The team led by Prof. Cenacchi, from the Department of Biomedical and Neuromotor Sciences at the University of Bologna, is one of the research groups seeking to achieve a treatment and / or cure for our Limb-girdle Muscular Dystrophy 1F.
We have just renewed the agreement by making a payment of € 6,000, which will cover the coming months of a job that is vital that it does not stop.
From the beginning, its main objective has been to clarify the role of Transportin 3 (TNPO3) (our mutated protein) in skeletal muscle both under normal conditions and in the case of our Limb-girdle Muscular Dystrophy 1F. The role of TNPO3 in muscle is unknown and several hypotheses have been made in recent years as to how the TNPO3 mutation causes LGMD1F / LGMDD2, but currently there is still no certainty as to how the disease is generated.
The main objective of this project is to create two study models of LGMD1F: one in vitro with cell cultures and a second in vivo with an animal model of zebrafish. In this way we will have clearer TNPO3 activity in the skeletal muscle that will help us understand how the defect in Transportin 3 originates LGMD1F. Deepening knowledge of the underlying disorders of LGMD 1F / LGMDD2 pathogenesis will be a crucial step for future studies and the development of therapeutic perspectives for LGMD1F / LGMDD2 patients who are still absent.
Specifically, with our new financing for the project called “Morphofunctional Characterization of Transportin3 (TNPO3) in the pathogenesis of LGMD1F / LGMDD2”, Prof. Cenacchi tells us that:
“In this next phase of our research project, which has also received a grant from AFM-Telethon, we are working on generating an animal model, using Danio Rerio or zebrafish, and a cell model capable of mimic the disease to clarify the functional role of TNPO3 in the muscle pathogenesis of LGMDD2 (formerly known as LGMD1F **). Deepening knowledge will also help develop specific therapeutic strategies that are lacking so far.
We have cloned the sequence corresponding to the cDNA of the human mutated gene product into a suitable expression vector. Subsequently, the mRNA sequence obtained in vitro will be microinjected into zebrafish embryos and we will study the stages of muscle differentiation. Research on the cell model will be carried out on the murine C2C12 cell line and on human satellite cells. Cells will be transfected with mRNA molecules corresponding to the mutated TNPO3 cDNA.
Another objective of the current project is to develop a stable silencing of the gene responsible for LGMDD2 through the CRISPR-CAS9 system. The C2C12 cell line and the control lines (HeLa) have been genetically engineered using this technique to obtain gene knockdown, which will be evaluated, after the selection of the cell clones obtained, with genetic and molecular analysis.
Achieving both objectives of the project will allow us to obtain valid study models of this disease also for an evaluation of the possible participation of other proteins in the TNPO3 pathway and, therefore, in the pathogenic mechanism of specific muscle disorders of LGMDD2. ”
As always, we emphasize that the times of science are not as fast as we all would like. But they have some necessary steps so that the work done is not in vain and that it can be applied to humans with confidence in the results and without danger to them.
The payment of the aforementioned amount serves to cover, mainly, the salary of the coming months of a biotechnologist, who is in charge of working exclusively on our disease. We want to thank Prof. Cenacchi and her entire group for all their work and dedication inside and outside the laboratory. As well as his personal involvement beyond professional and all his efforts towards research, even in times of crisis that we are experiencing because of the global pandemic caused by COVID-19. Thanks also to all the Conquistando Escalones partners, donors and collaborators for helping us raise funds and pay for research. In these times when, among other things, it will be difficult to carry out events in the short term, it is more important than ever.
** With all the Limb-girdle Muscular Dystrophies (LGMD) discovered in recent years, the letters to be assigned have run out (something like the change in the numbering of car license plates) and some have changed their nomenclature, although the disease obviously it is the same. Therefore, ours, LGMD1F, in the new nomenclature is LGMDD2.