Abstract 3199: Nano gelasomes: Novel lamellar metal-protein suprastructures for nucleic acid delivery

Abstract There is considerable interest in the development of self-assembled charged nanocarriers that form organized, predictable structures to enhance stable packaging and delivery of nucleic acid materials in vivo. However, the synthesis of stably-charged, organized nanostructures is highly challenging, requiring a controlled balance of nanomaterial components, stabilizers, and hydrophobic forces. Herein, we report a significant step forward in the controlled lamellar assembly of a charged complex between ultra-small 2 nm gold nanoparticles and a lipidoid within an organic matrix to form Gelasomes. These spherical suprastructures feature an intricate 3D concentric arrangement with a constant interlayer distance (4nm). The cationic lipidoid within this arrangement aids in the highly efficient packaging of nucleic acid materials in the nanoparticle. To demonstrate the potential biomedical applications of Gelasomes, we have loaded siRNA (AXL or YAP), mRNA (EGFP), or CRISPR components (Cas9 mRNA and guide RNA) and validated their functionality by in vitro or in vivo studies. Gelasomes with AXL siRNA (~20 bp) demonstrated rapid uptake (<2 hrs), successful endosomal escape, remarkable transfection efficiency (99.87 ± 0.04 %), gene downregulation (97 ± 2.8 %), and disassembly within the cell (<24 hrs). In vivo downregulation studies in Lung Cancer PDX (Gelasomes with AXL siRNA) and CDX (Gelasomes with YAP siRNA) models confirm the gene-silencing capability (91% or 38 % respectively) of Gelasomes underscoring their potential in advancing nanomedicine. Similarly, Gelasome loaded with EGFP mRNA (1 kb) showed 47 ± 8.54 % expression efficiency within 48 h of treatment in vitro. We also show the packaging of larger nucleic acids such as CRISPR Cas9 mRNA (5 kb) along with the guide RNA (100 bp) in vitro for generating knockouts, that achieved an editing efficiency of ~6%. Together, the results from this study pave the way for the development of adaptable, biodegradable, and functionally versatile supraparticles for medical applications. Citation Format: Agasthya Suresh Babu, Dhananjay Suresh, Zhaohui Li, Anandhi Upendran, Raghuraman Kannan. Nano gelasomes: Novel lamellar metal-protein suprastructures for nucleic acid delivery [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3199.