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Published on: May 2, 2018
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Pathologic Tissue Injury and Inflammation in Mice Immunized with Plasmid DNA-Encapsulated DOTAP-Based Lipid
Shasha Peng1,2, Yifan Zhang1,3, Xin Zhao1
1Institute of Medical Biology, Chinese Academy of Medical Sciences and Peking Union Medical College, Kunming 650118, China.
Bioconjugate Chemistry
|December 10, 2024
Summary
Microfluidic techniques create smaller DOTAP-based lipid nanoparticles (LNPs) for effective SARS-CoV-2 DNA delivery. While showing promise for lung and spleen targeting, these LNPs caused dose-dependent inflammation and injury in muscles, heart, and liver.
Area of Science:
- Biomedical Engineering
- Nanotechnology
- Vaccine Development
Background:
- Ionizable cationic lipids like DOTAP are crucial for lipid nanoparticle (LNP) delivery systems.
- DOTAP, despite toxicity concerns, aids LNP tissue localization and enhances T-cell responses when prepared via microfluidics.
- Traditional lipid-film methods for DOTAP-based LNPs are associated with adverse events, necessitating improved preparation techniques.
Purpose of the Study:
- To evaluate the tissue localization and adverse toxicity of DOTAP-based LNPs prepared using microfluidic techniques for SARS-CoV-2 DNA delivery.
- To compare the efficacy, biodistribution, and safety profile of microfluidically prepared LNP-DNA complexes against traditional methods.
- To investigate the dose-dependent effects of DOTAP-based LNPs on immune response and tissue pathology.
Main Methods:
- Encapsulation of SARS-CoV-2 receptor-binding domain (RBD) plasmid DNA within LNPs using DOTAP, DOPE, cholesterol, and DMG-PEG2000 via microfluidic mixing.
- Assessment of LNP size, delivery efficacy, biodistribution, inflammatory response, and pathological injury in various tissues post-immunization.
- Comparative analysis of LNP-DNA complexes versus dissociative DNA in terms of tissue presence and immune induction.
Main Results:
- Microfluidically prepared LNP-RBDs were smaller than those from lipid-film methods.
- Effective delivery and expression of LNP-DNA in muscle tissue induced specific antibodies.
- Sustained presence of LNPs and DNA in lungs and spleen was observed, with dose-dependent inflammatory responses and pathological injuries in muscle, heart, and liver.
Conclusions:
- Microfluidic preparation of DOTAP-based LNPs offers advantages for targeting the lungs and spleen and enhancing gene expression.
- The observed inflammatory responses and pathological injuries are dose-dependent, suggesting careful dose optimization is required.
- These findings support the development of advanced LNP delivery systems utilizing DOTAP for improved vaccine and therapeutic applications.
