Why to create a Center for Cell Therapy?
We propose a program for basic and clinical research to isolate, culture, and characterize embryonic, somatic, and neoplastic stem cells in order to understand their biology and apply this knowledge to therapy. We will focus on questions that derive from both our past experience and the present bottlenecks in the field, moving to a more “applied” research in normal and abnormal stem cells, without neglecting our strong basis in cell biology. A great deal of effort in stem cell research has been lost in trying to bring therapeutic tests prematurely to the bedside, based on untested, unproved, or unclear premises. Thus, our research program is firmly grounded in the analysis of mechanisms and pathways of normal and abnormal stem cells to serve as targets or to help manipulate cells for therapy.
The research project comprises four lines: pluripotent stem cells, which include both embryonic and induced pluripotent stem cells (SC); somatic stem cells, such as hematopoietic SC, mesenchymal SC, endothelial SC, and cancer SC; general mechanisms involved in maintaining “stemness”, focusing on the epithelial-mesenchymal, mesenchymal-epithelial, and endothelial-mesenchymal transitions; and clinical stem cell applications. These are complex and inter-related issues that can only be approached by an inter-disciplinary team working together in long-term cooperation.
The first strong argument for the existence of this center is that it consolidates the close relationship and collaboration between basic and clinical researchers already established, fostering a research program that is truly “bench-to-bedside” and the reverse. There is a balanced proportion of clinicians, who attend in wards and outpatient clinics, working together in the same laboratories with cell and molecular biologists, biochemists, and geneticists; there are many examples of MSc and PhD theses of basic scientists who were supervised by clinicians, as well as papers authored jointly. The second relevant point of this project is that it brings together research in normal and abnormal stem cells. The research proposed for the bone marrow failure syndromes is an example of an approach largely used for normal stem cell research (generation of iPS cells) applied to the investigation of basic mechanisms of a group of diseases (aplastic anemia, Fanconi anemia, dyskeratosis congenita) in which the hematopoietic stem cell fails; at the same time, the question of the integrity and dynamics of telomeres, which are relevant in these diseases, as demonstrated recently by one of us, , is essentially involved in the maintenance of “stemness” and SC senescence and differentiation, as well as a key player in genomic instability and leukemogenesis, as one of us has recently demonstrated.
Another example of the need to link the study of normal and cancer stem cells, and basic and clinical research is the project proposed for acute promyelocytic leukemia (APL) as a disease model for cancer studies. Based on the knowledge derived from the identification of murine hematopoietic stem cells, we have isolated and characterized APL stem cells in the transgenic mouse model for APL that contains the hCG-PML/RARA fusion gene. We now propose to apply this knowledge to isolate and characterize stem cells of human APL samples by xenotransplants into NSG mice of candidate APL cell populations. The intensive basic research on APL biology and creation of the International Consortium on APL (IC-APL) led by our Center resulted in obvious benefit to patients; one-year overall survival for patients with APL in Brazil dramatically increased from below 50% to above 75% after the IC-APL was implemented. From the bedside back to the bench, the IC-APL enabled the collection of more than 400 APL samples and we now propose to use these samples to evaluate, for instance, changes in the p73 pathway as a modulator of disease severity and the molecular mechanisms responsible for the coagulopathy associated with APL.
Most of the research proposed by the Center can only be carried out in the long run. This can be illustrated by the use of mesenchymal SC for treatment or prophylaxis of immunologic diseases or reactions (graft-versus-host disease, type 1 diabetes mellitus); the results from bench work isolating and expanding mesenchymal SC for clinical use need to be carried out in a GMP Tissue Culture facility, then tested in a clinical setting, the material obtained from the patients need again to be reviewed in the laboratory, results guide both new experimental work and changes in the protocol to prepare the cells (in this case, for instance, mesenchymal SC priming with interferon to prevent or treat immunologic reactions), and finally going back to the clinical tests. This complete cycle takes years instead of months. Likewise, genome sequencing of neoplastic cells to identify relevant abnormalities, as for instance proposed here for myelodysplastic syndromes, requires selection of clinical samples, followed by sequencing and bioinformatics work, and again access to the patients and patient follow-up to extract meaningful results to correlate basic with clinical findings. This research cycle needs long lasting basic-clinical cooperation.
Also, the center is necessary to give long-term continuity to an in-depth approach to a particular issue, instead of selection of unrelated publications. Thus, the clinical applications of the present project reflect the past history of the Center, focused mainly on the basic biology of mesenchymal stem cells. On the other hand, the basic component of the present project has a much broader basis, and encompasses ES and iPS cells, tissue specific progenitors, and abnormal stem cells; only if this group of basic researchers and clinicians are kept together, with access to the facilities to separate and expand cells in GMP conditions and to clinical wards to test them in patients, the results of this basic research can be brought to the bedside.
One of the bottlenecks for the development of Brazil and the State of São Paulo is the education of human resources in quantity and quality. We have dedicated a lot of effort for these purposes in the past, and as a consequence we have organized a graduate program that begins to enroll its first students this fall. For the success of this graduate program, the core of this group will have to be kept together with access to laboratories and funding in order to reach our goal, which is to double the numbers of MSc, PhD, and postdoctoral fellows trained. The counterpart offered by USP includes two new tenure-track faculty positions and three high level technicians to the group, which will strengthen this educational component of project.
The technology transfer project also takes into account that the development of biotechnology and health-focused products and process usually demands long-term initiatives, which are compatible with the time frame proposed for the Center. The educational project is firmly based on the success of the previous decade; it depends both on a long-term commitment of a dedicated team, which takes time to be put together, and infrastructure compatible with the proposed program.
Finally, this kind of long-term project involves international and national collaboration, including the exchange of researchers, training of young researchers, organization of workshops and meetings, but especially by carrying on complementary tasks as part of a research project (the table below lists active collaborations with research groups from Brazil or abroad; see also http://ctcusp.org/?page_id=437 ). For instance, in the collaboration with the French group we are responsible for the DNA analysis of over 900 samples from patients collected in four European countries.
|Project||Foreign coordinator||Financial support|
|Overexpression of EMT genes in endothelial cells during endothelial mesenchymal transition (DT Covas)||Robert Weinberg, MIT, USA||FAPESP (Brazil, 10/51962-9) & MIT|
|Telomere dysfunction as a mechanism of human stem cell disease; induced pluripotent stem cells to model telomere regulation and cell therapy (RT Calado)||Neal Young, NIH, Bethesda, USA||NIH Center for Regenerative Medicine Pilot Projects for Clinical Applications (2011-2012)|
|The role of Dido locus in the differentiation of stem cells and in myeloid neoplasias (DT Covas and MA Zago)||Carlos Martínez, National Center of Biotechnology, Madrid, Spain||CNPq (Brazil, 560884/2010-9 ) & MICINN (Spain)|
|Prognostic association of genetic polymorphisms of drug metabolism and innate immune response on umbilical cord blood transplantation outcomes (MA Zago)||Eliane Gluckman, Eurocord, Hôpital Saint Louis, Paris|
|Generation of iPS cells from patients with Fanconi anemia belonging to different complementation groups, to study the DNA-repair mechanisms in the hematopoietic stem cells (RT Calado and DT Covas)||Ricardo Pasquini, UFPR, Curitiba, Brazil|
|Impact of RNA processing and regulation in human normal and cancer stem cell differentiation (WA Silva Jr)||Gene Yeo, Cellular & Molecular Medicine, UCSD, USA|
|Molecular Genetics of Prostate Cancer: Role of Stem Cells (WA Silva Jr)||Gerhard A Coetzee, Preventive Med, Norris Cancer Center, USC, USA|
|Generation of a hiPSC library of the Brazilian population (LV Pereira)||Jeanne Loring, Scripps Institute, La Jolla, California, USA||BNDES (Brazil) and FINEP (Brazil)|
|Introducing embryonic and somatic mitochondria on parthenogenesis development in vitro (FV Meirelles)||FAPESP (Brazil, 2010/13384-3) & INCT (Brazil)|
|iPS reprogramming prior to cloning (FV Meirelles)||Konrad Hochedlinger, Harvard University, Boston, MA||FAPESP (Brazil, 2011/08376-4)|