Lecithin nano-liposomal particle as a CRISPR/Cas9 complex delivery system for treating type 2 diabetes

Abstract

BackgroundProtein-based Cas9 in vivo gene editing therapeutics have practical limitations owing to their instability and low efficacy. To overcome these obstacles and improve stability, we designed a nanocarrier primarily consisting of lecithin that can efficiently target liver disease and encapsulate complexes of Cas9 with a single-stranded guide RNA (sgRNA) ribonucleoprotein (Cas9-RNP) through polymer fusion self-assembly.ResultsIn this study, we optimized an sgRNA sequence specifically for dipeptidyl peptidase-4 gene (DPP-4) to modulate the function of glucagon-like peptide 1. We then injected our nanocarrier Cas9-RNP complexes directly into type 2 diabetes mellitus (T2DM) db/db mice, which disrupted the expression of DPP-4 gene in T2DM mice with remarkable efficacy. The decline in DPP-4 enzyme activity was also accompanied by normalized blood glucose levels, insulin response, and reduced liver and kidney damage. These outcomes were found to be similar to those of sitagliptin, the current chemical DPP-4 inhibition therapy drug which requires recurrent doses.ConclusionsOur results demonstrate that a nano-liposomal carrier system with therapeutic Cas9-RNP has great potential as a platform to improve genomic editing therapies for human liver diseases.Electronic supplementary materialThe online version of this article (10.1186/s12951-019-0452-8) contains supplementary material, which is available to authorized users.

Drug Delivery System in the Treatment of Diabetes Mellitus

“…Lecithin liposome was designed as a nano carrier to encapsulate complexes of Cas9-RNP through polymer fusion self-assembly for target delivery to liver. In this delivery system, the sgRNA was specifically optimized for dipeptidyl peptidase-4 gene (DPP-4) to inhibit the degradation of glucagon-like peptide 1 to enhance the secretion of insulin ( Cho et al, 2019 ). Results showed that T2DM mice injected with nano carrier Cas9-RNP complexes exhibited the remarkably down-regulation of DPP-4 gene, accompanied by euglycemia, insulin response, and alleviated liver and kidney damage.…”

Can CRISPR/Cas Technology Be a Felicitous Stratagem Against the COVID-19 Fiasco? Prospects and Hitches

“…CRISPR/Cas9-engineered β-cells have also been explored to characterize the pharmacology of compounds with activity at both glucose-dependent insulinotropic polypeptide receptor (GIPR) and glucagon-like peptide-1 receptor (GLP-1R) ( Naylor et al, 2016 ). Similarly, injection of lecithin based nanoliposomal carrier of CRISPR/Cas9 directly into type 2 diabetes mellitus (T2DM) db/db mice was reported to disrupt expression of dipeptidyl peptidase-4 (DPP-4) gene, consequently lowering the DPP-4 enzyme activity, complemented by normalized blood glucose levels, insulin response, and lessened organ destruction ( Cho et al, 2019 ). In another pertinent work, Maxwell et al (2020) had resorted to CRISPR/Cas system to rectify a Wolfram syndrome 1 ( WFS1 ) (characterized by a gamut of symptoms including diabetes, optic atrophy, etc.)…”

CRISPR/Cas System for Genome Editing: Progress and Prospects as a Therapeutic Tool

“…Furthermore, up to 80% genome modification was observed with Cas9:sgRNA complexes compared with DNA transfection (Zuris et al, 2015). In a recent study, Cho et al (2019) used nanoliposomes prepared using lecithin to deliver cas9-sgRNA RNPs directed against the DPP-4 gene to modulate the function of glucagon-like peptide 1. In vivo delivery of these complexes in type 2 diabetes mellitus db/db mice disrupted DPP-4 gene expression, leading to a decline in DPP-4 enzyme activity that resulted in normalized blood glucose levels, a declined in insulin resistance, and negligible impact on liver and kidney function (Cho et al, 2019).…”

“…In a recent study, Cho et al (2019) used nanoliposomes prepared using lecithin to deliver cas9-sgRNA RNPs directed against the DPP-4 gene to modulate the function of glucagon-like peptide 1. In vivo delivery of these complexes in type 2 diabetes mellitus db/db mice disrupted DPP-4 gene expression, leading to a decline in DPP-4 enzyme activity that resulted in normalized blood glucose levels, a declined in insulin resistance, and negligible impact on liver and kidney function (Cho et al, 2019). In another study, Kim et al complexed cas9 RNPs with lipofectamine 2000 delivered subretinally for the treatment of wet age-related macular degeneration (Kim et al, 2017b).…”