01. Gene therapy Boulikas.pdf - Gene therapy & Molecular Biology

lipoprotein
receptor
(VLDL-R)
HSV
TK+ganciclo
vir (GCV)
HSV TK
plus
ganciclovir
HSV TK
plus
ganciclovir
Boulikas: An overview on gene therapy
olemia (FH) cholesterol by approximately 50% at
days 4 and 9; marked reduction in the
intermediate density lipoprotein/low
density lipoprotein (IDL/LDL)
Gliomas Retr Treatment of rats with cerebral glioma;
intratumoral stereotaxic injection of
murine fibroblasts (G418-selected)
producing a retroviral vector with the
HSV TK gene
prostate cancer Adenovi
rus
mouse
adenocarcinom
a
TH PD Cat
lipos rat
rat glial cell
line-derived
neurotrophic
factor
(rGDNF),
Ganciclovir is converted by HSV TK
into its triphosphate form which is then
incorporated into the DNA of
replicating mammalian cells leading to
inhibition in DNA replication and cell
death; it is only viral TK, not the
mammalian enzyme, that can use
efficiently phosphorylated ganciclovir
as a substrate
Carcinoembryonic antigen-producing
human lung cancer cells
Overexpress TH to alleviate
degeneration of dopaminergic
nigrostriatal neurons (DNN) in PD rat
models
PD AAV To protect nigral dopaminergic neurons
in the progressive Sauer and Oertel 6hydroxydopamine
(6-OHDA) lesion
model of Parkinson's disease (backlabeled
fluorogold-positive neurons in
the substantia nigra)
Factor IX Hemophilia B Adenovi
rus mice
Factor IX Hemophilia B Adenovi
rus mice
in vivo
Factor IX Hemophilia B Retrovir
us dogs
in vivo
Factor IX Hemophilia B Adenovi
rus dogs
in vivo
Correction of FIX defect by injection of
recombinant adenovirus hosting the
canine FIX gene into hind leg muscle of
mice
92
Tretament with GCV 5 days
postinjection resulted in complete
regression of gliomas
Subcutaneous tumors induced by
injection of RM-1 (mouse prostate
cancer) cells followed by injection of
HSV TK and treatment with
ganciclovir for 6 days showed
reduction in tumor volume (16% of
control) and higher apoptotic index in
tumor cells
Cell type-specific expression of
herpes simplex virus thymidine
kinase gene
Direct injection of lipofectin-TH
expression plasmid on nigra-lesioned
side generated L-DOPA locally and
decreased contralateral rotations.
94% cell protection; 85% of tyrosine
hydroxylase-positive cells
Establishment of factor IX levels in
the blood in nude mice for 300 days;
only for 10 days in normal mice;
transduced cells were removed by
cell-mediated as well as humoral
immune responses;
cyclophosphamide or cyclosporin A
immunosuppression maintained FIX
protein for 5 months
Correction of the factor IX gene Therapeutic plasma levels of factor
IX in mice
Expression of factor IX gene in liver in
hemophilia B dogs
Establishment of the blood serum factor
IX levels in hemophilic B dogs by
infection with recombinant adenovirus
vectors delivering the Factor IX gene
and treatment with cyclosporin A
1% of hepatocytes were transduced
with a retroviral vector carrying the
canine factor IX gene injected into
the portal vein (2/3 hepatectomy was
required); low therapeutic levels of
factor IX
Treatment with cyclosporin A
suppressed T cell activation; T cells
attack the adenovirus-transduced
cells eliminating them from the body;
Cycl A tretment led to prolonged
Factor IX levels in the blood of
hemophilic dogs
Factor IX Hemophilia B AAV Successful transduction of the mouse
liver in vivo after a single
administration; persistent, curative
concentrations of functional human
factor IX can be achieved
Gene target
or delivered
Human
disease
Culver et al, 1992
Eastham et al, 1996
Osaki et al, 1994
Cao et al, 1995
Mandel et al, 1997
Dai et al, 1995
Smith et al, 1993
Kay et al, 1993
Kay et al, 1994; Fang et
al, 1995
Snyder et al, 1997
Method Goal/rationale Results Reference