Reinforced plant-derived lipid nanoparticles for oral precise epigenome editing in colonic diseases
Qiang Gao1,2, Yunxia Gao3, Yingui Cao1
1State Key Laboratory of Resource Insects, College of Sericulture, Textile, and Biomass Sciences, Southwest University, Beibei, Chongqing 400715, China.
Abstract:
The clinical application of CRISPR-Cas9 remains limited by delivery challenges, particularly for oral administration. Lysine-specific demethylase 1 (Lsd1) plays a key role in colonic inflammation and tumorigenesis. Here, we developed an oral genome-editing platform (TPGS-RNP@LNP), where Lsd1-targeting ribonucleoproteins (RNPs) were encapsulated in mulberry leaf lipid nanoparticles (LNPs) and formulated with d-α-tocopherol polyethylene glycol succinate (TPGS). TPGS reinforced the lipid bilayer of LNPs, enhanced gastrointestinal stability, and facilitated colonic mucus penetration. Upon the galactose receptor-mediated endocytosis of TPGS-RNP@LNPs by macrophages, their fusion with the endosomal membrane and the presence of nuclear localization signals ensured the nuclear delivery of RNPs. TPGS-RNP@LNPs achieved 59.7% Lsd1 editing efficiency in macrophages, surpassing the commercial CRISPRMAX (43.0%). Oral TPGS-RNP@LNPs promoted H3K4 methylation to modulate epigenetic states, achieving inflammation mitigation, epithelial barrier restoration, and retardation of colitis and its associated tumorigenesis. As an LNP-based oral RNP delivery system, TPGS-RNP@LNPs provide a promising platform for precise treatment of colorectal diseases.
Insights
This study introduces a novel oral genome-editing platform using lipid nanoparticles (LNPs) for precise CRISPR-Cas9 delivery. The system effectively targets Lsd1 in macrophages, mitigating colon inflammation and tumorigenesis for potential colorectal disease treatment.
Area of Science:
- Biotechnology
- Molecular Biology
- Genomics
Background:
- CRISPR-Cas9 gene editing faces delivery challenges, especially for oral administration.
- Lysine-specific demethylase 1 (Lsd1) is implicated in colonic inflammation and cancer development.
Purpose of the Study:
- To develop an oral genome-editing platform for targeting Lsd1 in the colon.
- To enhance CRISPR-Cas9 delivery and efficacy for treating colitis and associated tumorigenesis.
Main Methods:
- Development of TPGS-RNP@LNP, an oral platform encapsulating Lsd1-targeting ribonucleoproteins (RNPs) in lipid nanoparticles (LNPs) with TPGS.
- Utilizing galactose receptor-mediated endocytosis for macrophage targeting and ensuring nuclear delivery of RNPs.
- Assessing Lsd1 editing efficiency, epigenetic modulation (H3K4 methylation), and therapeutic effects in a colitis model.
Main Results:
- TPGS-RNP@LNPs demonstrated superior Lsd1 editing efficiency (59.7%) compared to commercial CRISPRMAX (43.0%).
- Oral administration modulated epigenetic states, reduced inflammation, restored epithelial barriers, and inhibited colitis-associated tumorigenesis.
- The platform showed enhanced gastrointestinal stability and colonic mucus penetration.
Conclusions:
- TPGS-RNP@LNPs represent a promising LNP-based oral RNP delivery system for precise colorectal disease treatment.
- This platform overcomes oral delivery challenges for CRISPR-Cas9 applications in treating inflammatory and cancerous conditions of the colon.
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