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

of rejecting tumour cells from the patient, especially on
immunoresponsive malignancies such as melanomas,
colorectal carcinomas, and renal cell carcinomas
Table 5. Ex vivo studies on animal models
Boulikas: An overview on gene therapy
Gene target Human
disease
ADA SCID (severe Retrovir Immunodeficiient mice were injected
combined us with peripheral blood lymphocytes
immunodeficie
ncy)
from ADA- patients transduced with a
retroviral vector for human ADA
bcl-2 prostate cancer bcl-2 expressing LNCaP human
prostate cancer cells are rendered
highly resistant to apoptotic stimuli
Factor IX hemophilia B Injection of transduced primary
myoblasts into the muscle
88
(Uchiyama et al, 1993; Chang et al, 1996; Finke et al,
1997; Das Gupta et al, 1997; Mahvi et al, 1997).
Method Animal model, objective, and method Results Reference
Factor IX hemophilia B retr Transplantation of retrovirustransduced
keratinocytes
Factor IX hemophilia B retroviru
s
Factor VIII Hemophilia A transferr
in
Factor VIII Hemophilia A Retrovir
us
Growth
hormone
(human)
Growth
hormone
(human,
hGH)
none mice
electrop
oration
Mouse primary myoblasts were
infected with retrovirus expressing the
canine factor IX under control of mouse
muscle creatine kinase and human
CMV promoter; myoblasts were
injected into the hindlegs of recipient
mice; secreted canine factor IX was
monitored in the plasma
Transfection of fibroblasts and
myoblasts with B-domain-deleted
factor VIII gene followed by
implantation into mice
Mouse primary fibroblasts infected
with a recombinant retrovirus
containing factor VIII gene deleted at
the B domain
Ex vivo modified C2C12 cells with the
hGH gene under control of the
inducible UAS promoter and a
synthetic hybrid steroid receptor
(TAXI), activating transcription from
the inducible promoter after treatment
with the synthetic nontoxic drug
inducer RU486; transplanted in mouse
muscle
general retr Injection of genetically engineered
myoblasts into mouse muscle
HSV-tk glioma Retr To directly transfer HSV TK gene and
kill transduced proliferating brain
tumor cells with ganciclovir without
affecting nondividing normal cells
HSV-tk pancreatic
cancer
retroviru
s
BXPC3 primary human pancreatic
adenocarcinoma cells were transduced
with retroviral vector carrying the
HSV-tk gene driven by the CEA
promoter; engrafted subcutaneously
into nude mice eliciting pancreatic
tumors
Restoration of immune functions
(presence of human immunoglobulin
and antigen-specific T cells)
LNCaP-bcl-2 cells induced earlier,
larger, and hormone-refractory
prostate tumors in nude mice
Factor IX was being synthesized and
delivered to the circulation for over 6
months
Human factor IX was detected in the
bloodstream of nude mice in small
quantities for one week
Sustained expression of factor IX for
over six months without any apparent
adverse effects on the recipient mice;
however, the levels of the factor IX
protein secreted into the plasma (10
ng/ml for 107 injected cells) were not
sufficient to be of therapeutic value;
100 times higher amounts of factor
IX were needed
Therapeutic levels of factor VIII in
the blood of the animals for 24 hours
Therapeutic levels of factor VIII in
blood of animals for 1 week after
surgical implantation into the
peritoneal cavity in SCID mice of 15
million cells in the form of neoorgans
This model allows up to 100-fold
induction of the hGH gene and can be
finely tuned to lower levels of
induction
hGH could be detected in serum for 3
months; myoblasts were fused into
preexisting multinucleated myofibers
that were vascularized and innervated
Murine fibroblasts transduced ex
vivo with HSV TK retroviral vectors
caused complete regression of
gliomas in rat brain after intratumor
injection
Animals treated with 0.1 mg/Kg
ganciclovir exhibited a significant
reduction in tumor growth
Ferrari et al, 1991
Rafo et al, 1995
Dai et al, 1992
Gerrard et al, 1993,
Dai et al, 1992; Yao et al,
1994
Zatloukal et al, 1994
Dwarki et al, 1995
Delort and Capecchi,
1996
Dhawan et al, 1991
Culver et al, 1992
DiMaio et al, 1994
HSV-tk proliferative retroviru Traction retinal detachment results Significant inhibition of PVR (killing Kimura et al, 1996