Manuel Tena-Sempere1*

1Department of Cell Biology, Physiology & Immunology, University of Córdoba; Maimónides Institute of Biomedical Research of Córdoba (IMIBIC); CIBER Fisiopatología de la Obesidad y Nutrición (CIBEROBN), Instituto de Salud Carlos III, Córdoba, Spain; and FiDiPro Program, Institute of Biomedicine, Research Centre for Integrative Physiology and Pharmacology, University of Turku, Turku, Finland.

*Corresponding author:

Manuel Tena-Sempere, Department of Cell Biology, Physiology & Immunology, University of Córdoba; Maimónides Institute of Biomedical Research of Córdoba (IMIBIC); CIBER Fisiopatología de la Obesidad y Nutrición (CIBEROBN), Instituto de Salud Carlos III, Córdoba, Spain; and FiDiPro Program, Institute of Biomedicine, Research Centre for Integrative Physiology and Pharmacology, University of Turku, Turku, Finland. E-mail: fi1tesem@uco.es.

Puberty is crucial developmental event, driven by a large set of sophisticated regulatory networks, which are capable to integrate internal and peripheral signals to precisely and timely activate the brain centers governing the reproductive axis. In humans and other mammalian species, the timing of puberty is genetically determined. Yet, puberty is also highly sensitive to numerous internal and external cues, among which metabolic and nutritional signals are especially prominent. Compelling epidemiological evidence suggests that alterations of the age of puberty are becoming more frequent in humans, via as yet unknown mechanisms; yet, the escalating prevalence of obesity and other metabolic/ feeding disorders has been pointed out as major contributing factor. Notably, this phenomenon is likely to have translational implications, since alterations in the timing of puberty have been associated to adverse health outcomes later in life, including higher risk of earlier all-cause mortality. This urges for a better understanding of the neurohormonal basis of normal puberty and the mechanisms of its deviations.

In recently years, compelling evidence has documented the master role of hypothalamic neurons producing kisspeptins, which are encoded by the Kiss1 gene and operate via de G-protein coupled receptor, Gpr54, in the neuroendocrine pathways controlling puberty, with a particularly prominent function of Kiss1 neurons located in the arcuate nucleus (ARC). Other transmitters, such as tachykinins (e.g., Neurokinin B), opioids (e.g., dynorphin) and melanocortins, cooperate with kisspeptins in the precise regulation of puberty. In addition, Kiss1 neurons seemingly participate in transmitting at least part of the regulatory actions of the signals responsible for the metabolic control of puberty. In this context, recent work from our group has unveiled the important role of key cellular metabolic sensors, such as AMP-activated protein kinase (AMPK), the master cellular sensor activated in conditions of energy insufficiency, and the fuel-sensing deacetylase, Sirt1, as major components for the metabolic modulation of female puberty onset. Our data indicate that AMPK and Sirt1, acting in ARC Kiss1 neurons, are major molecular effectors for the metabolic control of Kiss1 and, thereby, puberty onset, whose alteration may contribute to the perturbations of pubertal timing frequently linked to conditions of metabolic stress, such as sub-nutrition and obesity.

Keywords: puberty; KISS1; kisspeptins; AMPK; Sirt1. Published May, 2019. Copyright: © 2019 Manuel Tena-Sempere. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Tena-Sempere, M. Novel molecular pathways for the central control of puberty: Basic aspects and translational implications. IBJ Plus 2019 S(3):e0001 doi: 10.24217/2531-0151.19v1s3.00001. Funding: The authors declare that no fundings. Competing Interests: The authors declare that no competing interests exit.

To sleep or not to sleep: how the brain decides to wake us up.

Luis de Lecea1*

1Department of Psychiatry and Behavioral Sciences, Standford University School of Medicine, Standford, CA. USA.

*Corresponding author:

Luis de Lecea, Department of Psychiatry and Behavioral Sciences, Standford University School of Medicine, Standford, CA. USA. E-mail: llecea@stanford.edu

The arousal construct underlies a spectrum of behaviors that include sleep, exploration, feeding, sexual activity and adaptive stress. Pathological arousal conditions include stress, anxiety disorders, and addiction. In the past few years we have used optogenetics to identify and interrogate neuronal circuits underlying transitions between arousal states. In the first presentation, I will talk about how the hypocretin system coordinates and makes the decisions about when to mark the transition between sleep and wakefulness. The dynamics between arousal state transitions are also modulated by norepinephrine neurons in the locus coeruleus, histaminergic neurons in the hypothalamus, dopaminergic neurons in the mesencephalon and cholinergic neurons in the basal forebrain. I will introduce our own model of sleep/wake dynamics using probabilistic estimates of neurotransmitter function based on optogenetic stimulations. I will also discuss how these models can help characterize dissociated arousal states and develop treatments for neuropsychiatric disorders.

Keywords: arousal; hypothalamus; optogenetics; hypocretin; sleep; wakefulness; addiction; neuropsychiatric disorders. Published May, 2019. Copyright: © 2019 Luis de Lecea. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: de Lecea, L. To sleep or not to sleep: how the brain decides to wake us up. IBJ Plus 2019 S(3):e0002 doi: 10.24217/25310151.19v1s3.00002. Funding: The authors declare that no fundings. Competing Interests: The authors declare that no competing interests exit.

Emerging genes and pathways in thyroid differentiation and tumorigenesis.

Pilar Santisteban1*

1Instituto de Investigaciones Biomédicas, CSIC-UAM and Ciberonc, ISCIII Madrid, Spain.

*Corresponding author:

Pilar Santisteban, Instituto de Investigaciones Biomédicas, CSIC-UAM and Ciberonc, ISCIII Madrid, Spain. E-mail: psantisteban@iib.uam.es

Thyroid cancer remains the most common endocrine malignancy worldwide and its incidence and mortality has increased steadily over the last four decades. In general, it has a good outcome; however, some patients develop aggressive forms of thyroid cancer that are untreatable and the molecular bases are poorly understood. These aggressive forms have lost NIS (Na/I Symporter) function, one of the most important hallmarks during thyroid cancer progression, as it leads to radioiodine-resistant metastatic disease. Our work has contributed to understand the mechanisms involved in iodide uptake repression and tumor progression. We have found that BRAF activation decreases NIS expression and impairs NIS trafficking to the membrane of follicular thyroid cells, and accordingly causes (RAI)-refractory metastatic disease in patients with papillary thyroid cancer. We have demonstrated that the mechanism by which BRAF impairs NIS function is mediated by a TGFβ autocrine loop. Furthermore, by next-generation sequencing and gene expression analysis we have identified a master miRNA (miR) regulatory network involved in essential biological process such as thyroid differentiation. Among those miRNAs, the most abundantly expressed in thyroid tumors is the miR-146b and we found that it binds to the 3’-UTR region of thyroid differentiation genes such PAX8 and NIS, leading to impaired protein translation and subsequently a reduction of iodide uptake. Besides, we show that miR-146b and PAX8 regulate each other sharing common target genes, thus highlighting a novel regulatory circuit that govern differentiated phenotype in thyroid tumors. Furthermore, we have shown that the overexpression of miR-146b induces an hyperactivation of the PI3K/AKT pathway, via PTEN suppression, leading to a more aggressive tumoral behavior. In summary, our work described molecular determinants that may be exploited therapeutically to modulate thyroid cell differentiation and iodide uptake for improved treatment of advanced thyroid cancer.

Keywords: thyroid cancer; iodide; NIS; braf gene; microRNA146b; PAX8; PTEN; signaling. Published May, 2019. Copyright: © 2019 Pilar Santisteban. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Santisteban, P. Emerging genes and pathways in thyroid differentiation and tumorigenesis. IBJ Plus 2019 S(3):e0003 doi: 10.24217/2531-0151.19v1s3.00003. Funding: Grants SAF2016-75531-R from MINECO, Spain (FEDER); B2017/BMD-3724 from CAM and GCB14142311CRES from AECC. Competing Interests: The authors declare that no competing interests exit.

Adipose tissue expandability, lipotoxicity and the metabolic syndrome.

Antonio Vidal-Puig1*

1University of Cambridge, Metabolic Research Laboratories, Wellcome Trust-MRC Institute of Metabolic Science, Addenbrooke’s Hospital, Cambridge, CB2 0QQ, United Kingdom.

*Corresponding author:

Antonio Vidal-Puig, University of Cambridge, Metabolic Research Laboratories, Wellcome Trust-MRC Institute of Metabolic Science, Addenbrooke’s Hospital, Cambridge, CB2 0QQ, United Kingdom. E-mail: ajv22@medschl.cam.ac.uk

The link between obesity and type 2 diabetes is clear on an epidemiological level, however the mechanism linking these two common disorders is not well defined. One hypothesis linking obesity to type 2 diabetes is the adipose tissue expandability hypothesis. The adipose tissue expandability hypothesis states that a failure in the capacity for adipose tissue expansion, rather than obesity per se is the key factor linking positive energy balance and type 2 diabetes. All individuals possess a maximum capacity for adipose expansion which is determined by both genetic and environmental factors. Once the adipose tissue expansion limit is reached, adipose tissue ceases to store energy efficiently and lipids begin to accumulate in other tissues. Ectopic lipid accumulation in non-adipocyte cells causes lipotoxic insults including insulin resistance, apoptosis and inflammation. This article discusses the links between adipokines, inflammation, adipose tissue expandability and lipotoxicity. Finally, we will discuss how considering the concept of allostasis may enable a better understanding of how diabetes develops and allow the rational design of new anti diabetic treatments.

Keywords: adipogenesis; obesity; type 2 diabetes; lipotoxicity; insulin resistance; immunometabolism ; inflammation. Published May, 2019. Copyright: © 2019 Antonio Vidal-Puig. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Vidal-Puig, A. Adipose tissue expandability, lipotoxicity and the metabolic syndrome. IBJ Plus 2019 S(3):e0004 doi: 10.24217/2531-0151.19v1s3.00004. Funding: The authors declare that no fundings. Competing Interests: The authors declare that no competing interests exit.

Regulation of tissue macrophage functions by LXR nuclear receptor dependent transcriptional pathways.

Antonio Castrillo1*

1Instituto de Investigaciones Biomédicas “Alberto Sols” CSIC-UAM, Madrid, Spain.

*Corresponding author:

Antonio Castrillo, Instituto de Investigaciones Biomédicas “Alberto Sols” CSIC-UAM, Madrid, Spain. E-mail: antonio.castrillo@iib.uam.es

Macrophages are professional phagocytic cells that play crucial roles in immune processes, but they also perform other important functions in the regulation of metabolism and maintenance of tissue homeostasis. A simple definition of the term “macrophage” is currently a challenge due to the continuous advances in the field, with the classification of their multiple origins, the study of their reprogramming capacities during homeostasis and disease. Indeed, exciting research findings have emerged during the last several years, in which macrophage heterogeneity is now believed to be determined by a combination of signals governed by the cellular origin and others coming from the environment. We have studied the transcriptional regulatory circuits that control macrophage behaviour in response to different physiological or pathological situations. A fraction of the transcriptional control of macrophage functions is achieved by the Liver X receptors, LXRα and LXRβ, which are transcription factors that belong to the nuclear hormone receptor superfamily. LXRs are involved in the regulation of cholesterol, fatty acid and phospholipid metabolism. In addition to their role in sterol metabolism, LXRs are important for the immune response against microbial pathogens. LXRα and LXRβ are highly similar in sequence and most of their reported functions are substantially overlapping. In this symposium, we will discuss our recent advances in LXR biology. With a combination of functional assays, expression profiling and ChIPsequencing data, we will shed light on our understanding of LXR´s cell-specific targets and the specific actions of LXRα and LXRβ in macrophage immune responses. Collectively we will discuss the current view of LXR transcription factors in immune processes and their impact in macrophage biological processes.

Keywords: LXR; liver X receptor; nuclear receptor; gene expression; transcription; inflammation; macrophage; immunology. Published May, 2019. Copyright: © 2019 Antonio Castrillo. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Castrillo, A. Regulation of tissue macrophage functions by LXR nuclear receptor dependent transcriptional pathways. IBJ Plus 2019 S(3):e0005 doi: 10.24217/2531-0151.19v1s3.00005. Funding: The authors declare that no fundings. Competing Interests: The authors declare that no competing interests exit.

Genetic determinants of thyroid function: novel insights and new approaches.

Marco Medici1*

1Department of Internal Medicine, Erasmus Medical Center, Rotterdam. The Netherlands.

*Corresponding author:

Marco Medici, Department of Internal Medicine, Erasmus Medical Center, Rotterdam. The Netherlands. E-mail: m.medici@erasmusmc.nl

In the last few years studies have shown that subtle variations in thyroid function, including subclinical thyroid dysfunction, and even variation in thyroid function within the normal range, are associated with morbidity and mortality. It is estimated that 40-65% of the inter-individual variation in serum TSH and FT4 levels is determined by genetic factors. To identify these factors, various linkage and candidate gene studies have been performed in the past, which have identified only a few genes. In the last decade, genome-wide association studies identified >100 genetic variants, while whole exome and whole genome sequencing studies are expected to further clarify the genetic basis of thyroid function in the near future. The identification of these genes has paved the way for various lines of research. Examples of followup in vitro characterization studies of these genes include the identification of SLC17A4 as a novel thyroid hormone transporter and AADAT as a novel thyroid hormone metabolizing enzyme. Furthermore, mendelian randomization studies are expected to clarify whether the observed associations between minor variations in thyroid function and clinical endpoints are causal or not, which is key when considering treatment for these mild variations in thyroid function. Finally, these genetic markers have been associated with normal range thyroid function as well as thyroid dysfunction, thereby possibly serving as predictive markers for the individual thyroid setpoint and for thyroid disease.

Keywords: thyroid; TSH; FT4; genome-wide association studies; SLC17A4. Published May, 2019. Copyright: © 2019 Marco Medici. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Medici, M. Genetic determinants of thyroid function: novel insights and new approaches. IBJ Plus 2019 S(3):e0006 doi: 10.24217/2531-0151.19v1s3.00006. Funding: The authors declare that no fundings. Competing Interests: The authors declare that no competing interests exit.

The role of endocrine disruptors in the etiology of diabetes mellitus.

Ángel Nadal1*

1Instituto de Investigación, Desarrollo e Innovación en Biotecnología Sanitaria de Elche. Universidad Miguel Hernández de Elche y CIBERDEM, Alicante, Spain.

*Corresponding author:

Ángel Nadal, Instituto de Investigación, Desarrollo e Innovación en Biotecnología Sanitaria de Elche. Universidad Miguel Hernández de Elche y CIBERDEM, Alicante, Spain. E-mail: nadal@umh.es

Keywords: Published May, 2019. Copyright: © 2019 Ángel Nadal. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Nadal, A. The role of endocrine disruptors in the etiology of diabetes mellitus. IBJ Plus 2019 S(3):e0007 doi: 10.24217/25310151.19v1s3.00007. Funding: The authors declare that no fundings. Competing Interests: The authors declare that no competing interests exit.

Mónica Marazuela1*

1Hospital Universitario La Princesa, Facultad de Medicina UAM, Madrid, Spain.

*Corresponding author:

Mónica Marazuela, Hospital Universitario La Princesa, Facultad de Medicina UAM, Madrid, Spain. E-mail: monica.marazuela@uam.es

Autoimmune thyroid disease (AITD), which includes Hashimoto’s thyroiditis (HT) and Graves’ disease (GD) are one of the most prevalent organ-specific autoimmune diseases which are characterized by circulating antibodies and lymphocyte infiltration of the thyroid gland. The etiology of AITD is multifactorial due to a complex interplay of specific susceptibility genes and environmental exposures and is characterized by reactivity to self-thyroid antigens due to autoreactive lymphocytes escaping tolerance. Different adhesion molecules, cytokines and chemokines help lymphocyte migration to the thyroid in AITD. Although humoral and cellular mechanisms have been classically considered separately in AITD, recent research suggests a close reciprocal relationship between these two immune pathways. The simple dichotomy Th1/Th2 has been outweighed by the recently described T cells subtypes Treg and Th17 have an essential role in the pathogenesis of AITD. Although several types of CD4 Treg cells (Foxp3, CD69, Tr1), which are able to prevent the appearance of autoimmune diseases, are found in peripheral blood and thyroid tissue from patients with AITD, these cells are apparently unable to put down the autoimmune process. In addition, many reports indicate the involvement of different Th17 cells in AITD. Recently, specific miRNA have been involved in the disregulation of immune mechanisms in AITD probably through differential Treg and Th17 differentiation. Nowadays, both Treg and Th17 cells must be considered as key elements in the pathogenesis of AITD as well as plausible potential targets for the next generation of therapeutic options of this condition.

Keywords: autoimmune thyroid disease; lymphocyte differentiation; immunity; immunometabolism; inflammation. Published: May, 2019. Copyright: © 2019 Mónica Marazuela. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Marazuela, M. Pathogenesis of autoimmune thyroid diseases: role of cellular immunity. IBJ Plus 2019 S(3):e0008 doi: 10.24217/2531-0151.19v1s3.00008. Funding: The authors declare that no fundings. Competing Interests: The authors declare that no competing interests exit.

Vítor Ferreira1,2*, Patrícia Rada1,2, Diana Grajales1,2, Irma García-Martinez1,2, Ángela M. Valverde1,2

1IIBm Alberto Sols (CSIC-UAM), Madrid, Spain. 2CIBER de Diabetes y Enfermedades Metabólicas Asociadas (CIBERDEM), ISCIII, Spain.

*Corresponding author:

Vitor Ferreira, Madrid, Spain. E-mail: vdasilva@iib.uam.es

Schizophrenia is a chronic mental disorder that affects approximately 50 million people worldwide. Also, it is associated with psychotic experiences that not allow patients to have a normal life. Fortunately, treatments used can suppress the symptomatology and lead to a productive life and integration in the society. According to the current guidelines, the first line in schizophrenia’s therapy are the second generation antipsychotic drugs (SGA). Besides being D2 receptors antagonists/ partial agonists, the SGAs have the ability to interact also with serotonergic, histaminergic and other receptors; minimizing the symptomatology and not resulting in neurological side effects (contrarily to first generation antipsychotic drugs). Nevertheless, recent clinical observations show a variety of metabolic dysfunctions in patients under SGAs treatment, such as abnormal gain weight, hyperglycemia and dyslipidemia. However, the molecular mechanisms behind these alterations are still very poorly understood. Having this in consideration, the objective of this work was to study the effect of Olanzapine administration in wild-type (WT) and PTP1B-deficient (KO) mice on glucose homeostasis and the mechanisms involved. Of interest, PTP1B is the main tyrosine phosphatase of the insulin receptor, and PTP1B KO mice are protected against insulin resistance and development of type 2 diabetes mellitus. After administration of 10 mg/kg/day of Olanzapine for 8 weeks to WT and PTP1B-KO mice, the glucose homeostasis tests suggest that Olanzapine induced insulin resistance only in WT mice. Also, primary hepatocytes from WT mice treated with Olanzapine showed lower glucose uptake than those from non-treated WT mice. In contrast, Olanzapine treatment of PTP1B KO mice significantly increased hepatocyte glucose uptake, suggesting that Olanzapine-induced insulin resistance is at least in part dependent on PTP1B. These results support the relevance of PTP1B in Olanzapine-induced insulin resistance in hepatocytes.

Keywords: Insulin sensitivity; Insulin resistance; Liver; Second generation antipsychotics. Published: May, 2019. Copyright: © 2019 Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Ferreira V., Rada P., Grajales D., García-Martinez I., Valverde A.M., PTP1B deficiency protects mice against metabolic dysfunction in glucose homeostasis upon Olanzapine intraperitoneal treatment. IBJ Plus 2019 S(3):e0025 doi: 10.24217/2531-0151.19v1s3.00025. Funding: European Union's EU Framework Programme for Research and Innovation Horizon 202. ITN-TREATMENT under Grant Agreement No GA 721236. Competing Interests: The authors declare that no competing interests exit.

TGFβ3 as a novel target in obesity-associated kidney failure.

Elia Escasany1, Adriana Izquiero-Lahuerta1, Patricia Corrales1, Almudena García-Carrasco1, Lucía Torres1, Yamila López1,2, Paul Petrus3, Estela Maldonado2, Concha Martínez Álvarez2, Manuel Ros1, Peter Arner3, Miakel Ryden3, Gema Medina-Gómez1

1Universidad Rey Juan Carlos. Dpto. de Ciencias Básicas de la Salud. Avda. de Atenas s/n. 28922. Alcorcón. Madrid. Spain 2Departamento de Anatomía y Embriología Humanas, Facultad de Medicina, Universidad Complutense. 28040. Madrid. Spain. 3Department of Medicine, Karolinska Institutet, Karolinska University Hospital, Huddinge, 141 86, Stockholm, Sweden

*Corresponding author:

Gema Medina-Gómez, Universidad Rey Juan Carlos. Dpto. de Ciencias Básicas de la Salud. Avda. de Atenas s/n. 28922. Alcorcón. Madrid. Spain. E-mail: gema.medina@urjc.es

Obesity is associated with development of lipotoxicity in metabolically important organs such as the kidney leading to apoptosis and dedifferentiation, which contributes to fibrosis and chronic kidney diseases progression. Glomerulosclerosis and tubulointerstitial fibrosis are the reversible common final pathological features of chronic kidney diseases (CKD), regarding the aetiology making it an appealing target of study for both finding new biomarkers and developing therapeutic strategies for CKD. Transforming growth factor β (TGFβ) is involved in differentiation, apoptosis and fibrosis. TGFβ1 has long been associated with renal fibrosis; however, the role for TGFβ3 is not so clear.

Our aim was to unravel the role of TGFβ3 in the pathophysiology of the kidney. We have used in vivo and in vitro lipotoxic models. Regarding the in vitro model, we have used heterozygous (HZ) TGFβ3 mice, due to lethality of the homozygous TGFβ3 mutant mice. These mice were subjected to either a control (CD) or a high fat diet (HFD) and sacrificed at 16 weeks of age.

In HZ mice, lower levels of TGFβ3 did not affect body weight; however, under HFD, HZ mice showed glucose intolerance but no insulin resistance, with higher basal levels of glucose and insulin. Despite similar renal hypertrophy index, glomerular area or blood pressure, HZ mice under CD already showed decreased 24h urine volume and a tendency towards higher albumin/creatinine ratio under both diets, which was aggravated under HFD.

Histological analysis (PAS and α-SMA IHC) showed renal fibrosis in HZ mice under both diets confirmed by increased fibronectin, osteopontin and N-cadherin and decreased E-cadherin and β-catenin mRNA levels. Partial lack of TGFβ3 lead to higher oxidative stress (ROS), thickening of the glomerular basement membrane (GBM), foot process effacement and loss of podocytes, as observed by electronic microscopy and flow cytometry.

Kidneys of HZ mice showed altered expression of lipid metabolism genes, with an increase in lipid synthesis genes (ACC and SREBP1) and a decrease in β-oxidation genes (PPARα, CPT1, PGC1α and PGC1β). This translated into an accumulation of lipids inside the kidney as observed histologically (Oil Red) and by flow cytometry experiments (Bodipy staining).

Altogether, these data suggest a tight link between TGFβ3 and lipid metabolism at the renal levels as well as a possible important role of TGFβ3 as anti-fibrotic in the development of renal fibrosis under lipotoxic conditions.

Keywords: kidney, fibrosis, TGFβ3, lipotoxicity, podocytes, CKD, obesity Published: May, 2019. Copyright: © 2019 Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Escasany E., Izquiero-Lahuerta A., Corrales P., García-Carrasco A., Torres L., López Y., Petrus P., Maldonado E., Martínez Álvarez C., Ros M., Arner P., Ryden M., Medina-Gómez G., TGFβ3 as a novel target in obesity-associated kidney failure. IBJ Plus 2019 S(3):e0026 doi: 10.24217/2531-0151.19v1s3.00026. Funding: BFU2016-78951-R, B2017BMD-3684, BFU2017-90578-REDT and Karolinska Institutet. Competing Interests: The authors declare that no competing interests exit.

Initial age-associated metabolic alterations in white and brown fat depots: effect of long-term caloric restriction.

Patricia Corrales1, Yurena Vivas1, Adriana Izquierdo-Lahuerta1, Daniel Horrillo1, Patricia Seoane-Collazo2, Ismael Velasco1, Lucia Torres1, Yamila Lopez1, Carmen Martínez1, Miguel López2, Manuel Ros1, Maria Jesús Obregón3, Gema Medina-Gómez1.

1Universidad Rey Juan Carlos, Alcorcón, Madrid, Spain. 2Universidad Santiago de Compostela-Instituto de Investigación Sanitaria, Santiago de Compostela, Spain. 3Instituto de Investigaciones Biomédicas (IIB), Centro Mixto CSIC-UAM, Madrid, Spain.

*Corresponding author:

Patricia Corrales, Universidad Rey Juan Carlos, Alcorcón, Madrid, Spain. Email: patricia.corrales@urjc.es

Changes in the body composition such as high adiposity and sarcopeny occur during aging. These changes are usually associated to metabolic alterations like insulin resistance, type 2 diabetes and metabolic syndrome. Adipose tissue is not homogeneous and dysfunction of white adipose tissue (WAT) and brown adipose tissue (BAT) can develop differently during aging. Our aim is to study different WAT and BAT depots at middle-age to detect the initial signals of alteration. Moreover, we have studied the effect of the long-term caloric restriction (CR) and its benefits.

Sv129 mice 3 and 12-month-old mice fed ad libitum and 12-month-old mice fed with a 20% CR (3m, 12m and 12mCR, respectively) have been used. RT-PCR has been employed to study lipid metabolism gene expression in eWAT, scWAT and BAT. Moreover, circulating serum and BAT thyroid hormones levels were measured by radioimmunoassay (RIA).

12m animals showed insulin resistance and lower adiponectin levels in plasma compared with 3m. Expression of insulin pathway (Glut4, Irs1), lipogenesis (Fas, Scd1) and lipolysis (Hsl, Atgl, Pparα) genes was decreased in scWAT of 12m animals, but not in eWAT. Furthermore, UCP-1 expression was significantly lower in scWAT of aged mice compared with younger mice, suggesting a decrease in beige adipose tissue associated to aging. On the other hand, we found decreased expression of some BAT-selective genes12m animals such as Ucp1, βAr. CR recovered the expression levels to those of 3m mice. Thyroxine (T4) in plasma did not reach significance among middle-aged and young animals, however triiodothyronine (T3) levels were lower in 12m than 3m and 12mCR mice. Dio2 expression, T4 and T3 concentrations in BAT followed a similar pattern, a decrease in 12m mice and an increase in CR mice.

Although eWAT has been described as the most dangerous tissue from the metabolic point of view, our data could suggest an earlier metabolic alteration in scWAT than in eWAT. Our findings prompt that CR could revert the initial agerelated alterations in scWAT. In agreement, we point to alteration of thyroid axis status with age is an important factor contributing to BAT dysfunction at middle-age, which can be ameliorated by CR.

Keywords: aging, caloric restriction, adipose tissue, thyroid hormone. Published: May, 2019. Copyright: © 2019 Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Corrales P., Vivas Y., Izquierdo-Lahuerta A., Horrillo D., Seoane-Collazo P., Velasco I., Torres L., Lopez Y., Martínez C., López M., Ros M., Obregón MJ., Medina-Gómez G., Initial age-associated metabolic alterations in white and brown fat depots: effect of longterm caloric restriction. IBJ Plus 2019 S(3):e0027 doi: 10.24217/2531-0151.19v1s3.00027. Funding: BFU2016-78951-R, B2017BMD-3684, BFU2017-90578-REDT, Fundacion Mapfre, CAMS210/BMD-2433. Competing Interests: The authors declare that no competing interests exit.

Microsomal Prostaglandin E Synthase-1 (mPGES-1) mediates the development of metabolic and cardiovascular alterations associated to obesity.

Ballesteros-Martinez C1, Rodrigues-Diez R1,2,3, Gonzalez-Amor M1, Salaices M1,2,3, Briones AM1,2,3

1Department of Pharmacology and Therapeutics, Faculty of Medicine, Universidad Autónoma de Madrid, Madrid, Spain. 2Hospital La Paz Institute for Health Research (IdiPAZ), Madrid, Spain. 3CIBER Cardiovascular Diseases, Madrid, Spain.

*Corresponding author:

Briones AM1,2,3, Madrid, Spain. E-mail: ana.briones@uam.es

Introduction: Obesity is linked to chronic low-grade inflammation. Adipose tissue responds to metabolic signals, being also metabolically active secreting peptides, hormones and adipokines that, under obesity situation, participate in the development of cardiovascular and metabolic alterations such as endothelial dysfunction, arterial stiffness and insulin resistance. Microsomal prostaglandin E Synthase-1 (mPGES-1) is an inducible isomerase responsible for the production of prostaglandin E2 (PGE2) under inflammatory conditions. PGE2 is a key lipid mediator implicated in the regulation of adipose tissue homeostasis. It is also involved in vascular remodeling in different inflammatory pathologies.

Objective: To evaluate the role of mPGES-1 in the development of obesity and in the metabolic and cardiovascular alterations associated.

Methods: We have developed a model of high fat diet (HFD, 60% fat)-induced obesity in mPGES-1-/- and their control littermates mPGES-1+/+ mice. Body weight was measured twice a week and organs and adipose tissue weights were measured at the end of the experiment. The glycemic profile was studied by the glucose tolerance test (GTT) and insulin tolerance test (ITT). Arterial pressure was measured by tail-cuff plethysmography. Aortic vascular function and structural and mechanical properties of mesenteric resistance arteries where evaluated by isometric and perfusion myographs respectively. Histological analysis was also performed. Changes in mRNA expression were studied by q-RT- PCR.

Results: Our results show that mPGES-1-/- mice fed with a HFD are protected against body weight gain and present reduced adiposity with less grade of macrophage infiltration. In addition, mPGES-1 deficient mice have better glycemic profile compared to wild type mice. At cardiovascular level, mPGES-1+/+ but not mPGES-1-/- mice on HFD, develop an increase in the size of cardiomyocytes, which results in cardiac hypertrophy, show aortic endothelial dysfunction and vascular insulin resistance, and show structural alterations in resistance arteries.

Conclusion: Our data suggest that the lack of mPGES-1 protects against some of the cardiovascular and metabolic alterations associated to obesity.

Keywords: obesity, mPGES-1, vascular function, adipose tissue. Published: May, 2019. Copyright: © 2019 Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Ballesteros-Martinez C., Rodrigues-Diez R., Gonzalez-Amor M., Salaices M., Briones AM., Microsomal Prostaglandin E Synthase-1 (mPGES-1) mediates the development of metabolic and cardiovascular alterations associated to obesity. IBJ Plus 2019 S(3):e0028 doi: 10.24217/2531-0151.19v1s3.00028. Funding: This project was supported by the Spanish Ministerio de Economía, Industria y Competitividad (SAF2016-80305P); Instituto de Salud Carlos III (PI13/01488; CIBER de Enfermedades Cardiovasculares, CB16/11/00286) and Comunidad de Madrid (B2017/BMD- 3676). Competing Interests: The authors declare that no competing interests exit.

Lack of improvement in sperm quality and changes in reproductive hormones in obese mice and patients males after obesity surgery and diet.

Casado ME1,2, Calderón B2,4, Huerta L1, Botella-Carretero JI2,3 and Martín-Hidalgo A1,2

1Servicio de Bioquímica-Investigación, Hospital Universitario Ramón y Cajal, Instituto Ramón y Cajal de Investigación Sanitaria (IRyCIS), Madrid, Spain. 2CIBER de la Fisiopatología de la Obesidad y Nutrición (CIBERobn), ISCIII, Madrid, Spain. 3Servicio de Endocrinología y Metabolismo, Hospital Universitario Ramón y Cajal, E-28034 Madrid, Spain. 4Instituto Tecnológico de Santo Domingo (INTEC), Santo Domingo, Republica Dominicana.

*Corresponding author:

Martín-Hidalgo A, Servicio de Bioquímica-Investigación, Hospital Universitario Ramón y Cajal, Instituto Ramón y Cajal de Investigación Sanitaria (IRyCIS), Ctra. Colmenar Km 9.1, 28034-Madrid, Spain. E-mail: antonia.martinl@hrc.es

Obesity in men is associated with a decrease in sperm concentration, motility, alterations in sperm morphology, chromatin integrity and hormonal profile that leads to infertility. Even though obesity surgery normalizes circulating testosterone concentrations in males with obesity-associated secondary hypogonadism, its impact on spermatogenesis remains controversial. We aimed to evaluate sperm characteristics and changes in reproductive hormones in obese men after bariatric surgery and obese mice after low fat diet.

Methods: Twenty severely obese men (body mass index (BMI) ≥ 35 kg/m2) were evaluated before and 2 years after bariatric surgery. We assayed insulin, leptin, kisspeptin, and inhibin B in the serum of the patients. The obese mice model included the following experimental groups: 1) LFD 10 weeks; 2) HFD 10 weeks; 3) LFD 18 weeks; 4) HFD 18 weeks; 5) HFD 10 weeks and LFD 8 weeks. The seminal fluid was get from epididymis for spermatozoa counts and motility. We studied the mice at 10 and 18 weeks after the different diets. We assayed glucose, insulin and testosterone in plasma. Homeostasis model assessment of insulin resistance (HOMA-IR) was estimated. We used World Health Organization reference values for sperm analysis.

Results: Before surgery, serum total testosterone, calculated free testosterone, leptin, glucose and fasting insulin, HOMA-IR, increased in obese patients. After surgery, serum total testosterone, calculated free testosterone, inhibin B, and kisspeptin increased, whereas fasting insulin, HOMA-IR, and leptin concentrations decreased in the patients. Despite these improvements, sperm volume showed a small decrease after surgery, while the rest of sperm characteristics remained mostly unchanged. Abnormal sperm concentration persisted in 60% of the obese patients. In mice, plasma glucose and insulin increased in HFD compared with LFD at 10 and 18 weeks. These values decreased in the group of HFD+LFD at 18 weeks. The HFD mice have insulin resistance compared with LFD mice. Plasma testosterone decreased in HFD compared to LFD at 10 and 18 weeks; and remained low in HFD+LFD. There was not reversion of plasma testosterone levels with the change of the diet. Spermatozoa concentration decreased in HFD compared to LFD mice, and there was not reversion of sperm concentration after the change of HFD to LFD at 18 weeks.

Conclusions: Sperm characteristics may not improve after bariatric surgery despite the beneficial changes of reproductive hormones in obese patients. There is not improvement in sperm quality and testosterone in obese mice after diet.

Keywords: Obesity, bariatric surgery, testosterone, kisspeptin, leptin, semen analysis, male infertility. Published: May, 2019. Copyright: © 2019 Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Casado ME., Calderón B., Huerta L., Botella-Carretero JI. and Martín-Hidalgo A., Lack of improvement in sperm quality and changes in reproductive hormones in obese mice and patients males after obesity surgery and diet. IBJ Plus 2019 S(3):e0029 doi: 10.24217/2531-0151.19v1s3.00029. Funding: CIBEROBN. Financed by PI16/00154. Instituto de Salud Carlos III. Ministerio de Economía y Competitividad, Spain. Competing Interests: The authors declare that no competing interests exit.

The short-term dietary impact on the central and circulating IGF systems in rats.

Santiago Guerra-Cantera1,2,3*, Francisca Díaz2,3, Purificación Ros1,4, Alejandra Freire-Regatillo1,2,3, Laura M. Frago1,2,3, Vicente Barrios2,3, Jesús Argente1,2,3,5, Julie Chowen2,3,5

1Department of Pediatrics, Universidad Autónoma de Madrid, Madrid, Spain. 2Department of Endocrinology, Hospital Infantil Universitario Niño Jesús, Research Institute “La Princesa”, Madrid, Spain. 3Centro de Investigación Biomédica en Red de la Fisiopatología de la Obesidad y Nutrición (CIBEROBN), Instituto de Salud Carlos III, Madrid, Spain. 4Hospital Universitario Puerta de Hierro-Majadahonda, Madrid, Spain. 5IMDEA Food Institute, CEI UAM + CSIC, Madrid, Spain.

*Corresponding author:

Santiago Guerra-Cantera, Madrid, Spain. E-mail: santiguerra8@gmail.com

The insulin-like growth factor system (IGF) exerts a wide range of functions in the organism, participating in linear bone growth, metabolism and neuroprotection. Poor nutrition and obesity can alter this system; however, little is known regarding the effects of short-term dietary changes on the IGF axis. Our aim was to determine if a 1-week high-fat diet (HFD) or a low-fat diet (LFD) modifies the central and peripheral IGF systems.

Adult male and female Wistar rats were given a HFD (60% fat, 8.9% sucrose, 5.1 kcal/g), LFD (10% fat, 33.1% sucrose, 3.76 kcal/g) or standard rodent chow (3.1% fat, 0.9% sucrose, 3.41 kcal/g) for 1 week. Serum hormone levels were measured by ELISA and tissue gene expression by RT-qPCR. Hepatic glucose-6-phosphate dehydrogenase (G6PDH) activity was evaluated (Sigma-Aldrich) and malic enzyme activity was determined as described by Geer et al (1).

HFD increased body weight (p<0.05) exclusively in males. In serum, males had higher levels of free and total IGF1, IGF-binding protein (IGFBP) 3, IGFBP5, insulin, leptin and triglycerides compared to females (p<0.001) and pregnancyassociated plasma protein-A2 (PAPP-A2) was higher in females (p<0.05), with no dietary effect on these parameters. In females, serum IGF2 levels increased on the LFD (p<0.01). Hepatic IGF1 and IGFBP3 mRNA levels were affected by both sex and diet (p<0.01) and IGF2 mRNA levels were increased in response to HFD, reaching significance in females (p<0.01). Hepatic IGFBP2 mRNA levels were increased in response to HFD in both sexes (p<0.001) with sex differences, as levels were higher in females than males (p<0.001). Liver G6PDH activity was higher on the LFD compared to chow in both sexes (p<0.001), whereas in females HFD also increased G6PDH activity. Liver malic enzyme activity was increased in response to LFD compared to chow in both sexes (p<0.001).

Hypothalamic mRNA levels of IGF2 (p<0.01) and IGFBP2 (p<0.001) increased after LFD intake, but only in males. No effects were found on the remaining members of the IGF system. In males, LFD also increased the mRNA levels of neuropeptide-Y (p<0.01) and Agouti-related peptide (p<0.05), with no changes in proopiomelanocrotin or cocaine- and amphetamine-regulated transcript mRNA levels.

In conclusion, a short-term dietary change can modify both the systemic and central IGF systems, with these modifications differing between males and females. Thus, this system could be involved in the sex difference in weight gain in response to poor dietary habits. Further investigation is warranted to determine the role of IGF2 and IGFBP2 in metabolism.

(1) Geer et al., Comp. Biochem. Physiol., 1980. Vol. 65B, pp. 25-34.

Keywords: Obesity, IGF1 system, neuropeptides, HFD, LFD. Published: May, 2019. Copyright: © 2019 Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Guerra-Cantera S., Díaz F., Ros P., Freire-Regatillo A., Frago L.M., Barrios V., Argente J., Chowen J., The short-term dietary impact on the central and circulating IGF systems in rats. IBJ Plus 2019 S(3):e0030 doi: 10.24217/2531-0151.19v1s3.00030. Funding: This work was funded by CIBEROBN, PI16/00485, and BFU2017-82565-C2-1-R, fondos FEDER. Competing Interests: The authors declare that no competing interests exit.

Myeloid SAPKs in the control of obesity and BAT thermogenesis.

Maria Crespo1, Ivana Nikolic1, Aránzazu Pintor1, Alfonso Mora1, Elena Rodríguez1, Luis Leiva-Vega1, Magdalena Leiva1*, Guadalupe Sabio1*.

1Centro Nacional de Investigaciones Cardiovasculares (CNIC), Melchor Fernandez Almagro 3, Madrid, Spain

*Corresponding author:

Magdalena Leiva, Centro Nacional de Investigaciones Cardiovasculares (CNIC), Madrid, Spain. E-mail: magdalena.leiva@cnic.es Guadalupe Sabio, Centro Nacional de Investigaciones Cardiovasculares (CNIC), Madrid, Spain. E-mail: guadalupe.sabio@cnic.es

Chronic inflammation of white adipose tissue during obesity is a main driver of insulin resistance and type 2 diabetes (T2D). Among this inflammatory scenario, macrophages play important roles. Stress activated kinases (SAPKs) are key members of the signaling pathways implicated in inflammatory response, function and cytokine production in macrophages. Brown adipose tissue (BAT) is a potential therapeutic target for obesity because of its capacity to burn fat during thermogenesis, thus increasing energy expenditure. M1 (pro-inflammatory or classically activated) and M2 (anti-inflammatory or alternatively activated) macrophages are important for the control of BAT thermogenic activity, by blocking or promoting it, respectively. Using a mice model lacking the main upstream activators of some SAPKs in the myeloid lineage we found that this pathway is crucial for M2, but not M1 macrophage polarization. Moreover, this mice model is more sensitive to high-fat diet (HFD)-induced obesity as they have more body weight and liver steatosis, they are more insulin resistant. This correlates with the less energy expenditure, BAT thermogenic activity and subcutaneous browning found in this mice model. Accordingly with this, we found there is a significant depletion of M2 macrophage population in BAT from these mice. All this data suggests the crucial role of myeloid SAPKs in the protection against obesity and in the control of BAT thermogenesis during HFD.

Figure 1. The lack of SAPKs in the myeloid lineage leads to impaired BAT thermogenesis and browning of white adipose tissue and contributes to the development of obesity during HFD.

Keywords: macrophages, SAPKs, obesity, BAT, thermogenesis. Published: May, 2019. Copyright: © 2019 Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Crespo M., Nikolic I., Pintor A., Mora A., Rodríguez E., Leiva-Vega L., Leiva M., Sabio G., Myeloid SAPKs in the control of obesity and BAT thermogenesis. IBJ Plus 2019 S(3):e0031 doi: 10.24217/2531-0151.19v1s3.00031. Funding: FPI 2018 Ministerio de Ciencia, Innovación y Universidades. Competing Interests: The authors declare that no competing interests exit.

Intermittent Hypoxia a powerful stimulus of the endocrine system: Anabolic/Catabolic Hormone response in Athletes.

Fernández-Lázaro Diego1, Fernández-Lázaro César Ignacio1,2, Mielgo-Ayuso Juan3, Caballero García Alberto4, Pascual Fernández Jorge5, Córdova Martínez Alfredo3

1Department of Cell Biology, Histology and Pharmacology, Faculty of Physiotherapy, University of Valladolid, Campus de Soria, Soria. (Spain). 2Department of Preventive Medicine and Public Health, Faculty of Medicine, University of Navarra, Pamplona (Spain). 3Department of Physiology, Faculty of Physiotherapy, University of Valladolid, Soria Campus, Soria (Spain) 4Department of Anatomy, Faculty of Physiotherapy, University of Valladolid, Soria Campus, Soria (Spain) 5García Orcogen Hospital, Estella, Navarra (Spain)

*Corresponding author:

Dr. Diego Fernández Lázaro, Department of Cell Biology, Histology and Pharmacology Faculty of Physiotherapy, University of Valladolid, Campus of Soria, Spain. E-mail: diego.fernandez.lazaro@uva.es

Introduction: Exposure to intermittent hypoxia (IH), that is used as a complement to training to increase sports performance, affects the function on the hypothamulus-pitutary-adrenal axis, increases the corticotroph number and the levels of plasma ACTH and stimulates the expression of the steroidogenic acute regulatory protein and enhances the secretion of glucocorticoids as cortisol (C) (1). Also, the extreme physical effort induces a powerful stimulus of the endocrine system, that generates a hormonal response (2,3). Therefore, we investigated the effects of acute short duration IH along a macrocycle of training, over variations and behavior of anabolic/catabolic hormones. In this context we wont to evaluate also the influence of IH on specific test of performance. Material and Methods: We investigated the effects of IH along 4 weeks training on endocrine response by anabolic/catabolic hormones, Testosterone (T) and C. We studied professional male athletes exposed (PA) (n=12) or not (n=12) to IH, at three different moments: baseline (M1) (i.e., under resting conditions immediately before start the study); after two weeks of IH (M2); and at the end of the 4-week IH exposure period (M3). Physical performance was assessed by individual test: aerobic power, lactic power and speed were performed in the athletics track, over distances of 1000 meters (m), 400m and 60m respectively. For the strength test, force of the quadriceps with a dynamometer was measured. The tests were performed on M1 and M3. Results: Significant differences were observed throughout the study (M1 to M3) in T and T/C between groups. In PA group T increase significantly (P<0.001), however the tendency of C was to increase but no significantly. With respect T/C ratio increase (p<0.01) along of study (M1 to M3). In CG both, T and T/C were showed a significant decrease along the study (p<0.05), and T/C ratio decrease significantly in M2 and M3 with respect baseline (M1). The stepwise discriminant function analysis resulted indicated athletes who use hypoxia (centroid = 1,793) had higher Δ T values, whereas the athletes who use normoxia (centroid = -1,569) had lower Δ T values. The results, of physical parameters, were better in PA group that in CG, being significant the differences in anaerobic (P=0.003) and aerobic (P>0.001) power between both groups. Conclusion: We have observed that IH might potentially stimulate performance through an anabolic effect. The testosterone is a determinant informative to evaluate the answer to exercise in PA. For all this we recommended performing the resistance training under the intermittent hypoxia to induce anabolic hormone responses and after improve the physical performances.

References:

1.Kujala UM, Alen M, Huhtaniemi IT. Gonadotrophin-releasing hormone and human chorionic gonadotrophin tests reveal that both hypothalamic and testicular endocrine functions are suppressed during acute prolonged physical exercise. Clin Endocrinol (Oxf). 1990; 33: 219-225.

2.Cordova A, Fernandez-Lazaro D, Mielgo J, Seco J, Caballero A. Effect of magnesium supplementation on muscular damage markers in basketball players during a full season. Magnes Res. 2017; 1; 30: 61-70.

3.Navarrete-Opazo A, Mitchell GS. Therapeutic potential of intermittent hypoxia: A matter of dose. Am J Physiol Regul Integr Comp Physiol. 2014; 307: 1181-1197.

Keywords: Intermittent Hypoxia, endocrine system, athletes. Published: May, 2019. Copyright: © 2019 Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Fernández-Lázaro D., Fernández-Lázaro César I., Mielgo-Ayuso J., Caballero García A., Pascual Fernández J., Córdova Martínez A., Intermittent Hypoxia a powerful stimulus of the endocrine system: Anabolic/Catabolic Hormone response in Athletes. IBJ Plus 2019 S(3):e0032 doi: 10.24217/2531-0151.19v1s3.00032. Funding: No conflict of interest authors has related to the work presented. Competing Interests: The authors recognize the Institute of Studies of Health Sciences of Castilla y León (ICSCYL) for their support and collaboration throughout the research process of this study.

Influence of malnutrition during pregnancy on maternal and fetus plasmatic corticosterone concentrations.

Andrea Gila-Diaz1* , Cynthia Guadalupe Reyes-Hernández1, David Ramiro-Cortijo1, Pilar Rodríguez-Rodríguez1, Angel Luis López de Pablo1, Mª del Rosario López-Giménez2, Mª del Carmen González1, Silvia M. Arribas1.

1Department of Physiology, Faculty of Medicine, University Autónoma of Madrid, Spain. 2Department of Epidemiology, Public Health & Microbiology, Faculty of Medicine, University Autónoma of Madrid, Spain.

*Corresponding author:

Andrea Gila Díaz, Department of Physiology, Madrid, Spain. E-mail: andrea.gila@estudiante.uam.es

Introduction: Exposure to stress factors during gestation can negatively influence fetal development leading to low birth weight (LBW), which has been associated with later development of cardiometabolic diseases (fetal programming). Malnutrition and exposure to excess glucocorticoids during gestation are some of the stress factors that negatively influence fetal development. In situations of maternal undernutrition, the stress induced by reduced nutrient availability may elevate maternal cortisol levels. Under normal conditions, cortisol concentrations in the fetus are low due to the placental enzymatic barrier (11-β-HSD2) that converts cortisol to inactive cortisone. However, this barrier may be inhibited in malnutrition situations.

Hypothesis: Maternal undernutrition during gestation may induce a poor fetal development, in part due to increased access of cortisol to the fetus.

Objective: To analyze if undernutrition during gestation in the rat increases fetal corticosterone levels and to assess the relationship with fetal growth.

Material and methods: Dams were fed ad libitum (Control) or with 50% of the control diet (Maternal undernutrition, MUN) during the second half of gestation. At the end of gestation (gestational day 20) a blood sample was obtained from the dam and the fetuses were extracted. The fetuses were weighted, and blood was collected after decapitation. Plasma corticosterone levels were assessed by ELISA; maternal protein levels by Bradford and glucose levels with a glucometer. Mann-Whitney U and Student’s t test were used for statistical analysis, as appropriate.

Results: In MUN dams, plasma protein levels were significantly lower, glycaemia was not statistically different and corticosterone levels were significantly higher compared to control dams. MUN fetuses exhibited significantly lower weight and length compared to controls, while plasma corticosterone was significantly higher.

Conclusions: Nutritional restriction during gestation in the rat produces a situation of stress that: 1) increases maternal plasma corticosterone, which can contribute to low protein availability through its catabolic effects; 2) increases passage of corticosterone to the fetus, which may contribute to the deficient fetal growth. Evaluation of 11-β-HSD2 enzymatic activity or protein expression in the placenta is needed to elucidate if these alterations are related to an alteration of this enzymatic barrier in situations of malnutrition.

Keywords: corticosterone, fetal programming, low birth weight, malnutrition, pregnancy, rat. Published: May, 2019. Copyright: © 2019 Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Gila-Diaz A., Reyes-Hernández C.G., Ramiro-Cortijo D., Rodríguez-Rodríguez P., López de Pablo A.L., López-Giménez M.R., González M.C., Arribas S.M., Influence of malnutrition during pregnancy on maternal and fetus plasmatic corticosterone concentrations. IBJ Plus 2019 S(3):e0033 doi: 10.24217/2531-0151.19v1s3.00033. Funding: Plan Nacional I+D+I (FEM2015-63631-R) co-financed with FEDER funds. Competing Interests: The authors declare that no competing interests exist.

Maternal cortisol in early pregnancy as potential biomarker of fetal complications.

David Ramiro-Cortijo1, Andrea Gila-Diaz1, Pilar Rodríguez-Rodríguez1, Ángel L. López de Pablo1, Mª Carmen González1, María de la Calle2, Rubén Gómez-Rioja3, Silvia M. Arribas1.

1Department of Physiology; Faculty of Medicine; Universidad Autónoma de Madrid, C/ Arzobispo Morcillo 2, Madrid, Spain. 2Obstetrics and Gynecology servicie, Hospital Universitario La Paz, Paseo de la Castellana, 261, Madrid, Spain. 3Laboratory Medicine, Hospital Universitario La Paz, Paseo de la Castellana, 261, Madrid, Spain.

*Corresponding author:

Dr. David Ramiro-Cortijo, Department of Physiology, Faculty of Medicine, Universidad Autónoma de Madrid, Spain. E-mail: david.ramiro@uam.es

Introduction: Maternity pattern has changed in Spain, leading to an increase in the age of first pregnancy and the used of assisted reproduction techniques (ART). A relevant consequence is the increase in fetal complications, such as small for gestational age (SGA) and preterm birth, both of which lead to low birth weight (LBW). LBW has a negative impact for both neonatal and long term health, being a risk factor for cardiometabolic disease. Delayed maternity and ART are associated with maternal psychological stress. We have evidenced that stress during pregnancy is associated with LBW. A possible mechanism could be an excess in the level of cortisol access to the fetus that may be deleterious for fetal growth. We hypothesized that maternal plasma cortisol at the beginning of pregnancy, associated with fetal complications and LBW and, therefore, it can be used as early diagnostic tool.

Material and Methods: The study included 200 healthy pregnant women, being attended at the Obstetrics and Gynecology Service from La Paz University Hospital (Madrid, Spain). Women were appointed for blood sample extraction at 10th week of gestation and followed-up until delivery, recording: 1) development of fetal complications including SGA (growth <p3th or <p10th with hemodynamic alterations by Echo-Doppler) and preterm birth (<37gestational age) and 2) offspring birth weight. Blood samples were centrifuged to obtain plasma and cortisol was assessed by competitive immunoassay using a direct chemiluminescent assay. Data were expressed as mean +- standard error of mean or relative frequencies (%). Statistical differences were assessed by Student´s t test with Confidence Interval at 95%. Pearson test (r) was used for correlation analysis, stablishing significance differences at p-value<0.05.

Results: Age of the pregnant women was 34.4+-0.33 years old. The prevalence of fetal complications was 26.5% and LBW was 28.8%. Both were significantly higher in older women (fetal complications=95% CI [-3.25; -0.18]; LBW=95% CI [-3.03; -0.11]). Maternal cortisol, in the first trimester of pregnancy, exhibited a positive correlation with maternal age (r=0.225; p=0.008) and a negative correlation with birth weight (r=-0.326; p=0.006). Plasma cortisol was significantly higher in women who developed fetal complications (95% CI [-5.24; -0.16]) and also in women with premature delivery (95% CI [-6.61; -1.44]).

Conclusions: Maternal plasma cortisol at the beginning of pregnancy is related to the age of maternity and fetal complications leading to LBW. Plasma cortisol could be used as potential biomarker to predict fetal complications particularly in delayed maternity context.

Keywords: birth weight, maternal plasma cortisol, pregnant, preterm birth, small for gestational age. Published: May, 2019. Copyright: © 2019 Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Ramiro-Cortijo D., Gila-Diaz A., Rodríguez-Rodríguez P., López de Pablo A.L., González M.C., De la Calle M., Gómez-Rioja R., Arribas S.M., Maternal cortisol in early pregnancy as potential biomarker of fetal complications. IBJ Plus 2019 S(3):e0034 doi: 10.24217/2531-0151.19v1s3.00034. Funding: This work was supported by Ministerio de Economía y Competitividad (FEM2015-63631-R). Competing Interests: The authors declare that no competing interests exist.

Metabolic response to diets enriched with sunflower seed oil or palm seed oil.

María Valencia-Avezuela1, Victor Naranjo2, Ana Conreras2, Santiago Guerra-Cantera1,3, Sandra Canelles1,3, Jesús Argente1,3,4, Nuria Del Olmo2, Laura

M. Frago1,3, Mariano Ruiz-Gayo2, Julie Chowen1,3,4

1Department of Pediatrics, Universidad Autónoma of Madrid, Madrid, Spain. Department of Endocrinology, Hospital Infantil Universitario Niño Jesús, Instituto de Investigación La Princesa, Madrid, Spain. 2Department of Pharmaceutical and Health Sciences. Universidad CEU-San Pablo, Madrid, Spain 3Centro de Investigación Biomédica en Red de la Fisiopatología de la Obesidad y Nutrición (CIBEROBN), Instituto de Salud Carlos III, Madrid, Spain. 4IMDEA Food Institute, CEI UAM + CSIC, Carretera de Cantoblanco 8. 28049 Madrid, Spain.

*Corresponding author:

J. Chowen Madrid, Spain. E-mail: julieann.chowen@salud.madrid.org

The hypothalamus is the main integrating center for metabolic control, regulating the response to nutrients and other metabolic signals to maintain energy homeostasis. This response includes not only neuronal circuits that control appetite, but also glial cells that are involved in nutrient processing and inflammatory responses to high fat intake. However, it is now clear that the metabolic response is different depending on the type of fatty acid consumed. Our aim was to analyze the hypothalamic response to the consumption of a diet enriched with sunflower seed oil or palm seed oil.

Adult male C57/BL6J mice were given ad libitum access to a normal rodent chow diet enriched with sunflower seed oil with high oleic content (SO) or with palm seed oil (PO), or to a standard rodent chow (SD) for 8 weeks. Energy intake and body weight were monitored weekly. Relative protein expression was analyzed by Western blotting and relative gene expression by RT-PCR.

Both the SO and PO diets increased body weight and energy intake compared to SD (p<0.01). In the hypothalamus neuropeptide Y (NPY) mRNA levels were lower in mice consuming the SO diet compared to SD (p<0.05). No dietary effects were found on the neuropeptides Agouti-related peptide (AgRP) or pro-opiomelanocortin (POMC). Expression of the leptin receptor did not change. There was no change in the relative protein levels of ionized calcium-binding adaptor molecule 1 (Iba 1), a marker of microglia, or of glial fibrillary acidic protein (GFAP), a marker of astrocytes. We found no significant effect on hypothalamic mRNA levels of the cytokine interleukin-6 (IL-6), although there was a clear tendency to increase on the PO diet. There was also no change in the cytokine TNFα or the cell stress marker DNA damageinducible transcript 3 (CHOP). No activation of c-Jun N-terminal kinases (JNK) was found. To explore hypothalamic fatty acid metabolism, we analyzed the relative expression of fatty acid synthase (FASn) and carnitine palmitoyltransferase (Cpt1a). PA decreased Cpt1a mRNA expression compared to SD (p<0.01), with no change in FASn.

In conclusion, although both diets enriched in fat induce weight gain, the central response to these diets differs. The mRNA levels of the orexigenic neuropeptide NPY were lower on the SO diet, which may suggest that this diet could be more effective in reducing food intake, at least at this time-point. In contrast, the PO diet decreased Cpt1a mRNA expression, indicating a possible reduction in hypothalamic fatty acid oxidation that could lead to stress of the system. Further studies are necessary to determine the secondary complications as a result of these specific high fat diets.

Keywords: Obesity, fatty acids, hypothalamus, Published: May, 2019. Copyright: © 2019 Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Valencia-Avezuela M., Naranjo V., Conreras A., Guerra-Cantera S., Canelles S., Argente J., Del Olmo N., Frago L.M., Ruiz- Gayo M., Chowen J. Metabolic response to diets enriched with sunflower seed oil or palm seed oil. IBJ Plus 2019 S(3):e0035 doi: 10.24217/2531-0151.19v1s3.00035. Funding: This work was funded by CIBEROBN, PI16/00485, and BFU2017-82565-C2-1-R, BFU2016-78556R and fondos FEDER. Competing Interests: The authors declare that no competing interests exist.

Skeletal muscle kinases in the regulation of energy balance.

Leticia Herrera-Melle1, Alfonso Mora1, Juan Antonio López2, Sara Cogliati3, María Elena Rodríguez1, Luis Leiva-Vega1, Daniel Beiroa4, Cintia Folgueira4, José Antonio Enríquez3, Jesús Vázquez2, Rubén Nogueiras4, Guadalupe Sabio1*

1Stress kinases in diabetes, cancer and cardiovascular disease Lab, Myocardial Pathophysiology Area, Centro Nacional de Investigaciones Cardiovasculares (CNIC), C/Melchor Fernández Almagro 3, 28029 Madrid, Spain. 2Cardiovascular Proteomics Unit, CNIC, C/Melchor Fernández Almagro 3, 28029 Madrid, Spain. 3Functional genetics of the oxidative phosphorylation system Lab, Myocardial Pathophysiology Area, CNIC, C/Melchor Fernández Almagro 3, 28029 Madrid, Spain. 4Department of Physiology, CiMUS, University of Santiago de Compostela-Instituto de Investigación Sanitaria, Santiago de Compostela 15782, Spain; CIBER Fisiopatología de la Obesidad y Nutrición (CIBERobn), 15706 Santiago de Compostela, Spain.

*Corresponding author:

Guadalupe Sabio, Stress kinases in diabetes, cancer and cardiovascular disease Lab, Myocardial Pathophysiology Area, CNIC, 28029 Madrid, Spain. E-mail: guadalupe.sabio@cnic.es

Obesity is a well-known risk factor for several chronic diseases. Physical inactivity and changes in dietary habits have increased the incidence and prevalence of obesity in current societies, which has become a major health problem. Although several pharmacological treatments to lose weight through a reduction in food intake have been proposed, understanding the molecular mechanisms that lead to increased energy expenditure may help to find new therapies. In fact, exercise has the capacity to improve the metabolic status in obesity and type 2 diabetes, being skeletal muscle an important regulator of glucose homeostasis and an essential contributor to exercise-induced changes in metabolism.

In this work, using a conditional mouse model lacking a kinase in striated muscle, we have attempted to increase knowledge about the role of this protein both at the local and the systemic level. Our results show that the deficiency of this kinase in striated muscle decreases body weight and protects mice against high-fat diet (HFD)-induced obesity by increasing energy expenditure. The molecular alterations caused by this deficiency lead to skeletal muscle metabolic remodeling, increasing mitochondrial oxidative metabolism. Altogether, these molecular and metabolic changes in skeletal muscle are physiologically manifested as enhanced locomotor activity as well as improved glucose homeostasis, decreasing the risk of developing diabetes and liver steatosis, therefore linking local and systemic manifestations of the deficiency of this protein.

Keywords: skeletal muscle, obesity, kinase, energy balance. Published: May, 2019. Copyright: © 2019 Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Herrera-Melle L., Mora A., López J.A., Cogliati S., Rodríguez M.E., Leiva-Vega L., Beiroa D., Folgueira C., Enríquez J.A., Vázquez J., Nogueiras R., Sabio G., Skeletal muscle kinases in the regulation of energy balance. IBJ Plus 2019 S(3):e0036 doi: 10.24217/25310151.19v1s3.00036. Funding: L.H-M. is a Ministry of Education, Culture and Sport (MECD) fellow. This study was funded by the following grants to G.S: ERC 260464, EFSD 2030, MICINN/SAF1305, Fundación BBVA, AstraZeneca and Comunidad de Madrid S2010/BMD-2326. The CNIC is supported by the Ministerio de Ciencia, Innovación y Universidades (MCNU) and the Pro CNIC Foundation, and is a Severo Ochoa Center of Excellence (SEV-2015-0505). Competing Interests: The authors declare that no competing interests exist.

Role of pleiotrophin in hepatic metabolism: characterization in an aging model.

Julia Ramírez-Moya1,2, León Wert-Lamas1,3, Garcilaso Riesco-Eizaguirre1,2,3, Pilar Santisteban1,2.

1Instituto de Investigaciones Biomédicas "Alberto Sols"; Consejo Superior de Investigaciones Científicas (CSIC); Universidad Autónoma de Madrid (UAM), Madrid, Spain. 2Centro de Investigación Biomédicas en Red de Cáncer (CIBERONC), Instituto de Salud Carlos III (ISCIII), Madrid, Spain. 3Departamento de Endocrinología y Nutrición, Hospital Universitario de Móstoles, Madrid, Spain.

*Corresponding author:

Pilar Santisteban, Instituto de Investigaciones Biomédicas "Alberto Sols"; Consejo Superior de Investigaciones Científicas (CSIC); Universidad Autónoma de Madrid (UAM), Madrid, Spain; Centro de Investigación Biomédicas en Red de Cáncer (CIBERONC), Instituto de Salud Carlos III (ISCIII), Madrid, Spain. E-mail: psantisteban@iib.uam.es

Thyroid cancer is a common endocrine malignancy that has rapidly increased its global incidence in recent decades. Currently the important role of microRNAs (miRNAs) in cancer has been described, including thyroid neoplasms. It is known that miRNAs are important modulators of cancer progression and are globally downregulated in several human tumor types, through mechanisms that remain largely undefined. Functional maturation of most miRNAs requires processing of the primary transcript by DICER1, an RNAse III-type enzyme essential for both mammalian development and cell differentiation. Here we identified DICER1 and miRNA downregulation as sentinel events in thyroid cancer, providing a clear cut evidence of the molecular mechanisms underlying this effect. We demonstrated that this key miRNA-processing enzyme is a target of the most upregulated miRNAs in thyroid cancer. Specifically, miR-146b reduced DICER1 expression and DICER1 overexpression inhibited all the miR-146b-induced aggressive phenotypes, both in cells and in tumor models. Our analysis of The Cancer Genome Atlas revealed a general decrease in DICER1 expression in thyroid cancer and suggested a clinical association between DICER1 and patient prognosis. Notably, DICER1 downregulation promoted proliferation, migration, invasion and epithelial-mesenchymal transition in thyroid cancer cell lines, whereas it suppressed the expression of pro-differentiation transcription factors. Interestingly, these factors transcriptionally upregulated DICER1 expression, supporting the existence of a positive feedback loop. Finally, administration of the small molecule enoxacin to promote DICER1-complex activity reduced cell aggressiveness in vitro and tumor growth in vivo. Overall, our data establish DICER1 as a new tumor suppressor in thyroid cancer, and highlight a potential therapeutic approach of RNA-based therapies including antagomiRs and restoration of the biogenesis machinery may provide treatments for thyroid cancer and other cancers.

Keywords: Thyroid cancer, miRNAs, DICER1, miR-146b, antagomiR, Enoxacin. Published: May, 2019. Copyright: © 2019 Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Ramírez-Moya J., Wert-Lamas L., Riesco-Eizaguirre G., Santisteban P., DICER1 downregulation and impaired miRNA processing in thyroid tumorigenesis. IBJ Plus 2019 S(3):e0045 doi: 10.24217/2531-0151.19v1s3.00045. Funding: There are no fundings to declare. Competing Interests: The authors declare that no competing interests exist.

Adrián Acuña-Ruiz1,2, Miguel Angel Zaballos1,2, Garcilaso Riesco-Eizaguirre2,3, Piero Crespo 2,4, Pilar Santisteban1,2,*.

1Instituto de Investigaciones Biomédicas “Alberto Sols”, Madrid, Spain. 2Instituto de Salud Carlos III (ISCIII), CIBERONC, Madrid, Spain. 3Hospital Universitario de Móstoles, Móstoles, Spain. 4Instituto de Biomedicina y Biotecnología de Cantabria, Santander, Spain.

*Corresponding author:

Pilar Santisteban, Instituto de Investigaciones Biomédicas “Alberto Sols”, Madrid, Spain. E-mail: psantisteban@iib.uam.es

Thyroid cancer is the most prevalent human carcinoma of endocrine origin and its incidence is continuously increasing. Around 80% of all the somatic mutations are accumulated in only five driver genes (BRAF, H/N/K-RAS and RET) which converge in the hyperactivation of the MAPK signalling pathway. Given the central role of this pathway in thyroid tumorigenesis, thyroid cancer research is focused on the identification of new drugs that inhibit the different kinases of the pathway. DEL22379 is an inhibitor of ERK dimerization previously characterized in melanoma and colorectal cancer-derived cell lines as a new approach to partially inhibit ERK activation. It has been reported that ERK dimerization contributes to the activation of ERK cytoplasmic effectors, such as RSK, and enhances malignant cell features.

Our objective was to analyse the antitumor effect of the inhibitor of ERK dimerization DEL22379 as a possible therapeutic compound in thyroid cancer.

In vitro assays were made with different thyroid cancer cell lines with mutations in BRAF, H/N/K-RAS or RET. To evaluate cell proliferation, migration and invasion; we have used cell cycle, wound healing and matrigel invasion assays, respectively. We also have developed an orthotopic mouse model of anaplastic thyroid cancer by the generation of the 8505c-luc cell line. In vivo tumour progression was monitored by measuring luciferase activity.

Treatment with DEL22379 impairs ERK dimers formation in BRAF-like thyroid cells whereas Ras-like cells were unaffected. Unlike melanoma, phosphorylated ERK levels were substantially reduced after treatment with the inhibitor. Inhibition of ERK dimerization and phosphorylation correlated with diminished RSK phosphorylation. We also observed a decrease in cell proliferation, migration and invasion in the anaplastic thyroid cell lines 8505c and OCUT2. DEL22379 treatment significantly reduced tumour growth and formation of lung metastases in an orthotopic mouse model.

We propose DEL22379 as a new effective molecule for the inhibition of MAPK signalling and the reduction of cell malignant behaviour, tumour growth and metastasis in vivo, specifically in BRAF-like thyroid tumours. It promises to be another step forward in the achievement to cure thyroid cancer.

Keywords: Thyroid cancer, MAPK signaling, ERK, BRAF. Published: May, 2019. Copyright: © 2019 Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Acuña-Ruiz A., Zaballos MA., Riesco-Eizaguirre G., Crespo P., Santisteban P., Antitumor activity of the ERK inhibitor DEL22379 against BRAF-like cells in thyroid cancer. IBJ Plus 2019 S(3):e0046 doi: 10.24217/2531-0151.19v1s3.00046. Funding: This work was supported by grants SAF2016-75531-R from Ministerio de Economía y Empresa (MINECO), Spain and PI14/01980 from ISCIII, Spain (FEDER) and GCB14142311CRES from Asociación Española contra el Cancer (AECC). Competing Interests: The authors declare that no competing interests exist.

Role of the IQGAP1 scaffold protein in thyroid cancer.

Carlos Carrasco-López1, 2, Miguel Angel Zaballos1, 2, Pilar Santisteban1, 2, *

1Instituto de Investigaciones Biomédicas, C/ Arturo Duperier 4, Madrid, Spain. 2Instituto Carlos III, CIBERONC, Madrid, Spain.

*Corresponding author:

Pilar Santisteban, Instituto de Investigaciones Biomédicas, Madrid, Spain. E-mail: psantisteban@iib.uam.es

The IQGAP family of scaffold proteins comprises three members which have been described to interact with numerous proteins in several biological processes and participate in the signaling cascade of multiple pathways. Among these proteins IQGAP1 has been the most deeply studied, having characterized its function as a scaffold protein in the MAPK pathway and its interaction with actin leading to regulation of the cytoskeleton. Because of these functions it has been studied in multiple malignancies, where it has been assigned a tumorigenic effect.

We have studied IQGAP1 expression using different stablished cell lines of human thyroid carcinomas. Our results indicate that these carcinoma cell lines have variable levels of IQGAP1 but in general these levels are higher when compared to the normal thyroid cell line control. The results obtained are in agreement with the TCGA and Oncomine databases. Using IQGAP1 silenced cell lines we tried to elucidate how this protein was affecting tumor hallmarks such as proliferation, invasion, cell migration and epithelial mesenchymal transition. The effects of silencing IQGAP1 were different depending on the cell line and the oncogenic signature. While in some of them silencing produced an increase in the migration rate of the cells, others had the opposite behavior. We also found these differences when studying EMT markers such as E-cadherin and N-cadherin. Furthermore, cell viability was unaffected by IQGAP1 silencing and consequently the size of the primary tumor was not altered when cells were xenografted in a chicken embryo model. Importantly IQGAP1 silencing greatly decreased the ability of the tumor cells to intravasate and metastasize to distant organs such as lungs and brain.

Further studies will focus on trying to discover the underlying mechanisms responsible for the differences observed between cell lines. Our results indicate a role forIQGAP1 in thyroid tumorigenesis and underscore the importance of defining the oncogenic signature and particularities of a given tumor type in order to design effective therapeutic strategies.

Keywords: IQGAP1, Thyroid, Cancer Published: May, 2019. Copyright: © 2019 Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Carrasco-López C., Zaballos MA., Santisteban P., Role of the IQGAP1 scaffold protein in thyroid cancer. IBJ Plus 2019 S(3):e0047 doi: 10.24217/2531-0151.19v1s3.00047. Funding: Funding This work was supported by grants SAF2016-75531-R from MINECO, Spain and PI14/01980 from ISCIII, Spain (FEDER); B2017/BMD-3724 from CAM and GCB14142311CRES from AECC. Competing Interests: The authors declare that no competing interests exist.

Proteomic analysis of fine-needle aspirates (FNA) for the molecular characterization of non-diagnostic follicular neoplasia.

Vanessa Arosa1, Mikel Azkargorta2, Leire Isasa1, Carmen Fernández1, Leire Pérez1, María Angeles Antón1, Gonzalo Maldonado1 .

1Servicio de Endocrinología y Nutrición, Hospital Universitario Araba, Vitoria-Gasteiz. 2CIC bioGUNE-Proteomics Platform, ProteoRed---ISCIII, Derio, Bizkaia.

*Corresponding author:

Vanessa Arosa, Servicio de Endocrinología y Nutrición, Hospital Universitario Araba, Vitoria-Gasteiz. E-mail: vanessa.arosa@osakidetza.com

Thyroid cancer is the most prevalent malignant endocrine pathology. Differentiated thyroid cancer represents more than 90% of these cases, and 12% of these are follicular carcinomas. Currently, the diagnosis of thyroid nodules is based on the cytologic analysis of fine-needle aspirates (FNA) aided by echography. Around 10% of the cases provided are categorized as IV in the Bethesda system. Since malignancy cannot be ruled out, these cases are subjected to local surgery. The post-operatory analysis of these samples reveals that in 70-85% of them this intervention was not necessary. Given this, additional tools for sample categorization would be welcome.

Mass spectrometry-based proteomics provides a number of tools for the large-scale analysis of proteins in biological samples. Current technology allows the analysis of the presence and/or relative abundance of hundreds/thousands of proteins in a given sample; thus quantitative proteomics allows the comparison of protein levels between conditions (i.e.: healthy vs. disease). Further bioinformatics analyses help deepen into the molecular functions and pathways these proteins are involved in, providing additional information for the understanding of the disease. Thus, proteomics provides a very valuable tool for the molecular characterization of a disease, as well as for biomarker discovery.

In this work we present preliminary results on the analysis of FNA samples by proteomics. The aspirates were centrifuged, and both the resulting supernatant and the pellet containing aspirated cells and debris were processed and loaded into an LTQ Orbitrap XL-ETD mass spectrometer for protein identification. Identified proteins were further characterized using bioinformatic tools. The methods and results presented in this poster lay foundation for coming analyses in the context of the molecular characterization of these poorly defined cases, being our main goal the search for FNA markers that help distinguish follicular adenomas from carcinomas and thus avoid surgery.

Keywords: mass spectrometry, fine-needle aspirate (FNA), biomarker discovery, functional analysis. Published: May, 2019. Copyright: © 2019 Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Arosa V., Azkargorta M., Isasa L., Fernández C., Pérez L., Antón MA., Maldonado G., Santisteban P., Proteomic analysis of fine-needle aspirates (FNA) for the molecular characterization of non-diagnostic follicular neoplasia. IBJ Plus 2019 S(3):e0048 doi: 10.24217/2531-0151.19v1s3.00048. Funding: No fundings to declare. Competing Interests: The authors declare that no competing interests exist.

Radioiodide therapy in ovarian cancer.

Mielu Lidia Mirela1,2, Fajardo-Delgado Dánae2, Hortigüela Rafael2, Diego-Hernández Cristina3, García-Jiménez Custodia4, Martín-Duque Pilar5, and De la Vieja Antonio1,2

1CiberOnc (ISCIII), Spain 2Endocrine Tumors Unit. UFIEC (ISCIII), Majadahonda, Spain 3Hospital Universitario de Móstoles (HUM), Móstoles, Spain 4Universidad Rey Juan Carlos, Alcorcón, Spain 5Instituto Aragonés de Ciencias de la Salud, Spain

*Corresponding author:

Lidia Mirela Mielu, CiberOnc, Madrid, Spain, E-mail: mielulidia@gmail.com

Introduction: Ovarian cancer is the most lethal gynecological malignancy. Early diagnosis has a survival rate of 90%. Unfortunately, more than 70% of cases are diagnosed when the cancer has already metastasized, and survival rates do not exceed 30% in these cases. The sodium iodide symporter (NIS) mediates active transport of iodide into the thyroid as a fist steps in thyroid hormone biosynthesis, but also in others tissues like salivary gland, lactating mammary gland and stomach. Our group has demonstrated that NIS is expressed in ovarian surface epithelium and is overexpressed in human epithelial ovarian cancer, establishing NIS as a tumor marker. The aim of this study is to determine whether overexpression of NIS in ovarian cancer can be use as therapeutic tool using radioiodide therapy (RAI) in ovarian tumors. Materials and Methods: serous ovarian cancer cell line (SKOV3) was transfected permanently with exogenous NIS (SKOV3hNIS) and in vitro characterized by different techniques (western-blot, PCR, flow cytometry, immunofluorescence, iodide uptake and proliferation, migration, invasion, and adhesion assays). In vivo, NIS transfected cells and no transfected was injected into the flanks of nude and NSGs mice. Tumor growth was monitored by measurements with caliber and IVIS. The expression of NIS in tumors was analyzed by different molecular biology techniques and NIS functionality in animal models was measured by SPECT-CT. Results: PCR and western-blot show NIS expression in both in vitro cancer cells and in vivo with xenotransplanted cells in animal models. Immunofluorescence and immunohistochemistry show that NIS expression occurs in plasma membrane, and iodide uptake assays show that the expression of NIS in plasma membrane is functional in vitro and in SPECT-CT assay in vivo. Additionally, NIS presence alters in vitro and in vivo proliferation, migration, invasion and adhesion of tumor cell properties. Conclusion: NIS expression in human ovarian cancer cell lines is functional in vitro and in vivo, targeted to the plasma membrane and able to accumulate iodide. The high expression levels are correlated with worse prognosis pointing NIS as a future therapeutic approach in treatment of ovarian cancer.

Keywords: ovarian cancer, sodium/iodide symporter (NIS), Radioioide therapy (RAI), SPECT-CT Published: May, 2019. Copyright: © 2019 Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Mielu Lidia M., Fajardo-Delgado D., Hortigüela R., Diego-Hernández C., García-Jiménez C., Martín-Duque P., and De la Vieja A., Radioiodide therapy in ovarian cancer. IBJ Plus 2019 S(3):e0049 doi: 10.24217/2531-0151.19v1s3.00049. Funding: This work was supported by grants from the Ministerio de Economía y Competitividad and European Regional Development Fund (FEDER) SAF2015-69964-R and CiberOnc from the ISCIII. Competing Interests: The authors declare that no competing interests exist.

Metabolic adaptations in spontaneously immortalized PGC-1α knock-out mouse embryonic fibroblasts increase their oncogenic potential.

Ignacio Prieto1, Carmen Rubio-Alarcón1, Raquel García-Gómez1, Rebeca Berdún2, Manuel Portero2, Reinald Pamplona2, Antonio Martínez-Ruiz3,4, José Ignacio Ruiz-Sanz5, Mª Begoña Ruiz-Larrea5, Mariona Jove2, Sebastian Cerdán1, María Monsalve1* .

1Instituto de Investigaciones Biomédicas “Alberto Sols” (CSIC-UAM). Arturo Duperier 4. 28029-Madrid (Spain). 2Institut de Recerca Biomédica Lleida. Avda. Alcalde Rovira Roure 80, 25198-Lleida (Spain). 3Unidad de Ivestigación, Hospital Universitario Santa Cristina, Instituto de Investigación Sanitaria Princesa (IIS-IP). Maestro Vives 3, 28009 – Madrid (Spain). 4Centro de Investigación Biomédica en Red de Enfermedades Cardiovasculares (CIBERCV), Spain. 5Departamento de Fisiología, Facultad de Medicina y Enfermería. Universidad del País Vasco/Euskal Herriko Unibertsitea. Barrio Sarriena s/n. 48940-Leioa (Spain).

*Corresponding author:

María Monsalve, Instituto de Investigaciones Biomédicas Alberto Sols (CSIC-UAM), Madrid, Spain. E-mail: mpmonsalve@iib.uam.es

PGC-1α controls, to a large extent, the capacity of cells to respond to changing nutritional requirements and energetic demands. The key role of metabolic reprogramming in tumor development has highlighted the potential role of PGC-1α in cancer. To investigate how loss of PGC-1α activity in primary cells impacts the oncogenic characteristics of spontaneously immortalized cells, and the mechanisms involved, we used the classic 3T3 protocol to generate spontaneously immortalized mouse embryonic fibroblasts (iMEFs) from wild-type (WT) and PGC-1α knockout (KO) mice and analyzed their oncogenic potential in vivo and in vitro. We found that PGC-1α KO iMEFs formed larger and more proliferative primary tumors than WT counterparts, and fostered the formation of lung metastasis by B16 melanoma cells. These characteristics were associated with the reduced capacity of KO iMEFs to respond to cell contact inhibition, in addition to an increased ability to form colonies in soft agar, an enhanced migratory capacity, and reduced growth factor dependence. The mechanistic basis of this phenotype is likely associated with the observed higher levels of nuclear β-catenin and c-myc in KO iMEFs. Evaluation of the metabolic adaptations of the immortalized cell lines identified a decrease in oxidative metabolism and an increase in glycolytic flux in KO iMEFs, which were also more dependent on glutamine for their survival. Furthermore, glucose oxidation and tricarboxylic acid cycle forward flux were reduced in KO iMEF, resulting in the induction of compensatory anaplerotic pathways. Indeed, analysis of aminoacid and lipid patterns supported the efficient use of tricarboxylic acid cycle intermediates to synthesize lipids and proteins to support elevated cell growth rates. All these characteristics have been observed in aggressive tumors and support a tumor suppressor role for PGC-1α, restraining metabolic adaptations in cancer.

Keywords: PGC-1α, metabolism, cancer, tumor, metastasis. Published: May, 2019. Copyright: © 2019 Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Prieto I., Rubio-Alarcón C., García-Gómez R., Berdún R., Portero M., Pamplona R., Antonio Martínez-Ruiz A., Ruiz-Sanz J.I., Ruiz- Larrea MB., Jove M., Cerdán S., Monsalve M., Santisteban P., Metabolic adaptations in spontaneously immortalized PGC-1α knock-out mouse embryonic fibroblasts increase their oncogenic potential. IBJ Plus 2019 S(3):e0050 doi: 10.24217/2531-0151.19v1s3.00050. Funding: This work was funded by grants from the Spanish “Ministerio de Economía Industria y Competitividad” (MINEICO) and ERDF/FEDER funds, SAF2012-37693, SAF2015-63904-R, SAF2015-71521-REDC, to M.M., SAF2017-83043-R and B2017/BMD-3724 to S.C., PI15/00107 to A.M.R, the University of the Basque Country UPV/EHU grant GIU16/62) to J.l.R.S. and M.B.R.L., and the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement 721236-TREATMENT to M.M. Competing Interests: The authors declare that no competing interests exist.

Gene regulatory and phenotypic effects of calcitriol and canonical WNT in human colon fibroblasts.

Gemma Ferrer-Mayorga1,2,3, Núria Niell1,4,^, Ramón Cantero2,5, José Manuel González-Sancho1,3,4, Luis del Peso1,2,4,6, Alberto Muñoz1,2,3,* &

María Jesús Larriba1,2,3,*.

1Instituto de Investigaciones Biomédicas “Alberto Sols”, Consejo Superior de Investigaciones Científicas, Universidad Autónoma de Madrid, Madrid, Spain. 2Instituto de Investigación Sanitaria Hospital Universitario La Paz, Madrid, Spain. 3CIBERONC, Instituto de Salud Carlos III, Madrid, Spain. 4Departamento de Bioquímica, Facultad de Medicina, Universidad Autónoma de Madrid, Madrid, Spain. 5Servicio de Cirugía General, Hospital Universitario La Paz, Madrid, Spain. 6CIBERES, Instituto de Salud Carlos III, Madrid, Spain. ^Present address: Departamento de Biología, Facultad de Ciencias, Universidad Autónoma de Madrid, Madrid, Spain.

*Corresponding authors:

Alberto Muñoz, Instituto de Investigaciones Biomédicas “Alberto Sols”, Consejo Superior de Investigaciones Científicas, Universidad Autónoma de Madrid, Madrid, Spain. amunoz@iib.uam.es María Jesús Larriba, Instituto de Investigaciones Biomédicas “Alberto Sols”, Consejo Superior de Investigaciones Científicas, Universidad Autónoma de Madrid, Madrid, Spain. mjlarriba@iib.uam.es

Vitamin D3 (cholecalciferol) is synthesized in the skin by action of solar UV radiation or incorporated via diet, and is the inactive precursor of 1α,25-dihydroxyvitamin D3 (calcitriol), a major pleiotropic hormone in the human organism. By binding and activation of vitamin D receptor (VDR), a member of the superfamily of nuclear receptors, calcitriol regulates the expression of hundreds of genes in a tissue- and cell-type specific manner by transcriptional and posttranscriptional mechanisms.

Human WNT factors are a family of 19 members of secreted proteins that regulate crucial processes in many tissues and organs during development and adult life. Some (canonical) WNTs act by modulating the transcriptional activity of β-catenin whereas other (non-canonical) WNTs affect different signal transducers (Ca2+, Rho, JNK...) independently of β-catenin.

Colorectal cancer (CRC) is one of the most important neoplasias worldwide in terms of incidence and mortality. The key role of the constitutive activation of the WNT/β-catenin signaling pathway promoting CRC and the protective effect of VDR agonists, in part by antagonizing the WNT/β-catenin pathway, have been widely described. However, the effects of calcitriol and canonical WNTs on stromal fibroblasts, which are important players in CRC with protumorigenic action, remain mostly unknown.

We have studied the effect of calcitriol and WNT3A on the human CCD-18Co colonic fibroblast cell line and on primary colon fibroblasts isolated from CRC patients. Data from global transcriptomic analyses by RNA-sequencing and also functional assays indicate that calcitriol and WNT3A exert a wide regulatory action on the gene expression and phenotype of colonic fibroblasts. Remarkably, a complex interplay exists between the two agents: while calcitriol and WNT3A act cooperatively in some effects, antagonistic actions are found in others.

Our results show that calcitriol and WNT3A are important modulators of human colon fibroblasts, with potential implications in colon homeostasis and neoplastic transformation.

Keywords: Calcitriol, vitamin D, WNT, colon fibroblasts, colorectal cancer Published: May, 2019. Copyright: © 2019 Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Cite as: Ferrer-Mayorga G., Niell N., Cantero R., González-Sancho J.M., Del Peso L., Muñoz A. & Larriba MJ., Gene regulatory and phenotypic effects of calcitriol and canonical WNT in human colon fibroblasts. IBJ Plus 2019 S(3):e0051 doi: 10.24217/25310151.19v1s3.00051. Funding: Ministerio de Ciencia, Innovación y Universidades (SAF2016-76377-R, Nurcamein2) and Instituto de Salud Carlos III (CIBERONC) of Spain - Fondo Europeo de Desarrollo Regional. Competing Interests: The authors declare that no competing interests exist.