Swimming Towards a Cure in Primary Ciliary Dyskinesia: A Zebrafish Morpholino Model to Assess an LNP-mRNA Therapy Potential

December 5, 2024

Primary Ciliary Dyskinesia (PCD) is a rare genetic disorder characterised by dysfunctional motile cilia, leading to chronic respiratory infections, fertility complications, and organ laterality defects. PCD affects approximately 1:7,500 individuals and is associated with mutations in over 50 genes. Given its genetic heterogeneity and the current lack of causative treatments, there is a need for personalised therapies. Zebrafish, sharing 70% of their genes with humans, including PCD-associated genes and with easily accessible multiciliated cells that resemble human respiratory epithelium in their olfactory pit, offer an excellent model for studying PCD. This dissertation aimed to create a morpholino-induced PCD zebrafish disease model and evaluate the potential of an innovative LNP-mRNA therapy in restoring ciliary motion and functionality. Ciliary Beat Frequency and Active Area analyses demonstrated significant improvements in ciliary motion and activity following therapy administration. Notably, the area displaying hyperkinetic ciliary movements increased from approximately 10% in morpholino-injected embryos to around 80% in those treated with 240pg of LNP-mRNA. Additionally, this improvement showed a dose-dependent response. However, the functional analysis, using Directed Particle Tracking, did not show significant improvements, suggesting a complex relationship between ciliary structure, motion and functionality. The overall findings suggest that this LNP-mRNA therapy has the potential to recover cilia motility in PCD. According to previous studies, restoration of function in 20-25% of cilia may be sufficient for effective mucociliary clearance, thus, the observed improvement is promising. While further validation is needed, our Proof-of-Concept study supports the development of personalised therapies to improve PCD patients' quality of life.

Authors in the community:

• 

  C

  Carolina Folques Cardoso

  ist195925

Supervisors of this institution:

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  A

  Arsénio do Carmo Sales Mendes Fialho

  ist12833

industrial-biotechnology - Industrial Biotechnology

eng - English

Instituto Superior Técnico