ThalLab: past, present and future
(Release 1, September, 2009)Roberto Gambari
Full Professor of Biochemistry (BIO/10)
Ferrara University,
Director of ThalLab
The aim of this commentary article is to present the results obtained by the ThalLab during the past few years; recently several research project have been completed and relative publications submitted and accepted by important international journals (Blood, J Biol Chem, J Med Chem, Eur J Hematol, Am J Hematol, Apoptosis). Several milestones have been reached and novel fields of investigation have been added to the main topics of ThalLab. Some of these novel research areas should be in the future object of great efforts by the researchers at ThalLab.
At present, ThalLab is constituted by a scientific group composed by more than 12 researchers (mainly PhD students and research fellows). The most important external collaborators of ThalLab are Prof. Stefano Rivella (Cornell University, New York) and Prof. Eitan Fibach (Hadassah Hospital, Jerusalem, Israel). The scientific activity of at ThalLab is funded, at it will be described, by Telethon, by Foundation Cassa di Risparmio di Padova e Rovigo and by Veneta Association for the Fight against Thalassemia, AVLT.
THE ThalLab AND YOUNG INVESTIGATORS
First of all, I would like to underline the contribution of our young investigators (PhD fellows, PhD students, research fellows), which was extremely important to reach the objectives of ThalLab. The researchers who worked at ThalLab (*, still present and active) have been: Dr. Giulia Breveglieri (*,ER-GenTech fellow), Dr. Francesca Salvatori (*, research fellow), Dr. Monica Borgatti (*, FIRB fellow), Dr. Laura Breda, Dr. Sara Gardenghi, Dr. Alessia Finotti (*, research fellow supported by FIRB), Dr. Irene Mancini (*, PhD student), Dr. Ilaria Lampronti (*, AVLT, fellow supported by Ferrara Ricerche), Dr. Elisabetta Lambertini (*, research fellow), Dr. Letizia Penolazzi (*, technician, UniFE), Dr. Enrica Fabbri (*, research fellow), Dr. Laura Piccagli (*, AVLT, research fellow), Dr. Eleonora Brognara (*, PhD student), Dr. Cristina Zuccato (*, research fellow), Dr. Chiara Multineddu (*, fellow supported by Regione Sardegna), Dr. Mahmud Tareq Hassan Khan, Dr. Antoine Saab, Dr. Teresa Gagliano, Dr. Marina Tommasetti, Dr. Cristina Rutigliano, Dr. Cristiano Chiarabelli.
In these years the ThalLab was able to help several young investigators to improve their scientific background and technical skills; these researchers of course should be considered as a high value and unique resource for the ThalLab. Unfortunately the budget restrictions and the difficulties to program for all of them a scientific and academic career should introduce a debate and the need of a clear strategy which will maintain a critical number of researchers working at the ThalLab. This will be one of the major issue in the next future.
TECHNOLOGY TRANSFER: THE PATENTS.
We have submitted the following patents, most of which concern the induction of fetal hemoglobin as experimental strategy for the treatment of thalassemia: (a) patent PCT/EP01/02804, March 2001: "Synthetic oligonucleotides as inducers of erythroid differentiation", (b) Patent n. T02001A000633, July 2 2001: "Novel use of heterocycle and bezoheterocycle polyamides structurally correlated to the natural antibiotic distamycin A"; (c) Patent PCT/IB02/02628, July 1 2002: "Use of heterocyclic and benzoheterocyclic polyamidines structurally related to the natural antibiotic distamycin A for the treatment of beta-thalassemia"; (d) Patent PCT/IB03/02632 July 3 2003: "A novel use of rapamycin and structural analogues thereof"; (e) Patent PCT/IB03/03462 July 30 2003: "A novel use of angelicin and structural analogues thereof"; (f) Patent n.TO2004A000099 "Synthetic double stranded oligonucleotides for the treatment of osteopenic diseases", February 30 2004; (g) Patent n. VE2005A000054, 4-11-2005: "Drug for the treatment of thalassemia, sickle cell disease and related pathologies, method for drug activation, pharmaceutical composition with the drug as the active molecule and photochemotherapic approach using the drug".
TECHNOLOGY TRANSFER: THE ThalLab AND THE BIOTECH INDUSTRY.
The interactions with the industrial biotech companies are among the most important issues of ThalLab. In order to facilitate the interactions with small and medium enterprises, we have produced a DOSSIER incorporating all the information on the projects and the results of the ThalLab, which we consider more important and suitable for biomedical applications. This document, in collaboration with Dr. Michele Lipucci, Responsible of External Relations of ThalLab, is the basis for the activation of novel collaborative efforts and, possibly, for the consideration of possible clinical trials.
PARTICIPATION TO NETWORKS: THE ITHANET PROJECT.
Shortcomings in cross-border communication, collaborative development of treatments and research projects, dissemination to a broad base of clinicians and researchers, and education of members of the public across European and partner countries did not go unnoticed, and prompted the conception of Ithanet (Electronic Infrastructure for Thalassaemia Research Network), co-funded by the Information Society Technology priority of the European-Commission (EC) Framework Program 6. Ithanet as a network of researchers, clinicians, educators, and patient organizations has addressed many of the issues impairing the transfer of skills, expertise, and databases both, between EU members, and between the EU and its partner countries. Ithanet comprises 26 organizations from 16 countries, which function as 11 medical, 16 diagnostic, and 16 research centers, five institutions of scientific education, three bodies in patient relations, and four developers of electronic infrastructures.
The ITHANET Project (eInfrastructure for Thalassemia Research Network) has been carried out with the participation of several european partners, among which the Cyprus Institute of Neurology and Genetics (CING), the CESNET, the Thalassaemia International Federation (TIF), the European Genetics Foundation (EGF), the Oxford Radcliffe Hospitals NHS Trust ORH NHS Trust, ST Barts and the Royal London Hospital, the University of Cagliari (Thal Unit Cagliari), the University of Malta (UoM), the Asclepion Genetics, the National e Kapodistrian University of Athens (NKUA), the Hadassah Medical Organization, Hadassah (Israel), the University of Ferrara (ThalLab, UniFE), the Ministry of Health, Makarios III Hospital (NAMIII), the Laikon General Hospital (KMA-LAIKON), the University of Cairo (Egypt), the Chronic Care Center (CCC), the Suleiman Demirel University (SDU), the Erasmus University Medical Center (Erasmus MC), the Leiden University Medical Center (Leiden University), the AHEPA University Hospital (AHEPA Univ. Hospital), the University of Bucharest (IGUB), the Bogazici University Bebek (TURKEY B.U.), the Research Laboratory of Molecular Biology, l’EGE University (EU), the Hopital Henri Mondor (HHM), the Hospital Clinico San Carlos (HCSC).
Of central importance to the workings of the network are the seven main activity themes of Ithanet, specifically Electronic Infrastructure (CESNET), Clinical and Molecular Research Tools (TIF and ORH Trust, respectively), Training and Knowledge Transfer (EGF), development of the Ithanet Portal, Dissemination, and Coordination and Administration (CING).
The ThalLab has been involved in the Workpackage il WP4, concerning molecular studies on thalassemias. This WP had as the major object the development of methodologies and shared protocols for the study of thalassemia. Data-banks have been developed on results, methods (including analytical methods), advances studies, personnel involved in specific fields of research on thalassemia. We think that this activity will facilitate the constitutions of one or more groups for the preparation and submission of research proposal within the European Union FP7.
We have developed a Ithanet WEB site on thalassemia and other hemoglobinopathies. ThalLab is working on the realization of a WEB site (HbF-NET) on the induction of fetal hemoglobin as a clinical option to cure thalassemia.
An important role in the worldwide disseminations of the activities of ITHANET has been played by TIF (the International Federation on Thalassemia), which is sustaining the activity of 59 countries with high frequency of hereditary hemoglobinopathies (for instance thalassemia, sickle cell anemia, HbE). The dissemination of the results is an activity of great interest, considering the worldwide distribution of thalassemia. Thalassaemia is among the major health problems in developing countries, where affected patients and healthy carriers are numerous, mainly due to the absence of genetic counseling and prenatal diagnosis. Several developing countries are unable to efficiently sustain the high-cost clinical management of β-thalassaemia patients requiring regular transfusion regimen, chelating therapy and advanced hospital facilities. It is well known that, in addition of “direct costs”, blood transfusions requires accurate monitoring of the blood safety, by using expensive technologies, some of which are based on multiple PCR covering all the possible hematological infectious diseases. As far as alternative therapeutic approaches are concerned, gene therapy and bone-marrow transplantation are very promising strategies, but they are expected to be useful only for a minority of patients, selected on the basis of biological/genetic parameters and the economic possibility to afford these therapies.
PARTICIPATION TO NETWORKS: THE ITALY-EGYPT PROJECT.
The Project “FETAL HEMOGLOBIN INDUCERS FOR THE TREATMENT OF BETA-THALASSEMIA” has been recently approved and is ongoing since November 2008 within the Programma Esecutivo della Cooperazione Scientifica e Tecnologica between Italy and Egypt (2008-2010). The Project is based on a strict collaboration between Prof. Roberto Gambari (ThalLab, Ferrara University, Department of Biochemistry and Molecular Biology, Molecular Biology Section) and Prof. Amal El Beshlawy (Cairo University, El Monira University Hospital, Pediatric Hematology Dept., Cairo). The first objective is to reinforce the scientific collaboration between the two institutions, by establishing common experimental protocols and approaches. The second objective is to establish a comprehensive data bank of the undergoing therapeutic treatments of β-thalassaemia patients in Egypt, delivering a list of the HbF inducers employed. The third objective is to perform joint experiments using all the HbF inducers developed by the Italian partner and the HbF inducers used in therapy by the Egyptian partner with the main objective of the identification and validation of HbF inducers to be used for therapy of β-thalassemia.
FUND RAISING.
One of the major activity of ThalLab is to obtain additional research funds from external sponsor (both institutional and private organizations); this activity is very important for the following reasons: (a) allows the increase of the number of researchers participating to the ThalLab projects; (b) facilitates synergic interactions with other research groups; (c) allows to increase the funds, with a parallel increase of activated novel sub-projects and specific tasks. I would like to underline the most recent and important Projects, such as the already mentioned European Project ITHANET, the recent PRIN Project on the use of PNA useful for targeting micro RNAs and pathological RNAs (DESIGN, SYNTHESIS AND BIOMOLECOLAR CHARACTERIZATION OF PEPTIDE NUCLEIC ACIDS ANALOGUES WITH THERAPEUTIC AND DIAGNOSTIC PROPERTIES), the TELETHON Project “Modifiers of globin gene expression for therapy of beta−thalassemia” (GGP07257), the already mentioned Italy-Egypt Project, the Project “NEW FRONTIERS IN THE DEVELOPMENT OF DIAGNOSTIC AND THERAPEUTIC APPROACHES OF BETA-THALASSEMIA”, funded by Fondazione CARIPARO. The PRRIITT ER-GenTech Project has been included in the recent BioPharmaNet Project, approved by Regione Emilia-Romagna. The mentioned funds allowed ThalLab to implement the instrumentations available for the Projects and maintain the possibility to sustain an important number of fellowships and research positions.
I would like to mention that a very important role in the activity of Fund Raising ha been carried on by AVLT, with Dr. Elio Zago’s contribution.
THE NETWORK ThalLab-FONDAZIONE CARIPARO AND AVLT.
The Research Project “NEW FRONTIERS IN MOLECULAR DIAGNOSIS AND EXPERIMENTAL THERAPY OF BETA THALASSEMIA” is based on a long-term agreement of collaboration between the ThalLab, the Department of Biochemistry and Molecular Biology, Ferrara University, Fondazione Cassa di Risparmio di Padova e Rovigo (CARIPARO) and Veneta Association for the Fight against Thalassemia (AVLT, Rovigo, Italy).
This project, active since July 2005 will end in July 2010 and is sustained with 500.000 Euro funding by Fondazione CARIPARO. The general objective of the research is to develop novel experimental models and to identify novel molecules of possible use for the therapy of thalassemia. Moreover, the project aims to develop novel diagnostic methodologies, aimed at the non-invasive prenatal diagnosis.
The experimental model and the research method. The experimental model closest to patients is represented by erythroid precursor cells which are isolated from the peripheral blood of the patients through a very simple blood isolation. Just before blood transfusion, 20 ml of blood are taken from the patient and the researchers, following a special treatment, start the in vitro cell culture and after ten days the cells are treated with erythropoietin (a molecule able to specifically stimulate proliferation and differentiation of erythroid cells). These cells start therefore to produce high levels of hemoglobin and, if treated with inducers of fetal hemoglobin, it can be determined whether they start to produce, following this treatment, HbF.
The Network ThalLab-Fondazione CARIPARO-AVLT. The project funded by Fondazione CARIPARO has among the major scientific objectives, the characterization of novel inducers of erythroid differentiation, including the screening of medicinal plants (among which we underline rapamycin, mithramycin, resveratrol), the development of novel experimental models for the identification of inducers of fetal hemoglobin, the production of in vitro model systems mimicking the gene mutations causing the most frequent types of beta-thalassemia, the development of thalassemic mice, studies of transcriptomics and proteomics, development of PNA/DNA hybrids for decoy experiments finalized to the reactivation of the expression of gamma-globin genes, vehiculation and delivery of decoy molecules, development of Lab-on-a-chip platforms for the molecular diagnosis, induction of apoptosis in osteoclasts.
NOVEL ACTIVITIES AT ThalLab.
The objective of ThalLab in the next future is to maintain the commitment in the “historical” projects (induction of fetal hemoglobin, studies on gene expression, development of decoy molecules, gene therapy). At the same time, the ThalLab will start novel projects for which we have already obtained very promising preliminary results.
Development of transgenic mice carrying the most important mutations of the human β-globin gene. These animal models are expected to be very important for the validation of the therapeutic potential of drugs designed for specific mutations leading to β-thalassemia. We are proud of confirming that the first class of homozygous mice (for the IVS-1-6 mutations) has been recently obtained and are now under molecular characterization.
Gene Therapy. The ThalLab will consider the possibility of sustaining a research task aimed at demonstrating the pre-clinical efficiency of novel therapeutic lentivirus able to induce ex novo HbA production in cells isolated from β-thalassemia patients. These novel vectors have been developed by Dr. Stefano Rivella (Cornell University, New York).
Stem Cells. One of the expected major objectives of ThalLab is to develop adult stem cells, retrodifferentiated stem cells, and, at the same time, employ human embryonic stem cell lines (according to the legislation) with the aim to determine the activity to modulate their differentiation state with the very high numbers of HbF inducers we have recently reported (many of which are patented). The second objective is to compare the gene expression of human stem cells induced to differentiation with retrodifferentiated adult cells (iPS, induced pluripotent cells), with the aim to combine the research on pluripotent cells with molecular gene therapy approaches employing modified retrovirus, lentivirus and short DNA and RNA molecules, including analogues. I believe that this experimental research filed is one of the most important, able to contribute to more deep interpretation of gene therapy.
Cellular models and biosensors. We have developed cellular biosensors (some data sheet have been included in the BioPharmaNet web-site) useful for the screening of novel molecules of possible application in therapeutic trials. This research field, in addition to the scientific importance, might be useful for novel collaborative efforts with international research groups interested in using these cellular systems.
Non-invasive prenatal diagnosis. At present, no method does exist for non-invasive prenatal diagnosis. This issue is very important. This research filed will be considered in the future activities of ThalLab.
FINAL COMMENTS.
The coming months will be very important for the ThalLab’s future activity. The results of the research projects conducted in the past few years have produced a high number of scientific publications, of patents, of protocols, of proprietary cellular models and novel animal models for the study of thalassemia.
However, my impression is that it will be very difficult to sustain, in the future, all the activities that have been the feature of ThalLab and AVLT. The costs more relevant, that have been sustained in these years, are devoted to (a) personnel, (b) instrumentation, (c) reagents and disposable material and (d) patents.
The implementation of the activity at ThalLab, or its reduction, unfortunately depends not only by us and by the ThalLab team, but also by the commitment of external sponsors to sustain our research activity.
In all these years our research projects, our results, our fundings, our general activity have been made available at our WEB site talassemiaricerca.it. In all these years we tried very hard to maintain our activity efficient and productive in all the mentioned fields.
We have to realize that one of the possible necessity will be, if funding will not be available to us, a significant reduction of the objectives, at it will be decided following the strategy established by the Council of ThalLab.
Our hope is that this reduction of our activity will be on one hand temporary, on the other limited, in order to be able to continue along the road that will bring the basic and applied research to important therapeutic protocols finalized to cure thalassemia.
Professor Roberto Gambari
Department of Biochemistry and Molecular Biology,
Section of Molecular Biology,
Ferrara University,
Ferrara, Italy
Tel: 0532-424443;
Mobile: 348-5521942;
Fax: 0532-424500;
E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it
