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Delivery of Therapeutic siRNA to the CNS Using Cationic and Anionic Liposomes
Published on: July 23, 2016
Treatment of Ulcerative Colitis by Cationic Liposome Delivered NLRP3 siRNA
Jing Huang1, Mengmeng Dai1, Mingxia He1
1Joint National Laboratory for Antibody Drug Engineering, The First Affiliated Hospital, Henan University, Kaifeng, Henan Province, 475004, People's Republic of China.
Purpose:
The abnormal activation of NLRP3 inflammasome is related to the occurrence and development of ulcerative colitis (UC). However, the ideal drug and delivery system remain important factors limiting the targeting of NLRP3 inflammasome in UC therapy. Gene therapy by delivering siRNA is effective in treating various diseases. Therefore, delivering siNLRP3 using an ideal vector for UC treatment is necessary.
Materials And Methods:
Nanoparticles delivering siNLRP3 were developed based on cationic liposome (CLP/siNLRP3). Their ability to inhibit NLRP3 inflammasome activation was monitored using Western blot (WB) and Enzyme-linked Immunosorbent Assay (ELISA). The ASC oligomerization in LPS-primed peritoneal macrophages (PMs) was detected by WB and immunofluorescence. Moreover, we assessed the role of CLP/siNLRP3 on dextran sodium sulfate (DSS)-induced UC by examining NLRP3 levels, pro-inflammatory cytokines expression, and disease-associated index (DAI). Flow cytometry (FCM) was used to detect the contents of macrophages and T cells. Finally, we assessed the safety of CLP/siNLRP3.
Results:
The prepared CLP was spherical, with a small particle size (94 nm) and low permeability. The CLP could efficiently protect siNLRP3 from degradation and then deliver siNLRP3 into PMs, inhibiting NLRP3 inflammasome activation. Also, the CLP/siNLRP3 could inhibit the secretion of mature IL-1β and IL-18 from PMs, thereby achieving a favorable anti-inflammation effect. In vivo, CLP/siNLRP3 could effectively alleviate intestinal injury in UC mice, which was attributed to down-regulating levels of IL-1β and IL-18, inhibiting infiltration of macrophages and other immune cells, and the polarization of M1 macrophages. Finally, pathological testing of tissue sections and blood biochemical tests showed no significant toxic effects of CLP/siNLRP3.
Conclusion:
We introduced a prospective approach for the efficient delivery of siRNA in vitro and in vivo with high safety and stability, which was found to have great potential in treating NLRP3-driven diseases in an RNA-silencing manner.
Insights
Cationic liposome nanoparticles effectively deliver siNLRP3 to inhibit the NLRP3 inflammasome, reducing inflammation and intestinal injury in ulcerative colitis models with good safety.
Area of Science:
- Biomedical Engineering
- Molecular Medicine
- Immunology
Background:
- NLRP3 inflammasome overactivation is implicated in ulcerative colitis (UC) pathogenesis.
- Targeting NLRP3 inflammasome in UC therapy is challenged by limitations in drug and delivery systems.
- Gene therapy using small interfering RNA (siRNA) offers potential for disease treatment.
Purpose of the Study:
- To develop an efficient and safe delivery system for siNLRP3 to target NLRP3 inflammasome in UC.
- To evaluate the therapeutic efficacy of the developed system in preclinical models of UC.
Main Methods:
- Development of cationic liposome nanoparticles encapsulating siNLRP3 (CLP/siNLRP3).
- In vitro assessment of NLRP3 inflammasome inhibition in macrophages using Western blot and ELISA.
- In vivo evaluation in dextran sodium sulfate (DSS)-induced UC mouse models, assessing disease activity, cytokine levels, and immune cell infiltration.
- Safety assessment through pathological and biochemical analyses.
Main Results:
- CLP/siNLRP3 nanoparticles were spherical, small (94 nm), and protected siNLRP3 from degradation.
- CLP/siNLRP3 effectively inhibited NLRP3 inflammasome activation and reduced pro-inflammatory cytokines (IL-1β, IL-18) in vitro.
- In vivo, CLP/siNLRP3 alleviated DSS-induced UC, reducing intestinal injury, cytokine levels, and immune cell infiltration.
- No significant toxicity was observed in safety assessments.
Conclusions:
- CLP/siNLRP3 represents a promising strategy for targeted RNA silencing of NLRP3 inflammasome.
- This delivery system demonstrates high safety and stability for in vitro and in vivo applications.
- The developed system holds significant potential for treating NLRP3-driven diseases like ulcerative colitis.
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