(AAV), Herpes Simplex Virus (HSV-1), et al.
Non-virus method: Cyclodextrin Polymers, Lipids, Peptide, Antibodies, Aptamers,
and small molecules
■ Background
■ Background
Hybrid virus vector
HSV/AAV Ad/EBV HSV/EBV Ad/AAV Ad/retrovirus
Virus based transduction
■ Background
siRNA-based gene therapy
■ Background
Chemical modification
……
■ Background
Gene therapy Definition: Gene therapy is the use of DNA as a drug to treat
disease by delivering therapeutic DNA into a patient's cells.
Non-virus method
Can deliver both siRNA and plasmid DNA
■ Background
Conjugate delivery
Non-virus method
Dynamic PolyConjugates (DPC)
First reported in 2007 PEG: shielding effect GalNAc ligand was essential for both uptake by hepatocytes and in vivo silencing activity. Other targeting ligands has been explored, including peptides, antibodies, small molecules, glycans, lectins and nucleic acids.
■ Background
Technical problem: Barriers to the target
DNA/RNA is unstable in the bloodstream, can be immunogenic and does not readily cross membranes to enter cells. Nuclease, renal filtration
■ Background
■ Background
Different treatments to the diseases
Chemical drugs Surgery Physical Therapy Transplantation therapy Immunotherapy Regenerative medicine
Gene therapy
99% knockdown of liver genes after a single 0.2 mg per kg dose in non-human primates, with the effect lasting nearly 7 weeks
■ Background
Conjugate delivery
Triantennary GalNAc–siRNA
Non-virus method
ASGPR, on hepatocytes
Both subcutaneous and intravenous administration of this conjugate revealed great accumulation of siRNA in the liver and improved knockdown of the target gene.
※ Using DNA to express functional protein ※ Using siRNA or shRNA to attenuate abnormal gene
expression in mRNA level ※ Genome editing to correct mutant gene sequence: ZFN,
■ Background
Lentivirus:
Virus based transduction
RNA virus derived from HIV Can infect non-dividing cell Low Immunogenic Long expression period
Insertion mutation by random integration and oncogenicity Low virus titer
■ Background
Adenovirus:
Virus based transduction
DNA virus High virus titer (up to 1014VP/ml) and high infection efficiency Wide host range and large transgene capacity ( ~37kb ) Infect dividing and non-dividing cell Low integration level, exist as episome in host cell No insertion mutation by random integration and oncogenicity Short expression period (5-20 days) Complex procedure and manipulation Potential immunogenic and inflammatory response
※ The first approved gene therapy case in the United States took place on 14 September 1990.
※ There are more than 2000 clinical trials have being launched in the past seven years.
Adeno-associated virus (AAV)
DNA virus without pathogenicity Specific host range Infect dividing and non-dividing cell Low integration level, exist as episome in host cell No insertion mutation by random integration and oncogenicity (site-specific integrate into 19 chromosome) Long expression period No immunogenic and inflammatory response Small transgene capacity ( ~3kb ) Low virus titer (1012 VP/ml) Complex procedure and manipulation Host range limitation
■ Background
Herpes Simplex Virus-1
Virus based transduction
DNA virus High infection efficiency Infect dividing and non-dividing cell Neurotropic virus Large transgene capacity ( up to ~150kb ) Long expression period High immunogenic and inflammatory response and necrosis
Non-virus method
Can deliver both siRNA and plasmid DNA■ Bac源自groundLiposome
Protect entrapped oligonucleotides from nuclease degradation and renal clearance Promote cellular uptake and endosomal escape They include the use of cationic or ionizable lipids, shielding lipids, cholesterol and targeting ligands
Gene Therapy
■ Background
Human beings fight against all kind of diseases
Cancer CCVD: Cardio-Cerebrovascular Diseases
Viral Disease Genetic Disease: more than 2000 diseases CNS: Central Nervous System Disease Immune Diseases Diabetes ……
Poor selectivity and inefficiency of enrichment in the target cell or tissue.
■ Background
Different delivery tools
Virus Vector: Retrovirus, Lentivirus, Adenovirus, Adeno-associated virus
TALEN, CRISPR-Cas9…
■ Background