Beruflich Dokumente
Kultur Dokumente
Department Of Biotechnology,
Presented by,
Avinash V
• Introduction
• Gene therapy overview
• Suicide gene therapy
• In vitro and In vivo mechanism
• Advantages
• Limitations
• Future
• Conclusion
Suicide – to kill oneself
Birth of Gene therapy approaches –
frequency of occurrence , lack of
efficacy of available oncogenetic
treatment, diverse genetic
background of different malignant
diseases
Suicide gene therapy
Genetic manipulation
Gene Therapy- original concept
70’s – utility of DNA as Theurapetic agent
1971 – First unauthorized gene transfer into
humans by stanfield Rogers
80’s –various preliminary studies
1990 (s) – first approved gene treatment
treatment for trial, better and safer vector
development, increase in understanding of many
diseases
2000 – First patients cured with aid of gene
therapy
2003 – first gene therapy protocol approved for
clinical practice in china against head and neck
squamous cell carcinoma
Target disease Gene delivered Phase of
Clinical
Inherited disorders development
Hemophelia FIX or FVIII I
Cystic fibrosis CFTR I
Chronic granulomatous p47phox I
disease
Acquired diseases
Cancer
head and neck squamous cell p53 Approved
carcinoma
Glioma HSV-TK I/II
Alzheimer’s disease NGF I
Lower limb ischemia VEGF II
Features of optimal vectors – Ability to
transduce cells of different tissue, target the
vectors to certain tissue, And a stable,
sufficiently long lasting and regulated
transgene expression in target tissue.
Problems – side effects due to hazardous
interaction and immunological reactions
Viral vectors – adeno, retro, lenti, herpes
viruses
Non viral vectors – less toxic, low
immunogenicity , low efficacy, unlimited
transgene capacity. – cationic liposome's,
cellular gene delivery, stem cells
66.4 percent gene therapies are
aimed against cancer
Focus on 3 major themes – discover
new means of killing, improvement of
gene delivery and vector systems,
translation of preclinical studies to
clinical protocols and trials
Melanoma, leukemia, prostrate,
ovary, squamous cell carcinoma
Gene therapy strategy Example gene
p53
Tumor suppressor gene BRCA1
RB
ERBB2
Oncogene
KRAS
Anti-angiogenesis VEGF
Immunotherapy IL-2
Chemo-protective therapy 1
Adeno virus
Virotherapy, oncolysis
Herpes virus
HSV-tk
Suicide gene therapy
CD
Original concept
Basic strategy
herpes simplex virus thymidine
kinase (HSV-tk) gene coupled with
the pro-drug ganciclovir (GCV)
cytosine deaminase (CD) gene
coupled with the pro-drug 5'
fluorouracil (5-FU)
xanthine guanine phosphoribosyl
transferase (XGPRT)
purine nucleoside phosphorylase
(Besnard et al., 1987, Mroz and
Moolten., 1993).
Thymidine kinase
HSV-tk – ability to phosphorylate
broad range of guanosine analogues
like Ganciclovir, acylovir, buciclovir,
penciclovor
Molten observed
Culver and coworkers – first noticed
GCV-p from HSV-tk positive cells to
wild type
Touraine and co-workers – relation
b/w gap junctions and Bystanders
effect
Other routes also proposed by many
other researchers
Initial
suggestions from studies
Bi and coworkers (1993)- used
radiolabelled GCV
Gap junctions
In vivo Mechanism
rapidly replicating tumor cells are
more susceptible to impairment of
DNA synthesis
chemotherapy resistant tumors can
be made sensitive when genetically
modified
HSV-tk/GCV-treated tumor cells have
the ability to kill neighboring tumor
cells through the bystander effect
showed enhanced growth, invasiveness,
resistance to chemotherapy of tumors transduced
with HSV-TK.
GCV uptake and its low affinity to HSV-TK may
also limit the clinical efficacy of this treatment
form
GCV might not be the best substrate for HSV-TK
due to its inadequate transport into the cells as
well as the low levels of GCV phosphorylation.
It has also been noticed that sensitivity to GCV
and the Bystanders effect varies between
different tumor cells lines,
Showed that GCV uptake increased along with the
percentage of HSV-TK expressing cells, which was
combine traditional cancer treatment methods with
gene therapy.
Enhanced therapeutic effect has also been observed
by combining prodrug therapies.
combining two widely used suicide genes
HSV-TK in combination with other genes has
demonstrated increased efficacy
Immune system related gene has been combined with
HSV-TK
In addition to the combinations of different genes and
HSV-TK/GCV therapy, there are a number of other
interesting treatment combinations.