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

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