Introduction

Gene therapy offers tremendous potential for treating a broad spectrum of diseases by modifying or replacing faulty genes. Historically viral vectors have been the primary means of delivery but non-viral approaches are now gaining traction due to their potential safety advantages and manufacturing scalability. This news article highlights some of the innovative non-viral gene delivery methods being developed which were recently showcased at the American Society of Gene & Cell Therapy (ASCGT) 2024 meeting.

Lipid Nanoparticles (LNPs)

LNPs have emerged as a leading non-viral delivery system especially for nucleic acid therapeutics. Researchers are continuously innovating to improve their efficacy and tissue-targeting abilities:

ReNAgade Therapeutics

ReNAgade Therapeutics has developed novel immune-tropic LNPs that can efficiently deliver mRNA to T cells in non-human primates. Their high-throughput LNP discovery engine identified formulations achieving ~60% delivery to splenic T cells and ~50% to peripheral blood T cells. Read more

Nanite Inc.

Nanite Inc. is using machine learning approaches to design polymeric nanoparticles for mRNA delivery to T cells. Their iterative prediction and testing process allows rapid identification of effective carriers with tunable biodegradation properties. Read more

Johns Hopkins University

Researchers at Johns Hopkins University have created ligand-conjugated poly(beta-amino ester) nanoparticles that show enhanced mRNA delivery to primary T cells. By incorporating multiple T cell-targeting ligands they achieved significantly higher transfection rates compared to non-targeted particles. Read more

Polymeric Nanoparticles

Polymeric nanoparticles offer versatility in design and the ability to incorporate various functional elements:

The Pennsylvania State University

A team at The Pennsylvania State University developed a biomimetic surface coating using DNA-alginate templates to display aptamers on cell surfaces. This approach showed greatly improved aptamer stability and enhanced cell recognition capabilities compared to conventional methods. Read more

University of Alberta

Researchers at the University of Alberta evaluated lipopolymers as siRNA delivery agents for leukemia treatment. Their formulations showed promising gene silencing and anti-tumor effects in primary patient samples. Read more

Cell-Penetrating Peptides

Peptide-based delivery systems are gaining attention for their ability to facilitate cellular uptake of therapeutic cargo.

Spotlight Therapeutics

Spotlight Therapeutics has engineered a novel class of non-viral gene editors called eRNPs (engineered ribonucleoproteins). These combine Cas nucleases with cell-penetrating and nuclear-localizing peptides to enable efficient in vivo gene editing particularly for ophthalmic disorders. Read more

Entrada Therapeutics Inc.

Entrada Therapeutics Inc. has developed the Endosomal Escape Vehicle (EEV™) technology which uses cyclic cell-penetrating peptides (CPPs) to deliver oligonucleotides to skeletal and cardiac muscles. These peptides efficiently penetrate cell membranes and are resistant to proteolytic degradation. The EEV platform has demonstrated its potential in preclinical models of Duchenne muscular dystrophy (DMD) showing enhanced exon skipping and broad dystrophin protein expression in skeletal and cardiac muscles. Read more

Université Laval and Feldan Therapeutics

Researchers at Université Laval and Feldan Therapeutics have developed a method to improve the intracellular delivery of phosphorodiamidate morpholino oligomers (PMOs) to airway epithelial cells. By conjugating S10 peptides to PMOs they achieved homogeneous delivery throughout the lung airway epithelia in mice. This method could enable the effective treatment of respiratory diseases such as asthma and cystic fibrosis. Read more

Engineered Extracellular Vesicles

Extracellular vesicles (EVs) are emerging as promising natural nanocarriers for gene therapy.

Lonza Cell and Gene

Lonza Cell and Gene has developed a platform for producing engineered EVs with enhanced therapeutic payload delivery capabilities. Their approach includes protein scaffolds and chemical linkers to effectively load various biomolecules onto or into EVs. Read more

Physical Methods

Physical techniques like electroporation continue to be refined for gene delivery applications.

Kytopen

Kytopen has implemented a custom particle swarm-differential evolution optimization algorithm to enhance plasmid DNA transfection in T cells using their Flowfect® microfluidic electroporation technology. Read more

CellFE

CellFE has developed a microfluidics-based cell compression system for sequential delivery of multiple gene-modifying payloads. This method enables multiplex genome editing while maintaining cell viability. Read more

MxT Biotech and Korea University

MxT Biotech and Korea University have introduced a microfluidic droplet-enabled cell mechanoporator for high-efficiency genome editing of human primary immune cells. This platform co-encapsulates cells and genetic materials in droplets achieving high delivery efficiency and scalability. Read more

Shanghai Cell Therapy Group

Shanghai Cell Therapy Group has developed the JL transposon system a novel tool for gene and cell therapy with enhanced transposition efficiency and lower cytotoxicity. Read more

MxT Biotech and Korea University

MxT Biotech and Korea University have developed the Hydroporator® a non-viral gene delivery platform for precision T cell engineering. Read more

Avectas

Avectas has introduced Solupore® an automated non-viral transfection system for manufacturing gene-modified T cells. Read more

DNA/RNA Nanostructures

Researchers are exploring the use of nucleic acid-based nanostructures for gene delivery:

Kookmin University

A team at Kookmin University has created self-assembled functional RNA nanoparticles for delivering CRISPR-Cas9 components. Their system utilizes rolling circle transcription on magnetic nanoparticle surfaces to generate RNA shells encapsulating the gene editing machinery. Read more

Inorganic Nanoparticles

While less common inorganic nanoparticles are also being investigated for gene therapy applications:

German Cancer Research Center

Researchers at the German Cancer Research Center have developed S/MAR DNA nanovectors using scaffold/matrix-associated regions for stable genetic modification of hematopoietic stem and progenitor cells. These episomally maintained vectors showed efficient and persistent transgene expression without the risk of insertional mutagenesis. Read more

Peptide-Based Nanoplatform

Clemson University researchers have developed a peptide-based nanoplatform for delivering Cas9 Ribonucleoprotein (RNP) complexes for gene editing. The DIV3W peptide facilitates endosomal escape and protects RNA cargo showing efficient internalization and gene knockout in HEK293 cells. This delivery system holds promise for advanced gene editing applications. Read more

Conclusion

In conclusion, the field of non-viral gene therapy is rapidly advancing with innovative approaches to overcome delivery challenges. From smart nanoparticle designs to novel physical methods researchers are pushing the boundaries to create safer and more effective genetic medicines. As these technologies continue to mature we can expect to see more non-viral gene therapies entering clinical trials and eventually reaching patients in need.