Cell Cycle Checkpoint and Cancer

R4 / VS

 Introduction and Cell Cycle DNA damage and Checkpoint Checkpoint and Carcinogenesis Checkpoint and Drug Therapy

Introduction and Cell Cycle

Cancer predisposing gene

Oncogene
Tumor Suppressor Apoptosis

Nature Medicine 2004;10:789-799

Control of cell cycle by cdk-cyclin complexes

S / M phase kinase : cdk + cyclin

Molecular cell biology

Regulation of Cell Cycle

INK4

(RB)

Cell 2004;116:221-34

Cell 2004;116:235-46

DNA damage and checkpoint

Theory of checkpoint ATM protein

Checkpoint
A control loop that makes the initiation of one
event in the cell cycle dependent on the successful completion of an earlier event
A cell must replicate every sequence of DNA once and only once The mass of the cell must double

Gene VIII p845-6

Checkpoint
Initiation of late events is dependent on
the completion of early events
Substrate-product order If then Complement cascade, apoptosis cascade Give enough substrate, and yield product Control mechanism Checkpoint unless Found some chemical / mutant relieve a
dependent relationship Relief of dependence
Hartwell and Weinerg, Science 1989;246:629-633

Molecular biology of the cell 2002

p53 and checkpoint

p53 mutant let DNA damaged cell enter S phase but not impact G2 checkpoint
Molecular cell biology

RAD9 system in yeast

cdc9 (DNA ligase) mutation DNA
replicated but unligated arrest at S phase cdc9-rad9 double mutant:
Enter into mitosis Cell die much more rapidly Loss chromosome spontaneously 21X than wild-type

Science 1989;246:629-633

Stopped by next checkpoint

Damage of checkpoint

Molecular cell biology

ATM (Ataxia Telangiectasia mutated)

Heterozygous for AT mildly manifest cancer
predisposition and radiation sensitivity Cell of AT patients have a reduced life-span, higher requirements for serum growth factor, defect in cytoskeleton Inhibition of semiconservative DNA synthesis and cell cycle progression by ionizing irradiation is reduced in AT cell
Science 1995;26:1749-53

ATM
AT cell failed to induced p53 in response
to irradiation
ATM act upstream to p53

Identify ATM gene from patient with AT Similarity between ATM and rad3
rad3 protein is required for the G2-M checkpoint, it monitor the completion of DNA damage repair

Similarity between ATM and PI3-kinase

Cell 1992;71:587-95 Science 1995;26:1749-53

mTOR, p110 TIBS 1995;20:426-30

p53

p53

TIBS 1995;20:426-30

Nature 2004;432:316-23

Trends in Cell Biology 2002;12:509-16

Trends in Cell Biology 2002;12:509-16

ATM

Cancer cell 2003;3:421-429

Checkpoint in S. cerevisiae

Science 2002;297:552-7

Human DNA damage response pathway

DNA damage Replication block
Signals Common intermediate

ATM or ATR Rad17/RFC

Sensors

Hus1/Rad1/Rad9

Transducers

Chk1 BRCA1 Others

p53

Effectors

Cell cycle Apoptosis arrest

DNA repair

Nature 2000;408:433-9

Checkpoint and Carcinogenesis

Checkpoint and carcinogenesis

Nature 2004;432:316-23

Adenoma-Carcinoma Sequence

 Why the patient so unlucky? Dose each oncogene mutate randomly

and independently? Dose tumor progress steadily?

Science 2002;297:565-9

Oncogene induced a oncogenic stress (cyclin E), which activate DNA repair response (ATM, pChk2)

U-2-OS cell Cyclin E expressed in a Tetracyclin-repressible manner Mcm7: loading control Nature 2005;434:864-70

pT-Chk2 level in bladder cancer

Absent Low level Medium level High level

Nature 2005;434:864-70

Checkpoint damage in cancer (bladder cancer)

Nature 2005;434:864-70

Genome instability during cancer progression

Nature 2005;434:907-13

TUNEL stain: hallmark of apoptosis

Nature 2005;434:907-13

Summary
Oncogene mutation hyperplasia DNA replication stress Double strand break Activate checkpoint cell cycle arrest, DNA
repair, or apoptosis no tumor progress Defect of checkpoint (ATM-Chk2-p53) Genomic instability and cancer

Nature 2005;434:864-70 Nature 2005;434:907-13

A model for carcinogenesis

Nature 2005;434:907-13

Science 2008;319:1352-5

P.I.: proliferation index

DDR.I.: DNA damage reponse index A./S.I.: apoptosis or senescence index
Science 2008;319:1352-5

Checkpoint and Drug Therapy

Chemotherapy with intact checkpoint Chemotherapy with defective checkpoint Targeting checkpoint

Mechanism of alkylating agent

Cancer Principles and proctice of Oncology, 7ed p339

Therapy with intact checkpoint

DNA damage activate DNA repair and
checkpoint pathway ATM->Chk1/2->p53->p21 Cell cycle arrest or death? Therapy shift arrest to death

Arrest after SN-38

(CPT-11) JCO 2005;23:4499-4508

Flavopiridol
CDK inhibitor Inhibit bcl-2, XIAP, p21,
mcl-1, cyclin D1,30 and phospho-survivin Inhibit p21-induced cell cycle arrest at G2 checkpoint

JCO 2005;23:4499-4508

JCO 2005;23:4499-4508

Therapy with defective checkpoint

Response to Flavopiridol according to
p53 status Resistance to anti-CD33 in AML cell unable to activate Chk1/Chk2
JCO 2005;23:4499-4508

Blood 2003;101:4589-97

Targeting checkpoint
Loss of ATM sensitized p53-deficient cell to
topoisomerase poisons and antimetabolites Annals of Oncology 2003;14:938-45 SN-38 followed by UCN-01 (inhibit Chk1 /Chk2) enhance apoptosis in cell with mutated p53 but not intact p53 Under development: CHIR124, inhibitor of Chk1, and 17-AAG: suppresses Chk1expression JCO 2005;23:4499-4508

Flavopiridol inhibit cell cycle arrest

Apoptosis

Chk1/p21

UCN-01

Flavopiridol

Molecular cell biology

A model for targeting checkpoint

Up stream p53 inhibit G2 checkpoint

Down stream p53 inhibit G1/S checkpoint

So Simple?
JCO 2005;23:4499-4508

To be defined
What determines DNA repair,
cell cycle arrest, or apoptosis? Some drug induced cell cycle arrest, some apoptosis, and some necrosis? Different cell line had different response? Undefined pathway? p53?
Cell 2005;122:21-32

?
Cancer cell 2003;3:421-429

Thanks for your attention

Mitotic checkpoint

Nature review cancer 2005;5:773-85

Mitotic checkpoint defect in cancer

Nature review cancer 2005;5:773-85

p53 in checkpoint

Cell 2005;122:21-32

p53 in checkpoint

Cell 2005;122:21-32