ROLE OF TRANSGENIC ANIMAL

IN TARGET VALIDATION

SUBMITTED BY
SYAMA. J.S
SWATYY
SHANI

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 TRANSGENIC ANIMALS
 Transgenic animals are animals that have a foreign gene deliberately
inserted or deleted from their genome.
 For drug discovery and development, genetically engineered mice and
genetically engineered rat models are constructed
standard techniques for insertion arev:
random introduction of a foreign gene(transgenic)
precise targeting to eradicate(knock out)an endogenous
gene

Transgenic animals are constructed either by

 Microinjection or systemic gene delivery
 Embryonic stem cell
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 Retroviral vector method
Another major new approach is the generation of

Conditional Transgenic Genetically Engineered Mouse

 Such animals bear engineered genes that include a tissue-
specific promoter.
 They are controlled by a ligand-inducible enhancing or
repressing element.
 Which limits the impact of the modified gene to a given
developmental stage or organ, or even to a given region or cell
type within an organ.

The utility of transgenic models in drug discovery and development

programs depends greatly on the extent to which their genotypic and
phenotypic characteristics have been mapped 3
In discovery projects, almost all engineered lines are newly created.

 It require substantial analysis before they can be used for anything more
than basic mechanistic experiments.

Transgenic animals can be used broadly in discovery of new medicine

Transgenic system helps
 deliver drug targets
new treatment opportunities
to maximize the quality of new chemical compounds

The main utility of transgenic animals during target validation phase, i.e.
increasing the possibility that a specific target is a safe and efficient drug target
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Knock out animals have a major role in
 analyzing the safety
specificity

metabolism and efficacy of future drug candidates

Identify and validating new drug targets

The animal welfare issues associated with transgenic animals are fundamentally
not different from other animals used in biomedical research

The development and use of transgenic animals in research is subject to

stringent control including government regulations.

The 3R principles and ethical review are rigorously applied. 5

 In preclinical development efforts to elucidate efficacy or toxicity of new
therapeutic candidates ,the use of engineered animals depends on the entity
being tested

 Small molecules typically evaluated using genetically engineered mice models

available from mutant mouse resource centers or commercial vendors

 The use of engineered animals in drug discovery and development is limited to

inventing and testing novel agents rather than to producing drugs

 Some important roles of transgenic animals in drug discovery and development

are
Identification of new drug targets
Disease modeling and drug development
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Replacing non-human primate (NHP) models
USES OF TRANSGENIC ANIMALS IN DRUG DISCOVERY
1.BASIC RESEARCH GENOTOXICITY
2.APPLIED RESEARCH
CARCINOGENICITY

The first and most common paradigm in discovery projects is to generate models
of de novo by engineering a novel gene of interest in hopes of achieving a useful
phenotype

The focus for transgenic animals in such programs is to either define the role of a
gene in the pathogenesis of a disease process or to evaluate the functions of
previous uncharacterized genes

Alternatively, an existing model for sufficient phenotypic analysis has been

defined may be evaluated.eg; genetically engineered animals have been used to
assess the mechanisms, mutagenic, and carcinogenic properties and specificity of
xenobiotics 7
 The choice between transgenic models and existing one in drug discovery
development is also found on financial and time constraints

 Most drug discovery developments use novel GEM and GER to gain and protect
proprietary intellectual property concerning molecular targets of interest and they
will continue to do so regardless of time and expense

 The time required for complete assessment is measured in months to years

 The drug discovery development choice is predicted on

 ready accessibility of well characterized animals (reduces the time required
to fully develop a new in-house model)
 growing literatures regarding their responses to xenobiotics challenge and
their increasing acceptance at some regulatory agencies.
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BASIC RESEARCH
Target identification
Target Validation Efficacy Evaluation
Compound validation
Specificity evaluation
 to explore the essential molecular mechanisms whereby a gene regulates the
balance between health and disease

 Characterize and validate target, and to confirm proposed therapeutic strategy will
be effective

 In most firms, they are performed using newly generated proprietary models
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 detailed knowledge of physiological machinery that engenders a positive
phenotype, as such understanding is necessary before a model can be used to
make predictions suitable for guiding drug discovery development work.

1.Target identification: Identifying novel genes that are potential drug targets.
 Target interaction: Interaction with a perfect target should result in prevent or
regression of disease
 Not lead in serious target related side effects
 Target should belongs to a class of molecules that could be
affected by small orally available molecule
This technology have challenges for in vivo use including
Delivery
Ability to penetrate cell membrane 10

Specificity
 Therefore the most widely used in vivo technology is transgenic technology

 Compound Validation: It is the possibility of test compound and potential

future medicine.
 Efficacy Evaluation :If target is not properly linked to disease
or that the compound has not been tested in relevant efficacy
models
 e.g.;
possible to measure parameters in healthy animals(BP,
Stomach ulcer)
 Specificity Evaluation : Knock out animals can be used both
to understand target related toxicity and off target toxicity

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Target identification
eg; To evaluate the impact of long term treatment in engineered mice to lack β-
secretase , the brain enzyme proposed as the primary molecule responsible for
the generation of neurotoxic amyloid beta fibrils in patients with Alzheimer's
disease

Suppose we identify disease ie Alzheimer

Translated this disease model into animals.

Animal model of Alzheimer disease( Streptozocin is used to induce beta amyloid

protein in hippocampus)

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Target identification by proteomics and genomics. Over expression of APP
protein is responsible for disease.
 Target Validation

 Validate that really the APP is responsible for AD

 Deliver mutant APP gene into animal

 Progenies having mutant APP gene over expressed

 Target validated

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APPLIED RESEARCH
 Modern drug discovery development programs for small molecule drugs and
chemicals exploit several commercially available transgenic models during
late stage preclinical research to assess the capacity for xenobiotic induced
genetic damage

 2 principle applications are bioassays for mutagenicity and carcinogenicity

 Mutagenicity- several transgenic models can be purchased to evaluate

mutagenic activity of xenobiotics in vivo

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 The high cost and time required for such tests in terms of labor, money, and
time suggests the rodent bioassays will be used to supplement rather than
replace the rapid bacteria based screening assays

 The standard models used for in vivo mutagenicity testing in drug

discovery development target either endogenous genes for enzymes that
participate in DNA repair or bacterial transgenes that encode marker
proteins

 Knock out mice were used in which the purine salvage pathway required
for DNA repair has been disrupted by eliminating one copy of gene
encoding either adenine phosphoribosyl transferase or hypoxanthine
phosphoribosyl transferase

 In the absence of xenobiotic exposure, the spontaneous mutation rate for

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such endogenous genes is low, affected by the animals age

 Carcinogenicity- Transgenic models used for carcinogenicity
testing in drug discovery development bears
 induced mutations on tumor suppressor gene
 an oncogene is over expressed
 both suppressor and oncogene is altered

 These engineered defects provide a built in predisposition to xenobiotic

induced genetic damage in the form of pre existing ‘first hit’ in all tissues.

 Thus the rationale for using transgenic models in carcinogenicity risk

assessment is that the nature of their induced mutations resembles the main
spontaneous alterations that form genetic basis for many human neoplasms 16
 Transgenic models provide a readout of a drug candidates carcinogenic
potential of greater sensitivity but in much less time

 An advantage of transgenic carcinogenic models is the ability to harvest

information regarding the action between specific tumor related genes
and new chemical entity

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 CONCLUSION

 In vivo target validation is a key step in drug development program to

decipher gene functions.

 Gene based biomedical research offers best hopes yet for curing the major
diseases which will still affect mankind .

 The use of transgenic animal is central to realising that hope.

 The appropriate use of transgenic animal is a positive development with

demonstrated significant medical benefits
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 REFERENCE

 Blake RA, 2007 “Target validation in drug discovery” Journal of Methods

Mol Biology,Pg 367-77.
 The use and usefulness of transgenic systems in the discovery of new
medicines by professor Jan Tornell, 2004

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