FFay Lecture Gene Therapy Non-Viral Vectors PDF

GENE THERAPY NON VIRAL VECTORS François Fay Institut Galien Paris-Saclay, UMR CNRS 8612 [email protected]  Add DNA Add mRNA React with protein Remove DNA Destroy mRNA (prevent translation) Replace DNA Modify mRNA (modif...

GENE THERAPY NON VIRAL VECTORS François Fay Institut Galien Paris-Saclay, UMR CNRS 8612 [email protected]  Add DNA Add mRNA React with protein Remove DNA Destroy mRNA (prevent translation) Replace DNA Modify mRNA (modify translation) Natural barriers? Figure from http://www.bonac.com/global/en/nucleic/about/ Natural barriers Physiological barriers Yin et al Nature Reviews Genetics 2014 Natural barriers DNA / RNA Biochemistry NIH Natural barriers DNA / RNA Biochemistry Aspirin 0,18 kda RNA ? Antibodies 150 kda NIH Natural barriers DNA / RNA Biochemistry Aspirin 0,18 kda iRNA ∼10 kDa Antibodies 150 kda NIH Acides nucléiques à visées thérapeutiques Aspirine 0,18 kda mRNA ∼400 kDa Antibodies 150 kda Adapted from R Robinson et al PLoS Biol 2004 NIH Adapted from U. Elia et al ACS Nano 2021 Natural barriers DNA / RNA Biochemistry Low stability High molecular weight Negative charge Hydrophilicity NIH Natural barriers DNA / RNA Biochemistry Low stability High molecular weight Negative charge Hydrophilicity → Need modification and/ or vectorisation! NIH DNA / RNA Biochemistry DNA / RNA Biochemistry Duarte Moreno DOI:10.3389/fchem.2014.00087 Vectors Viral Vectors Viral Vectors Strengths High transfection efficiency Natural tropism (ability to infect different cells) Evolved mechanisms for endosomal escape Natural transportation mechanism of DNA into nucleus Weaknesses Strong immune reactions against viral proteins prohibit multiple administrations Possibility of chromosomal insertion and protooncogene activation Complicated synthesis process Limitation on gene size Toxicity, contamination of live virus Non-viral vectors Design criteria ? Verbeke et al Nanotoday https://doi.org/10.1016/j.nantod.2019.100766 Yin et al Nature Reviews Genetics 2014 viral and non-viral vectors Some design criteria Packaging of large DNA/RNA Cell Internalization quantities Endolysosomal escape Easy administration Nuclear transport Serum stability Efficient unpackaging Targetability to specific cell types Infection of non-dividing cells Inexpensive synthesis Safety Easy purification Non-toxic Robustness/stability Non-immunogenic Non-pathogenic Non-viral vectors Huang https://doi.org/10.3390/pharmaceutics14081586 Non-viral vectors Invitrogen / Polyplus Non-viral vectors Huang https://doi.org/10.3390/pharmaceutics14081586 Lipid NanoParticles Ionisable lipid Cholesterol % molar ratio : 30-50% % molar ratio : 30-50% - pKa