Title:Help Comes from Unexpected Places: How a Tiny Fairy and a Tropical Fish may help us Model Mucopolysaccharidoses
Volume: 24
Issue: 16
Author(s): Sofia Carvalho, Luciana Moreira, Juliana Inês Santos, Paulo Gaspar, Mariana Gonçalves, Liliana Matos, Hugo David, Marisa Encarnação, Diogo Ribeiro, Ana Joana Duarte, Olga Amaral, Hugo Rocha, Luísa Diogo, Sara Ferreira, Constança Santos, Esmeralda Martins, Teresa Neuparth, Joana Soares, Marta Ribeiro, Brígida Ribeiro Pinho, Nuno Oliveira, Jorge Miguel Ascenção Oliveira, Maria João Prata, Miguel Santos, Sandra Alves and Maria Francisca Coutinho*
Affiliation:
- Research and Development Unit, Department of Human Genetics, INSA
- Center for the Study of Animal Science, CECA-ICETA, University of Porto
- Associate Laboratory for Animal and Veterinary
Sciences, AL4 Animals
Keywords:
Lysosomal Storage Disorders; Mucopolysaccharidoses; Disease Models; Cell models; Dental Pulp Stem Cells; Animal models; Zebrafish
Abstract:
Introduction: When it comes to disease modeling, countless models are available for
Lysosomal Storage Diseases (LSD). Historically, two major approaches are well-established: in
vitro assessments are performed in patient fibroblasts, while in vivo pre-clinical studies are performed
in mouse models. Still, both platforms have a series of drawbacks. Thus, we implemented
two alternative and innovative protocols to mimic a particular sub-group of LSDs, the Mucopolysaccharidoses
both in vitro and in vivo.
Methods: The first one relies on a non-invasive approach using dental pulp stem cells from deciduous
teeth (SHEDs). SHEDs are multipotent neuronal precursors that can easily be collected.
The second uses a state-of-the-art gene editing technology (CRISPR/Cas9) to generate zebrafish
disease models.
Results: Even though this is an ongoing project, we have already established and characterized
two MPS II and one MPS VI SHED cell models. These cells self-maintain through several passages
and can give rise to a variety of cells including neurons. Furthermore, all MPS-associated
sub-cellular phenotypes we have assessed so far are easily observable in these cells. Regarding
our zebrafish models, we have successfully knocked down both naglu and hgsnat and the first
results we got from the behavioral analysis are promising ones, as we can observe altered activity
and sleep patterns in the genetically modified fish. For this particular approach we chose MPS III
forms as our target disorders, since their neurological features (hyperactivity, seizures and motor
impairment) and lifespan decrease would be easily recognizable in zebrafish.
Conclusion: Now that these methods are well-established in our lab, their potential is immense.
On one hand, the newly developed models will be of ultimate value to understand the mechanisms
underlying MPS sub-cellular pathology, which have to be further elucidated. On the other
hand, they will constitute an optimal platform for drug testing in house. Also noteworthy, our
models will be published as lab resources and made available for the whole LSD community.
