Lipid nanoparticle-encapsulated Dnai1 mRNA rescues ciliary activity in primary ciliary dyskinesia mouse cell models
Amanda J Smith1,2, Patrick R Sears1, Mirko Hennig3
1Marsico Lung Institute/Cystic Fibrosis Research and Treatment Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
Abstract:
Primary ciliary dyskinesia (PCD) is a rare, genetically heterogenous disorder resulting from dysfunctional motile cilia that is characterized by chronic, progressive lung disease with currently no corrective therapies available. Here, we test the efficacy of selective organ targeting lipid nanoparticles (SORT-LNPs) that were optimized for potency and delivery to respiratory cells containing an mRNA encoding an axonemal protein to rescue ciliary activity in a murine culture model of PCD. Utilizing murine nasopharyngeal epithelial cell (mNPEC) cultures isolated from a conditional Dnai1 knockout mouse model of the known human PCD-associated gene DNAI1 homolog, we tested if SORT-LNPs containing an optimized Dnai1 mRNA could rescue ciliary activity. Treatment of differentiating and well-differentiated Dnai1 knockout mNPECs with SORT-LNP-Dnai1 mRNA led to a dose-dependent increase in levels of DNAI1 protein and incorporation into ciliary axonemes, resulting in rescued ciliary activity with normal ciliary beat frequency that persisted for over 3 weeks. These data support further clinical development of an mRNA-based therapeutic with LNP-mediated delivery as a treatment for individuals with PCD with disease-causing DNAI1 mutations.
Insights
Researchers developed a novel mRNA therapy delivered via lipid nanoparticles to restore ciliary function in a mouse model of Primary Ciliary Dyskinesia (PCD). This treatment successfully rescued ciliary activity, offering hope for a new therapeutic approach for PCD patients with DNAI1 mutations.
Area of Science:
- Biochemistry
- Genetics
- Respiratory Medicine
Background:
- Primary ciliary dyskinesia (PCD) is a rare genetic disorder causing chronic lung disease due to dysfunctional motile cilia.
- Current treatments for PCD are limited, lacking corrective therapies for the underlying ciliary defect.
Purpose of the Study:
- To evaluate the efficacy of selective organ targeting lipid nanoparticles (SORT-LNPs) delivering DNAI1 mRNA to restore ciliary function in a mouse model of PCD.
- To assess the potential of mRNA-based therapy with LNP delivery as a treatment for PCD.
Main Methods:
- Utilized murine nasopharyngeal epithelial cell (mNPEC) cultures from a Dnai1 knockout mouse model.
- Treated mNPECs with SORT-LNPs containing optimized Dnai1 mRNA.
- Quantified DNAI1 protein levels, axonemal incorporation, and ciliary beat frequency.
Main Results:
- SORT-LNP-Dnai1 mRNA treatment dose-dependently increased DNAI1 protein levels in knockout mNPECs.
- Restored DNAI1 incorporation into ciliary axonemes, rescuing ciliary activity.
- Achieved normal ciliary beat frequency that persisted for over 3 weeks.
Conclusions:
- mRNA-based therapy delivered by SORT-LNPs can effectively rescue ciliary function in a preclinical model of PCD.
- This approach shows promise for clinical development as a treatment for individuals with PCD caused by DNAI1 mutations.
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