , Senescencia Celular , Células Epiteliales/metabolismo , Células Epiteliales/patología , Femenino , Fibroblastos/citología , Fibroblastos/metabolismo , Humanos , Masculino , Proteína Oncogénica p21(ras)/metabolismo , Proteínas Proto-Oncogénicas B-raf/metabolismo , Proteínas Proto-Oncogénicas c-myc/genética , Proteínas Proto-Oncogénicas c-myc/metabolismo , Transducción de Señal , Neoplasias Cutáneas/metabolismo , Neoplasias Cutáneas/patología , Telomerasa/metabolismo , Telómero/química , Telómero/metabolismo , Homeostasis del Telómero
4.
Hum Mol Genet ; 25(11): 2324-2330, 2016 06 01.
Artículo en Inglés | MEDLINE | ID: mdl-26936823

RESUMEN

Leukocyte telomere length (LTL), which reflects telomere length in other somatic tissues, is a complex genetic trait. Eleven SNPs have been shown in genome-wide association studies to be associated with LTL at a genome-wide level of significance within cohorts of European ancestry. It has been observed that LTL is longer in African Americans than in Europeans. The underlying reason for this difference is unknown. Here we show that LTL is significantly longer in sub-Saharan Africans than in both Europeans and African Americans. Based on the 11 LTL-associated alleles and genetic data in phase 3 of the 1000 Genomes Project, we show that the shifts in allele frequency within Europe and between Europe and Africa do not fit the pattern expected by neutral genetic drift. Our findings suggest that differences in LTL within Europeans and between Europeans and Africans is influenced by polygenic adaptation and that differences in LTL between Europeans and Africans might explain, in part, ethnic differences in risks for human diseases that have been linked to LTL.


Asunto(s)
Leucocitos/citología , Homeostasis del Telómero/genética , Acortamiento del Telómero/genética , Telómero/genética , Adolescente , Adulto , Negro o Afroamericano/genética , Anciano , Anciano de 80 o más Años , Alelos , Población Negra/genética , Niño , Femenino , Flujo Genético , Humanos , Masculino , Persona de Mediana Edad , Polimorfismo de Nucleótido Simple , Población Blanca/genética
5.
EMBO J ; 31(13): 2839-51, 2012 Jun 29.
Artículo en Inglés | MEDLINE | ID: mdl-22569128

RESUMEN

In normal human somatic cells, telomere dysfunction causes cellular senescence, a stable proliferative arrest with tumour suppressing properties. Whether telomere dysfunction-induced senescence (TDIS) suppresses cancer growth in humans, however, is unknown. Here, we demonstrate that multiple and distinct human cancer precursor lesions, but not corresponding malignant cancers, are comprised of cells that display hallmarks of TDIS. Furthermore, we demonstrate that oncogenic signalling, frequently associated with initiating cancer growth in humans, dramatically affected telomere structure and function by causing telomeric replication stress, rapid and stochastic telomere attrition, and consequently telomere dysfunction in cells that lack hTERT activity. DNA replication stress induced by drugs also resulted in telomere dysfunction and cellular senescence in normal human cells, demonstrating that telomeric repeats indeed are hypersensitive to DNA replication stress. Our data reveal that TDIS, accelerated by oncogene-induced DNA replication stress, is a biological response of cells in human cancer precursor lesions and provide strong evidence that TDIS is a critical tumour suppressing mechanism in humans.


Asunto(s)
Senescencia Celular/fisiología , Oncogenes/fisiología , Telómero/fisiología , Línea Celular , Transformación Celular Neoplásica/efectos de los fármacos , Transformación Celular Neoplásica/genética , Senescencia Celular/efectos de los fármacos , Senescencia Celular/genética , Replicación del ADN/efectos de los fármacos , Replicación del ADN/genética , Replicación del ADN/fisiología , Humanos , Oncogenes/efectos de los fármacos , Oncogenes/genética , Inhibidores de la Síntesis de la Proteína/farmacología , Puromicina/farmacología , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Transducción de Señal/fisiología , Telómero/efectos de los fármacos , Telómero/genética
6.
J Med Genet ; 52(3): 157-62, 2015 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-25624462

RESUMEN

BACKGROUND: Leucocyte telomere length (LTL), which is fashioned by multiple genes, has been linked to a host of human diseases, including sporadic melanoma. A number of genes associated with LTL have already been identified through genome-wide association studies. The main aim of this study was to establish whether DCAF4 (DDB1 and CUL4-associated factor 4) is associated with LTL. In addition, using ingenuity pathway analysis (IPA), we examined whether LTL-associated genes in the general population might partially explain the inherently longer LTL in patients with sporadic melanoma, the risk for which is increased with ultraviolet radiation (UVR). RESULTS: Genome-wide association (GWA) meta-analysis and de novo genotyping of 20 022 individuals revealed a novel association (p=6.4×10(-10)) between LTL and rs2535913, which lies within DCAF4. Notably, eQTL analysis showed that rs2535913 is associated with decline in DCAF4 expressions in both lymphoblastoid cells and sun-exposed skin (p=4.1×10(-3) and 2×10(-3), respectively). Moreover, IPA revealed that LTL-associated genes, derived from GWA meta-analysis (N=9190), are over-represented among genes engaged in melanoma pathways. Meeting increasingly stringent p value thresholds (p<0.05, <0.01, <0.005, <0.001) in the LTL-GWA meta-analysis, these genes were jointly over-represented for melanoma at p values ranging from 1.97×10(-169) to 3.42×10(-24). CONCLUSIONS: We uncovered a new locus associated with LTL in the general population. We also provided preliminary findings that suggest a link of LTL through genetic mechanisms with UVR and melanoma in the general population.


Asunto(s)
Proteínas Portadoras/genética , Leucocitos/citología , Melanoma/genética , Homeostasis del Telómero/genética , Alelos , Proteínas Portadoras/biosíntesis , Proteínas Portadoras/sangre , Regulación de la Expresión Génica , Estudio de Asociación del Genoma Completo , Humanos , Melanoma/sangre , Melanoma/patología , Factores de Riesgo , Telómero/genética
7.
Nucleic Acids Res ; 41(10): 5321-40, 2013 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-23571757

RESUMEN

Telomere integrity is critical for telomere function and genomic stability. We previously demonstrated that non-erythroid α-spectrin (αIISp) is present in mammalian cell nuclei where it is important in repair of DNA interstrand cross-links (ICLs) and chromosome stability. We now demonstrate that αIISp is also important for telomere maintenance after ICL damage. It localizes to telomeres in S phase after ICL damage where it has enhanced association with TRF1 and TRF2 and is required for recruitment of the ICL repair protein, XPF, to damage-induced foci at telomeres. In telomerase-positive normal cells depleted of αIISp by siRNA or in Fanconi anemia, complementation group A (FA-A) cells, where αIISp levels are 35-40% of normal, ICL damage results in failure of XPF to localize to telomeres, markedly increased telomere dysfunction-induced foci, followed by catastrophic loss of telomeres. Restoration of αIISp levels to normal in FA-A cells corrects these deficiencies. Our studies demonstrate that αIISp is critical for repair of DNA ICLs at telomeres, likely by facilitating the recruitment of repair proteins similar, but not identical, to its proposed role in repair of DNA ICLs in genomic DNA and that this function in turn is critical for telomere maintenance after DNA ICL damage.


Asunto(s)
Daño del ADN , Reparación del ADN , Espectrina/fisiología , Telómero/fisiología , Línea Celular , Aberraciones Cromosómicas , Reactivos de Enlaces Cruzados/toxicidad , Proteínas de Unión al ADN/análisis , Proteína del Grupo de Complementación D2 de la Anemia de Fanconi/análisis , Humanos , Fase S/genética , Espectrina/antagonistas & inhibidores , Espectrina/metabolismo , Telómero/química , Telómero/metabolismo , Proteína 1 de Unión a Repeticiones Teloméricas/metabolismo , Proteína 2 de Unión a Repeticiones Teloméricas/metabolismo
8.
Proc Natl Acad Sci U S A ; 107(20): 9293-8, 2010 May 18.
Artículo en Inglés | MEDLINE | ID: mdl-20421499

RESUMEN

Telomeres are engaged in a host of cellular functions, and their length is regulated by multiple genes. Telomere shortening, in the course of somatic cell replication, ultimately leads to replicative senescence. In humans, rare mutations in genes that regulate telomere length have been identified in monogenic diseases such as dyskeratosis congenita and idiopathic pulmonary fibrosis, which are associated with shortened leukocyte telomere length (LTL) and increased risk for aplastic anemia. Shortened LTL is observed in a host of aging-related complex genetic diseases and is associated with diminished survival in the elderly. We report results of a genome-wide association study of LTL in a consortium of four observational studies (n = 3,417 participants with LTL and genome-wide genotyping). SNPs in the regions of the oligonucleotide/oligosaccharide-binding folds containing one gene (OBFC1; rs4387287; P = 3.9 x 10(-9)) and chemokine (C-X-C motif) receptor 4 gene (CXCR4; rs4452212; P = 2.9 x 10(-8)) were associated with LTL at a genome-wide significance level (P < 5 x 10(-8)). We attempted replication of the top SNPs at these loci through de novo genotyping of 1,893 additional individuals and in silico lookup in another observational study (n = 2,876), and we confirmed the association findings for OBFC1 but not CXCR4. In addition, we confirmed the telomerase RNA component (TERC) as a gene associated with LTL (P = 1.1 x 10(-5)). The identification of OBFC1 through genome-wide association as a locus for interindividual variation in LTL in the general population advances the understanding of telomere biology in humans and may provide insights into aging-related disorders linked to altered LTL dynamics.


Asunto(s)
Leucocitos/fisiología , Receptores CXCR4/fisiología , Proteínas de Unión a Telómeros/fisiología , Telómero/fisiología , Estudios de Cohortes , Estudio de Asociación del Genoma Completo , Genotipo , Humanos , Leucocitos/química , Polimorfismo de Nucleótido Simple/genética , Receptores CXCR4/genética , Telómero/genética , Proteínas de Unión a Telómeros/genética
9.
Cell Genom ; 3(4): 100293, 2023 Apr 12.
Artículo en Inglés | MEDLINE | ID: mdl-37082139

RESUMEN

Although oncogene-induced senescence (OIS) is a potent tumor-suppressor mechanism, recent studies revealed that cells could escape from OIS with features of transformed cells. However, the mechanisms that promote OIS escape remain unclear, and evidence of post-senescent cells in human cancers is missing. Here, we unravel the regulatory mechanisms underlying OIS escape using dynamic multidimensional profiling. We demonstrate a critical role for AP1 and POU2F2 transcription factors in escape from OIS and identify senescence-associated chromatin scars (SACSs) as an epigenetic memory of OIS detectable during colorectal cancer progression. POU2F2 levels are already elevated in precancerous lesions and as cells escape from OIS, and its expression and binding activity to cis-regulatory elements are associated with decreased patient survival. Our results support a model in which POU2F2 exploits a precoded enhancer landscape necessary for senescence escape and reveal POU2F2 and SACS gene signatures as valuable biomarkers with diagnostic and prognostic potential.

10.
Aging Cell ; 20(5): e13344, 2021 05.
Artículo en Inglés | MEDLINE | ID: mdl-33939265

RESUMEN

Aging leads to a progressive functional decline of the immune system, rendering the elderly increasingly susceptible to disease and infection. The degree to which immune cell senescence contributes to this decline remains unclear, however, since markers that label immune cells with classical features of cellular senescence accurately and comprehensively have not been identified. Using a second-generation fluorogenic substrate for ß-galactosidase and multi-parameter flow cytometry, we demonstrate here that peripheral blood mononuclear cells (PBMCs) isolated from healthy humans increasingly display cells with high senescence-associated ß-galactosidase (SA-ßGal) activity with advancing donor age. The greatest age-associated increases were observed in CD8+ T-cell populations, in which the fraction of cells with high SA-ßGal activity reached average levels of 64% in donors in their 60s. CD8+ T cells with high SA-ßGal activity, but not those with low SA-ßGal activity, were found to exhibit features of telomere dysfunction-induced senescence and p16-mediated senescence, were impaired in their ability to proliferate, developed in various T-cell differentiation states, and had a gene expression signature consistent with the senescence state previously observed in human fibroblasts. Based on these results, we propose that senescent CD8+ T cells with classical features of cellular senescence accumulate to levels that are significantly higher than previously reported and additionally provide a simple yet robust method for the isolation and characterization of senescent CD8+ T cells with predictive potential for biological age.


Asunto(s)
Envejecimiento/inmunología , Linfocitos T CD8-positivos/citología , Senescencia Celular/inmunología , beta-Galactosidasa/metabolismo , Linfocitos T CD8-positivos/enzimología , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Diferenciación Celular , Separación Celular , Células Cultivadas , Senescencia Celular/genética , Inhibidor p16 de la Quinasa Dependiente de Ciclina/metabolismo , Citometría de Flujo , Expresión Génica , Humanos , Telómero
11.
Sci Rep ; 11(1): 5115, 2021 03 04.
Artículo en Inglés | MEDLINE | ID: mdl-33664422

RESUMEN

Recent hypotheses propose that the human placenta and chorioamniotic membranes (CAMs) experience telomere length (TL)-mediated senescence. These hypotheses are based on mean TL (mTL) measurements, but replicative senescence is triggered by short and dysfunctional telomeres, not mTL. We measured short telomeres by a vanguard method, the Telomere shortest length assay, and telomere-dysfunction-induced DNA damage foci (TIF) in placentas and CAMs between 18-week gestation and at full-term. Both the placenta and CAMs showed a buildup of short telomeres and TIFs, but not shortening of mTL from 18-weeks to full-term. In the placenta, TIFs correlated with short telomeres but not mTL. CAMs of preterm birth pregnancies with intra-amniotic infection showed shorter mTL and increased proportions of short telomeres. We conclude that the placenta and probably the CAMs undergo TL-mediated replicative aging. Further research is warranted whether TL-mediated replicative aging plays a role in all preterm births.


Asunto(s)
Senescencia Celular/genética , Membrana Corioalantoides/metabolismo , Placenta/fisiología , Homeostasis del Telómero/genética , Adulto , Envejecimiento/genética , Membrana Corioalantoides/crecimiento & desarrollo , Daño del ADN/genética , Replicación del ADN/genética , Femenino , Edad Gestacional , Humanos , Hibridación Fluorescente in Situ , Recién Nacido , Placenta/metabolismo , Placentación , Embarazo , Nacimiento Prematuro/genética , Nacimiento Prematuro/patología , Telómero/genética
12.
Clin Cancer Res ; 27(24): 6800-6814, 2021 12 15.
Artículo en Inglés | MEDLINE | ID: mdl-34593527

RESUMEN

PURPOSE: To investigate the therapeutic role of a novel telomere-directed inhibitor, 6-thio-2'-deoxyguanosine (THIO) in gliomas both in vitro and in vivo. EXPERIMENTAL DESIGN: A panel of human and mouse glioma cell lines was used to test therapeutic efficacy of THIO using cell viability assays, flow cytometric analyses, and immunofluorescence. Integrated analyses of RNA sequencing and reverse-phase protein array data revealed the potential antitumor mechanisms of THIO. Four patient-derived xenografts (PDX), two patient-derived organoids (PDO), and two xenografts of human glioma cell lines were used to further investigate the therapeutic efficacy of THIO. RESULTS: THIO was effective in the majority of human and mouse glioma cell lines with no obvious toxicity against normal astrocytes. THIO as a monotherapy demonstrated efficacy in three glioma cell lines that had acquired resistance to temozolomide. In addition, THIO showed efficacy in four human glioma cell lines grown as neurospheres by inducing apoptotic cell death. Mechanistically, THIO induced telomeric DNA damage not only in glioma cell lines but also in PDX tumor specimens. Integrated computational analyses of transcriptomic and proteomic data indicated that THIO significantly inhibited cell invasion, stem cell, and proliferation pathways while triggering DNA damage and apoptosis. Importantly, THIO significantly decreased tumor proliferation in two PDO models and reduced the tumor size of a glioblastoma xenograft and a PDX model. CONCLUSIONS: The current study established the therapeutic role of THIO in primary and recurrent gliomas and revealed the acute induction of telomeric DNA damage as a primary antitumor mechanism of THIO in gliomas.


Asunto(s)
Neoplasias Encefálicas , Glioma , Animales , Neoplasias Encefálicas/tratamiento farmacológico , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/patología , Línea Celular Tumoral , Desoxiguanosina/análogos & derivados , Glioma/tratamiento farmacológico , Glioma/genética , Glioma/patología , Humanos , Ratones , Nucleósidos/uso terapéutico , Proteómica , Tionucleósidos , Ensayos Antitumor por Modelo de Xenoinjerto
13.
J Gerontol A Biol Sci Med Sci ; 75(2): 230-235, 2020 01 20.
Artículo en Inglés | MEDLINE | ID: mdl-30388200

RESUMEN

Telomeres progressively shorten with age, and it has been proposed that critically short and dysfunctional telomeres contribute to aging and aging-associated diseases in humans. For many years it was thought that telomere erosion was strictly a consequence of the "end replication problem," or the inability of replicative polymerases to completely duplicate linear DNA ends. It is becoming increasingly evident, however, that telomere shortening of cultured human cells is also caused because of other replication defects in telomeric repeats, those that cause fragile telomeres and other aberrant telomeric structures that can be detected on metaphase chromosomes. Whether these replication defects contribute to telomere erosion also in human tissues is currently unknown. By analyzing peripheral blood mononuclear cells from a total of 35 healthy subjects ranging in age from 23 to 101 years, we demonstrated that telomeres increasingly display aberrant structures with advancing donor age. Although the percentages of fragile telomeres increased only until adulthood, the percentages of chromosomes displaying sister telomere loss and sister telomere chromatid fusions increased consistently throughout the entire human life span. Our data, therefore, suggest that telomeric replication defects other than the end replication problem contribute to aging-associated telomere erosion in humans.


Asunto(s)
Envejecimiento/patología , Telómero/patología , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Voluntarios Sanos , Humanos , Hibridación Fluorescente in Situ , Leucocitos Mononucleares , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Acortamiento del Telómero
14.
Nat Cell Biol ; 22(10): 1286-1288, 2020 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-32939047

RESUMEN

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

15.
Nat Cell Biol ; 22(7): 842-855, 2020 07.
Artículo en Inglés | MEDLINE | ID: mdl-32514071

RESUMEN

Senescent cells affect many physiological and pathophysiological processes. While select genetic and epigenetic elements for senescence induction have been identified, the dynamics, epigenetic mechanisms and regulatory networks defining senescence competence, induction and maintenance remain poorly understood, precluding the deliberate therapeutic targeting of senescence for health benefits. Here, we examined the possibility that the epigenetic state of enhancers determines senescent cell fate. We explored this by generating time-resolved transcriptomes and epigenome profiles during oncogenic RAS-induced senescence and validating central findings in different cell biology and disease models of senescence. Through integrative analysis and functional validation, we reveal links between enhancer chromatin, transcription factor recruitment and senescence competence. We demonstrate that activator protein 1 (AP-1) 'pioneers' the senescence enhancer landscape and defines the organizational principles of the transcription factor network that drives the transcriptional programme of senescent cells. Together, our findings enabled us to manipulate the senescence phenotype with potential therapeutic implications.


Asunto(s)
Senescencia Celular , Cromatina/metabolismo , Epigénesis Genética , Fibroblastos/citología , Regulación de la Expresión Génica , Histonas/metabolismo , Factor de Transcripción AP-1/metabolismo , Transcriptoma , Animales , Cromatina/genética , Femenino , Fibroblastos/metabolismo , Histonas/genética , Humanos , Ratones Endogámicos C57BL , Factor de Transcripción AP-1/genética
16.
Aging Cell ; 17(6): e12838, 2018 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-30244523

RESUMEN

Cells that had undergone telomere dysfunction-induced senescence secrete numerous cytokines and other molecules, collectively called the senescence-associated secretory phenotype (SASP). Although certain SASP factors have been demonstrated to promote cellular senescence in neighboring cells in a paracrine manner, the mechanisms leading to bystander senescence and the functional significance of these effects are currently unclear. Here, we demonstrate that TGF-ß1, a component of the SASP, causes telomere dysfunction in normal somatic human fibroblasts in a Smad3/NOX4/ROS-dependent manner. Surprisingly, instead of activating cellular senescence, TGF-ß1-induced telomere dysfunction caused fibroblasts to transdifferentiate into α-SMA-expressing myofibroblasts, a mesenchymal and contractile cell type that is critical for wound healing and tissue repair. Despite the presence of dysfunctional telomeres, transdifferentiated cells acquired the ability to contract collagen lattices and displayed a gene expression signature characteristic of functional myofibroblasts. Significantly, the formation of dysfunctional telomeres and downstream p53 signaling was necessary for myofibroblast transdifferentiation, as suppressing telomere dysfunction by expression of hTERT, inhibiting the signaling pathways that lead to stochastic telomere dysfunction, and suppressing p53 function prevented the generation of myofibroblasts in response to TGF-ß1 signaling. Furthermore, inducing telomere dysfunction using shRNA against TRF2 also caused cells to develop features that are characteristic of myofibroblasts, even in the absence of exogenous TGF-ß1. Overall, our data demonstrate that telomere dysfunction is not only compatible with cell functionality, but they also demonstrate that the generation of dysfunctional telomeres is an essential step for transdifferentiation of human fibroblasts into myofibroblasts.


Asunto(s)
Transdiferenciación Celular , Miofibroblastos/patología , Telómero/patología , Transdiferenciación Celular/efectos de los fármacos , Senescencia Celular/efectos de los fármacos , Daño del ADN , Humanos , Miofibroblastos/efectos de los fármacos , Miofibroblastos/metabolismo , NADPH Oxidasa 4/metabolismo , Comunicación Paracrina/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Proteína smad3/metabolismo , Telomerasa/metabolismo , Telómero/metabolismo , Factor de Crecimiento Transformador beta1/farmacología , Proteína p53 Supresora de Tumor/metabolismo
17.
Clin Cancer Res ; 24(19): 4771-4784, 2018 10 01.
Artículo en Inglés | MEDLINE | ID: mdl-29563139

RESUMEN

Purpose: Telomerase promoter mutations are highly prevalent in human tumors including melanoma. A subset of patients with metastatic melanoma often fail multiple therapies, and there is an unmet and urgent need to prolong disease control for those patients.Experimental Design: Numerous preclinical therapy-resistant models of human and mouse melanoma were used to test the efficacy of a telomerase-directed nucleoside, 6-thio-2'-deoxyguanosine (6-thio-dG). Integrated transcriptomics and proteomics approaches were used to identify genes and proteins that were significantly downregulated by 6-thio-dG.Results: We demonstrated the superior efficacy of 6-thio-dG both in vitro and in vivo that results in telomere dysfunction, leading to apoptosis and cell death in various preclinical models of therapy-resistant melanoma cells. 6-thio-dG concomitantly induces telomere dysfunction and inhibits the expression level of AXL.Conclusions: In summary, this study shows that indirectly targeting aberrant telomerase in melanoma cells with 6-thio-dG is a viable therapeutic approach in prolonging disease control and overcoming therapy resistance. Clin Cancer Res; 24(19); 4771-84. ©2018 AACR See related commentary by Teh and Aplin, p. 4629.


Asunto(s)
Desoxiguanosina/análogos & derivados , Melanoma/tratamiento farmacológico , Regiones Promotoras Genéticas/genética , Telomerasa/genética , Tionucleósidos/farmacología , Animales , Línea Celular Tumoral , Desoxiguanosina/farmacología , Resistencia a Antineoplásicos/efectos de los fármacos , Humanos , Melanoma/genética , Melanoma/patología , Ratones , Mutación , Telómero/efectos de los fármacos , Telómero/genética
18.
Mech Ageing Dev ; 128(1): 36-44, 2007 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-17116315

RESUMEN

Cellular senescence, a stress induced growth arrest of somatic cells, was first documented in cell cultures over 40 years ago, however its physiological significance has only recently been demonstrated. Using novel biomarkers of cellular senescence we examined whether senescent cells accumulate in tissues from baboons of ages encompassing the entire lifespan of this species. We show that dermal fibroblasts, displaying markers of senescence such as telomere damage, active checkpoint kinase ATM, high levels of heterochromatin proteins and elevated levels of p16, accumulate in skin biopsies from baboons with advancing age. The number of dermal fibroblasts containing damaged telomeres reaches a value of over 15% of total fibroblasts, whereas 80% of cells contain high levels of the heterochromatin protein HIRA. In skeletal muscle, a postmitotic tissue, only a small percentage of myonuclei containing damaged telomeres were detected regardless of animal age. The presence of senescent cells in mitotic tissues might therefore be a contributing factor to aging and age related pathology and provides further evidence that cellular senescence is a physiological event.


Asunto(s)
Envejecimiento/fisiología , Senescencia Celular/fisiología , Fibroblastos/fisiología , Mitosis/fisiología , Animales , Células Cultivadas , Fibroblastos/citología , Papio
19.
Methods Mol Biol ; 1534: 69-78, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-27812868

RESUMEN

Expressing oncogenes in normal somatic human cells leads to cellular senescence after just a few cell division cycles. In cells that are more resistant to culture stresses, such as human dermal fibroblasts, this oncogene-induced senescence (OIS) is a result of a DNA damage response (DDR) that is activated due to the formation of DNA lesions at both non-telomeric and telomeric DNA sequences. DNA lesions can be visualized as DDR foci by immunofluorescence microscopy using antibodies against a number of DDR factors, including ϒ-H2AX and 53BP1. Over time and as cells remain arrested in OIS, non-telomeric DDR foci progressively become resolved, while telomeric DDR foci, also called dysfunctional telomeres, persist. Here we describe a protocol to detect dysfunctional telomeres in cultured human cells, to monitor a temporal enrichment of dysfunctional telomeres in cells that had undergone OIS, and to detect dysfunctional telomeres in paraffin-embedded and formalin-fixed human tissue.


Asunto(s)
Senescencia Celular/genética , Oncogenes/genética , Telómero/genética , Biomarcadores , Línea Celular Tumoral , Daño del ADN , Fibroblastos/metabolismo , Técnica del Anticuerpo Fluorescente , Expresión Génica , Vectores Genéticos/genética , Humanos , Hibridación Fluorescente in Situ , Retroviridae/genética , Transducción Genética
20.
Mech Ageing Dev ; 127(1): 16-24, 2006 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-16229875

RESUMEN

After a limited number of divisions, most eukaryotic cells grown in culture will undergo a terminal growth arrest called cellular senescence. This growth arrest is thought to be a consequence of progressive telomere shortening that occurs due to incomplete DNA replication of the chromosome ends. In addition, cellular senescence can also be induced by a number of environmental stresses and signaling imbalances which are independent of telomere shortening. The cyclin dependent kinase inhibitors p21 and p16(INK4a) have been shown to execute and maintain the cell cycle arrest in senescence but the nature of the signals that cause upregulation of these inhibitors in senescent cells are only now starting to be discovered. Here we will review the current literature that leads us to propose a model how independent signals activate distinct signaling pathways to regulate p21 and p16(INK4a) levels in senescent cells.


Asunto(s)
Senescencia Celular/fisiología , Telómero/patología , Animales , Proliferación Celular , Inhibidor p16 de la Quinasa Dependiente de Ciclina/metabolismo , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/metabolismo , Humanos , Telómero/metabolismo , Regulación hacia Arriba
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