Nanocarriers targeting senescent cells

Ageing and multiple chronic diseases in humans associate with an aberrant accumulation of senescent cells in a diversity of tissues, which contributes to organ dysfunction and drives their pathological manifestations. It emerges that elimination of senescent cells ameliorates and even reverts the pro- gression of age-related disorders, thereby extending healthspan and lifespan in preclinical mouse models. Development of delivery carriers specifically targeting senescent cells offers innovative thera- peutic and diagnostic applications in translational medicine.The laboratory of D.M.-E. is funded by Cancer Research UK (CRUK), the CRUK Cambridge Centre Early Detection Programme, the Medical Research Council (MRC), and the Royal Society

Nuclear localization signals in phage terminal proteins provide a novel gene delivery tool in mammalian cells

Terminal proteins (TPs) of bacteriophages prime DNA replication and become covalently linked to the genome ends. Unexpectedly, we have found functional eukaryotic nuclear localization signals (NLSs) within the TP sequences of bacteriophages from diverse families and hosts. Given the role of bacteriophages as vehicles for horizontal gene transfer (HGT), we postulated that viral genomes that have covalently linked NLS-containing terminal proteins might behave as vectors for HGT between bacteria and the eukaryotic nucleus. To validate this hypothesis, we profited from the in vitro Φ29 amplification system that allows the amplification of heterologous DNAs producing linear molecules of DNA with TP covalently attached to both 5' ends. Interestingly, these in vitrogenerated TP-DNA molecules showed enhanced gene delivery in mammalian cells, supporting a possible role in HGT by transferring genes between prokaryotes and eukaryotes. Moreover, these TP-DNA molecules are a useful tool to amplify and subsequently deliver genes efficiently into the eukaryotic nucleus. Here, we suggest various possible applications and further developments of the technique with biotechnological and therapeutic purposesThis work was supported by Grant BFU 2011-23645 and Consolider-Ingenio Grant 2010 24717 from the Spanish Ministry of Economy and Competitiveness (MINECO) and by an institutional grant from Fundacion Ramon Areces to the Centro de Biologia Molecular “Severo Ochoa.

Targeting senescent cells in translational medicine.

Organismal ageing is a complex process driving progressive impairment of functionality and regenerative potential of tissues. Cellular senescence is a state of stable cell cycle arrest occurring in response to damage and stress and is considered a hallmark of ageing. Senescent cells accumulate in multiple organs during ageing, contribute to tissue dysfunction and give rise to pathological manifestations. Senescence is therefore a defining feature of a variety of human age-related disorders, including cancer, and targeted elimination of these cells has recently emerged as a promising therapeutic approach to ameliorate tissue damage and promote repair and regeneration. In addition, in vivo identification of senescent cells has significant potential for early diagnosis of multiple pathologies. Here, we review existing senolytics, small molecules and drug delivery tools used in preclinical therapeutic strategies involving cellular senescence, as well as probes to trace senescent cells. We also review the clinical research landscape in senescence and discuss how identifying and targeting cellular senescence might positively affect pathological and ageing processes

Temperature Inversions due to warm air advections at low levels in the Vega Media del Segura river (Region of Murcia)

En el Sureste peninsular son frecuentes las situaciones anticiclónicas alimentadas en invierno por la dorsal subtropical marítima. Bajo estas condiciones, las cuencas y valles fluviales intrabéticos son bastante proclives al desarrollo de inversiones térmicas. Un claro ejemplo lo constituye la Vega Media del Segura en los meses de invierno, especialmente cuando el enfriamiento nocturno del suelo aparece acompañado por movimientos de aire anticiclónico subsidente, advecciones cálidas de componente SO a niveles bajos atmosféricos y flujos fríos catabáticos procedentes de la Sierra de Carrascoy. En el presente artículo se analizan tales factores y las situaciones de inversión térmica más importantes registradas en esta Vega durante el período 2000-2011. Para ello se han utilizado los datos de las estaciones meteorológicas y de los sondeos aerológicos disponibles en el área, en particular los relativos a régimen de temperaturas, estratificación térmica vertical e indicadores de estabilidad atmosférica.Anticyclonic situations fed by the maritime subtropical dorsal in winter are frequent in the South-east Spain. Under these conditions, the intrabaetic depressions and fluvial valleys are prone to thermal inversions. A clear example is the Middle Valley of the Segura River during winter months, especially when cooling at night appears to be accompanied by anticyclonic subsident air, warm advection SW flows at low atmospheric levels and cold katabatic winds proceeding from Sierra de Carrascoy. Such factors and the most temperature inversion situations registered in the study area during the period 2000-2011 are analyzed in this work. For this purpose data of meteorological stations and aerological soundings (e.g. temperature regime, vertical thermal stratification, indicators of atmospheric stability) were used

Dual-Specificity Phosphatase 1 (DUSP1) Has a Central Role in Redox Homeostasis and Inflammation in the Mouse Cochlea.

Stress-activated protein kinases (SAPK) are associated with sensorineural hearing loss (SNHL) of multiple etiologies. Their activity is tightly regulated by dual-specificity phosphatase 1 (DUSP1), whose loss of function leads to sustained SAPK activation. Dusp1 gene knockout in mice accelerates SNHL progression and triggers inflammation, redox imbalance and hair cell (HC) death. To better understand the link between inflammation and redox imbalance, we analyzed the cochlear transcriptome in Dusp1-/- mice. RNA sequencing analysis (GSE176114) indicated that Dusp1-/- cochleae can be defined by a distinct profile of key cellular expression programs, including genes of the inflammatory response and glutathione (GSH) metabolism. To dissociate the two components, we treated Dusp1-/- mice with N-acetylcysteine, and hearing was followed-up longitudinally by auditory brainstem response recordings. A combination of immunofluorescence, Western blotting, enzymatic activity, GSH levels measurements and RT-qPCR techniques were used. N-acetylcysteine treatment delayed the onset of SNHL and mitigated cochlear damage, with fewer TUNEL+ HC and lower numbers of spiral ganglion neurons with p-H2AX foci. N-acetylcysteine not only improved the redox balance in Dusp1-/- mice but also inhibited cytokine production and reduced macrophage recruitment. Our data point to a critical role for DUSP1 in controlling the cross-talk between oxidative stress and inflammation

Runoff Water as A Resource in the Campo de Cartagena (Region of Murcia): Current Possibilities for Use and Benefits

The scarcity of water in the Campo de Cartagena has limited its exploitation, which is why, historically, runoff water has been used through sustainable traditional practices which have been dismissed by technological advances. In order to demonstrate the potential of this resource at present, an analysis by interpolation of rainfall distribution in the sub-basin of the Rambla de Fuente Álamo-Albujón was carried out (for the intense rainfall episodes of 2012 and 2016) as well as hydraulic modelling of the estimation of surface runoff. In addition, taking into account the future climate scenarios, a projection of the total runoff in the study area was made up to the year 2100. The bibliographic review and the press analysis showed that the traditional use of runoff water has remained in disuse, although there are infrastructures to collect water from floods but with an eminently sanitary purpose. The current model of agricultural and touristic exploitation is giving rise to serious socio-environmental conflicts which manifest in obsolescence. Therefore, the increase in the availability of water with the use of a specific endogenous resource may lead to a decrease in the pressures exerted on the study area.La escasez hídrica del Campo de Cartagena ha limitado su explotación, por ello, históricamente, se han aprovechado las aguas de escorrentía mediante prácticas tradicionales sostenibles que han sido desterradas por los avances tecnológicos. Con el propósito de demostrar el potencial de este recurso en la actualidad, se ha llevado a cabo (para los episodios pluvimétricos intensos de 2012 y 2016), un análisis por interpolación de la distribución de precipitaciones en la subcuenca de la Rambla de Fuente Álamo-Albujón, así como una modelización hidráulica de la estimación de la escorrentía superficial. Además, atendiendo a los escenarios climáticos futuros, se ha realizado una proyección de la escorrentía total en el área de estudio hasta el año 2100. La revision bibliográfica y el análisis de prensa ha puesto de manifiesto que el aprovechamiento tradicional de las escorrentías ha quedado en desuso aunque existen infraestructuras de captación de arroyadas pero con una finalidad eminentemente sanitaria. El modelo actual de explotación agrícola y túristica está dando lugar a conflictos socioambientales graves que manifiestan su obsolescencia por lo que el aumento de la disponibilidad de agua con el aprovechamiento de un recurso endógeno puntual, supondría una disminución de las presiones ejercidas sobre el área de estudio

Cellular senescence in cancer: from mechanisms to detection

Senescence refers to a cellular state featuring a stable cell‐cycle arrest triggered in response to stress. This response also involves other distinct morphological and intracellular changes including alterations in gene expression and epigenetic modifications, elevated macromolecular damage, metabolism deregulation and a complex pro‐inflammatory secretory phenotype. The initial demonstration of oncogene‐induced senescence in vitro established senescence as an important tumour‐suppressive mechanism, in addition to apoptosis. Senescence not only halts the proliferation of premalignant cells but also facilitates the clearance of affected cells through immunosurveillance. Failure to clear senescent cells owing to deficient immunosurveillance may, however, lead to a state of chronic inflammation that nurtures a pro‐tumorigenic microenvironment favouring cancer initiation, migration and metastasis. In addition, senescence is a response to post‐therapy genotoxic stress. Therefore, tracking the emergence of senescent cells becomes pivotal to detect potential pro‐tumorigenic events. Current protocols for the in vivo detection of senescence require the analysis of fixed or deep‐frozen tissues, despite a significant clinical need for real‐time bioimaging methods. Accuracy and efficiency of senescence detection are further hampered by a lack of universal and more specific senescence biomarkers. Recently, in an attempt to overcome these hurdles, an assortment of detection tools has been developed. These strategies all have significant potential for clinical utilisation and include flow cytometry combined with histo‐ or cytochemical approaches, nanoparticle‐based targeted delivery of imaging contrast agents, OFF‐ON fluorescent senoprobes, positron emission tomography senoprobes and analysis of circulating SASP factors, extracellular vesicles and cell‐free nucleic acids isolated from plasma. Here, we highlight the occurrence of senescence in neoplasia and advanced tumours, assess the impact of senescence on tumorigenesis and discuss how the ongoing development of senescence detection tools might improve early detection of multiple cancers and response to therapy in the near future

A Two-Photon Probe Based on Naphthalimide-Styrene Fluorophore for the In Vivo Tracking of Cellular Senescence

Cellular senescence is a state of stable cell cycle arrest that can negatively affect the regenerative capacities of tissues and can contribute to inflammation and the progression of various aging-related diseases. Advances in the in vivo detection of cellular senescence are still crucial to monitor the action of senolytic drugs and to assess the early onset or accumulation of senescent cells. Here, we describe a naphthalimide-styrene-based probe (HeckGal) for the detection of cellular senescence both in vitro and in vivo. HeckGal is hydrolyzed by the increased lysosomal β-galactosidase activity of senescent cells, resulting in fluorescence emission. The probe was validated in vitro using normal human fibroblasts and various cancer cell lines undergoing senescence induced by different stress stimuli. Remarkably, HeckGal was also validated in vivo in an orthotopic breast cancer mouse model treated with senescence-inducing chemotherapy and in a renal fibrosis mouse model. In all cases, HeckGal allowed the unambiguous detection of senescence in vitro as well as in tissues and tumors in vivo. This work is expected to provide a potential technology for senescence detection in aged or damaged tissues

Expression of sterol regulatory element-binding proteins in epicardial adipose tissue in patients with coronary artery disease and diabetes mellitus: preliminary study

[Abstract] Objectives: Sterol regulatory element-binding proteins (SREBP) genes are crucial in lipid biosynthesis and cardiovascular homeostasis. Their expression in epicardial adipose tissue (EAT) and their influence in the development of coronary artery disease (CAD) and type-2 diabetes mellitus remain to be determined. The aim of our study was to evaluate the expression of SREBP genes in EAT in patients with CAD according to diabetes status and its association with clinical and biochemical data. Methods: SREBP-1 and SREBP-2 mRNA expression levels were measured in EAT from 49 patients with CAD (26 with diabetes) and 23 controls without CAD or diabetes. Results: Both SREBPs mRNA expression were significantly higher in patients with CAD and diabetes (p<0.001) and were identified as independent cardiovascular risk factor for coronary artery disease in patients with type-2 diabetes (SREBP-1: OR 1.7, 95%CI 1.1-2.5, p=0.02; SREBP-2: OR 1.6, 95%CI 1.2-3, p=0.02) and were independently associated with the presence of multivessel CAD, left main and anterior descending artery stenosis, and higher total and LDL cholesterol levels, and lower HDL cholesterol levels, in patients with CAD and diabetes. Conclusions: SREBP genes are expressed in EAT and were higher in CAD patients with diabetes than those patients without CAD or diabetes. SREBP expression was associated as cardiovascular risk factor for the severity of CAD and the poor lipid control. In this preliminary study we suggest the importance of EAT in the lipid metabolism and cardiovascular homeostasis for coronary atherosclerosis of patients with diabetes and highlight a future novel therapeutic target.Instituto de Salud Carlos III; PI13/02542Instituto de Salud Carlos III; PI11/01661Red de Investigación Cardiovascular; RD12/0042/003

Robust, universal biomarker assay to detect senescent cells in biological specimens.

Cellular senescence contributes to organismal development, aging, and diverse pathologies, yet available assays to detect senescent cells remain unsatisfactory. Here, we designed and synthesized a lipophilic, biotin-linked Sudan Black B (SBB) analogue suitable for sensitive and specific, antibody-enhanced detection of lipofuscin-containing senescent cells in any biological material. This new hybrid histo-/immunochemical method is easy to perform, reliable, and universally applicable to assess senescence in biomedicine, from cancer research to gerontology