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Somatostatin, Cortistatin, Ghrelin and Their Receptors: from an Endocrine System to a Pleiotropic System of Patho- Physiologic Relevance

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Somatostatin, Cortistatin, Ghrelin and Their Receptors: from an Endocrine System to a Pleiotropic System of Patho- Physiologic Relevance DEPARTAMENTO DE BIOLGÍA CELULAR, FISIOLOGÍA E INMUNOLOGÍA Somatostatin, cortistatin, ghrelin and their receptors: From an endocrine system to a pleiotropic system of patho- physiologic relevance Manuel D. Gahete Ortiz Córdoba 2010 TITULO: Somatostatin, cortistatin, ghrelin and their receptors: from an endocrine system to a pleitropic system of pathophysiologic relevance AUTOR: MANUEL D. GAHETE ORTIZ © Edita: Servicio de Publicaciones de la Universidad de Córdoba. 2010 Campus de Rabanales Ctra. Nacional IV, Km. 396 14071 Córdoba www.uco.es/publicaciones [email protected] ISBN-13: 978-84-693-2990-0 DEPARTAMENTO DE BIOLGÍA CELULAR, FISIOLOGÍA E INMUNOLOGÍA Somatostatin, cortistatin, ghrelin and their receptors: From an endocrine system to a pleiotropic system of patho- physiologic relevance Memoria de Tesis Doctoral presentada por Manuel D. Gahete Ortiz, Licenciado en Biología, para optar al grado de Doctor en Ciencias. Los Directores Dr. Justo P. Castaño Fuentes Dr. Raúl Luque Huertas Profesor Titular de Biología Celular Investigador Ramón y Cajal de la Universidad de Córdoba Universidad de Córdoba En Córdoba, a 8 de Marzo de 2010 DEPARTAMENTO DE BIOLGÍA GRUPO DE CELULAR, FISIOLOGÍA E ENDORINOLOGÍA INMUNOLOGÍA CELULAR Y MOLECULAR Dº Justo Pastor Castaño Fuentes y Dº Raúl Luque Huertas, Profesor Titular e Investigador Contratado del Departamento de Biología Celular, Fisiología e Inmunología de la Uniersidad de Córdoba, INFORMAN Que Dº Manuel David Gahete Ortiz, Licenciado en Biología, ha realizado bajo nuestra dirección el trabajo titulado “Somatostatin, cortistatin, ghrelin and their receptors: From an endocrine system to a pleiotropic system of patho-physiologic relevance” y que bajo nuestro juicio reúne los méritos suficientes para optar al Grado de Doctor en Ciencias Y para que conste, firmo la presente en Córdoba, a 8 de Marzo de 2010 Fdo.: Prof Dr. Justo Pastor Fdo.: Dr. Raúl Miguel Castaño Fuentes Luque Huertas Esta Tesis Doctoral ha sido realizada en el Departamento de Biología Celular, Fisiología e Inmunología de la Universidad de Córdoba, bajo la dirección de los Dres. Justo P. Castaño Fuentes y Raúl Luque Huertas. Dicho proyecto fue subvencionado mediante una beca del Programa Nacional de becas de FPU (FPU-AP20052473) concedida por el Ministerio de Educación y Ciencia. “Si he conseguido ver más lejos es porque me he aupado en hombros de gigantes” Isaac Newton “La potencia intelectual de un hombre se mide por la dosis de humor que es capaz de utilizar” Friedrich Nietzsche AGRADECIMIENTOS AGRADECIMIENTOS Cuando termina un ciclo parece inevitable echar la vista atrás y hacer balance; yo he tenido la oportunidad de hacer lo que realmente me gusta y por eso, en mi caso, la experiencia ha sido positiva, de hecho ya lo dijo Lee Smolin “en el fondo, los científicos somos gente con suerte; podemos jugar a lo que queramos durante toda la vida”. Además, he tenido el placer de compartir este camino con gente que me ha ayudado a crecer en todos los aspectos, con los que compartido grandes momentos y de los que conservaré un buen recuerdo. Mis primeras palabras de agradecimiento van dirigidas a mis directores de Tesis, Justo y Raúl, que hace ya algún tiempo me “convencieron” para iniciar esta aventura científica; ellos me han guiado durante todo este proceso y me han ayudado a terminar en tiempo y forma. Justo, tú me has enseñado a buscarle una finalidad a la ciencia, a mirar más allá, a no conformarme, a esforzarme al máximo, a dar lo mejor de mí. En definitiva me has enseñado que la ciencia no es sólo un trabajo sino una forma de entender la vida. Raúl, tú has sido un referente, un ejemplo a seguir. Tu apoyo y amistad han sido indispensables para mí. Gracias a ti he aprendido que un buen científico puede ser una persona leal, colaboradora, comprometida y solidaria. También gracias por tu apoyo Antonio, tú has sido como un director de Tesis para mí. Siempre he podido contar con tus consejos en el laboratorio y con tus historias en el comedor. Gracias también a Paco, María, Soco y Rafa por ayudarme a madurar científicamente y por mostrarme siempre otros puntos de vista. Compartir el día a día con vosotros, me ha enseñado mucho, a nivel personal y profesional. No quiero olvidarme de los doctores con los que he compartido mis visitas al extranjero; gracias a Rhonda por su hospitalidad y apoyo absoluto, a Stefan por enseñarme otra forma trabajar en el laboratorio y a Hubert, ejemplo de una vida dedicada a la ciencia. Obviamente, tengo mucho que agradecer a Alberto, que ha pasado de ser “aquel tipo raro de barbas que se sentaba al fondo del laboratorio” a un gran apoyo y amigo tanto dentro como fuera del laboratorio. Contigo he compartido todos los momentos, los buenos y los malos. Deseo que todo tu esfuerzo y dedicación tengan pronto su recompensa. También gracias a ti, Bea, por ayudarme a sobrellevar a tu novio en estos últimos años. Quiero hacer también una mención especial a Mario que me enseñó mucho y me guió durante momentos duros, a Ana por contagiarme su buen humor, a David con el que he compartido buenos ratos de ciencia y deporte, a Ester por aguantarme durante tantos años, a Jose por tu espontaneidad e inconformismo, y a Anabel, gracias por tu ayuda y por hacerme más ameno el día a día Gracias también a mi familia y amigos. Especialmente a mis padres y mi hermano, que siempre han estado a mi lado, dándome su apoyo, pendientes de cada cosa que he necesitado. Por supuesto, tengo muchísimo que agradecer a Almudena. Tú has sido la razón para seguir, la luz al final del túnel, quien ha estado siempre a mi lado, quien me ha dado fuerza en momentos de debilidad, quien ha disfrutando junto a mí de los buenos momentos, quien me ha dado su apoyo incondicional. Tú has sido mi baluarte, sin ti no lo hubiera conseguido. Por último, gracias a todas y cada una de las personas que, en algún momento, han compartido esta andanza conmigo. Todos vosotros formáis parte te este trabajo, sin vuestra ayuda no hubiera sido posible. A Almudena, mis padres y mi hermano LIST OF PUBLICATIONS LIST OF PUBLICATIONS This Thesis is based on the research articles listed bellow, which will be referred in the text by their Roman numerals. I. Raúl M. Luque, Manuel D. Gahete, Ute Hochgeschwender, and Rhonda D. Kineman. Evidence that endogenous SST inhibits ACTH and ghrelin expression by independent pathways. 2006. American Journal of Physiology: Endocrinology and Metabolism 291(2), 395–403. II. Manuel D. Gahete, José Córdoba-Chacón, Roberto Salvatori, Justo P. Castaño, Rhonda D. Kineman, Raúl M. Luque. Metabolic regulation of ghrelin O-acyl transferase (GOAT) expression in the mouse hypothalamus, pituitary, and stomach. 2010. Molecular and Cellular Endocrinology 317(1-2), 154-60. III. Manuel D. Gahete, Alicia Rubio, Mario Durán-Prado, Jesús Ávila, Raúl M. Luque, Justo P. Castaño. Expression of somatostatin, cortistatin, and their receptors, as well as dopamine receptors, but not of neprilysin, are reduced in the temporal lobe of Alzheimer disease patients. 2010. Journal of Alzheimer Disease. DOI: 10.3233/JAD-2010-1385 IV. Manuel D Gahete, Raúl M. Luque, Justo P. Castaño. A novel human ghrelin transcript containing intron-2 (In2-ghrelin) as well as Ghrelin-O-acyltransferase is over-expressed in breast cancer: pathophysiological relevance. Manuscript. V. Manuel D Gahete, Raúl M. Luque, Justo P. Castaño. The sst5 truncated isoform, sst5TMD4, is expressed in primary breast cancer samples and increases the malignancy of breast MCF-7 cells. Manuscript. VI. Manuel D. Gahete, Raúl M. Luque, Justo P. Castaño. Expression of the ghrelin and neurotensin systems is altered in the temporal lobe of Alzheimer disease patients. Manuscript. LIST OF ABBREVIATIONS LIST OF ABBREVIATIONS aa – amino acid ACTH – Adrenocorticotropic hormone AD – Alzheimer Disease CNS – Central nervous system CST – Cortistatin DIO – Diet-induced obesity GH – Growth hormone GHRL – Ghrelin/obestatin gene GHRH – Growth hormone releasing hormone GHRP-6 - Growth hormone releasing peptide 6 GHS-R – Ghrelin receptor GIT – Gastrointestinal tract GOAT – Ghrelin O-acyltransferase GPCR – G-protein coupled receptor HPT - hypothalamus MBOAT – Membrane-bound O-acyltransferase NMUR - Neuromedin receptor NTSR - Neurotensin receptor PC - Prohormone convertase Pit – Pituitary gland PRL - Prolactin SST – Somatostatin sst – Somatostatin receptor TMD – Transmembrane domain UAG – non-acylated ghrelin TABLE OF CONTENTS TABLE OF CONTENTS Table of contents 1.- Introduction ................................................................................................................................................................. - 1 - 2.- Aims of the study .................................................................................................................................................... - 13 - 3.- Results and general discussion ......................................................................................................................... - 17 - 3.1.- Regulation of SST, ghrelin and GOAT in metabolic conditions (Articles I & II)......................... - 17 - 3.2.- SST, ghrelin and breast cancer (Articles IV & V) .................................................................................... - 19 - 3.2.1- The role of sst5TMD4 in breast cancer (Article IV) ........................................................................... - 20 - 3.2.2- In2-ghrelin variant and breast cancer (Article V) .............................................................................. - 21 - 3.3.- SST,
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