Cancer therapy at protein level

1. CANCER
THERAPY AT
PROTEIN
LEVEL
Dr. P. Suganya
Assistant Professor
Departement of Biotechnology
Sri Kaliswari College
(Autonomous),Sivakasi

2. Cancer Therapy
• Cancer treatment is use of Surgery, radiation, Medications and other therapies to cure a cancer
• Shrink a cancer or stop the progression of a cancer
• New anti cancer therapies have accompanied the classical approaches of surgery and
radiotherapy, Chemotherapy
• Aim of the treatment to inhibit specific molecular targets namely altered or deregulated
proteins, which offer the possibility of individualized therapies
• The specificity and efficiency of these new approaches, however bring about a number of
challenges essential to specificity and quantify protein targets in tumour tissue for reasonable
use of targeted therapies

3. Expression
– The analysis of tumor specific protein expression profiles prior to therapy has
been suggested as the most effective way to predict possible therapeutic
results.

4. RPPA (Reverse Phase Protein
Microarray)
– Is a promising technology,
– Requirements while enable the quantitative measurement of proteins
– It may provide to quantify therapeutic targets and diagnostic markers in the near future
and reliably screen for new protein targets
– The deregulation of signalling pathways in tumors can lead to enhanced cancer cell
growth, proliferation, survival, invasion and metastasis or reduced apoptosis.

5. Kinase
– Receptor molecules or downstream regulators of signalling cascades
– E.g Human epidermal growth factor receptors 1 (EGFR) and (HER2)
– Both kinase are targets for anticancer drugs
– And to analysed for their expression by clinically approved tests, such as
ImmunoHistochemistry (IHC) and Fluorescence in situ hybridization (FISH)

6. EGFR
– Is gene amplification
– Essential found in human cancers in gliomas, this deregulation is often
associated with structural rearrangements leading to inframe detections in the
extracellular too domain of the receptor

7. HER2
– Is found in 25 to 30% of
– human breast cancer c
– an be mediated either by
– transcriptional activation
– or gene amplification

8. HER2
– Target for the treatment of Metastasized gastric cancers and thus, the
anticancer drug Herceptin was approved for the treatment of advanced gastric
carcinomas
– Recent evidence-that down stream signalling molecules may be better
predictors for a response to a HER2 directed therapy than the receptor itself

9. Drugs Currently used for targeted therapy
Drug Tumor Type Target Detection method
Trestuzumb
(herceptin)
Metastatic breast cancer,Gastric cancer HER2 IHC (Immunohistochemistr), FISH, IST
(Insitu)Hybridization)
Cetuximab Metastatic Clorectal cancer EGFR-Epidermal Growth factor receptor IHC, FISH
Imaatirub Mesylate (Glecvec) CML, GIST with acticvation receptor tyrosine
kinase, other sacromas
BCR labl, c-kit, PDGER chronic Myeloid
leukemia
IHC_ Immunohistochemistry
Bevacizamab Colorectal cancer VEGF- Vacular endothelial
growthfactr.homodimeria glycoptn
IHC-Immunohistochemistry
Gefitirib (Iressa) Non- small cell Lung cancer Mutant_EGFR Mutaational analysis, Immunoblotting

10. Drug Tumor Type Target Detection method
G3139 (Genta, Barkles) Hematologic malignaanearcies and malignant melanoma Antiapoptotic gene bcl-2 Immunophenoty-piug byIHC
Erlotinib (Traveca) Non-small-cell Lung cancer Mutant EGFR Mutational analysis, immunoblotting
Rapamycin
RADOO1
Breast, Prostate, renal cancer mTOR ELISA
BAY43-9006 Melanoma(skin) RAF kinase Mutational analysis
BMS354825 GIST- Gastrointestinal stromal tumor KIT IHC
Lapatinib Breast cancer EGFR, HER2 IHC, EISH, ISH
Sunitinib Renaal cell cancer VEGFR, PDGFR, CKIT, FIT-3 IHC
Pertuzumab
Dasatinib
Breast cancer
Breast cancer
HER2
Bcr labl
IHC, FISH,ISH
RT-PCR

11. Drug Tumor Type
Thyroglobulin Thyroid
Carcino embryoine antigen Colon breast lung
Cathepsin- specific Ag Prostate
CA19-9
CA195
CA50
Pancreaaas
Pancreatic oncofetal Ag
CA549-3
Breast
Breast cancer Mucin CA549 Breast
C-peptide Insulinoma
CA125 Ovary
Ovarian cancer antigen
CA72-4
Hydroxyproline
Stomach, Ovary, colon, breast, lung

12. – Re-analytical parameters influencing the quality of biological samples and
hindexing the quantitative analysis of potential biomarkers and drug targets
– Meddication before surgery and type of Anaesthesis- source of –Infkuenca on
gene or protein expression

13. RPPA
– Protein lysates- spotted nitrocellulose- coated –glass slides
– A) Single proteins (eg PAI) can be detected by an antibody assay- similar to western blotting
analysis
– Specific antibody binds to the spotted protein
– After binding of an enzyme- coupled secondary antibody, protein expression can be
measured by light- or- fluorescence- based as well as colorimetric methods

14. – B) Each ssample is spotted in triplicate and in a sex-step dilution series to
ensure the –quantitative measurement –of the target protein- in the linear
detection range
– The normalization of every antibody- detected on the slide is performed using
a total protein- stained slid (e.g Sypro-Ruby protein strain)

15. A)

16. B)

18. – RRPAs – can be – implemented in nearly all areas ---- regarding the field of –
personalized medicine
– Recent studies--------- in RRPAs technology is a very promising tool for signalling
pathway--- profiling of human tissue and cell lines -------------- produce valuable
information ----------- development of new therapeutics or Patient solution.
– Feinberg et al was the first to utilize a micropot technique to detect antigens in
serum
– In 2001 another study used RPPAs to show the activation of pro-survival
proteins and pathways during prostate cancer progression

19. – In addition to classical biomarkers screenings RPPAs is also adequate tool to
compare different types of samples regarding their protein expression profiles

20. The Integration of DNA, RNA and
protein profiling into a routine works
– Parallel analysis of the morphology as well as DNA, RNA
and protein expression profiles can provide a very
description of any given tumor entity, there by select the
patients for targeted, Indivialualized therapy.

21. Conculsion
– Scope of New targeted cancer therapy
– Approaches new technology
– Which meet the issue of fast and peruse target detection and quantification
are desperately needed
– RPPAs is a very promising tool in this regard

22. – THANK YOU