Papel de la citoquina LIGHT en la esteatosis hepática y la resistencia a insulina

[ES] La enfermedad del hígado graso no alcohólico (EHGNA) es una complicación metabólica que se asocia frecuentemente a la diabetes mellitus tipo 2 (DMT2) y a la disfunción del tejido adiposo. La progresión de la EHGNA está mediada, entre otras, por la activación de vías inflamatorias. En este estudio, se exploró el papel de la citoquina proinflamatoria LIGHT(TNFSF14) en el desarrollo de la EHGNA y la DMT2 en ratones. Con este fin, se evaluó la expresión de LIGHT y de sus receptores, el mediador de entrada del virus herpes (HVEM, del inglés herpes virus entry mediator) y el receptor de la linfotoxina β (LTβR), en ratones silvestres (WT) tratados con dieta control (DC) y con dieta alta en grasa y alta en colesterol (DAGAC), que induce hígado graso, durante 16 semanas. Por otro lado, se investigó el papel de LIGHT mediante la pérdida de función en ratones deficientes en Light (Light-/-) y en sus controles WT alimentados con DC o DAGAC durante 16 semanas. Además, se analizó la inactivación génica de Light en ratones parcialmente deficientes en Irs2 (Irs2+/-Light-/-), que es un mediador clave en la señalización de la insulina, y sus controles (Irs2+/-) alimentados con DAGAC durante 16 semanas. Para la caracterización de los ratones tratados con las diferentes dietas y condiciones se analizó la tolerancia a la glucosa, la sensibilidad a insulina, los parámetros determinantes de hígado graso, la inflamación tisular y sistémica y la expresión de genes metabólicos. Los ratones WT alimentados con DAGAC mostraron un incremento en los niveles proteicos de los receptores LTβR y HVEM, junto con un aumento del contenido hepático de triacilgliceroles (TG), de linfocitos T CD3+, de la fibrosis y de marcadores inflamatorios comparado con ratones WT alimentados con DC. Los resultados del análisis de los ratones alimentados con DC mostraron que la deficiencia genética de Light no altera el metabolismo, el hígado graso o la inflamación. Sin embargo, en condiciones de DAGAC, la inactivación de Light, ratones Light-/- e Irs2+/-Light-/-, mejoró la tolerancia a la glucosa. Además, los ratones Light-/- mostraron un aumento de la sensibilidad a insulina. En el análisis hepático se observó una disminución del hígado graso y del contenido de macrófagos F4/80+ en los ratones Light-/- e Irs2+/-Light-/-. Además, los ratones Light-/- mostraron también una reducción de linfocitos T CD3+ hepáticos y de la expresión del factor transcripcional Zbtb16, determinante en los linfocitos NKT. Consistente con el papel potencial del tejido adiposo en la homeostasis hepática, los ratones Light-/- e Irs2+/-Light-/- alimentados con DAGAC exhibieron un aumento de macrófagos antiinflamatorios F4/80+CD206+ y una reducción de macrófagos proinflamatorios F4/80+CD11c+ y de la secreción de las citoquinas proinflamatorias en el tejido adiposo. De manera similar, en el análisis de la inflamación sistémica se observó un aumento de los monocitos antiinflamatorios, una reducción de linfocitos proinflamatorios, y una disminución de los niveles de citoquinas proinflamatorias en los ratones Light-/- e Irs2+/-Light-/- comparado con los ratones WT e Irs2+/. El estudio de la expresión de genes hepáticos en los ratones Light-/- e Irs2+/-Light-/- mostró un patrón de expresión consistente con una mejora en la homeostasis hepática y una reducción en la expresión de genes previamente implicados en la transición esteatosis simple-esteatohepatitis no alcohólica (EHNA), en la activación de células estrelladas y en el carcinoma hepatocelular (CHC). Estos resultados indican que la deficiencia de Light mejora el metabolismo de carbohidratos, el hígado graso y la inflamación inducidos por una DAGAC. Por lo tanto, la señalización dependiente de LIGHT puede ser una diana terapéutica para restablecer la homeostasis hepática, moderar la EHGNA y aliviar la inflamación en condiciones de DAGAC.[CA] La malaltia del fetge gras no alcohòlic (MFGNA) és una complicació metabòlica que s'associa freqüentment a la diabetis mellitus tipus 2 (DMT2) i a la disfunció del teixit adipós. La progressió de l'EHGNA es regeix, entre d'altres, per l'activació de vies inflamatòries. En aquest estudi, es va explorar el paper de la citocina pro inflamatòria LIGHT(TNFSF14) en el desenvolupament de la MFGNA i la DMT2 en ratolins. Amb aquesta finalitat, es va avaluar l'expressió de LIGHT i dels seus receptors, el mediador d'entrada de l'herpes virus (HVEM, del anglès herpes virus entry mediator) i el receptor de la limfotoxina β (LTβR), en ratolins silvestres (WT) tractats amb dieta control (DC) i amb dieta alta en greixos i colesterol (DAGAC), que indueix fetge gras, al llarg de 16 setmanes. D'altra banda, es va investigar el paper de LIGHT mitjançant la pèrdua de funció en ratolins deficients en Light (Light-/-) i els seus controls WT alimentats amb DC o DAGAC al llarg de 16 setmanes. A més a més, es va analitzar la inactivació gènica de Light en ratolins parcialment deficients en lrs2 (lrs2+/-Light-/-), que és un mediador clau en la senyalització de la insulina, i els seus controls (lrs2+/-) alimentats amb DAGAC 16 setmanes. Per caracteritzar els ratolins tractats amb les diferents dietes i condicions es va analitzar la tolerància a la glucosa, la sensibilitat a la insulina, els paràmetres determinats del fetge gras, la inflamació tissular i sistèmica, i l'expressió de gens metabòlics. Els ratolins WT alimentats amb DAGAC van mostrar un increment als nivells proteics dels receptors LTβR i HVEM, junt amb un augment del contingut hepàtic de triacilglicerids (TG), de limfòcits T CD3+, de la fibrosi i de marcadors inflamatoris comparats amb ratolins WT alimentats amb DC. Els resultats de l'anàlisi dels ratolins alimentats amb DC van mostrar que la deficiència genètica de Light no altera el metabolisme, el fetge gras o la inflamació. Tanmateix, en condicions de DAGAC, la inactivació de Light, ratolins Light-/- i Irs2+/-Light-/-, millorà la tolerància a la glucosa. A més a més, els ratolins Light-/- mostraren un augment de la sensibilitat a insulina. A l'anàlisi hepàtic es va observar una disminució del fetge gras i del contingut de macròfags F4/80+ en ratolins Light-/- i Irs2+/-Light-/-, i els ratolins Light-/- van mostrar també una reducció de limfòcits T CD3+ hepàtics i de l'expressió del factor transcripcional Zbtb16, determinant als limfòcits NKT. Consistent amb el paper potencial del teixit adipós en l'homeòstasi hepàtica, els ratolins Light-/- i Irs2+/-Light-/- alimentats amb DAGAC exhibiren un augment de macròfags antiinflamatoris F4/80+CD206+ i una reducció de macròfags pro inflamatoris F4/80+CD11c+ i de la secreció de les citocines pro inflamatòries al teixit adipós. De manera semblant, l'anàlisi de la inflamació sistèmica va mostrar un augment dels monòcits antiinflamatoris, una reducció dels limfòcits pro inflamatoris, i una disminució dels nivells de citocines pro inflamatòries en ratolins Light-/- i Irs2+/-Light-/- en comparació amb els ratolins WT i Irs2+/-. L'estudi de l'expressió de gens hepàtics amb els ratolins Light-/- i Irs2+/-Light-/- va mostrar un patró d'expressió consistent amb una millora en l'homeòstasi hepàtica i una reducció en l'expressió de gens prèviament implicats en la transició esteatosi simple-esteatohepatitis no alcohòlica (EHNA), en l'activació de cèl·lules estrellades i en carcinoma hepatocel·lular (CHC). Aquests resultats indiquen que la deficiència de Light millora el metabolisme de carbohidrats, el fetge gras i la inflamació induïts per la DAGAC. Per tant, la senyalització depenent de LIGHT pot ser una diana terapèutica per restablir l'homeòstasi hepàtica, moderar la MFGNA i mitigar la inflamació en condicions de DAGAC.[EN] Non-alcoholic fatty liver disease (NAFLD) is frequently associated with type 2 diabetes mellitus (T2DM) and adipose tissue dysfunction. Progression of NAFLD is mediated, among others, by activation of inflammatory pathways. In the present study, the role of the proinflammatory cytokine LIGHT(TNFSF14) was explored in NAFLD and T2DM in mice. To this end, the expression of LIGHT and its receptors, herpes virus entry mediator (HVEM) and lymphotoxin β receptor (LTβR), was investigated in wild-type (WT) mice on a regular chow diet (RCD) and on a high-fat high-cholesterol diet (HFHCD) for 16 weeks to induce fatty liver. The role of LIGHT in NAFLD was as well investigated by loss-of-function approaches using Light-deficient (Light−/−) mice and WT controls which were fed either a RCD or HFHCD for 16 weeks. In addition, Light deficiency was also evaluated in mice partial deficient in Irs2, a major insulin effector which lack of function induces insulin resistance. Thus, Irs2+/-Light-/- and control Irs2+/- mice were fed a HFHCD for 16 weeks. To characterize mice, glucose tolerance, insulin sensitivity, non-alcoholic fatty liver (NAFL), systemic and tissue inflammation and metabolic gene expression were explored. HVEM and LTβR expression, hepatic triacyclglycerols (TG) and CD3+ T lymphocytes content, fibrosis and inflammatory markers were markedly increased in HFHCD-fed WT mice compared with RCD-fed WT. Consistent with the above findings, Light deficiency did not affect metabolism, NAFL and tissue and systemic inflammation in RCD-fed WT mice. However, under HFHCD, Light deficiency in WT mice and in Irs2+/- mice improved glucose tolerance. Moreover, Light-/- mice displayed improved insulin sensitivity. Fatty liver and hepatic F4/80+ macrophage content were decreased in both HFHCD-fed Light-/- and Irs2+/-Light-/- mice compared with their respective HFHCD-fed controls. A decrease in hepatic CD3+ T lymphocytes content and in expression levels of Zbtb16, the transcription factor essential for natural killer T (NKT) cell function, in HFHCD-fed Light−/− mice compared to its controls was also observed. Consistent with a potential role of adipose tissue in hepatic homeostasis, Light-/- and Irs2+/-Light-/- mice exhibited augmented anti-inflammatory F4/80+CD206+ adipose tissue macrophages, reduced proinflammatory F4/80+CD11c+ adipose tissue macrophages and diminished secretion of cytokines. Systemic inflammation showed augmented anti-inflammatory monocytes and a decrease in proinflammatory lymphocytes in Light−/− and Irs2+/-Light-/- mice compared to their controls. Analysis of hepatic gene expression in Light−/− and Irs2+/-Light-/- mouse livers showed an alteration in metabolic gene expression pattern consistent with improved hepatic homeostasis and a reduction in the expression of genes involved in NAFL-to-non-alcoholic steatohepatitis (NASH) transition, hepatic stellate cells activation and hepatocellular carcinoma (HCC). These results indicate that Light deficiency improves carbohydrate metabolism and reduces NAFL and inflammation induced by HFHCD. These results suggest that therapies to block LIGHT-dependent signalling might be useful to restore hepatic homeostasis, to restrain NAFLD and to alleviate inflammation in HFHCD conditions.La presente Tesis Doctoral ha sido financiada por las siguientes ayudas: Desarrollo preclínico nuevo tratamiento de la ateroesclerosis APOTIP/2018/02. Entidad financiadora: Generalitat Valenciana Conselleria de Investigació, Cultura i Esport. Investigadora principal: Herminia González Navarro. / Papel de los procesos inflamatorios asociados a la diabetes en la estabilidad de la placa de ateroma y estudio del uso potencial de estrategias terapéuticas PI16/00091. Entidad financiadora: Instituto de Salud Carlos III Acción Estratégica en Salud. Investigadora principal: Herminia González Navarro. / Role of the cytokine LIGHT (TNFSF14) in the development of IR and fatty liver Disease NN_2016_3. Entidad financiadora: European foundation for the study of diabetes (EFSD)/Novo Nordisk Programme for Diabetes Research in Europe. Investigadora principal: Herminia González Navarro. / Estudio de los mecanismos moleculares de la Diabetes Mellitus y su papel en el desarrollo de la aterosclerosis PI13/00834. Entidad financiadora: Instituto de Salud Carlos III Acción Estratégica en Salud. Investigadora principal: Herminia González NavarroHerrero Cervera, A. (2020). Papel de la citoquina LIGHT en la esteatosis hepática y la resistencia a insulina [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/137778TESI

Type 1 diabetic mellitus patientswith increased atherosclerosisriskdisplay decreased CDKN2A/2B/2BAS gene expression in leukocytes

Background: Type 1 diabetes mellitus (T1DM) patients display increased risk of cardiovascular disease (CVD) and are characterized by a diminished regulatory T (Treg) cell content or function. Previous studies have shown an association between decreased CDKN2A/2B/2BAS gene expression and enhanced CVD. In the present study the potential relationship between CDKN2A/2B/2BAS gene expression, immune cell dysfunction and increased cardiovascular risk in T1DM patients was explored. Methods: A cross-sectional study was performed in 90 subjects divided into controls and T1DM patients. Circulating leukocyte subpopulations analysis by flow cytometry, expression studies on peripheral blood mononuclear cell by qPCR and western blot and correlation studies were performed in both groups of subjects. Results: Analysis indicated that, consistent with the described T cell dysfunction, T1DM subjects showed decreased circulating CD4+CD25+CD127- Treg cells. In addition, T1DM subjects had lower mRNA levels of the transcription factors FOXP3 and RORC and lower levels of IL2 and IL6 which are involved in Treg and Th17 cell differentiation, respectively. T1DM patients also exhibited decreased mRNA levels of CDKN2A (variant 1 p16Ink4a), CDKN2A (p14Arf, variant 4), CDKN2B (p15Ink4b) and CDKN2BAS compared with controls. Notably, T1DM patients had augmented pro-atherogenic CD14++CD16+-monocytes, which predict cardiovascular acute events and enhanced common carotid intima-media thickness (CC-IMT). Conclusions: Decreased expression of CDKN2A/2B/2BAS in leukocytes associates with increased CC-IMT atherosclerosis surrogate marker and proatherogenic CD14++CD16+ monocytes in T1DM patients. These results suggest a potential role of CDKN2A/2B/2BAS genes in CVD risk in T1DM

Hacia la investigación basada en la evidencia

ORIGINAL ARTICLE Lund H, Brunnhuber K, Juhl C, Robinson K, Leenaars M, Dorch BF, Jamtvedt G, Nortvedt MW, Christensen R, Chalmers I. Towards evidence based research. BMJ. 2016;355:i5440. doi: 10.1136/bmj.i5440Publication date of the original article: October 21, 2016RIGHTS AND EXEMPTIONS FROM RESPONSIBILITIESTranslated with permission from BMJ Publishing Group Ltd. Lund H, Brunnhuber K, Juhl C, Robinson K, Leenaars M, Dorch BF, Jamtvedt G, Nortvedt MW, Christensen R, Chalmers I. Towards Evidence Based Research, BMJ 2016;355 :i5440 © 2016 BMJ Publishing Group Ltd.The original authors have not revised and verified the Spanish translation, and not necessary endorse it.BMJ takes no responsibility for the accuracy of the translation from the published English original and is not liable for any errors which may occur. No responsibility is assumed, and responsibility is hereby disclaimed, by BMJ for any injury and/or damage to persons or property as a matter of product liability, negligence or otherwise, or from any use or operation of methods, products, instructions or ideas presented in the original content.KEY MESSAGESEmbarking on research without reviewing systematically what is already known, particularly when the research involves people or animals, is unethical, unscientific, and wastefulA systematic review of relevant evidence can establish whether the proposed research is truly neededSome research funders now require applicants to refer to a systematic review of existing researchResearch waste can also be reduced by efficient production, updating, and dissemination of systematic reviewsARTÍCULO ORIGINALLund H, Brunnhuber K, Juhl C, Robinson K, Leenaars M, Dorch BF, Jamtvedt G, Nortvedt MW, Christensen R, Chalmers I. Towards evidence based research. BMJ. 2016;355:i5440. doi: 10.1136/bmj.i5440Fecha de publicación del artículo original: 21 de octubre de 2016DERECHOS Y EXENCIÓN DE RESPONSABILIDADESTraducido con el permiso de BMJ Publishing Group Ltd. Lund H, Brunnhuber K, Juhl C, Robinson K, Leenaars M, Dorch BF, Jamtvedt G, Nortvedt MW, Christensen R, Chalmers I. Towards Evidence Based Research, BMJ 2016;355 :i5440 © 2016 BMJ Publishing Group Ltd.Los autores originales no han revisado ni verificado la traducción del manuscrito al español, y no necesariamente están de acuerdo con su contenidoBMJ no se hace responsable de la exactitud de la traducción publicada ni de los errores que pueda contener. No se asume responsabilidad alguna por parte de BMJ por cualquier lesión o daño a personas o propiedad por responsabilidad del producto, negligencia o de otra forma, ni de cualquier uso u operación de métodos, productos, instrucciones o ideas presentadas en el contenido original.MENSAJES CLAVEAventurarse en investigación sin revisar de manera sistemática lo ya conocido, especialmente cuando la investigación implica personas o animales, no es ético, carece de rigor científico y es un desperdicio.Una revisión sistemática de pruebas imparciales (evidencias) relevantes puede determinar si la investigación propuesta es realmente necesaria.Algunos organismos de financiación ahora requieren que los solicitantes hagan referencia a revisiones sistemáticas de investigaciones existentes.El desperdicio en investigación también se puede reducir mediante la producción, actualización y difusión eficientes de revisiones sistemáticas

Pharmacogenetics in Neuroblastoma: What Can Already Be Clinically Implemented and What Is Coming Next?

Pharmacogenetics is one of the cornerstones of Personalized Precision Medicine that needs to be implemented in the routine of our patients' clinical management in order to tailor their therapies as much as possible, with the aim of maximizing efficacy and minimizing toxicity. This is of great importance, especially in pediatric cancer and even more in complex malignancies such as neuroblastoma, where the rates of therapeutic success are still below those of many other types of tumors. The studies are mainly focused on germline genetic variants and in the present review, state of the art is presented: which are the variants that have a level of evidence high enough to be implemented in the clinic, and how to distinguish them from the ones that still need validation to confirm their utility. Further aspects as relevant characteristics regarding ontogeny and future directions in the research will also be discussed

Treatment with tocilizumab or corticosteroids for COVID-19 patients with hyperinflammatory state: a multicentre cohort study (SAM-COVID-19)

Objectives: The objective of this study was to estimate the association between tocilizumab or corticosteroids and the risk of intubation or death in patients with coronavirus disease 19 (COVID-19) with a hyperinflammatory state according to clinical and laboratory parameters. Methods: A cohort study was performed in 60 Spanish hospitals including 778 patients with COVID-19 and clinical and laboratory data indicative of a hyperinflammatory state. Treatment was mainly with tocilizumab, an intermediate-high dose of corticosteroids (IHDC), a pulse dose of corticosteroids (PDC), combination therapy, or no treatment. Primary outcome was intubation or death; follow-up was 21 days. Propensity score-adjusted estimations using Cox regression (logistic regression if needed) were calculated. Propensity scores were used as confounders, matching variables and for the inverse probability of treatment weights (IPTWs). Results: In all, 88, 117, 78 and 151 patients treated with tocilizumab, IHDC, PDC, and combination therapy, respectively, were compared with 344 untreated patients. The primary endpoint occurred in 10 (11.4%), 27 (23.1%), 12 (15.4%), 40 (25.6%) and 69 (21.1%), respectively. The IPTW-based hazard ratios (odds ratio for combination therapy) for the primary endpoint were 0.32 (95%CI 0.22-0.47; p < 0.001) for tocilizumab, 0.82 (0.71-1.30; p 0.82) for IHDC, 0.61 (0.43-0.86; p 0.006) for PDC, and 1.17 (0.86-1.58; p 0.30) for combination therapy. Other applications of the propensity score provided similar results, but were not significant for PDC. Tocilizumab was also associated with lower hazard of death alone in IPTW analysis (0.07; 0.02-0.17; p < 0.001). Conclusions: Tocilizumab might be useful in COVID-19 patients with a hyperinflammatory state and should be prioritized for randomized trials in this situatio

Papel del eje inflamatorio LIGHT(TNSF14)/Receptor de la Linfotoxina beta en la apoptosis y la proliferación asociados a la aterosclerosis

[ES] El objetivo principal del presente proyecto es el estudio del eje inflamatorio LIGHT(TNSF14)/Receptor de la Linfotoxina &#946; (LT&#946;-R) en el desarrollo de la aterosclerosis, la apoptosis y la proliferación asociadas a dicho proceso. Con este fin se llevarán a cabo tres objetivos específicos: 1. Estudio de la expresión génica del eje inflamatorio LIGHT/LT&#946;-R y correlación con el desarrollo de la apoptosis, la proliferación y en la aterosclerosis asociada al envejecimiento en ratones apoE-/-. Para ello, se analizarán y compararán los parámetros a estudiar en ratones de 4 y 10 meses de edad. 2. Estudio del efecto del tratamiento con LIGHT en la proliferación, la apoptosis y la aterosclerosis in vivo en lesiones de ratones apoE-/-. Con este fin se tratarán ratones apoE-/- de 2 meses de edad con LIGHT durante 7 y 28 días y se comparará con ratones tratados con vehículo durante el mismo tiempo. Dado el papel de LIGHT en la activación de células inmunes también se caracterizará la inflamación sistémica asociada a los tratamientos con LIGHT. 3. Estudio del efecto de la citocina LIGHT en la apoptosis y la proliferación en células de la pared arterial: en células de músculo liso vascular de aorta humana y en macrófagos murinos.Herrero Cervera, A. (2016). Papel del eje inflamatorio LIGHT(TNSF14)/Receptor de la Linfotoxina beta en la apoptosis y la proliferación asociados a la aterosclerosis. http://hdl.handle.net/10251/68872TFG

Understanding the Impact of Dietary Cholesterol on Chronic Metabolic Diseases through Studies in Rodent Models

The development of certain chronic metabolic diseases has been attributed to elevated levels of dietary cholesterol. However, decades of research in animal models and humans have demonstrated a high complexity with respect to the impact of dietary cholesterol on the progression of these diseases. Thus, recent investigations in non-alcoholic fatty liver disease (NAFLD) point to dietary cholesterol as a key factor for the activation of inflammatory pathways underlying the transition from NAFLD to non-alcoholic steatohepatitis (NASH) and to hepatic carcinoma. Dietary cholesterol was initially thought to be the key factor for cardiovascular disease development, but its impact on the disease depends partly on the capacity to modulate plasmatic circulating low-density lipoprotein (LDL) cholesterol levels. These studies evidence a complex relationship between these chronic metabolic diseases and dietary cholesterol, which, in certain conditions, might promote metabolic complications. In this review, we summarize rodent studies that evaluate the impact of dietary cholesterol on these two prevalent chronic diseases and their relevance to human pathology

Therapies for the Treatment of Cardiovascular Disease Associated with Type 2 Diabetes and Dyslipidemia

Cardiovascular disease (CVD) is the leading cause of death worldwide and is the clinical manifestation of the atherosclerosis. Elevated LDL-cholesterol levels are the first line of therapy but the increasing prevalence in type 2 diabetes mellitus (T2DM) has positioned the cardiometabolic risk as the most relevant parameter for treatment. Therefore, the control of this risk, characterized by dyslipidemia, hypertension, obesity, and insulin resistance, has become a major goal in many experimental and clinical studies in the context of CVD. In the present review, we summarized experimental studies and clinical trials of recent anti-diabetic and lipid-lowering therapies targeted to reduce CVD. Specifically, incretin-based therapies, sodium-glucose co-transporter 2 inhibitors, and proprotein convertase subtilisin kexin 9 inactivating therapies are described. Moreover, the novel molecular mechanisms explaining the CVD protection of the drugs reviewed here indicate major effects on vascular cells, inflammatory cells, and cardiomyocytes, beyond their expected anti-diabetic and lipid-lowering control. The revealed key mechanism is a prevention of acute cardiovascular events by restraining atherosclerosis at early stages, with decreased leukocyte adhesion, recruitment, and foam cell formation, and increased plaque stability and diminished necrotic core in advanced plaques. These emergent cardiometabolic therapies have a promising future to reduce CVD burden

Dapagliflozin Does Not Modulate Atherosclerosis in Mice with Insulin Resistance

Type 2 diabetes mellitus (T2DM) increases morbimortality in humans via enhanced susceptibility to cardiovascular disease (CVD). Sodium-glucose co-transporter 2 inhibitors (SGLT2i) are drugs designed for T2DM treatment to diminish hyperglycaemia by reducing up to 90% of renal tube glucose reabsorption. Clinical studies also suggest a beneficial action of SGLT2i in heart failure and CVD independent of its hypoglycaemiant effect. In the present study, we explored the effect of SGLT2i dapagliflozin (DAPA) in the metabolism and atherosclerosis in Apoe&minus;/&minus;Irs2+/&minus; mice, which display accelerated atherosclerosis induced by insulin resistance. DAPA treatment of Apoe&minus;/&minus;Irs2+/&minus; mice, which were fed a high-fat, high-cholesterol diet, failed to modify body weight, plasma glucose or lipid. Carbohydrate metabolism characterisation showed no effect of DAPA in the glucose tolerance test (GTT) despite augmented insulin levels during the test. In fact, decreased C-peptide levels in DAPA-treated mice during the GTT suggested impaired insulin release. Consistent with this, DAPA treatment of Apoe&minus;/&minus;Irs2+/&minus; isolated islets displayed lower glucose-stimulated insulin secretion compared with vehicle-treated islets. Moreover, insulin-signalling experiments showed decreased pAKT activation in DAPA-treated adipose tissue indicating impaired insulin signalling in this tissue. No changes were seen in lesion size, vulnerability or content of macrophages, vascular smooth muscle cells, T cells or collagen. DAPA did not affect circulating inflammatory cells or cytokine levels. Hence, this study indicates that DAPA does not protect against atherosclerosis in insulin-resistant mice in hypercholesterolemic conditions

Dissecting abdominal aortic aneurysm is aggravated by genetic inactivation of LIGHT (TNFSF14)

Abdominal aortic aneurysm (AAA), is a complex disorder characterized by vascular vessel wall remodeling. LIGHT (TNFSF14) is a proinflammatory cytokine associated with vascular disease. In the present study, the impact of genetic inactivation of Light was investigated in dissecting AAA induced by angiotensin II (AngII) in the Apolipoprotein E-deficient (Apoe−/−) mice. Studies in aortic human (ah) vascular smooth muscle cells (VSMC) to study potential translation to human pathology were also performed. AngII-treated Apoe−/−Light−/− mice displayed increased abdominal aorta maximum diameter and AAA severity compared with Apoe−/− mice. Notably, reduced smooth muscle α-actin+ area and Acta2 and Col1a1 gene expression were observed in AAA from Apoe−/−Light−/− mice, suggesting a loss of VSMC contractile phenotype compared with controls. Decreased Opn and augmented Sox9 expression, which are associated with detrimental and non-contractile osteochondrogenic VSMC phenotypes, were also seen in AngII-treated Apoe−/−Light−/− mouse AAA. Consistent with a role of LIGHT preserving VSMC contractile characteristics, LIGHT-treatment of ahVSMCs diminished the expression of SOX9 and of the pluripotency marker CKIT. These effects were partly mediated through lymphotoxin β receptor (LTβR) as the silencing of its gene ablated LIGHT effects on ahVSMCs. These studies suggest a protective role of LIGHT through mechanisms that prevent VSMC trans-differentiation in an LTβR-dependent manner