Final Report Summary - RSM-MICROSPHERE-IPSC (Novel Strategies for Microsphere-Mediated Cellular Control - A Technology to generate induced pluripotent stem cells for Regenerative Medicine)
Regenerative medicine is an emerging interdisciplinary field that aims to replace or regenerate cells from tissues or organs that are structurally and functionally damaged. In the field of regenerative medicine, one of its ultimate goals is to generate specific tissues from pluripotent stem cells directly derived from a patient's adult tissue. The generation of patient-specific induced pluripotent stem (iPS) cells will have a significant impact on the study of human diseases and on the treatment of these diseases using “personalized” regenerative medicine. These cells have a potential therapeutic use as they offer an alternative to the controversial use of embryonic stem cells and, additionally, they avoid the issue of immune rejection. In 2007 Dr Yamanaka's team (2012 Nobel Prize in Medicine) had successfully reprogrammed mature, specialised cells (human adult skin cells) by introducing the appropriate genes - just 4 genes are needed - into immature cells (pluripotent stem cells), which are potentially capable of becoming any other human cell type including heart muscle cells, bone cells and neurons. To date, this reprogrammation has been achieved using virus-based or transfection methods to introduce these genes. This is problematic as it increases the risk that the viral or vectors used to express those genes in cells insert in genomic places that may trigger cancer or other problems (by the activation of oncogenes or desactivation of tumour suppressor genes) Other methods of transfection are currently in use such as liposomal and non liposomal agents but they present low efficiency and do not fully solve the problem of foreign DNA insertion. Consequently there is a need for a safe and efficient method for the production of pluripotent cells from human adult cells. In this context, this project bring us one step closer to obtain a safe and efficient method for the production of pluripotent cells from human adult cells by introducing the appropriate genes or proteins into adult cells. In this project, we have synthesized successfully microspheres and optimised specific strategies of conjugation of the bioactive cargo (DNA and proteins) to microspheres and evaluated them using a fluorescence-based in vitro model. We have optimized conditions to be able to transfect efficiently fibroblasts (our reprogrammmation model). Next step has been focused in the implementation of this technology to generate iPS cells using combination of reprogramming genes and human fibroblasts as an in vitro model. We have efficiently conjugated to the microspheres the Yamanaka´s transcription factors needed to generate iPS cells and efficiently delivery then into human fibroblasts.
The generation of iPS cells has a tremendous impact in the potential development of cell therapies replacement strategies for a wide range of human diseases. So far, with available transfection methods, the efficiency of reprogramming hasn´t been higher than 1 %, consequently, the development of a novel transfection agent that achieves higher efficiency of reprogramming is of big impact. The generation of patient-specific iPS cells will have a significant impact on the study of human diseases and on the treatment of these diseases using “personalized” regenerative medicine. For instance, skin cells can be obtained from patients with a particular disease, reprogrammed, and examined in the laboratory to determine how they differ from cells of healthy individuals. Such cells constitute invaluable tools for understanding disease mechanisms and so provide new opportunities to develop medical therapies. As a representative example of the huge number of important applications of iPS cells, the long waiting list of patients needing a organ transplant could be dramatically reduced by building the desired tissue from iPS cells obtained from the patient´s cells and, even more important, avoiding the huge problem of organ rejection by the patient. Moreover, iPS cells are also a great hope for drug development.
In general the development of a technology for the efficient delivery of DNA and protein is of huge interest for a huge scientific community. Many research groups of international recognized prestigious based their foundings in the use of an efficient and safe method of transfection to be able to express or silence specific genes involucrated in crucial cellular pathways related to severe diseases that our society is suffering. Additionally, research with iPS cells is one of the latest breakthroughs in biology and medicine. This research project has offered a new advance and step change in current cellular delivery techniques and as such offers significant benefit to European excellence and competitiveness. The benefit of carrying out this research is that it will allow the EU to gain a major foothold in the area of DNA and protein cellular delivery, an area which is undoubtedly now being dominated by the USA. Such technology is crucial to underpin new advances in the areas such as cellular re-programming, protein functional analysis and systems and synthetic biology.
The generation of iPS cells has a tremendous impact in the potential development of cell therapies replacement strategies for a wide range of human diseases. So far, with available transfection methods, the efficiency of reprogramming hasn´t been higher than 1 %, consequently, the development of a novel transfection agent that achieves higher efficiency of reprogramming is of big impact. The generation of patient-specific iPS cells will have a significant impact on the study of human diseases and on the treatment of these diseases using “personalized” regenerative medicine. For instance, skin cells can be obtained from patients with a particular disease, reprogrammed, and examined in the laboratory to determine how they differ from cells of healthy individuals. Such cells constitute invaluable tools for understanding disease mechanisms and so provide new opportunities to develop medical therapies. As a representative example of the huge number of important applications of iPS cells, the long waiting list of patients needing a organ transplant could be dramatically reduced by building the desired tissue from iPS cells obtained from the patient´s cells and, even more important, avoiding the huge problem of organ rejection by the patient. Moreover, iPS cells are also a great hope for drug development.
In general the development of a technology for the efficient delivery of DNA and protein is of huge interest for a huge scientific community. Many research groups of international recognized prestigious based their foundings in the use of an efficient and safe method of transfection to be able to express or silence specific genes involucrated in crucial cellular pathways related to severe diseases that our society is suffering. Additionally, research with iPS cells is one of the latest breakthroughs in biology and medicine. This research project has offered a new advance and step change in current cellular delivery techniques and as such offers significant benefit to European excellence and competitiveness. The benefit of carrying out this research is that it will allow the EU to gain a major foothold in the area of DNA and protein cellular delivery, an area which is undoubtedly now being dominated by the USA. Such technology is crucial to underpin new advances in the areas such as cellular re-programming, protein functional analysis and systems and synthetic biology.