Bobby Hodges, Elizabeth Ross, Natalie Simak, Keith Solvang and EliotVildaver
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
What Is Lung
Cancer?
•Types
•Risks
Predisposition Mutations
•EGFR
•Other
proteins
Angiogenesis
•VEGF
Your
Treatment Plan
h...
http://www.odec.ca/projects/2005/thog5n0/public_html/Tumour.gif
http://www.icowman.com/uploadfiles/2007072409225175.jpg http://www.icowman.com/uploadfiles/2007072409225175.jpg
NON SMALL CELL
 Squamous cell carcinoma
 Adenocarcinoma
 Large-cell carcinoma
SMALL CELL
 Bronchi
 Grows rapidly
 Sp...
http://www.taconichills.k12.ny.us/webquests/noncomdisease/lungcancerpic.jpg
 This year approximately
200,000 people will be
diagnosed with lung
cancer
 NSCLC accounts for 80%
of all lung cancers
h...
http://medicineworld.org/cancer/lung/ http://images.google.com/
While there is no known cure as of right
now, there ARE options!
http://dspace.mit.edu
Imatinib
Gefinitib
Cisplatin
http:/...
Nucleus
P
Protein
Cascade
P
P
AKT
P
P
Nucleus
Protein
Cascade
Protein
Cascade
http://rosenblumtv.files.wordpress.com
http://www.pharmgkb.org
http://academic.evergreen.edu
 Compact structures of
DNA
 Contains thousands of
genes
Centromere
Chromatid
Genes
http://staff.jccc.net/PDECELL/celldiv...
 Single variation in
nucleotide base pairs
 Alleles are a result of
SNPs within gene
sequence
 SNP’s are the basis for
...
 Variation across genomes
 Genotype cases along with
controls
 Find roles that genes play in
disease
 Large sample siz...
Centromere
p-arm
q-arm
6 RegionsAway
Region 5q6
5
http://images.google.com/
http://upload.wikimedia.org/ http://upload.wikimedia.org/
Nicotine binds and stimulates acetylcholine
ion channel-linked in neurons
http://www.jyi.org/articleimages/88/originals/im...
(Hung et al., 2008)
(Thorgeirson et al., 2008)
Cancerous Growth
http://images.google.com/i
(McKay et al., 2008)
P
P
AKT
P
P
Nucleus
Protein
Cascade
Protein
Cascade
P
EGF
Proliferation
Survival
Apoptosis
ERBB2
Nucleus
P
Protein
Cascade
ERBB2
How CanThis Lead to Cancer?
ERBB2
Nucleus
Proliferation
P
Protein
Cascade
(Chen et al., 2006)
ATP TKI
ATP binding cleft on the kinase
Tyrosine
Kinase
domain
(Chen et al., 2006)
(Pedersen et al., 2005)
(Pedersen et al., 2005)
(Chen et al., 2006)
P
P
AKT
P
P
Nucleus
Protein
Cascade
Protein
Cascade
WhatAre the Roles of Proteins
Associated with Lung Cancer?
http://www.taconichills.k12.ny.us/webquests/noncomdisease/lungc...
Recap
Nucleus
N-terminus
C-terminus
P
Serine
P
Threonine
Tyrosine
P
Leucine
Lysine
Alanine
ERBB2
P
PI3KSH2
Nucleus
P
Protein
Cascade
Change in
Gene
Expression
(Rikova et al. 2007)
Green and Blue circles = Proteins involved in cascade
Red lines = Interactions
Red circles = EGFR Receptor
(Jones et al. 2...
(Jones et al. 2006)
No.proteinreceptors
No.proteinreceptors
No.proteinreceptors
Affinity Threshold (nM) Affinity Threshold...
http://en.wikipedia.org/wiki/Proteins
(Rikova et al. 2007)
0
5
10
15
20
25
30
35
40
45
50
ROS ALK MET PDGFRα ErbB2 EGFR VEGFR-1
Tumor
Samples
Cell Lines
Chart configured from Table1...
(Engelman et al., 2007)
P
P
AKT
P
P
Nucleus
Protein
Cascade
Protein
Cascade
http://www.taconichills.k12.ny.us/webquests/noncomdisease/lungcancerpic.jpg
http://www.researchvegf.com/researchvegf/multimedia/index.m#type-slides
VEGF-A
VEGFR
Angiogenesis
Tumors
PDGF-β
PDGFR-β
P85-α
PIP2
P
P
PTEN
P
PIP3
P
P
P
P110-α
PI3K
AKT
SH2
P
P
Endothelial Cell
AKT
P
P
P
BAD
MDM2
mTOR
Growth,
Translation
Apoptosis
p53
P
Nucleus
Cancer Cell
Proto-Oncogene Oncogene
(Chen et al., 2009)
Giemsa-Stained Cells
(Chen et al., 2009)
P85-α
PIP2
P
P
PTEN
P
PIP3
P
P
P
P110-α
PI3K
AKT
SH2
P
P
VEGF-A
VEGFR
FLJ10540
Endothelial Cell
http://www.medicinescomplete.com/mc/martindale/2007/images/MRT9229C001.gif
Binds to ATP binding site of many enzymes:
p-PD...
http://www.medscape.com/pi/editorial/clinupdates/2000/583/art-tu02.fig04.jpghttp://www.nature.com/nm/journal/v7/n9/images/...
(Vlahovic et al. 2007)
Vlahovic et al. 2007
http://onctalk.com/wp-content/uploads/2008/01/angiogenesis-summary.jpg
Imatinib
P
P
AKT
P
P
Protein
Cascade
Protein
Cascade
Nucleus
(Rosenberg et al., 1969)
Nucleus
Simple Diffusion
Pt
NH3NH3
GG
P73
C-Abl
Cell Cycle
Arrest
&
Apoptosis
P53
HMG1
?
nucleus
Pt
Pt
Pt
Pt
Pt
Transporter
protein
Transporter
protein
Pt
(Hall et al., 2007)
crosslink
nucleus Pt
NH3
NH3
Cell Cycle Arrest
&
Apoptosis
G
G
1,2-intrastrand
d(GpG)adduct
(Ohndorf et al., 1999)
http://www.unc.edu/~shantanu/images/bkstcg_aacr07.png
Pt-DNA Adduct
Apoptosis
HMGB
HMGB
P73
P53
(Stross et al., 2002)
(Ohndorf et al., 1999)
*Nucleotides
crosslinked
Bold- nucleotides
contacted by the
protein
(Ohndorf et al., 1999)
Nucleus
?
Simple Diffusion
Pt
NH3NH3
GG
P73
C-Abl
Cell Cycle
Arrest
&
Apoptosis
P53
HMG1
?
tyrosine
kinase
mismatch
repair protein
cisplatin
homologue of
p53
Tumor
suppresor
(Gong et al., 1999)
P53 knockout= less sensitive to CDDP, survived better than wild type in
lower concentrations of CDDP
MLH1-/- & Abl= less s...
Nucleus
Simple Diffusion
Pt
NH3NH3
GG
P73
Cell Cycle
Arrest
&
Apoptosis
P53
ATM
Transporter
Proteins
C-AblP
Nucleus
P
Protein
Cascade
P
P
AKT
P
P
Nucleus
Protein
Cascade
Protein
Cascade
http://immunodefence.com/ii/gefitinib.gif
http://www.themesotheliomalibrary.com/cisplatin.jpg
http://www.specialityformula...
 New plasma membrane proteins have unique
mechanisms and effects on NSCLC
 Gefitinib effectively inhibits particular EGF...
 How does cisplatin enter the nucleus?
 Roles and mechanisms behind the various
mutated proteins involved in lung cancer...
Professor Sleiter
Schmooing yeast!!!
Dr. DebBurman
Mike Fiske
Abid MR, Guo S, Minami T, Spokes KC, Ueki K, et al. (2004) Vascular endothelial growth factor activates PI3K/Akt/forkhead ...
Johnson DH, Blot WJ, Carbone DP, et al. Cancer of the lung_Non-small cell lung cancer and small cell lung cancer. In: Abel...
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  • Keith:
    Thank you Michael Fiske for the wonderful introduc. My collegues and I are here, at the diseased genome hospital, to discuss a very prominent disease: LUNG CANCER.
  • Please follow us into examination room J200. Please Have a seat.
  • Let me pull up ur chart real quick.
    I first want to explain to you what’s going on. Here’s an xray of a healthy set of lungs. You can tell they’re healthy because they are clear.
  • I asked you to come in today, because I wanted to discuss something that is very concerning to us. Here is your chest xray from your last visit. What concerns us is this: CLICK. ARROW.
    There appears to be a mass of some sort, on your left lung.

    The symptoms you’ve previously described during your last visit
    Coughing
    wheezing
    shortness of breath
    chest pain
    difficulty swallowing
    hoarseness
    loss of appetite
    respiratory infection

    This has led my colleduges and I to speculate you have LC
    Before you come too upset, let me col. And I explain to you where we’re going to go with this apt today.
  • Today we will be discussing the different parts that could be wrong with ur body and could be causing this lc. We are hoping that this will provide you with a clear understanding of what you may be giong through and the future possiblities.
    First, we’ll start off today talking about what is lung cancer, the types there are the type that you may have. Also, the risks that led you to this disease.
    Then we r gonna talk about some mechs that could be going wrong in ur body. There are some poss genetic predis that could lead u to lc. Then we will talk about various mutations..
    Angiogenesis is key to what you may have, DO YOU KNOW WHAT THAT IS?

    Dr. Ross will take it from here.
  • Liz:
    Cancer starts out as a group of cells that are dividing continuisly. As the cancer progresses, cells have divided so frequently that they’ve begun to form solid masses known as tumors. That can be seen in ur pamphlet. Cells proliferate and invade other regions of tissue that theyre in. eventually these cells grow so much that they enter the blood suppy where they metastasize to other regions of the body. Right now u have a tumor in ur lung that has not yet metastasized but we should treat it before



    We’ll give you a roadmap about your disease…after this slide
    Lc background
    10 min intro
    Liz: egfr mutants
    angiogenesis: elliot
    Keith: EGFR +other proteins
    Bobby: chromosomal loci
    Natalie: cisplatin treatment

    Draw tumor epithelial cell with blood vessels coming in, nucleus containing chromosomes, efgr receptor + other proteins, met, vegf, megf, drugs on outside, killing cell
    End intro. 5 main parts of lc that we’ll be talking about today. Parts that involve the chromosome (highlight with box) this what bobby will talk to u about today. Theres also mutations on the egfr. In addition to egfr theres these proteins that well be talking about. How tumor cells get drug supply. And how drugs kill tumor cells?
    Each time u end a section show this image again and have summarizing words

    Dim old parts and highlight new parts

  • The distribution of cigarettes first began in World War II (1938-1944) to soldiers. In the 1940s and early '50s, cigarette smoking became more commonplace due to advertising. Cigarette ads usually featured people of a higher class or soldiers and groups laughing and having fun. People were also permitted to smoke anywhere they wanted, including nurse stations in hospitals, salons, barbershops, cafes and the grocery store.


    Lung cancer was first described by doctors in the mid-19th century. In the early 20th century it was considered relatively rare, but by the end of the century it was the leading cause of cancer-related death among men in more than 25 developed countries. In the United States it has surpassed breast cancer as the leading cause of death from cancer among women. This rapid increase is due mostly to the increased use of cigarettes since World War I.
  • 2 flavors: nclc and small cell. Nsc is what you have. Thankfully u have nsclc which is more treatable.

    Liz:
    Even the two main forms of lung cancer are diverse. There are three types of NSCLC: squamous cell carcinoma originates near the bronchus and which affects the squamos epithelial cells, adenocarcinoma which originates on the outer region of the lungs, & large-cell carcinoma which can originate at any part of the lung. NSCLC is classified by the cells in the different subtypes (either by size, shape, or chemical make up). Small cell lung cancer grows and spreads much more rapidly than NSCLC. This type of cancer is characterized by tiny cells that are referred to as OAT cells. These can be seen in the image on the right. So these are the cancers, but lets take a look at what causes the chaos at the cell and molecular level of nsclc.

    The development of lung cancer is not the result of a sudden, transforming event in the bronchial epithelium. Rather, it is a multi-step process involving sequential genetic and cellular changes, predominantly in recessive oncogenes
  • Eliot:
    Treatments
    Surgery, that’s very abrasive and very risky
    Radiation-Chemotherapy
    Various Drugs
  • This picture might seem very overwhelming, but what I want you to take away fro mthis is that there aer many poss in ur body that may be causing ur lc and my collegeues and I will be discussing these various poss.
    Dr. Hodges will be discussing the genetic predispositions
    Dr. Ross will be talking about mutations in EGFR protein kinase recep.
    Dr. Solvang will be talking further about RGFR and ERGG2, as well as other mutated proteins that associate with lc
    Dr. Vildaver will be meeting with u to discuss VEGF and its role in angiogenesis
    Finally, Dr. Simak present to u with various treatment options and find the proper course of treatment.
  • Big ques before this
    Are there regions of the chromosomes that might predis u to get lc
    Evidence y looking at this (slide before this)
    Main topic :CL- prev evidence ques (methods)resultssignificancemodel
  • Talk more about pic
    2 indiv from same species but regions where single nuc are changed (snp)
    Used for a wide variety of things what were gonna be talking about is how snps are related to ur susceptibility to lc
  • The way that they use snps to look at predis. To disease is called genome wide…
    Take entire genome sequence it and looks for snps, where they are and what genese theyre in and what role geneses are playing in the disease

    Snps that normal person doesn’t have maybe snps that are key to tlc

    Both might have one snps but not another
  • CITE
    good
  • Not made just for nicotine. FIGURE OUT
    For aceto…but nicotine can bind to nar receptors
  • Explain better
    Methods?
    Looking at ppl from diff regions
  • Ppl who have a certain gene are more likely to smoke
    Is it a snp?
  • How related to lc
  • This picture might seem very overwhelming, but what I want you to take away fro mthis is that there aer many poss in ur body that may be causing ur lc and my collegeues and I will be discussing these various poss.
    Dr. Hodges will be discussing the genetic predispositions
    Dr. Ross will be talking about mutations in EGFR protein kinase recep.
    Dr. Solvang will be talking further about RGFR and ERGG2, as well as other mutated proteins that associate with lc
    Dr. Vildaver will be meeting with u to discuss VEGF and its role in angiogenesis
    Finally, Dr. Simak present to u with various treatment options and find the proper course of treatment.
  • QUES: Are you aware of what the func of epidermal growth fac is?

  • First, a ligand (EGF) must bind to a receptor EGFR. A shape change occurs and EGFR is phos./activ.
    It then forms a heterodimer with another receptor named ERBB2. When this heterodimerization happens, they are both phos. Which provokes signalling cascade of spec. proteins which leads to the nuc.
    Here is an example of one specific pathway that leads to diff kinds of signaling.
  • EGFR can become mutated and when this happens, it has higher lvels of phos. Which means that the signalling cacscade is constantly turned on. you can imagine a light switch here that is constantly turned on.
  • There are many types of mutations that can occur on EGFR. Researchers have found that depending on where the EGFR mutation is located, there are diff phos. Found.
    Not one indiv. Tyr that must be affected

    Vector- Cell with the missing gene for EGFR…why no phosphorylation
    EGFR- Cells expressing normal amount of EGFR _no mutation
    Anti-Myc- the tag for EGFR- shows that it is present in the cells
    Rest are mutated EGFR cells. Whats important to know is that each of these mutated cell lines have a mutation at a different location along the EGFR receptor
    The cells were starved of EGF (the ligand) can be thought of as “food” for 12 hrs.

    Then they EGF was added (10ng/ml). The phosphorylations were observed at various amount of time after the EGF was added. It is expected to see stronger phos levels at 10 mins.

    Whats interesting is distinct patterns of try phos among the different mutated cell lines.

    This is a lottt to look at so I want to focus on just a few parts.

    Chen

    In response to EGF simulationuse a pic
    Bottom?
    Kinase domain with mutations but where not in kinase domain can be mut
  • There are many drugs that target TK by inhibiting the kinase domains.

    Treatment
    http://www.nature.com/nrc/journal/v6/n9/images/nrc1913-f4.jpg
  • Treatment

    Chen

    Show one or two
    Then clickkkkkkkkkkkk al these other mutations, don’t get signal transduction
    Put box around where theres no more phos.
    Left is control!!!
    As u move to right conc increasing
    Now lets look at all the other strains….similar story
    What is activity??? IC50 micromolar? Activity of pathway?
    Myoegfr, measure of egfr protein present

  • Treatment
    Pedersen

    An experiment was performed investigating the levels of EGFR/EGFRvIII and ERK phosphorylation in cells mock treated or treated with .01 or 2mM of gef and at the same time stimulated with 10nM EGF ofr 10 mins, 24, 48, 72 hours.

    Try1173 was chosen because to was found to be the most durable of the phosphorylation sites.

    Gef rapidlly and in a dose-dependent manner inhibits EGFR and ERK phosphorylations up to 72 h after EGF stimulation in both the high- and low-EGFR-expressing cell lines (A and B)

    Athe decrease in EGFR levels, in the absence of gef, in the low_EGFR-expressing cell line is due to receptor downregulation inducted by EGR, a mechanism that appears not to be functioning in the NR6W and NR6M lines.

    A concentration of .01 um gef seems to increase EGFRvIII phosphorylation on residue 1173 and ERK phosphorylation after 48 and 72h of treatment (c). Could explain why gef in low concentrations induce cell proliferation and anchorage-independent growth. However 2um of gef effectively inhibits both EGFRvIII and ERK phosph in the NR6M cell line for up to 72h. The high concen of gef seems to induce degradation of both EGFR and EGFRvIII independent of EGF. This is particulary evident in the low-EGFR expressing cell line where .01 um gef inhibit EGF-mediated downregulation, resumably due to its inhibtion of the Try1045 site, but fails to do so at a concentration of 2um (A).


    Delete?
    Stress the idea that no gef. Phos.
    As morve to the left increase phos.
  • Log to get the curve?
    Treatment

    Pedersen

    Here we have prolif of cells dep on conc of gef
    See that as u treat cells with gef their prolif rate decreases
    Black line-parental cell line…not mutated?
    Doesn’t work for all types of cancer…
  • Refer to phos levels in mutants

    Chen

    However, what is important to remember is that EGFR’s have many different mutations. The different mutations located in among various parts of the receptor were found to react differently to gefitinib. This is why Gef works for some and not others.


    In different EGFR variatn-expressing 32D cells, only cells harboring L858R, E746-A750 deletion, and G719S mutants were clearly more sensitive to gef than wild-type EGFR expressing cells. Cells expressing the L861Q mutant were also more sensitive, but to a less extent. Cells expressing S768I and E709G mutant were actually more resistant to gef than cells expressing the wild-type receptor. Interestingly, S768I and E709G were amont the mutants that lacked ubiquitination an dalso had sustained Try phosphorylation in response to EGF stimulation. It is unclear whether these properties are related to the relative resistance to gef in cells harboring these two mutants.

    Currently, unable to correlate particular mutations other than L858R and deletion mutants to treatment responses to gef due to the scarcity of those mutations in reported cases.
    How phos f mutants affects gef activ

    Used 32D cells for the assay

    Found that 32D cells expressing L858R, E746-A750 Deletion and G719s mutants were most sensitive to gefitinib. They were not viable at the lowest does (.05) of the inhibitor after 3 days.

    L861Q were more sensitive to gefitinib that then 32D cells harboring wild-type EGFR but to a lesser extent

    S7681 and E709G were slightly more resistant to gef than wild type

    Interestingly and for unknown reasons, at .05 had reproducible growth promoting ability in 32D cells expressing wild type, S7681 or E709G
  • This picture might seem very overwhelming, but what I want you to take away fro mthis is that there aer many poss in ur body that may be causing ur lc and my collegeues and I will be discussing these various poss.
    Dr. Hodges will be discussing the genetic predispositions
    Dr. Ross will be talking about mutations in EGFR protein kinase recep.
    Dr. Solvang will be talking further about RGFR and ERGG2, as well as other mutated proteins that associate with lc
    Dr. Vildaver will be meeting with u to discuss VEGF and its role in angiogenesis
    Finally, Dr. Simak present to u with various treatment options and find the proper course of treatment.
  • You might be asking urself..
  • But before we go on, I just wanted to make sure that you

    Show protein as beads on a string
    Lasrtt
    Which one can be phos?
    Yes! The phos of these can happen
    Imp to keep in mind
  • Protein cascade to the right
    Change in gene expression
  • Show old evidence that says these are typ found in lc
    Pic of lc
    Mutations in 2 genes
    This is y we wanna look at them
  • Even though its not the biggest contrib…
  • Green are proteins involved in cascade
    As increase conc activation of sig molec is increasing
    Add text
    These are the sig molc interacting with egfr recep
  • On Average, EGFR only binds around 7.2 proteins, but increases to about 50 proteins…same for ErbB2.Important that all of these proteins that are recruited were found to have that SH2 or PTB binding domain.
  • Label y axis
  • This picture might seem very overwhelming, but what I want you to take away fro mthis is that there aer many poss in ur body that may be causing ur lc and my collegeues and I will be discussing these various poss.
    Dr. Hodges will be discussing the genetic predispositions
    Dr. Ross will be talking about mutations in EGFR protein kinase recep.
    Dr. Solvang will be talking further about RGFR and ERGG2, as well as other mutated proteins that associate with lc
    Dr. Vildaver will be meeting with u to discuss VEGF and its role in angiogenesis
    Finally, Dr. Simak present to u with various treatment options and find the proper course of treatment.
  • Tumor cells’ vasculature is induced by continuous secretion of growth factorprotein (VEGF) affecting activity on nearby endothelial cells. In response to hypoxia, or deprivation of oxygen, malignant tumors turn on the “angiogenic switch” which tells them to secrete VEGF which binds to VEGF receptors on nearby blood vessels. The angiogenic switch causes rapid tumor growth and metastasis. Another protein that plays a role in angiogenesis is PDGF.
  • Normal angiogenesis is highly regulated, but this process is highly unregulated in tumor cells, especially in the case of NSCLC. Endothelial cells produce VEGF., which undergoes an autocrine signaling pathway in which a cell secretes a hormone or chemical messenger that binds to receptors on the same cell. VEGF is a potent proangiogenic protein that activates VEGF receptor (VEGFR) acts through receptor tyrosine kinase VEGFR-2 in angiogenesis. VEGF-A stimulates VEGFR-2 which is in high levels to promote angiogenesis, which in turn is required for metastasis of tumor cells. p-PDGFR-β Plays a role as well by acting on pericytes covering blood vessels, which secrete Ang-1 that contributes to blood vessel maturation and stability.
  • PI3-kinase is activated/phosphorylated by the majority of Receptor Tyrosine Kinases, including VEGFR-2. Phosphorylation of membrane-bound proteins can lead to the activation of these proteins’ kinase function, which sends a signal through the consequent proteins by phosphorylation. Specifically, these proteins are phosphorylated at tyrosine residues. This signal can then be sent through receptor protein/kinases such as PI3K. Activated PI3-kinase induces the phosphorylated of PIP2 to PIP3. The interaction of PIP3 with the PH domain of AKT likely induces conformational changes in AKT, thereby exposing the two main phosphorylation sites at T308 and S473. T308 and S473 phosphorylation by protein serine/threonine kinase 3'-phosphoinositide-dependent kinases 1 and 2 (PDK1 and PDK2) is required for maximal AKT activation. Activated AKT translocates to the nucleus and mediates the activation and inhibition of various targets resulting in cellular survival and cell growth and proliferation. PTEN is a specific PTPase that has no effect in the PI3K/Akt pathway in lung cancer. In total, all the proteins in the PI3K/Akt pathway must be phosphorylated to continue the signal. AKT is heavily implicated in angiogenesis.
  • Akt is a serine/threonine kinase. When Akt is phosphorylated, it inhibits the phosphorylation of certain proteins such as BAD, which have a role in apoptosis. Thus, apoptosis is inhibited and tumor cells continue to grow, especially with the activation of mTOR. P53 is related to cell-cycle arrest. Akt allows the phosphorylation of MDM2 to occur, which inhibits production of p53, resulting in cell cycle arrest being inhibited.
  • Ques: what is an oncogene? (Understand what it is precisely!!!)
    FLJ10504 is a protein that may function as an oncogene in the development of tumors. It is required for VEGF-A dependent signaling in the PI3K pathway. High levels of FLJ10540 help induce Akt phosphorylation and form a complex with PI3K. FLJ10540 is overexpressed in lung cancer tissue, and is in a positive correlation with VEGF-A overexpression. This leads to hyperactivation of the PI3K pathway. There are migratory and invasive qualities related to FLJ10540.
  • VEGF-A has a positive correlation with FLJ10540 in lung cancer cells. Stimulation with VEGF-A increased FLJ10540 protein expression, indicating that FLJ10540 has a role in enhancing complex formation with PI3K under VEGF-A stimulation. FLJ10540 overexpression associates with enhanced metastatic potential. CL1-0 cells are cancer cells. SU5416 is a VEGFR-2 tyrosine kinase inhibitor. Reduces expression of FLJ10540. FLJ10540 only shares expression patterns with VEGF-A, not VEGF-B.
  • Cancer cells exhibit more migratory and invasive abilities in those stimulated with VEGF-A alone than VEGF-A with SU5416 (Semaxinib). This effect was paralleled increase in FLJ10540 expression. Finally, the activation of PI3K/Akt is critical for FLJ10540-induced lung cancer cell migration and invasion upon VEGF-A stimulation. FLJ10540 could serve as a critical mediator of VEGF-A dependent activation of PI3K/Akt.
  • It has been observed that upon VEGF-A stimulation, increased levels of FLJ10540 can enhance AKT phosphorylation and that FLJ10540 can form a complex with PI3K. Increased levels of FLJ10540 can reinforce the association with p-110 and p-85. FLJ10540 stabilizes the PI3K complex and contributes to migration, invasion, and effects of angiogenesis prompted by VEGF-A stimulation in tumor cells.
  • IFP is marked by a reduced delivery of anticancer drugs. High levels of IFP are indicative of malignant tumors. Hypoxia is the deprivation of oxygen to parts of tumor cells that causes inability of chemotherapy and radiation effects to cure tumors. It also influences the activation of angiogenesis in tumors. Antitumor therapy cannot penetrate far enough into the tumor to destroy it and they only target rapidly-dividing cells. Tumour angiogenesis results in tortuous, leaky, and nonfunctional tumour vasculature, which contributes to tumour growth and metastasis.
  • It has been noted that PDGFR-B plays a crucial role in smooth muscle cell and pericyte recruitment. Involved in angiogenesis.
  • More recent data have suggested that abnormal vessel morphology, rather than vessel density, may be a better indicator of tumor angiogenesis.
  • Imatinib reduces tumor angiogenesis, presumably through simultaneous p-PDGFR-B inhibition and VEGF downregulation. Imatinib is an orally administered drug that disrupts the kinase activity of both p-PDGFR-B and VEGFR by binding to their respective ATP-binding sites on the protein.
  • This picture might seem very overwhelming, but what I want you to take away fro mthis is that there aer many poss in ur body that may be causing ur lc and my collegeues and I will be discussing these various poss.
    Dr. Hodges will be discussing the genetic predispositions
    Dr. Ross will be talking about mutations in EGFR protein kinase recep.
    Dr. Solvang will be talking further about RGFR and ERGG2, as well as other mutated proteins that associate with lc
    Dr. Vildaver will be meeting with u to discuss VEGF and its role in angiogenesis
    Finally, Dr. Simak present to u with various treatment options and find the proper course of treatment.
  • eliot
  • K
  • K
  • Immunoprecipitation

    1. 1. Bobby Hodges, Elizabeth Ross, Natalie Simak, Keith Solvang and EliotVildaver
    2. 2. http://blogs.usask.ca/medical_education/archive/ElksRehabFront%2520Door.jpg
    3. 3. http://1.bp.blogspot.com/_HI2mCx5P3H0/SLlEQoHj8AI/AAAAAAAAA8Q/My7tf78TNc8/s320/hospital-bed-786282.jpg
    4. 4. http://img.quamut.com/chart/9471/02_healthy_sick_x-ray.jpg
    5. 5. http://www.cardiothoracicsurgery.org/content/2/1/16/figure/F1?highres=y
    6. 6. What Is Lung Cancer? •Types •Risks Predisposition Mutations •EGFR •Other proteins Angiogenesis •VEGF Your Treatment Plan http://www.freefoto.com/images/21/16/21_16_2---Single-Carriageway-Road_web.jpg
    7. 7. http://www.odec.ca/projects/2005/thog5n0/public_html/Tumour.gif
    8. 8. http://www.icowman.com/uploadfiles/2007072409225175.jpg http://www.icowman.com/uploadfiles/2007072409225175.jpg
    9. 9. 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/852/202230 38.JPG http://images.google.com/imgres?imgurl=http://upload.wikimedia.org/ Lynch et al., (2004)
    10. 10. http://www.taconichills.k12.ny.us/webquests/noncomdisease/lungcancerpic.jpg
    11. 11.  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
    12. 12. http://medicineworld.org/cancer/lung/ http://images.google.com/
    13. 13. While there is no known cure as of right now, there ARE options! http://dspace.mit.edu Imatinib Gefinitib Cisplatin http://dspace.mit.edu
    14. 14. Nucleus P Protein Cascade
    15. 15. P P AKT P P Nucleus Protein Cascade Protein Cascade
    16. 16. http://rosenblumtv.files.wordpress.com http://www.pharmgkb.org http://academic.evergreen.edu
    17. 17.  Compact structures of DNA  Contains thousands of genes Centromere Chromatid Genes http://staff.jccc.net/PDECELL/celldivision/chromosome1.gif
    18. 18.  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
    19. 19.  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
    20. 20. Centromere p-arm q-arm 6 RegionsAway Region 5q6 5 http://images.google.com/
    21. 21. http://upload.wikimedia.org/ http://upload.wikimedia.org/
    22. 22. Nicotine binds and stimulates acetylcholine ion channel-linked in neurons http://www.jyi.org/articleimages/88/originals/img0.jpg
    23. 23. (Hung et al., 2008)
    24. 24. (Thorgeirson et al., 2008)
    25. 25. Cancerous Growth http://images.google.com/i
    26. 26. (McKay et al., 2008)
    27. 27. P P AKT P P Nucleus Protein Cascade Protein Cascade
    28. 28. P EGF Proliferation Survival Apoptosis ERBB2 Nucleus P Protein Cascade ERBB2
    29. 29. How CanThis Lead to Cancer?
    30. 30. ERBB2 Nucleus Proliferation P Protein Cascade
    31. 31. (Chen et al., 2006)
    32. 32. ATP TKI ATP binding cleft on the kinase Tyrosine Kinase domain
    33. 33. (Chen et al., 2006)
    34. 34. (Pedersen et al., 2005)
    35. 35. (Pedersen et al., 2005)
    36. 36. (Chen et al., 2006)
    37. 37. P P AKT P P Nucleus Protein Cascade Protein Cascade
    38. 38. WhatAre the Roles of Proteins Associated with Lung Cancer? http://www.taconichills.k12.ny.us/webquests/noncomdisease/lungcancerpic.jpg
    39. 39. Recap Nucleus N-terminus C-terminus P Serine P Threonine Tyrosine P Leucine Lysine Alanine
    40. 40. ERBB2 P PI3KSH2 Nucleus P Protein Cascade Change in Gene Expression
    41. 41. (Rikova et al. 2007)
    42. 42. Green and Blue circles = Proteins involved in cascade Red lines = Interactions Red circles = EGFR Receptor (Jones et al. 2006)
    43. 43. (Jones et al. 2006) No.proteinreceptors No.proteinreceptors No.proteinreceptors Affinity Threshold (nM) Affinity Threshold (nM) Affinity Threshold (nM)
    44. 44. http://en.wikipedia.org/wiki/Proteins
    45. 45. (Rikova et al. 2007)
    46. 46. 0 5 10 15 20 25 30 35 40 45 50 ROS ALK MET PDGFRα ErbB2 EGFR VEGFR-1 Tumor Samples Cell Lines Chart configured from Table1 of Rikova et al. 2007 Amountofoverphosphorylatedproteininsamples
    47. 47. (Engelman et al., 2007)
    48. 48. P P AKT P P Nucleus Protein Cascade Protein Cascade
    49. 49. http://www.taconichills.k12.ny.us/webquests/noncomdisease/lungcancerpic.jpg
    50. 50. http://www.researchvegf.com/researchvegf/multimedia/index.m#type-slides
    51. 51. VEGF-A VEGFR Angiogenesis Tumors PDGF-β PDGFR-β
    52. 52. P85-α PIP2 P P PTEN P PIP3 P P P P110-α PI3K AKT SH2 P P Endothelial Cell
    53. 53. AKT P P P BAD MDM2 mTOR Growth, Translation Apoptosis p53 P Nucleus
    54. 54. Cancer Cell Proto-Oncogene Oncogene
    55. 55. (Chen et al., 2009)
    56. 56. Giemsa-Stained Cells (Chen et al., 2009)
    57. 57. P85-α PIP2 P P PTEN P PIP3 P P P P110-α PI3K AKT SH2 P P VEGF-A VEGFR FLJ10540 Endothelial Cell
    58. 58. 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
    59. 59. 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
    60. 60. (Vlahovic et al. 2007)
    61. 61. Vlahovic et al. 2007
    62. 62. http://onctalk.com/wp-content/uploads/2008/01/angiogenesis-summary.jpg Imatinib
    63. 63. P P AKT P P Protein Cascade Protein Cascade Nucleus
    64. 64. (Rosenberg et al., 1969)
    65. 65. Nucleus Simple Diffusion Pt NH3NH3 GG P73 C-Abl Cell Cycle Arrest & Apoptosis P53 HMG1 ?
    66. 66. nucleus Pt Pt Pt Pt Pt Transporter protein Transporter protein Pt (Hall et al., 2007)
    67. 67. crosslink nucleus Pt NH3 NH3 Cell Cycle Arrest & Apoptosis G G 1,2-intrastrand d(GpG)adduct (Ohndorf et al., 1999)
    68. 68. http://www.unc.edu/~shantanu/images/bkstcg_aacr07.png
    69. 69. Pt-DNA Adduct Apoptosis HMGB HMGB P73 P53 (Stross et al., 2002)
    70. 70. (Ohndorf et al., 1999) *Nucleotides crosslinked Bold- nucleotides contacted by the protein
    71. 71. (Ohndorf et al., 1999)
    72. 72. Nucleus ? Simple Diffusion Pt NH3NH3 GG P73 C-Abl Cell Cycle Arrest & Apoptosis P53 HMG1 ?
    73. 73. tyrosine kinase mismatch repair protein cisplatin homologue of p53 Tumor suppresor (Gong et al., 1999)
    74. 74. P53 knockout= less sensitive to CDDP, survived better than wild type in lower concentrations of CDDP MLH1-/- & Abl= less sensitive to CDDP , survived better than wild type in lower concentrations of CDDP (Gong et al., 1999)
    75. 75. Nucleus Simple Diffusion Pt NH3NH3 GG P73 Cell Cycle Arrest & Apoptosis P53 ATM Transporter Proteins C-AblP
    76. 76. Nucleus P Protein Cascade
    77. 77. P P AKT P P Nucleus Protein Cascade Protein Cascade
    78. 78. http://immunodefence.com/ii/gefitinib.gif http://www.themesotheliomalibrary.com/cisplatin.jpg http://www.specialityformulations.com/pcat-gifs/products-small/imatinib.jpg
    79. 79.  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
    80. 80.  How does cisplatin enter the nucleus?  Roles and mechanisms behind the various mutated proteins involved in lung cancer  New drugs , secondary mutations
    81. 81. Professor Sleiter Schmooing yeast!!!
    82. 82. Dr. DebBurman
    83. 83. Mike Fiske
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