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Nuevas anomalías genéticas y epigéneticas en las células iPS, campo biomédico que bulle en investigaciones, se revelan en varios estudios que se publican hoy. Aún es pronto para determinar si esas cicatrices genómicas les impedirán desplegar todo su potencial terapéutico.
Tres nuevos estudios que se publican hoy en Nature se suman a otros dos recientes, aparecidos en Cell Stem Cell, que advierten sobre las anomalías genéticas y epigenéticas de las células pluripotenciales inducidas o iPS. Estos trabajos indican que el proceso de reprogramación celular, impulsado en 2006 por el científico japonés Shinya Yamanaka, de las universidades de Kioto y California, se asocia con aberraciones en la esencia celular que podrían dificultar su empleo biomédico.
A las alteraciones en la metilación de ADN del genoma de las iPS descubiertas en un primer trabajo que se difundió hace un mes, se añade una importante presencia de variaciones en el número de copias (CNV) y de mutaciones en genes codificadores de proteínas claves para las funciones celulares. ¿Un punto flaco que se superará o un escollo insalvable? El tiempo dará la respuesta.
Variaciones en el número de copias
La reprogramación de células somáticas para que adquieran capacidad pluripotencial embrionaria origina una presencia importante de variaciones en el número de copias (CNV) en las células iPS obtenidas.
Estas alteraciones del ADN por las que una determinada región cromosómica queda amplificada o desaparece son el resultado del propio proceso reprogramador, según sugiere un estudio coordinado por el laboratorio de Andras Nagy, en el Instituto de Investigación Samuel Lunenfeld, del Hospital Mount Sinai, de Toronto (Canadá), y por el grupo de Timo Otonkonski, del Centro Biomédico de Células Madre de la Universidad de Helsinki (Finlandia).
El trabajo se ha basado en el análisis de polimorfismos de un solo nucleótido (SNP) en varias líneas de células madre que ha comparado con los de células iPS humanas, tanto en una fase inicial como en un estadio intermedio de diferenciación. Las células inducidas presentaban más anomalías genéticas que las células somáticas de las que procedían y que las pluripotenciales embrionarias.
Con todo, bastó con dejar crecer en cultivo las células iPS durante unas semanas para seleccionar las líneas celulares que presentaban más mutaciones, y la mayoría de las anomalías genéticas se pudieron eliminar. “No obstante, algunas de esas mutaciones resultan beneficiosas para estas células y pueden sobrevivir mientras continúan en cultivo”, explica Otonkoski.
“Nuestro estudio apunta la necesidad de caracterizar rigurosamente las líneas de iPS generadas, en especial ahora que se está trabajando en potenciar la eficacia de la técnica reprogramadora”, asegura Samuel Hussein, primer autor del estudio. “Aumentar esa eficacia puede reducir la calidad de las células a largo plazo si no se asegura bien su integridad genómica”. (Nature 2011; 471: 58- 62).
Huellas genómicas permanentes
La posibilidad de que las células iPS presenten mutaciones genéticas debería tenerse en cuenta para establecer controles rutinarios en caso de que se empleen en la práctica clínica. Es la recomendación que se extrae de un trabajo llevado a cabo por varios de los grupos más prolíficos en la investigación sobre células pluripotenciales, entre ellos el del profesor de la Universidad de Wisconsin-Madison y descubridor de las células madre, James Thomson, y el equipo de Juan Carlos Izpisúa, del Instituto Salk (California) y del Centro de Medicina Regenerativa de Barcelona (CMRB).
El estudio se ha centrado en 22 líneas de células iPS humanas, obtenidas de siete de esos grupos de investigación, que emplearon diferentes medios de reprogramación en células de piel humana. En todas las líneas se hallaron mutaciones en genes codificadores de proteínas: unas seis mutaciones por exoma.
“Cada una de las células madre que observamos tenía mutaciones; esperábamos ver diez veces menos mutaciones de las que encontramos”, reconoce Kun Zhang, del Instituto de Medicina Genómica en la Universidad de California en San Diego, uno de los autores del trabajo.
Este trabajo ayuda a responder a la pregunta de si la reprogramación de células adultas humanas en iPS afecta a todo el genoma en el nivel esencial de un solo nucleótido. La respuesta es que lo hace. Zhang llama a esas mutaciones “cicatrices genómicas permanentes”.
Aunque algunas de estas mutaciones parecen ser silentes, la mayoría afectan a funciones específicas de las proteínas, incluidas las reguladas por genes que se asocian con efectos oncogénicos.
Este trabajo complementa a otro, publicado recientemente en Cell Stem Cell, en el que se advertía sobre la elevada presencia de aberraciones genéticas de las células iPS. Con el nuevo estudio se apunta a un daño genético concreto e importante: alteraciones en los nucleótidos únicos o en los pares de bases, que afectan a proteínas clave.
(Nature 2011; 471: 63- 67).
Alteraciones claves en la metilación
Un tercer trabajo, que ya adelantó la edición digital de Nature, pone al descubierto la existencia de alteraciones epigenéticas en las iPS. El estudio, dirigido por Joseph Ecker, del Laboratorio de Análisis Genómico en el Instituto Salk (California), y en el que también participa el grupo de James Thomson, analiza los perfiles de metilación de ADN -una de las marcas epigenéticas mejor conocidas- en todo el genoma de células madre embrionarias humanas y de iPS y sus progenitoras somáticas.
La reprogramación induce una reconfiguración en la metilación de ADN a lo largo del genoma completo, devolviendo dominios parcialmente metilados a un estado metilado completo y reinstalando la llamada metilación no CG, entre otros fallos. Existen también diferencias significativas en la metilación entre las células iPS y sus parientes próximas, las células madre embrionarias. En concreto, los autores hablaron de que las células reprogramadas en iPS guardaban en su memoria el recuerdo del tejido de origen.
O como explicaba Ecker: “Si las iPS se obtienen a partir de células de tejido adiposo, podemos ver que recuerdan algunas marcas de metilación de cuando eran adipocitos”. Este rastro que permanece en las iPS no se apreció en las otras células estudiadas: embrionarias y las del tejido de origen de las diferenciadas. Esta serie de cambios genómicos podrían comprometer a corto plazo la viabilidad de la diferenciación de las iPS.
(Nature 2011; 471: 68- 73).
Más dudas de certezas
Juan Carlos Izpisúa, director del CMRB y del Laboratorio de Expresión Génica, en el Instituto Salk, destaca que estos hallazgos han sido posibles gracias al desarrollo de técnicas de secuenciación de alto rendimiento y al estudio de la metilación de ADN a lo largo del genoma completo. Por eso confía en que los avances tecnológicos permitan determinar si es factible obtener células iPS prácticamente libres de errores genéticos. Sobre éstos considera que todavía hay que conocer sus consecuencias concretas: por ejemplo, no se sabe si las mutaciones en los exomas resultan o no en un fenotipo oncogénico. “Tampoco se ha determinado qué nivel de anomalías genéticas se presentará en las diferentes poblaciones celulares diferenciadas, ni hasta qué punto influyen los diversos protocolos en su aparición; se desconoce si las mutaciones generadas acortan la vida de las iPS, o si son indiferentes para la función celular una vez han madurado o se han diferenciado”. Más dudas despiertan las alteraciones epigenéticas. En definitiva, Izpisúa opina que es pronto para echar por tierra una técnica tan prometedora como desconocida.
Fuente: www.diariomedico.com
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