MicroRNA Dysregulation in Pulmonary Arteries from COPD: Relationships with Vascular Remodeling

Pulmonary vascular remodeling is an angiogenic-related process involving changes in smooth muscle cell (SMC) homeostasis, which is frequently observed in chronic obstructive pulmonary disease (COPD). MicroRNAs (miRNAs) are small, noncoding RNAs that regulate mRNA expression levels of many genes, leading to the manifestation of cell identity and specific cellular phenotypes. Here, we evaluate the miRNA expression profiles of pulmonary arteries (PAs) of patients with COPD and its relationship with the regulation of SMC phenotypic change. miRNA expression profiles from PAs of 12 patients with COPD, 9 smokers with normal lung function (SK), and 7 nonsmokers (NS) were analyzed using TaqMan Low-Density Arrays. In patients with COPD, expression levels of miR-98, miR-139-5p, miR-146b-5p, and miR-451 were upregulated, as compared with NS. In contrast, miR-197, miR-204, miR-485-3p, and miR-627 were downregulated. miRNA-197 expression correlated with both airflow obstruction and PA intimal enlargement. In an in vitro model of SMC differentiation, miR-197 expression was associated with an SMC contractile phenotype. miR-197 inhibition blocked the acquisition of contractile markers in SMCs and promoted a proliferative/migratory phenotype measured by both cell cycle analysis and wound-healing assay. Using luciferase assays, Western blot, and quantitative PCR, we confirmed that miR-197 targets the transcription factor E2F1. In PAs from patients with COPD, levels of E2F1 were increased as compared with NS. In PAs of patients with COPD, remodeling of the vessel wall is associated with downregulation of miR-197, which regulates SMC phenotype. The effect of miR-197 on PAs might be mediated, at least in part, by the key proproliferative factor, E2F1

Reduced alfa-MSH underlies hypothalamic ER-stress-induced hepatic gluconeogenesis

Alterations in ER homeostasis have been implicated in the pathophysiology of obesity and type-2 diabetes (T2D). Acute ER stress induction in the hypothalamus produces glucose metabolism perturbations. However, the neurobiological basis linking hypothalamic ER stress with abnormal glucose metabolism remains unknown. Here, we report that genetic and induced models of hypothalamic ER stress are associated with alterations in systemic glucose homeostasis due to increased gluconeogenesis (GNG) independent of body weight changes. Defective alpha melanocyte-stimulating hormone (α-MSH) production underlies this metabolic phenotype, as pharmacological strategies aimed at rescuing hypothalamic α-MSH content reversed this phenotype at metabolic and molecular level. Collectively, our results posit defective α-MSH processing as a fundamental mediator of enhanced GNG in the context of hypothalamic ER stress and establish α-MSH deficiency in proopiomelanocortin (POMC) neurons as a potential contributor to the pathophysiology of T2D

Mitochondrial cristae-remodeling protein OPA1 in POMC neurons couples Ca2+ homeostasis with adipose tissue lipolysis

Appropriate cristae remodeling is a determinant of mitochondrial function and bioenergetics and thus represents a crucial process for cellular metabolic adaptations. Here, we show that mitochondrial cristae architecture and expression of the master cristae-remodeling protein OPA1 in proopiomelanocortin (POMC) neurons, which are key metabolic sensors implicated in energy balance control, is affected by fluctuations in nutrient availability. Genetic inactivation of OPA1 in POMC neurons causes dramatic alterations in cristae topology, mitochondrial Ca2+ handling, reduction in alpha-melanocyte stimulating hormone (α-MSH) in target areas, hyperphagia, and attenuated white adipose tissue (WAT) lipolysis resulting in obesity. Pharmacological blockade of mitochondrial Ca2+ influx restores α-MSH and the lipolytic program, while improving the metabolic defects of mutant mice. Chemogenetic manipulation of POMC neurons confirms a role in lipolysis control. Our results unveil a novel axis that connects OPA1 in POMC neurons with mitochondrial cristae, Ca2+ homeostasis, and WAT lipolysis in the regulation of energy balance

Dietary α‐Linolenic Acid, Marine ω‐3 Fatty Acids, and Mortality in a Population With High Fish Consumption: Findings From the PREvención con DIeta MEDiterránea (PREDIMED) Study

Background: Epidemiological evidence suggests a cardioprotective role of α‐linolenic acid (ALA), a plant‐derived ω‐3 fatty acid. It is unclear whether ALA is beneficial in a background of high marine ω‐3 fatty acids (long‐chain n‐3 polyunsaturated fatty acids) intake. In persons at high cardiovascular risk from Spain, a country in which fish consumption is customarily high, we investigated whether meeting the International Society for the Study of Fatty Acids and Lipids recommendation for dietary ALA (0.7% of total energy) at baseline was related to all‐cause and cardiovascular disease mortality. We also examined the effect of meeting the society's recommendation for long‐chain n‐3 polyunsaturated fatty acids (≥500 mg/day). Methods and Results: We longitudinally evaluated 7202 participants in the PREvención con DIeta MEDiterránea (PREDIMED) trial. Multivariable‐adjusted Cox regression models were fitted to estimate hazard ratios. ALA intake correlated to walnut consumption (r=0.94). During a 5.9‐y follow‐up, 431 deaths occurred (104 cardiovascular disease, 55 coronary heart disease, 32 sudden cardiac death, 25 stroke). The hazard ratios for meeting ALA recommendation (n=1615, 22.4%) were 0.72 (95% CI 0.56–0.92) for all‐cause mortality and 0.95 (95% CI 0.58–1.57) for fatal cardiovascular disease. The hazard ratios for meeting the recommendation for long‐chain n‐3 polyunsaturated fatty acids (n=5452, 75.7%) were 0.84 (95% CI 0.67–1.05) for all‐cause mortality, 0.61 (95% CI 0.39–0.96) for fatal cardiovascular disease, 0.54 (95% CI 0.29–0.99) for fatal coronary heart disease, and 0.49 (95% CI 0.22–1.01) for sudden cardiac death. The highest reduction in all‐cause mortality occurred in participants meeting both recommendations (hazard ratio 0.63 [95% CI 0.45–0.87]). Conclusions: In participants without prior cardiovascular disease and high fish consumption, dietary ALA, supplied mainly by walnuts and olive oil, relates inversely to all‐cause mortality, whereas protection from cardiac mortality is limited to fish‐derived long‐chain n‐3 polyunsaturated fatty acids. Clinical Trial Registration URL: http://www.Controlled-trials.com/. Unique identifier: ISRCTN35739639

Bases moleculares del efecto del tungstato de sodio sobre la plasticidad pancreática y Tmem77

[spa] Esta tesis tiene dos partes bien diferenciadas. Una primera, en la que investigan las bases moleculares por las que el tratamiento con tungstato de sodio es capaz de inducir una regeneración del páncreas en los animales diabéticos. Y una segunda, que se centra en el papel de la proteína Tmem27 sobre los islotes pancreáticos y su posible uso como biomarcador de masa beta pancreática.Cada parte ha dado lugar a una publicación en una revista:A) "Molecular mechanisms of tungstate-induced pancreatic plasticity: a transcriptomics approach". BMC Genomics. 2009;10:406.ANTECEDENTES: El tungstato de sodio es un reconocido agente anti-diabético, capaz de incrementar la masa de células beta en modelos animales de diabetes. En la actualidad, los mecanismos moleculares por los que actúa y los genes que controlan la plasticidad pancreática son desconocidos. A través de una aproximación transcriptómica, nos proponemos descubrir lo mecanismos moleculares que participan en la recuperación de la función exocrina y endocrina de las ratas diabéticas-(STZ) tratadas con tungstato de sodio, determinando la contribución sobre este proceso del estado de hiperglicemia y el efecto directo/indirecto del tungstato. RESULTADOS: Las ratas diabéticas-(STZ) tratadas con tungstato presentaron una recuperación parcial de la función pancreática exocrina y endocrina, junto con un incremento en la masa de células beta. El análisis por microarrays del páncreas total definió tres grupos de genes: alterados por la diabetes, recuperados por el tratamiento e inducidos específicamente en los animales diabéticos tratados con tungstato. La contribución de la normalización de la hiperglicemia a estos cambios se estudió en animales diabéticos tratados con fluoricina, concluyendo que son independientes de la glicemia. Además, se estudió el efecto directo/indirecto del tungstato en células INS-1E tratadas con tungstato de sodio y con el suero de los diferentes animales, observándose que los cambios detectados eran debidos a efectos directos e indirectos del propio tratamiento. Por último, se examinó la vía de las MAPK, concluyéndose que era responsable del aumento de la proliferación observado sobre las células beta, a través de un mecanismo de modulación directo e indirecto.CONCLUSIONES: El tratamiento con tungstato mejora la función pancreática de forma directa e indirecta, a través de una combinación de vías independientes de la glicemia, donde la vía de las MAPK juega un papel clave sobre el incremento de la proliferación de las células beta.B) "The role of transmembrane protein 27 (TMEM27) in islet physiology and its potential use as a beta cell mass biomarker". Diabetologia.2010;53:1406-14.ANTECEDENTES: La proteína Transmembrana 27 (TMEM27) ha sido descrita en las células beta, donde estaría implicada en procesos de secreción de insulina y/o de proliferación. Además, ha sido propuesta como un marcador de masa de células beta. RESULTADOS: Los islotes provinentes de donantes pancreáticos con diabetes presentan niveles de ARNm de TMEM27 inferiores de los donantes sin patología, observándose una correlación significativa con el gen ESNAPINA en islotes humanos sin patología. En las ratas no-diabéticas tratadas con tungstato de sodio, que presentan un incremento en la proliferación de las células beta y una disminución en la secreción de insulina, la expresión de Tmem27 se haya disminuida. En células INS-1 832/13 e islotes, la sobrexpresión de Tmem27 da lugar a un incremento significativo de la secreción de glucosa inducida por glucosa con un incremento nulo o muy pequeño en la proliferación. Finalmente, Tmem27 puede ser escindido por células beta y células renales proximales tubulares.CONCLUSIONES: Los datos presentados apoyan la idea que Tmem27 está implicado en procesos de secreción de insulina inducidos por glucosa, pero no en procesos de proliferación. Además, el hecho que pueda ser escindido en células renales, invalida su posible uso como biomarcador de masa de células beta.[eng] The content of this thesis is summarised in the articles:a) Molecular mechanisms of tungstate-induced pancreatic plasticity: a transcriptomics approach. BMC Genomics. 2009;10:406.BACKGROUND: Molecular mechanisms which recover the pancreatic function of (STZ)-diabetic rats treated with tungstate are unknown. RESULTS: Tungstate treated (STZ)-diabetic rats showed a partial recovery of exocrine and endocrine function and increased beta-cell. Microarray analysis of the pancreases identified genes altered due to diabetes, restored by the treatment, and specifically induced by tungstate in diabetic animals. Hyperglycemia contribution was studied in STZ-diabetic rats treated with phloridzin, and direct effect of tungstate was determined in INS-1E cells treated with tungstate or serum from untreated or treated STZ-rats, observing that tungstate action in the pancreas takes places via hyperglycemia-independent pathways and a combination of tungstate direct and indirect effects. Finally, MAPK pathway was evaluated, observing that it has a key role in the tungstate-induced increase of beta-cell proliferation.CONCLUSIONS: Tungstate improves pancreatic function through a combination of hyperglycemia-independent pathways and through its own direct and indirect effects, with MAPK pathway playing a key role in the tungstate-induced increase of beta-cell proliferation.b) The role of transmembrane protein 27 (TMEM27) in islet physiology and its potential use as a beta cell mass biomarker. Diabetologia. 2010;53:1406-14.BACKGROUND: Transmembrane protein 27 (TMEM27) is a membrane protein cleaved with possible roles in insulin exocytosis and cell proliferation, although its function in beta-cells remains controversial. RESULTS: TMEM27 mRNA levels in islets are lower in human diabetic donors than in controls. Its gene expression correlates with that of insulin and SNAPIN in human islets. TMEM27 expression is downregulated in islets of tungstate-treated rats, which exhibit decreased insulin secretion and increased proliferation. TMEM27 overproduction in a beta-cell line and islets significantly enhanced glucose-induced insulin secretion, with modest or no effects on proliferation. Finally, TMEM27 is cleaved and shed by renal proximal tubular cells and pancreatic islets.CONCLUSIONS: Our data support a role for TMEM27 in glucose-induced insulin secretion but not in cell proliferation. The finding that its cleavage is not specific to beta-cells challenges the current support for its use as a potential beta-cell mass biomarker

Food craving-like episodes during pregnancy are mediated by accumbal dopaminergic circuits

Preparation for motherhood requires a myriad of physiological and behavioural adjustments throughout gestation to provide an adequate environment for proper embryonic development1. Cravings for highly palatable foods are highly prevalent during pregnancy2 and contribute to the maintenance and development of gestational overweight or obesity3. However, the neurobiology underlying the distinct ingestive behaviours that result from craving specific foods remain unknown. Here we show that mice, similarly to humans, experience gestational food craving-like episodes. These episodes are associated with a brain connectivity reorganization that affects key components of the dopaminergic mesolimbic circuitry, which drives motivated appetitive behaviours and facilitates the perception of rewarding stimuli. Pregnancy engages a dynamic modulation of dopaminergic signalling through neurons expressing dopamine D2 receptors in the nucleus accumbens, which directly modulate food craving-like events. Importantly, persistent maternal food craving-like behaviour has long-lasting effects on the offspring, particularly in males, leading to glucose intolerance, increased body weight and increased susceptibility to develop eating disorders and anxiety-like behaviours during adulthood. Our results reveal the cognitively motivated nature of pregnancy food cravings and advocates for moderating emotional eating during gestation to prevent deterioration of the offspring's neuropsychological and metabolic health.© 2022. The Author(s), under exclusive licence to Springer Nature Limited

Dietary inflammatory index and all-cause mortality in large cohorts: The SUN and PREDIMED studies

[Background]: Inflammation is known to be related to the leading causes of death including cardiovascular disease, several types of cancer, obesity, type 2 diabetes, depression-suicide and other chronic diseases. In the context of whole dietary patterns, the Dietary Inflammatory Index (DII®) was developed to appraise the inflammatory potential of the diet. [Objective]: We prospectively assessed the association between DII scores and all-cause mortality in two large Spanish cohorts and valuated the consistency of findings across these two cohorts and results published based on other cohorts.[Design]: We assessed 18,566 participants in the “Seguimiento Universidad de Navarra” (SUN) cohort followed-up during 188,891 person-years and 6790 participants in the “PREvencion con DIeta MEDiterránea” (PREDIMED) randomized trial representing 30,233 person-years of follow-up. DII scores were calculated in both cohorts from validated FFQs. Higher DII scores corresponded to more proinflammatory diets. A total of 230 and 302 deaths occurred in SUN and PREDIMED, respectively. In a random-effect meta-analysis we included 12 prospective studies (SUN, PREDIMED and 10 additional studies) that assessed the association between DII scores and all-cause mortality.[Results]: After adjusting for a wide array of potential confounders, the comparison between extreme quartiles of the DII showed a positive and significant association with all-cause mortality in both the SUN (hazard ratio [HR] = 1.85; 95% CI: 1.15, 2.98; P-trend = 0.004) and the PREDIMED cohort (HR = 1.42; 95% CI: 1.00, 2.02; P-trend = 0.009). In the meta-analysis of 12 cohorts, the DII was significantly associated with an increase of 23% in all-cause mortality (95% CI: 16%–32%, for the highest vs lowest category of DII).[Conclusion]: Our results provide strong and consistent support for the hypothesis that a pro-inflammatory diet is associated with increased all-cause mortality. The SUN cohort and PREDIMED trial were registered at clinicaltrials.gov as NCT02669602 and at isrctn.com as ISRCTN35739639, respectively.Supported by the official funding agency for biomedical research of the Spanish Government, Instituto de Salud Carlos III (ISCIII), through grants provided to research networks specifically developed for the trial (RTIC G03/140, to R.E.; RTIC RD 06/0045, to Miguel A. Martínez-González) and through Centro de Investigación Biomédica en Red de Fisiopatología de la Obesidad y Nutrición (CIBERobn), and by grants from Centro Nacional de Investigaciones Cardiovasculares (CNIC 06/2007), Fondo de Investigación Sanitaria–Fondo Europeo de Desarrollo Regional (Proyecto de Investigación (PI) 04-2239, PI 05/2584, CP06/00100, PI07/0240, PI07/1138, PI07/0954, PI 07/0473, PI10/01407, PI10/02658, PI11/01647, P11/02505, PI13/00462, PI13/00615, PI13/01090, PI14/01668, PI14/01798, PI14/01764), Ministerio de Ciencia e Innovación (Recursos y teconologia agroalimentarias(AGL)-2009-13906-C02 and AGL2010-22319-C03 and AGL2013-49083-C3-1- R), Fundación Mapfre 2010, the Consejería de Salud de la Junta de Andalucía (PI0105/2007), the Public Health Division of the Department of Health of the Autonomous Government of Catalonia, Generalitat Valenciana (Generalitat Valenciana Ayuda Complementaria (GVACOMP) 06109, GVACOMP2010-181, GVACOMP2011-151), Conselleria de Sanitat y, PI14/01764 AP; Atención Primaria (CS) 2010-AP-111, and CS2011-AP-042), and Regional Government of Navarra (P27/2011).). Drs. Shivappa and Hébert were supported by grant number R44DK103377 from the United States National Institute of Diabetes and Digestive and Kidney Diseases