lung cancer: the breathtaking battle of tkis and egfr mutants
DESCRIPTION
Bobby Hodges, Elizabeth Ross, Natalie Simak , Keith Solvang and Eliot Vildaver. Lung Cancer: The Breathtaking Battle of TKIs and EGFR Mutants. Diseased Genome Hospital. http:// blogs.usask.ca/medical_education/archive/ElksRehabFront%2520Door.jpg. - PowerPoint PPT PresentationTRANSCRIPT
Lung Cancer: The Breathtaking Battle
of TKIs and EGFR MutantsBobby Hodges, Elizabeth Ross, Natalie Simak, Keith Solvang and
Eliot Vildaver
DiseasedGenomeHospital
http://blogs.usask.ca/medical_education/archive/ElksRehabFront%2520Door.jpg
http://1.bp.blogspot.com/_HI2mCx5P3H0/SLlEQoHj8AI/AAAAAAAAA8Q/My7tf78TNc8/s320/hospital-bed-786282.jpg
http://img.quamut.com/chart/9471/02_healthy_sick_x-ray.jpg
http://www.cardiothoracicsurgery.org/content/2/1/16/figure/F1?highres=y
Where Are We Headed?What Is Lung Cancer?
• Types
• Risks
Predisposition
Mutations
• EGFR
• Other proteins
http://www.freefoto.com/images/21/16/21_16_2---Single-Carriageway-Road_web.jpg
Cancer
http://www.odec.ca/projects/2005/thog5n0/public_html/Tumour.gif
History of Lung Cancer
http://www.icowman.com/uploadfiles/2007072409225175.jpg http://www.icowman.com/uploadfiles/2007072409225175.jpg
Pathology
NON SMALL CELL
Squamous cell carcinoma
Adenocarcinoma Large-cell carcinoma
SMALL CELL
Bronchi Grows rapidly Spreads quickly
http://images.google.com/imgres?imgurl=http://www.dkimages.com/discover/previews/852/20223038.JPG
http://images.google.com/imgres?imgurl=http://upload.wikimedia.org/Lynch et al., (2004)
Risk Factors
http://www.taconichills.k12.ny.us/webquests/noncomdisease/lungcancerpic.jpg
You Are Not Alone
This year approximately 200,000 people will be diagnosed with lung cancer
NSCLC accounts for 80% of all lung cancers
http://www.cdc.gov/cancer/lung/statistics/http://seo2.0.onreact.com/wp-content/uploads/2008/09/you-re-not-alone.jpg
Epidemiology
http://medicineworld.org/cancer/lung/ http://images.google.com/
What Next?
While there is no known cure as of right now, there ARE options!
http://dspace.mit.edu
ImatinibGefinitib
Cisplatin
http://dspace.mit.edu
Cisplatin
Imatinib
Gefitinib
EGF ER
BB2
EGFR
Nucleus
P
Protein Cascade
?
?
P
P
EGF ER
BB2
EGFR
PI3K
AKT
P
P
MET
P
VEG
F-A
Cisplatin
Imatinib Gefiti
nib
Nucleus
Protein Cascade
PDGFR
Protein Cascade
Implications of Chromosomal Loci in the Development of Lung Cancer
http://rosenblumtv.files.wordpress.com
http://www.pharmgkb.org
http://academic.evergreen.edu
Chromosomes
Compact structures of DNA
Contains thousands of genes
Centromere
Chromatid
Genes
http://staff.jccc.net/PDECELL/celldivision/chromosome1.gif
Single Nucleotide Polymorphisms (SNPs)
Single variation in nucleotide base pairs
Alleles are a result of SNPs within gene sequence
SNP’s are the basis for lung cancer susceptibility
http://www.dnalandmarks.com/english/pictures/what_are_snps.jpg
Genome-wide Association Studies
Variation across genomes
Genotype cases along with controls
Find roles that genes play in disease
Large sample sizes
http://www.blog.speculist.com/archives/dna2two
Identifying Regions On Chromosomes
Centromere
p-arm
q-arm
6 Regions Away
Region 5q6
5http://images.google.com/
Three Regions of Association
http://upload.wikimedia.org/ http://upload.wikimedia.org/
Nicotinic Acetylcholine Receptor Genes Found on 15q24 and 15q25
Nicotine binds and stimulates acetylcholine ion channel-linked in
neurons
http://www.jyi.org/articleimages/88/originals/img0.jpg
How were these regions discovered?
(Hung et al., 2008)
15q24 highly linked with smoking behavior
(Thorgeirson et al., 2008)
TERT gene found on 5p15.33
Cancerous Growth
http://images.google.com/i
5p15.33 shows similar results
(McKay et al., 2008)
P
P
EGF ER
BB2
EGFR
PI3K
AKT
P
P
MET
P
VEG
F-A
Cisplatin
Imatinib Gefiti
nib
Nucleus
Protein Cascade
PDGFR
Protein Cascade
How Do Cells Proliferate?
EGFR/ERBB2 Forms a Heterodimer
EG
FR
P
EG
FR
EGF
ProliferationSurvival
Apoptosis
ERBB2
Nucleus
EG
FR
P
Protein Cascade
ERBB2
How Can This Lead to Cancer?
Mutated EGFR Leads to Cancer
ERBB2EG
FR
Nucleus
Proliferation
P
Protein Cascade
Mutated EGFRs have Distinct Patterns of Tyr Phosphorylation
(Chen et al., 2006)
Tyrosine Kinase Inhibitors (TKIs)Gefitinib
EG
FR
ATP TKI
ATP binding cleft on the kinase
TyrosineKinasedomain
Gefitinib Suppresses Phosphorylation of Most EGFR
Mutants
(Chen et al., 2006)
What About the Proteins in the
Signaling Pathway?
Effectiveness of Gefitinib is Dependent on Concentration
(Pedersen et al., 2005)
High Doses of Gefitinib Decreases Proliferation of Mutated EGFR Cells
(Pedersen et al., 2005)
Gefitinib has Variable Growth-Inhibitory Effects
(Chen et al., 2006)
P
P
EGF ER
BB2
EGFR
PI3K
AKT
P
P
MET
P
VEG
F-A
Cisplatin
Imatinib Gefiti
nib
Nucleus
Protein Cascade
PDGFR
Protein Cascade
What Are the Roles of Proteins Associated with Lung Cancer?
http://www.taconichills.k12.ny.us/webquests/noncomdisease/lungcancerpic.jpg
Recap
Nucleus
N-terminus
C-terminus
P
Serine
P
Threonine
Tyrosine
P
Leucine
Lysine
Alanine
How Does The Pathway Work?
Domain Attraction
ERBB2 P
PI3KSH2
Nucleus
P
Protein Cascade
Change in Gene
Expression
Why are EGFR and ERBB2 Special?
Main Contributors to NSCLC
(Rikova et al. 2007)
Link between Concentration and Interaction
Green and Blue circles = Proteins involved in cascade
Red lines = Interactions
Red circles = EGFR Receptor
(Jones et al. 2006)
Other ERBB2 Associations
(Jones et al. 2006)
No
. p
rote
in r
ece
pto
rs
No
. p
rote
in r
ece
pto
rs
No
. p
rote
in r
ece
pto
rs
Affinity Threshold (nM) Affinity Threshold (nM) Affinity Threshold (nM)
Are there other types of Proteins?
http://en.wikipedia.org/wiki/Proteins
Other Contributors to NSCLC
(Rikova et al. 2007)
ROS ALK MET PDGFRα ErbB2 EGFR VEGFR-10
5
10
15
20
25
30
35
40
45
50
Tumor Samples
Cell Lines
Chart configured from Table1 of Rikova et al. 2007
Specifically, some proteins more than others
Am
ount
of
over
phos
phor
ylat
ed p
rote
in in
sam
ples
Previous Research, Amplification of MET
(Engelman et al., 2007)
P
P
EGF ER
BB2
EGFR
PI3K
AKT
P
P
MET
P
VEG
F-A
Cisplatin
Imatinib Gefiti
nib
Nucleus
Protein Cascade
PDGFR
Protein Cascade
What is the Role of Angiogenesis in Lung Cancer?
http://www.taconichills.k12.ny.us/webquests/noncomdisease/lungcancerpic.jpg
Angiogenesis and VEGF
http://www.researchvegf.com/researchvegf/multimedia/index.m#type-slides
Tumor Angiogenesis Is Unregulated
VEGF-A
VEGFR
Angiogenesis
Tumors
PDGF-β
PDGFR-β
PI3K / Akt Pathway in Lung Cancer
P85-α PIP2
P
P
PTEN
P
PIP3P
P
P
P110-α
PI3KAKT
SH2
PP
Endothelial Cell
Akt is a Downstream Regulator
AKT
P
P
P
BAD
MDM2
mTORGrowth,
Translation
Apoptosis
p53
P
Nucleus
FLJ10540 Is An Oncogene
Cancer Cell
Proto-Oncogene Oncogene
VEGF-A Upregulates FLJ10540
(Chen et al., 2009)
FLJ10540 Mediates Cell Migration and Invasion Through PI3K/Akt
Giemsa-Stained Cells
(Chen et al., 2009)
Recap: FLJ10540 Missing Link in PI3K/Akt
P85-α PIP2
P
P
PTEN
P
PIP3P
P
P
P110-α
PI3K
AKT
SH2
PP
VEGF-A
VEGFR
FLJ10540
Endothelial Cell
Imatinib is a TKI
http://www.medicinescomplete.com/mc/martindale/2007/images/MRT9229C001.gif
Binds to ATP binding site of many enzymes:
p-PDGFR-β Inhibitor Reduced IFP, Hypoxia
Decreased VEGF Expression
Imatinib Targets IFP and Hypoxia
http://www.medscape.com/pi/editorial/clinupdates/2000/583/art-tu02.fig04.jpghttp://www.nature.com/nm/journal/v7/n9/images/nm0901-987-F1.gif
Normal
Tumor
Imatinib Reduces p-PDGFR-β and VEGF Expression
(Vlahovic et al. 2007)
Imatinib Reduces Hypoxia and MVD
Vlahovic et al. 2007
Imatinib Treatment
http://onctalk.com/wp-content/uploads/2008/01/angiogenesis-summary.jpg
Imatinib
P
P
EGF ER
BB2
EGFR
PI3K
AKT
P
P
MET
P
VEG
F-A
Cisplatin
Imatinib Gefiti
nib
Protein Cascade
PDGFR
Protein Cascade
Nucleus
Treatments
Dr. Vildaver
• ImatinibDr. Ross
• Gefinitib
Discovery of Cisplatin
(Rosenberg et al., 1969)
Old Model of Cisplatin
Cisplatin
NucleusSimple Diffusion
Pt
NH3NH3
GGP73
C-Abl
Cell Cycle Arrest
&Apoptosis
P53
HMG1?HMG1
?
How Pt Drugs Enter Cells
nucleus
Pt
Pt
Pt
Pt
Pt
Transporter
protein
Transporter
protein
Pt
(Hall et al., 2007)
Crosslinks
crosslink
nucleus
Pt
NH3
NH3
Cell Cycle Arrest
&Apoptosis
GG
1,2-intrastrand d(GpG)adduct
(Ohndorf et al., 1999)
Pt-DNA adduct
http://www.unc.edu/~shantanu/images/bkstcg_aacr07.png
High Mobility Group Proteins
Pt-DNA Adduct
Apoptosis
HMGB
HMGB
P73
P53
(Stross et al., 2002)
(Ohndorf et al., 1999)
*Nucleotides
crosslinked
Bold- nucleotides contacted by the
protein
Phenylalanine Mutations
(Ohndorf et al., 1999)
Old Model of Cisplatin
Cisplatin
Nucleus
?Simple Diffusion
Pt
NH3NH3
GGP73
C-Abl
Cell Cycle Arrest
&Apoptosis
P53
HMG1?HMG1
?
2 Distinct Regulating Mechanisms
tyrosine kinase
mismatch repair protein
cisplatin
homologue of p53
Tumor suppresor
(Gong et al., 1999)
Role of p73 in Cell Death
P53 knockout= less sensitive to CDDP, survived better than wild type in lower concentrations of CDDPMLH1-/- & Abl= less sensitive to CDDP , survived better than wild type in lower concentrations of CDDP
(Gong et al., 1999)
New Model of Cisplatin
CisplatinNucle
usSimple Diffusion
Pt
NH3NH3
GG
P73
Cell Cycle Arrest
&Apoptos
is
P53
HMG1
HMG1
ATM
TransporterProteins
C-AblP
Cisplatin
Imatinib
Gefitinib
EGF ER
BB2
EGFR
Nucleus
P
Protein Cascade
??
P
P
EGF ER
BB2
EGFR
PI3K
AKT
P
P
MET
P
VEG
F-A
Cisplatin
Imatinib Gefiti
nib
Nucleus
Protein Cascade
PDGFR
Protein Cascade
Cocktail of Drugs
http://immunodefence.com/ii/gefitinib.gif
http://www.themesotheliomalibrary.com/cisplatin.jpg
http://www.specialityformulations.com/pcat-gifs/products-small/imatinib.jpg
?
Current & Future Hypotheses
New plasma membrane proteins have unique mechanisms and effects on NSCLC
Gefitinib effectively inhibits particular EGFR mutants
SiRNA could be used as a possible treatment for various mutated proteins
Future Studies
How does cisplatin enter the nucleus?
Roles and mechanisms behind the various mutated proteins involved in lung cancer
New drugs , secondary mutations
Acknowledgements
Professor Sleiter
?Schmooing yeast!!!
Acknowledgements
Dr. DebBurman
Acknowledgements
Mike Fiske
Abid MR, Guo S, Minami T, Spokes KC, Ueki K, et al. (2004) Vascular endothelial growth factor activates PI3K/Akt/forkhead signaling in endothelial cells. Arterioscler Thromb Vasc Biol 24: 294-300.
Agami, R., Blandino, G., Oren, M., and Shaul, Y. (1999). Interaction of c-Abl and p73alpha and their collaboration to induce apoptosis. Nature 399, 809-813.
Alberg AJ, Ford JG, Samet JM; American College of Chest Physicins. Epidemiology of lung cancer: ACCP evidence-based clinical practice guidelines (2nd edition). Chest. 2007; 132:29S-55S.
Alberts, B., Bray, D., Hopkin, K., Johnson, A., Lewis, J., Raff, M., Roberts, K., and Walter, P. 2004. Essential Cell Biology. Garland Science. New York, pp740.
Chanock, Stephen J., Hunter, David J. 2008. When the smoke clears. Nature. 452: 537-538.
Chen, C.H., Lai, J.M., Chou, T.Y., Chen, C.Y., Su, L.J., Lee, Y.C., Cheng, T.S., Hong, Y.R., Chou, C.K., Whang-Peng, J., Wu, Y.C., and Huang, C.Y. (2009). VEGFA upregulates FLJ10540 and modulates migration and invasion of lung cancer via PI3K/AKT pathway. PLoS ONE 4, e5052.
Chen, Y.R., Fu, Y.N., Lin, C.H., Yang, S.T., Hu, S.F., Chen, Y.T., Tsai, S.F., and Huang, S.F. (2006). Distinctive activation patterns in constitutively active and gefitinib-sensitive EGFR mutants. Oncogene 25, 1205-1215.
Engelman, J.A., Janne, P.A., Mermel, C., Pearlberg, J., Mukohara, T., Fleet, C., Cichowski, K., Johnson, B.E., and Cantley, L.C. (2005). ErbB-3 mediates phosphoinositide 3-kinase activity in gefitinib-sensitive non-small cell lung cancer cell lines. Proc. Natl. Acad. Sci. U. S. A. 102, 3788-3793.
Ferrara, N. and Kerbel, R.S. (2005). Angiogenesis as a therapeutic target. Nature 438, 967.
Franks, L.M. and Teich, N. (1986). Introduction to the Cellular and Molecular Biology of Cancer (New York:Oxford University Press).
Gao, B., Lee, S.M., and Fang, D. (2006). The tyrosine kinase c-Abl protects c-Jun from ubiquitination-mediated degradation in T cells. J. Biol. Chem. 281, 29711-29718.
Hall, M.D., Okabe, M., Shen, D.W., Liang, X.J., and Gottesman, M.M. (2008). The role of cellular accumulation in determining sensitivity to platinum-based chemotherapy. Annu. Rev. Pharmacol. Toxicol. 48, 495-535.
Helotera, H. and Alitalo, K. (2007). The VEGF Family, the Inside Story. Cell 130:591-592
Hung, Rayjean J. et al. 2008. A susceptibility locus for lung cancer maps to nicotinic acetylcholine receptor subunit genes on 15q25. Nature. 452: 633-637.
Huang, J.C., Zamble, D.B., Reardon, J.T., Lippard, S.J., and Sancar, A. (1994). HMG-domain proteins specifically inhibit the repair of the major DNA adduct of the anticancer drug cisplatin by human excision nuclease. Proc. Natl. Acad. Sci. U. S. A. 91, 10394-10398.
Jemal A, Siegel R, Ward E, Hao Y, Xu J, et al. (2008) Cancer statistics, 2008. CA Cancer J Clin 58: 71-96.
References
ReferencesJohnson DH, Blot WJ, Carbone DP, et al. Cancer of the lung_Non-small cell lung cancer and small cell lung cancer. In: Abeloff MD, Armitage JO, Niederhuber JE, Kastan MB, McKena WG. Clinical Oncology. 4th ed.
Philadelphia, Pa: Churchill Livingstone Elsevier: 2008: chap 76.
Jones, R.B., Gordus, A., Krall, J.A., and MacBeath, G. (2006). A quantitative protein interaction network for the ErbB receptors using protein microarrays. Nature 439, 168-174.
Kwak, E.L., Sordella, R., Bell, D.W., Godin-Heymann, N., Okimoto, R.A., Brannigan, B.W., Harris, P.L., Driscoll, D.R., Fidias, P., Lynch, T.J. et al. (2005). Irreversible inhibitors of the EGF receptor may circumvent acquired resistance to gefitinib. Proc. Natl. Acad. Sci. U. S. A. 102, 7665-7670.
McKay, James D. et al. 2008. Lung cancer susceptibility locus at 5p15.33. Nature Genetics. 40(12): 1404-1406.
Moyer JD, Barbacci EG, Iwata KK, Arnold L, Boman B, et al. (2006) Induction of apoptosis and cell cycle arrest by CP-358,774, and inhibitor of epidermal growth factor receptor tyrosine kinase. Cancer Res 57: 4838-4848.
Ohndorf, U., Rould, M., He, Q., Pabo, C., Lippard (1999). Basis for recognition of cisplatin-modified DNA by high-mobility-group proteins. Nature. 399: 708-712.
Pedersen, M.W., Pedersen, N., Ottesen, L.H., and Poulsen, H.S. (2005). Differential response to gefitinib of cells expressing normal EGFR and the mutant EGFRvIII. Br. J. Cancer 93, 915-923.
Pietras K, Ostman A, Sjoquist M, Buchdunger E, Reed RK, Heldin CH, Rubin K (2001) Inhibition of platelet-derived growth factor receptors reduces interstitial hypertension and increases transcapillary transport in tumors. Cancer Res 61: 2929-2934.
Poklar, N., Pilch, D.S., Lippard, S.J., Redding, E.A., Dunham, S.U., and Breslauer, K.J. (1996). Influence of cisplatin intrastrand crosslinking on the conformation, thermal stability, and energetics of a 20-mer DNA duplex. Proc.Natl. Acad. Sci. U. S. A. 93, 7606-7611.
Rikova, K., Guo, A., Zeng, Q., Possemato, A., Yu, J., Haack, H., Nardone, J., Lee, K., Reeves, C., Li, Y. et al. (2007). Global Survey of Phosphotyrosine Signaling Identifies Oncogenic Kinases in Lung Cancer. Cell 131, 1190-1203.
Ruddon, R.W. (2007). Cancer Biology: Fourth Edition (New York: Oxford University Press).
Schiller JH, Harrington D, Belani CP, Langer C, Sandler A, Krook J, Zhu J, Johnson DH (2002) Comparison of four chemotherapy regimens for advanced non-small-cell lung cancer. N Engl J Med 346: 92-98.
Shilo, B., Yarden, Y. (2007). Snap Shot: EGFR Signaling Pathway. Cell.131, 1018e1-1018e2. Stros, M., Ozaki, T., Bacikova, A., Kageyama, H., and Nakagawara, A. (2002). HMGB1 and HMGB2 cell-specifically down-regulate the p53- and p73-dependent sequence-specific transactivation from the human Bax gene
promoter. J. Biol.Chem. 277, 7157-7164.
Thorgeirson, Thorgeir E. et al. 2008. A variant associated with nicotine dependence, lung cancer and peripheral arterial disease. Nature. 452: 638-642.
Vlahovic G, Rabbani ZN, Herndon II JE, Dewhirst MW, Vujaskovic Z (2006) Treatment with Imatinib in NSCLC is associated with decrease of phosphorylated PDGFR-beta and VEGF expression, decrease in interstitial fluid pressure and improvement of oxygenation. Br J Cancer 95: 1013– 1019.
Yarden, Y., and Sliwkowski, M.X. (2001). Untangling the ErbB signalling network. Nat. Rev. Mol. Cell Biol. 2, 127-137.
Yuan, Z.M., Shioya, H., Ishiko, T., Sun, X., Gu, J., Huang, Y.Y., Lu, H., Kharbanda, S., Weichselbaum, R., and Kufe, D. (1999). p73 is regulated by tyrosine kinase c-Abl in the apoptotic response to DNA damage. Nature 399, 814-817.
Yun, C.H., Mengwasser, K.E., Toms, A.V., Woo, M.S., Greulich, H., Wong, K.K., Meyerson, M., and Eck, M.J. (2008). The T790M mutation in EGFR kinase causes drug resistance by increasing the affinity for ATP. Proc. Natl. Acad. Sci. U. S. A. 105, 2070-2075.