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G M Institute of Technology

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
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
 2000 – First patients cured with aid of gene
 2003 – first gene therapy protocol approved for
clinical practice in china against head and neck
squamous cell carcinoma
Target disease Gene delivered Phase of
Inherited disorders development
Hemophelia FIX or FVIII I
Cystic fibrosis CFTR I
Chronic granulomatous p47phox I
Acquired diseases
head and neck squamous cell p53 Approved
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
 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
Tumor suppressor gene BRCA1
Anti-angiogenesis VEGF

Immunotherapy IL-2

Chemo-protective therapy 1

Adeno virus
Virotherapy, oncolysis
Herpes virus
Suicide gene therapy
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,
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
Other routes also proposed by many
other researchers
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
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
 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
 In addition to the combinations of different genes and
HSV-TK/GCV therapy, there are a number of other
interesting treatment combinations.

promising solution to kill tumors

 Even though with all these limitations and hurdles that this
suicide gene therapy is facing, there seems to be a lot of
research and scientific works going on to improve the
efficiency and effectiveness of this way of treating cancer.
May be in a few years the hurdles will be overcome and the
suicide gene therapy would come to the markets as the
most efficient and effective therapy for Cancer and other
 TIINA WAHLFORS: Enhancement of HSV-TK/GCV suicide gene therapy
of cancer
 Tumor killing using the HSV-tk suicide gene
 Rajagopal Ramesh1, Aizen J. Marrogi1 and Scott M. Freeman2,3
 1. Department of Surgery and Gene Therapy Program, LSU School of
Medicine, New Orleans, Louisiana, USA.
 2. Department of Pathology, Tulane University School of Medicine, New
Orleans, Louisiana, USA.
 Pasanen T., Karppinen A., Alhonen L., Jnne J. and Wahlfors J. Polyamine
biosynthesis inhibition enhances HSV-1 thymidine kinase/ganciclovir-
mediated cytotoxicity in tumor cells. Int J Cancer (2003) 104, 380-388
 Pasanen T., Hakkarainen T., Timonen P., Parkkinen J., Tenhunen A.,
Loimas S. and Wahlfors J. TK-GFP fusion gene virus vectors as tools for
studying the features of HSV- TK/ganciclovir cancer gene therapy in
vivo. Int J Mol Med (2003) 12, 525-531
 Wahlfors T., Hakkarainen T., Jnne J., Alhonen L., and Wahlfors J. In vivo
enhancement of Herpes simplex virus thymidine kinase/ganciclovir
cancer gene therapy with polyamine biosynthesis inhibition. Int J
Cancer, in press.
 Wahlfors T., Karppinen A., J?nne J., Alhonen L. and Wahlfors J.
Polyamine depletion and cell cycle manipulation in combination with
HSV thymidine kinase/ganciclovir cancer gene therapy. Int J Oncol, in