HOMOL. RECOMB. HESC · Generation of human disease models by homologous recombination in human embryonic stem cells
FP6 — Marie Curie Actions (Human Resources and Mobility)
- Duration
- 2006-12-01 → 2008-11-30
- EU contribution
- €80,000
- Participants
- 1
- Scheme
- IRG
Lines connect the coordinator with its partners.
Results in brief
Final Activity Report Summary - HOMOL. RECOMB. HESC (Generation of human disease models by homologous recombination in human embryonic stem cells)
A new laboratory specific for stem cell culture has been set up in the research unit of the Hospital de Cruces. This cell culture facility is physically separated from the general tissue culture room where untested biopsy material is manipulated. The stem cell culture room is cleaned daily and disinfected once a week. To work in this "clean" room, lab coats and shoe covers are worn. Using this facility we have implemented the thawing, culture and propagation of human embryonic stem cells (hESCs), using mouse embryonic fibroblasts as feeders. The production of a stock of hESC vials at low passages has been achieved; these vials provide a sufficient source of hESC for this project. Optimisation of the differentiation into hESC-cardiomyocytes has been completed. And genetic manipulation has been successfully optimised and will be used to introduce a point mutation associated with dilated cardiomyopathy into the genomic sequence of the LMNA gene. These goals focus on the generation of an experimental model of a human disorder associated with mutations in the LMNA gene, specifically dilated cardiomiopathy. hESCs are regarded as the most promising tool for cellular/tissue repair because their indefinite replicative capacity makes them a renewable cell source and highly suitable for culturing in large quantities. Moreover the possibility of modelling human disease where the animal model has proven inadequate would open several areas of research to study etiologies and pathologies of these difficult disorders. hESCs are also an excellent tool for discovering and testing new drugs that could cure or delay the development of these diseases. We also intend to study the effects of protease inhibitor treatments similar to those given to HIV patients on stem cells and which alter the maturation process of LMNA gene. Preliminary results suggest that human stem cells subjected to these protease inhibitor treatments show alterations in proliferation rates, morphology, mortality and chromatin organisation. Given the similar effects of lypodystrophic LMNA mutations on these cell characteristics, these stem cell models could provide an important tool in analysing the effects of protease inhibitor treatments, as well as the effects of potential ameliorative drug treatments for HIV patients.
Data: CORDIS, © European Union
Project objective
Homologous recombination technique has been applied to murine embryonic stem cells improving significantly our knowledge on the functionality of several genes. Human disease models are very valuable tools for the study of the phenotype as well as potential therapy for genetic diseases.The derivation of mutant mice using the technique of homologous recombination has been essential in the generation of models for human diseases. Despite the success of homologous recombination in mouse embryonic stem cells to study many human pathologies, the morphological, developmental and genetic differences between human and mice pose significant limitations for the use of these mutant animals as models for human disorders.In many cases, the mouse phenotype differs from t hat observed in humans. Human embryonic stem cells are non-transformed cells with the capacities of self-renewal and pluripotency, allowing them to differentiate into a wide range of cell types. Via genetic manipulation, human ES cells can also be used to model human diseases to study the disorder's etiology and pathology.This is especially important where the in vivo animal models have failed to fully recapitulate the phenotype in the human diseases. We propose to generate a model of dilated cardiomyopathy using hES cells by homologous recombination. A single base change mutation will be introduced in the LMNA gene, which encodes the lamin A and lamin C proteins, components of the inner nuclear lamina.This mutation that results in the change of one aminoacid has been associated with a high percentage of sudden death (45%) in affected members of a family, even though all the affected individuals are heterozygous for the mutation. Defining how lamin mutations alter cardiac cell biology will ultimately increase our understanding of the pathophysiology of dilated cardiomyopathy.
Original text from CORDIS.
Participants
- FUNDACIóN VASCA DE INNOVACION E INVESTIGACION SANITARIAS · SONDIKA - BIZKAIACoordinatorSpain
Links
Data: CORDIS, © European Union