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Specific Rearrangements and New Molecular Markers in Acute Myeloid Leukemia: Study of MYBL2 Genetic Alterations and Its Involvement in Leukemogenesis

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Specific Rearrangements and New Molecular Markers in Acute Myeloid Leukemia: Study of MYBL2 Genetic Alterations and Its Involvement in Leukemogenesis Facultad de Ciencias Biológicas Departamento de Bioquímica y Biología Molecular Señalización Celular y Patologías Asociadas TESIS DOCTORAL CON MENCIÓN INTERNACIONAL Specific rearrangements and new molecular markers in acute myeloid leukemia: study of MYBL2 genetic alterations and its involvement in leukemogenesis Doctoranda SANDRA DOLZ GIMÉNEZ Directoras Dra. Eva Barragán González Dra. Paloma García Rodríguez EVA BARRAGÁN GONZÁLEZ, doctora en Bioquímica, Biología Molecular y Biomedicina; facultativo especialista de Bioquímica Clínica en el Hospital Universitario y Politécnico La Fe, PALOMA GARCÍA RODRÍGUEZ, doctora en Bioquímica y Ciencias Biológicas; Investigadora Senior en Cáncer, Biología Celular y Biología Molecular en el Instituto de Investigación Biomédica de la Universidad de Birmingham, INFORMAN que el presente trabajo de investigación titulado “Specific rearrangements and new molecular markers in acute myeloid leukemia: study of MYBL2 genetic alterations and its involvement in leukemogenesis” y realizado por SANDRA DOLZ GIMÉNEZ, licenciada en Farmacia por la Universidad de Valencia y con título de Facultativo Especialista de Análisis Clínicos concedido por el Hospital Universitario y Politécnico La Fe, bajo nuestra supervisión en el laboratorio de Biología Molecular del Servicio de Análisis Clínicos del Hospital Universitario y Politécnico La Fe, en el marco del Instituto de Investigación Sanitaria La Fe, y en los laboratorios del Instituto de Investigación Biomédica de la Universidad de Birmingham, reúne los requisitos para su presentación y defensa en forma de Tesis Doctoral con Mención Internacional ante un tribunal. Y para que así conste a los efectos oportunos firman la presente en Valencia, octubre 2015. Fdo: Eva Barragán Fdo: Paloma García Este estudio se ha realizado con el apoyo económico de los proyectos de investigación PI06/0657 y PS09/01828, ambos financiados por el Instituto de Investigación en Salud Carlos III. Sandra Dolz Giménez ha disfrutado del Contrato de Investigación Post- Residente 2010/0258 concedido por el Instituto de Investigación Sanitaria La Fe y la Fundación Bancaja, y del Contrato de Investigación Río Hortega CM13/00022 concedido por el Instituto de Investigación en Salud Carlos III, así como de una estancia de 3 meses en el Instituto de Investigación Biomédica de la Universidad de Birmingham (Birmingham, Reino Unido) concedida por el Instituto de Investigación Sanitaria La Fe (2010/0258). Este trabajo se ha realizado en los laboratorios del Servicio de Análisis Clínicos y del Servicio de Hematología del Hospital Universitario y Politécnico La Fe (Valencia, España), en el marco del Instituto de Investigación Sanitaria La Fe, bajo la dirección de la Dra. Eva Barragán González, así como en los laboratorios del Instituto de Investigación Biomédica de la Universidad de Birmingham (Birmingham, Reino Unido), bajo la dirección de la Dra. Paloma García Rodríguez. A partir de la presente tesis se han derivado las publicaciones científicas que se citan a continuación: 1. Dolz S, et al. A novel Real-Time Polymerase-Chain Reaction assay for simultaneous detection of recurrent fusion genes in acute myeloid leukemia. J Mol Diagn. 2013; 15: 678-686. 2. Dolz S, et al. Study of the S427G polymorphism and of MYBL2 variants in patients with acute myeloid leukemia. Leuk Lymphoma 2015; 19: 1-7. 3. Dolz S, et al. Analysis of MYBL2 downstream targets and its role in human hematopoietic stem cell development. (Manuscrito en elaboración). A mi yaya Ángeles A mis padres A Javi El mejor científico está abierto a la experiencia, y ésta empieza con un romance, es decir, la idea de que todo es posible. Ray Bradbury La duda es la madre del descubrimiento. Ambrose Bierce AGRADECIMIENTOS Este trabajo es el resultado de un proceso de aprendizaje en el que, con esfuerzo y constancia, he adquirido conocimientos del método científico, técnica y disciplina. Me siento muy afortunada de poder aportar mi pequeño grano de arena a la ciencia, a esa facultad cuyo avance permite mejorar la salud y calidad de vida de las personas. Durante esta trayectoria, he tenido la suerte de conocer a gente que me ha acompañado y/ó ayudado en la misma, a la que quería dedicar unas palabras de agradecimiento. En primer lugar, quería agradecer la dedicación y el apoyo de mis directoras de tesis, las Dras. Eva Barragán y Paloma García, ya que sin ellas este trabajo no podría haber salido adelante. A la Dra. Eva Barragán, gracias a la oportunidad que me brindó de formarme a su lado. Ha sido todo un lujo para mí poder adquirir conocimientos junto con una persona de gran talento, capacidad de proyección científica y dedicación, que me ha estimulado a aprender a mejorar cada día, a perfeccionar mi trabajo, a crecer y a madurar en el ámbito profesional. Gracias a tus buenos consejos y a todo lo que me has enseñado. A la Dra. Paloma García, gracias a la afectuosa acogida en su centro de investigación. Fue una gran suerte para mí poder aprender de la mano de una profesional del ámbito científico internacional, de gran talento, creativa y con gran capacidad de transmitir conocimiento. Gracias a ello, consigue “hacer fácil lo difícil” y he aprendido a superar adversidades, a comunicar y a crecer en el campo de la investigación. Gracias por estar siempre dispuesta a ayudarme, a tu ánimo y sonrisa. En segundo lugar, quería agradecer al profesor Miguel A. Sanz, un ejemplo de superación, la confianza que depositó en mi trabajo, a su cercanía y accesibilidad (a pesar de su escaso tiempo libre), a sus enseñanzas y a la buena voluntad que demuestra en ayudar a potenciar los conocimientos de la gente que le rodea. Ha sido un honor poder dedicar con tal eminencia de la Hematología mis horas de trabajo. De igual manera, también quería agradecer al Dr. Pascual Bolufer y a todo el equipo del laboratorio de Biología Molecular de Análisis Clínicos el apoyo que me prestaron, a sus enseñanzas, ayuda, a su colaboración y a sus buenos consejos. Gracias a ellos, he aprendido y he desarrollado mis conocimientos de Biología Molecular bajo la tutela de auténticos profesionales en la materia. A Óscar, Sarai, Inma, Marta, Rosa, Gema, Juan Carlos, Nuria, Virginia, Esperanza, Jorge, Teresa y Enrique, gracias a las horas de trabajo dedicadas y a los buenos momentos que pasamos juntos. A todo el equipo del laboratorio de Citogenética de Hematología, en especial a Jose, Esperanza, Inés, Irene, Mariam y María, gracias también por su apoyo, colaboración y a su trabajo. Así como a Lourdes del Servicio de Citometría, gracias por su ayuda, su dedicación, su agradable trato y amable colaboración, siempre dispuesta a facilitar y contribuir al trabajo de la mejor manera. Mi agradecimiento a la Dra. Dolors Sánchez, coordinadora de la Unidad de Genómica, por su colaboración, accesibilidad y dedicación al trabajo, por sus enseñanzas, su ayuda, sus consejos y su entusiasmo en las investigaciones. Gracias al equipo de investigación de la Dra. Paloma García de Birmingham, por la cálida acogida (a pesar del frío invierno) que recibí durante mi estancia allí. A Mary, Carl, Lozan, Giacomo, Emilie, Maelle, John, Stephanie y Roger, por su profesional trabajo, su disponibilidad y atención, su colaboración y ayuda, sus enseñanzas, su simpatía y por hacerme disfrutar de muy buenos momentos. Fue un lujo para mí poder trabajar junto a ellos en un centro de investigación de prestigio, que me maravilló desde el primer momento. Muchas gracias a Leyre, Alessandra y Banessa, por hacerme pasar buenos momentos, disfrutar de su compañía y hacerme sentir como en casa. Gracias a todos ellos guardo un entrañable y grato recuerdo, una experiencia que nunca olvidaré. Gracias al equipo de trabajo del Centro de Transfusiones de Valencia, en especial a Marga, Isabel, Mª Ángeles, Amparo y a la Dra. Pilar Solves, con quien iniciamos esta colaboración. Gracias a todos ellos por su dedicación, por su amable disposición y contribución a este trabajo. Finalmente, quería dedicar unas palabras de agradecimiento a las personas que más quiero, a toda mi familia y “apegaos”, gracias por estar ahí. En especial, a mis padres y a mi hermana, gracias por estar siempre conmigo, apoyándome en todo, por confiar en mí, por comprenderme, por animarme y por vuestro cariño. A los mejores padres que podría tener, gracias por la educación que me habéis dado y a vuestro esfuerzo, me siento muy afortunada por ello. De igual manera, me siento muy afortunada de tener a mi hermana, mi médico preferida, con quien he compartido tantas horas de estudio. Gracias Loren por estar siempre cerca de mí y ser la mejor hermana. Guardo bonitos recuerdos juntas en casa de la yaya, con mi tío Antonio siempre estudiando y mi tía Paqui tecleando la máquina de escribir, y así seguimos después sus pasos. Gracias a esos momentos y a mi querida yaya Ángeles, a quien tanto echamos de menos, una persona ejemplar que siempre ha sido joven, despierta, inteligente, avanzada a su tiempo, tolerante, luchadora, optimista, carismática, llena de vida y ganas de vivir, y sobre todo apasionada de su familia, a la que siempre nos ha dado sabios consejos, nos ha animado a estudiar, a esforzarnos cada día y por todo ello, su memoria siempre nos mantendrá unidos. Gracias yaya por cuidarme, por tu cariño y por tu recuerdo. Gracias a mis amigos por apoyarme, en especial a quien siempre fue y es mi mejor y más que amigo, mi novio Javi. Gracias por estar siempre conmigo, por tu paciencia, por compartir los momentos difíciles y también los mejores, gracias al apoyo de tu familia y a los buenos momentos juntos, gracias por tu comprensión, por tu esfuerzo en ayudarme en todos los sentidos, por hacerme reír, por animarme, por ser mi compañero y por quererme, sin ti este trabajo no habría tenido un mejor final. TABLE OF CONTENTS TABLE OF CONTENTS Page Achnowledgments………………………………………………………………....
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