Conquistando Escalones will inject €27,000 into the University of Valencia to advance research

From the Conquistando Escalones Association we continue working together with the Translational Genomics Laboratory of the University of Valencia, directed by Rubén Artero, in order to find treatments for Limb-Girdle Muscular Dystrophy 1F/D2 (LGMDD2).

As we mentioned in this note, Águeda Blázquez’s years of work, which we paid for from the association, in addition to fruits, left a great legacy. And the predoctoral fellow Alicia Novella Estellés worked side by side with her and the rest of the team for a long time and continued thanks to the funds already contributed so that the research process was not paused. Now, so that it can continue during 2024, we have agreed to make a payment of 27,000 euros, of which we have already deposited 16,000 and the rest will be transferred between the months of June and July.

On the other hand, thanks to the granting of funds from the Ministry of Science that we tell you about here, a postdoctoral fellow was able to join the laboratory months ago. Specifically, Dr. Javier Poyatos, who had been working on our research for years.

In addition to communicating the issues of contracting and payment to, as always, be transparent in transferring where everything that is collected is destined, we inform you in detail how the research in this laboratory is progressing and the future objectives.

Murine (mouse) model of LGMDD2

Regarding the murine model of LGMDD2, until now the clinical signs that characterize the patients had not been observed in mice. Recently, signs of muscle alterations have been observed at a histological level (tissues analyzed by microscope), but, given the importance of a mouse model of the disease, the laboratory continues to search for more phenotypes (visible symptoms of the disease) that allow these animals to be used for pre-clinical studies. On the one hand, they are generating mice that have the mutation in homozygosity (both copies of the mutation) to check if the presence of a wild allele is the cause of the absence of phenotype. On the other hand, they had initially focused on studying mice that were older, however, it is possible that the effects of the mutated protein manifest in younger individuals and that these are masked by the functions of other proteins over time. Therefore, a timeline is being established, collecting samples from mice from a very young age to older ages in order to study the expression pattern of the mutated transportin 3 (TNPO3) protein and its effects on the mouse phenotype. Thus, once the optimal age and genotype (genetic information) at which to study the mice are established, different therapeutic approaches can be tested on them, such as, for example, the candidate compound for drug repositioning.

In addition, high-throughput approaches will also be carried out, such as the study of transcriptomics (study of the RNA of cells) of mice, so that we can compare it with the cellular model of the disease (which was established from samples from a patient with the disease) and, thus, check if the protein expression pattern is similar. Finally, since alterations in cytokines have been described in the blood of patients, their levels will be evaluated in the mice.

Study of pathogenesis mechanisms (how the disease originates and develops)

A key aspect to be able to develop therapies against a disease is to understand its pathogenesis. Therefore, it is important not to neglect the study of the pathogenic mechanisms that lead to the appearance of the characteristic phenotypes of the pathology. For this reason, a series of plasmid DNA constructs (molecules found in bacteria and other microscopic organisms) containing transportin 3, wild or mutant, labeled with different fluorescent labels, have been developed in the laboratory. The next step is to introduce these constructions into the cellular model of the disease to be able to observe where these proteins are located subcellularly and how they interact with each other. This information will allow us to better understand what is happening molecularly in the disease and, therefore, search for therapies that try to rescue these alterations.

Development of therapies for LGMDD2

Regarding the search for possible therapies for the disease, the approach that has been developed in the laboratory in recent years, as has been reported since its inception, consists of the repositioning of drugs. This is because this is the way in which a possible treatment could reach patients most immediately. At the same time, the development of possible therapies based on oligonucleotides (short DNA and RNA sequences) has been launched, which would act specifically against the patients’ mutation. Thus, several different designs have been generated that are currently being tested in the cellular model of the disease. The constructions mentioned in the previous section will be of great help to understand the mechanisms of action of both the candidate drug to be repositioned and the new approaches.

In summary, from the Translational Genomics Laboratory of the University of Valencia the short-term objectives consist of continuing with the search for the phenotype of the murine model of the disease, studying the subcellular localization of TNPO3 in the cellular model through the use of constructions of Plasmid DNA containing the protein, wild or mutated, labeled with fluorescent labels and testing different oligonucleotide-based therapies in the cellular model.

And in addition to the laboratory level, work is being done at the bureaucratic level to advance the procedures to be able to carry out the first clinical trial in humans of the most promising candidate drug for a possible treatment of LGMDD2. As we already told you, this would have an approximate cost of €100,000 that would have to be obtained, added to all the costs that the association already has to bear, so we cannot stop continuing to raise funds and ask for your help.