Descrubrimientos sobre reprogramación nuclear merecen el premio Nobel en 2012 / Discoveries on nuclear reprogramming deserves the Nobel prize in 2012

Este artículo fue escrito para honrar a J.B Gurdon y S. Yamanaka, laureados con el Premio Nobel en Fisiología p Medicina 2012 "por el descubrimiento de que las células maduras pueden ser reprogramadas para volverse pluripotentes". Se presentan en forma concisa sus aportes científicos y reseñas biográficas. J.B. Gardon, en Inglaterra, demostró hace 50 años en anfibios que al trasplantar el núcleo de una célula intestinal a un huevo u ovocito enuncleado se obtiene una célula totipotente que se convierte en un embrión y se desarrolla hasta convertirse en una rana adulta, lo cual implica la conservación de genoma en el proceso de diferenciación y la resersibilidad de dicho proceso. Estos descubrimientos llevaron a que otros autores realizaran la clonación de mamiferos utilizando el núcleo de células somáticas y la obtención de células madre pluripotentes a partir de los embrines que se producen in vitro por el desarrollo de las células totipotentes. Se mencionan varias aplicaciones y las contribuciones de Gurdon para comprender el proceso de reprogramación. S. Yamanaka, en Japón, hace seis años, reprogramó al estado embrionario fibroblastos cutáneos de ratones y humanos adultos insertando mediante vectores retrovirales una combinación de los genes de cuatro factores de transcripción: Oct3/4, Sox2, Klf4 y c-Myc. Las células reprogramadas fueron denominadas células madre pluripotentes inducidas. Utilizando la técnica desarrollada por Yamanaka y otras surgidas a raiz de sus descrubrimientos, miles de personas obtienen ahora células madre pluripotentes inducidas a partir de muchas especies y tejidos, incluyendo seres humanos sanos y enfermos. Las células madre pluripotentes o sus derivadas tienen un amplio potencial de aplicación, entre ellas, estudios de embriología y fisiopatología, modelos de enfermedades, descubrimiento de drogas y terapias celulares

This paper was written to honor J.B Gurdon y S. Yamanaka, 2012 Nobel Prize laureates in Physiology or Medicine for "the discovery that mature cells can be reprogrammed to become pluripotent". Their main scientific contributions and biography are presented in a concise manner. JB Gurdon, in England, showed fifty years ago in amphibians that the transplantation of the nucleus of an intestinal cell to an enucleated egg or oocyte produces a totipotent cell that develops into an embryo and adult frog. This implies that cellular differentiation is reversible and the genome is conserved in that process. The discoveries led to the cloning of mammals by other authors using the nucleus of somatic cells and to obtain pluripotent stem cells in vitro from the embryos produced by development of the totipotent cells. Some applications are considered. Gurdon's contribution to the understanding of the reprogramming process is mentioned. S. Yamanaka six years ago in Japan reprogrammed skin fibroblastis from adult mice and humans to the embryonic state by introducing via retroviral vectors a combination of the genes of 4 transcription factors, Oct3/4. Sox2, Klf4 and c-Myc. The reprogammed cells were named induced pluripontent stem cells. Throusands of people are now producing induced pluripotent stem cells from many tissues and species, including healthy and ill humans, using Yamanaka's methods and other techniques stimulated by his work. Pluripotent stem cells or their derivatives have great potential for a wide range of applications including research in embryology and pathophysiology, disease modeling, drug discovery and cell transplantation therapies

Wnt-dependent assembly of supermolecular Dishevelled-3-based complexes.

Dishevelled-3 (Dvl3) is a multivalent scaffold protein that is essential to Wnt signaling during development. Although Dvl-based punctae have been visualized by fluorescence microscopy; the physical nature and dynamic character of the such complexes are enigmatic. We use steric-exclusion chromatography, affinity pull-downs, proteomics and fluorescence correlation microscopy to characterize supermolecular Dvl3-based complexes of totipotent mouse F9 cells. The molecular mass of the complexes ranges from that of homodimeric Dvl3 to well-defined peaks harboring supermolecular complexes of 0.4 to 2.0 MDa. Addition of Wnt3a stimulates the formation of Dvl3-based complexes of greater molecular mass within 30 minutes. The presence of DKK1 and knockdown of Dishevelled proteins block formation of the 2 MDa Dvl3-based complexes and also block Wnt3a stimulation of the canonical pathway. Fluorescent correlation microscopy identified supermolecular Dvl3-based complexes with a molecular mass >30 MDa in live cells; these complexes were provoked to form structures with even greater molecular mass by Wnt3a. We establish for the first time the physical and functional nature of very large, supermolecular Dvl3-based complexes.

Totipotent stem cells bearing del(20q) maintain multipotential differentiation in Shwachman Diamond syndrome.

SBDS/7q11 gene mutations underlie the congenital Shwachman Diamond syndrome (SDS), characterized by bone marrow failure and high risk of haematological malignancies. In two cases of SDS with bone marrow failure and isolated del(20q) interphase fluorescence in situ hybridization (I-FISH) found no abnormalities in FHIT/3p14.2, IKZF1/7p13, D7S486/7q31, PTEN/10q23.3, WT1/11p13, ATM/11q23, D13S25/13q14, TP53/17p13, NF1/17q11, SMAD2/18q21, RUNX1/21q22. Fluorescence immunophenotype combined with I-FISH found del(20q) in a totipotent haematopoietic stem cell (CD34(+), CD133(+)) and downstream myelocyte (CD33(+), CD14(+), CD13(+)), erythrocyte (Glycophorin A(+)) and lymphocyte lineages (CD19(+), CD20(+), CD3(+), CD7(+)). These findings and clinical follow-ups confirm the benign course of SDS with isolated del(20q).

Primitive stem cells residing in the skeletal muscle of adult pigs are mobilized into the peripheral blood after trauma.

This study was designed to determine if trauma causes the release of adult-derived blastomere-like stem cells (BLSCs) from skeletal muscle into the circulating blood of adult pigs. Experimental procedures followed the guidelines of Fort Valley State University's Institutional Animal Care and Utilization Committee. Pigs were traumatized by splenectomy followed by pancreatectomy. Blood samples and skeletal muscle biopsies were taken before and after trauma. Adult-derived BLSCs were isolated from skeletal muscle and blood samples following established procedures. Nontraumatized skeletal muscle contained approximately 277 million BLSCs per gram of muscle. After trauma, skeletal muscle contained approximately 2 million BLSCs per gram of muscle. Blood taken before trauma contained approximately 22 million BLSCs per milliliter, whereas approximately 512 million BLSCs per milliliter were present within the blood after trauma. Blood values were statistically significant with a P < 0.05. This report is the first demonstration that trauma causes the release of adult-derived BLSCs from skeletal muscle into blood. Further studies are required to elucidate the roles that adult-derived BLSCs play in the response to injury and in the healing process. Surgeons must take a role in this evolving field.

Germ cell lineage differentiation in non-seminomatous germ cell tumours.

Human germ cell tumours (GCTs) have long fascinated investigators for a number of reasons. Being pluripotential tumours, they can differentiate into both extra-embryonic and embryonic (somatic) tissues. However, it has never been shown convincingly that, in humans, these tumours are truly totipotent and can also give rise to the germ lineage, the third major differentiation lineage occurring early during embryonic life. Using a number of newly available, distinct, immunohistochemical markers, such as OCT3/4, VASA and TSPY, the occurrence of germ cells was investigated in a number of germ cell tumours. Development of germ cells was identified in three independent non-seminomas, including two pure yolk sac tumours and one mixed tumour composed of yolk sac tumour and immature teratoma. Our finding indicates a previously unknown totipotent potential of human GCTs and raises the question of whether, under certain culture conditions, primordial germ cells could be derived from human GCT cell lines.

Paneth cell carcinoma of the ampulla of Vater.

We describe a Paneth cell carcinoma arising within the ampulla of Vater in a 64-year-old man. The phenotype of virtually all neoplastic cells was consistent with that of Paneth cells, based on routine morphology and their strong positive immunostaining for lysozyme. Additional widespread positive immunostaining for carcinoembryonic antigen and CA 19.9 supports a totipotential cell as the origin of such neoplastic cells. This case, therefore, represents a true Paneth cell carcinoma, as opposed to inclusion of occasional neoplastic Paneth cells into a poorly differentiated adenocarcinoma. This pattern of differentiation is rare, and predictions regarding its ultimate biological behavior and malignant potential must be guarded.