cell cycle i molecular cell biology november 6, 2014

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Cell Cycle I Molecular Cell Biology November 6, 2014 Stephen Oh, M.D., Ph.D. Assistant Professor Division of Hematology

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Cell Cycle I Molecular Cell Biology November 6, 2014. Stephen Oh, M.D., Ph.D. Assistant Professor Division of Hematology. Outline. Overview of the cell cycle C ell cycle regulation – fundamental concepts Cancer as a fundamental disruption in cell cycle regulation. - PowerPoint PPT Presentation

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Page 1: Cell Cycle I Molecular Cell Biology November 6, 2014

Cell Cycle I

Molecular Cell BiologyNovember 6, 2014

Stephen Oh, M.D., Ph.D.Assistant Professor

Division of Hematology

Page 2: Cell Cycle I Molecular Cell Biology November 6, 2014

Outline

• Overview of the cell cycle

• Cell cycle regulation – fundamental concepts

• Cancer as a fundamental disruption in cell cycle regulation

Page 3: Cell Cycle I Molecular Cell Biology November 6, 2014

What is the basic function of the cell cycle?

• Accurately duplicate the vast amount of DNA in chromosomes

• Segregate the copies precisely into genetically identical daughter cells

Figure 17-2 Molecular Biology of the Cell, 4th Edition

Page 4: Cell Cycle I Molecular Cell Biology November 6, 2014

The phases of the cell cycle

Figure 17-3. Molecular Biology of the Cell, 4th Edition

• G1 – gap between M and S phases• S – DNA replication• G2 – gap between S and M phases• M - mitosis

• Interphase ~23 hours• M phase ~1 hour

Why are gap phases needed?

Page 5: Cell Cycle I Molecular Cell Biology November 6, 2014

What critical features are needed for proper guidance through the cell cycle?

Figure 17-13 Molecular Biology of the Cell, 4th Edition

Page 6: Cell Cycle I Molecular Cell Biology November 6, 2014

• A clock, or timer, that turns on each event at a specific time

• A mechanism for initiating events in the correct order

• A mechanism to ensure that each event is triggered only once per cycle

• Binary (on/off) switches that trigger events in a complete, irreversible fashion

• Backup mechanisms to ensure that the cycle can work properly even when parts of the system malfunction

• Adaptability so that the system's behavior can be modified to suit specific cell types or environmental conditions

Figure 17-13 Molecular Biology of the Cell, 4th Edition

What critical features are needed for proper guidance through the cell cycle?

Page 7: Cell Cycle I Molecular Cell Biology November 6, 2014

The cell cycle is primarily regulated by cyclically activated protein kinases

Figure 17-15, 17-16 Molecular Biology of the Cell, 4th Edition

Page 8: Cell Cycle I Molecular Cell Biology November 6, 2014

Malumbres M, Nature Reviews Cancer 2009

Evolution of cell cycle control: from yeast to humans

Page 9: Cell Cycle I Molecular Cell Biology November 6, 2014

Table 17-1. Molecular Biology of the Cell, 4th Edition

Overview of major cyclins and Cdks of vertebrates and yeast

Page 10: Cell Cycle I Molecular Cell Biology November 6, 2014

Bardin AJ, Nature Rev Mol Cell Biol 2001

Overview of major cyclins and Cdks of vertebrates and yeast

Page 11: Cell Cycle I Molecular Cell Biology November 6, 2014

Cdk activity is regulated by inhibitory phosphorylation and inhibitory proteins

Figure 17-18, 17-19. Molecular Biology of the Cell, 4th Edition

Why is cell cycle progression governed primarily by inhibitory regulation?

Page 12: Cell Cycle I Molecular Cell Biology November 6, 2014

Figure 17-20. Molecular Biology of the Cell, 4th Edition

Cell cycle control depends on cyclical proteolysis

Page 13: Cell Cycle I Molecular Cell Biology November 6, 2014

Mechanisms controlling S-phase initiation

Figure 17-30. Molecular Biology of the Cell, 4th Edition

Page 14: Cell Cycle I Molecular Cell Biology November 6, 2014

DNA damage leads to cell cycle arrest in G1

Figure 17-33. Molecular Biology of the Cell, 4th Edition

Page 15: Cell Cycle I Molecular Cell Biology November 6, 2014

Figure 17-34. Molecular Biology of the Cell, 4th Edition

Overview of the cell cycle control system

Page 16: Cell Cycle I Molecular Cell Biology November 6, 2014

Table 17-2. Molecular Biology of the Cell, 4th Edition

Summary of major cell cycle regulatory proteins

Page 17: Cell Cycle I Molecular Cell Biology November 6, 2014

Figure 17-41. Molecular Biology of the Cell, 4th Edition

Mitogens stimulate cell division

Page 18: Cell Cycle I Molecular Cell Biology November 6, 2014

Figure 17-42. Molecular Biology of the Cell, 4th Edition

Excessive stimulation of mitogenic pathways can lead to cell cycle arrest or cell death

Page 19: Cell Cycle I Molecular Cell Biology November 6, 2014

Figure 17-44. Molecular Biology of the Cell, 4th Edition

Extracellular Growth Factors Stimulate Cell Growth

Page 20: Cell Cycle I Molecular Cell Biology November 6, 2014

Figure 17-47. Molecular Biology of the Cell, 4th Edition

Extracellular Survival Factors Suppress Apoptosis

Page 21: Cell Cycle I Molecular Cell Biology November 6, 2014

Intracellular signaling networks related to cell proliferation and cancer

Hanahan and Weinberg, Cell 2011

Page 22: Cell Cycle I Molecular Cell Biology November 6, 2014

Primary myelofibrosis

Essential thrombocythemia

Polycythemia vera

JAK2 V617F

Myeloproliferative neoplasms are clonal disorders derived from hematopoietic stem/progenitor cells

Page 23: Cell Cycle I Molecular Cell Biology November 6, 2014

Proliferation/Survival

JAK2JAK2

STAT3/5P P

STAT3/5 STAT3/5P P

P P

STAT3/5

TPOG-CSF

JAK2V617F

STAT3/5 STAT3/5P P

JAK-STAT activation is a hallmark of myeloproliferative neoplasms

Page 24: Cell Cycle I Molecular Cell Biology November 6, 2014

Proliferation/SurvivalTNFαPIM1

JAK2JAK2

STAT3/5P P

STAT3/5 STAT3/5P P

P P

STAT3/5

TPOG-CSF

JAK2V617F

Rux

STAT3/5 STAT3/5P P

PI3K

AKT

S6K

S6

P P

SCFFLT-3L

LNK LNKSOCS

CBL

P PNFkB

Proliferation/SurvivalTNFα, GM-CSFIkBα

IkB degradation

IKKε

IKKγ P

P P

IkBαNFkB

TLRs

TBK1P

IKKα IKKβ

IkBαP

NFkBP P

PIM1BAD

CREB

RAS

RAF

MEK

ERK

Dysregulated signaling networks in myeloproliferative neoplasms

Cell cycle inhibition/Apoptosis

STAT1P

STAT3/5

JAK1JAK1

STAT1 STAT1P P

P P

Ifna

STAT1STAT1P P

STAT1STAT1P P

P

Page 25: Cell Cycle I Molecular Cell Biology November 6, 2014

Spectral limitations of flow cytometry can be overcome with elemental mass cytometry

>30 parameters with single cell

resolution

Labeled cells

Metal conjugatedantibodies

CyTOF2mass cytometer

Mass channel readout

Page 26: Cell Cycle I Molecular Cell Biology November 6, 2014

SPADE links related cell types in a multidimensional continuum of marker expression

Bendall et al Science 2011

How can we visualize data in 30+ dimensions?

Page 27: Cell Cycle I Molecular Cell Biology November 6, 2014

SPADE identifies relevant cell subsets including HSPC

CD34 median expression:

Low High

HSPC

Page 28: Cell Cycle I Molecular Cell Biology November 6, 2014

Cell cycle analysis via mass cytometry

Behbehani et al, Cytometry 2012

Page 29: Cell Cycle I Molecular Cell Biology November 6, 2014

Cell cycle analysis via mass cytometry

Behbehani et al, Cytometry 2012

Page 30: Cell Cycle I Molecular Cell Biology November 6, 2014

Malumbres M, Nature Reviews Cancer 2001

Cell cycle regulators are frequently disrupted in cancer

Page 31: Cell Cycle I Molecular Cell Biology November 6, 2014

Malumbres M, Nature Reviews Cancer 2009

Overview of CDK inhibitors in clinical development for cancer therapy

Results thus far have been somewhat disappointing – why?

Page 32: Cell Cycle I Molecular Cell Biology November 6, 2014

Suggested reading• Alberts et al., Molecular Biology of the Cell, 4th Edition, Garland.

Updated 2001. Chapter 17.– http://www.ncbi.nlm.nih.gov/books/bv.fcgi?rid=mboc4.TOC&depth=2

• Malumres M, Barbacid M. Cell cycle, CDKs and cancer: a changing paradigm. Nat Rev Cancer. 2009 Mar;9(3):153-66.– http://www.nature.com/nrc/journal/v9/n3/full/nrc2602.html

• Hanahan and Weinberg. Hallmarks of Cancer: The Next Generation. Cell. 2011 Mar 4;144(5):646-74.– http://www.sciencedirect.com/science/article/pii/S0092867411001279

• Anand S, Huntly BJ. Disordered signaling in myeloproliferative neoplasms. Hematol Oncol Clin North Am. 2012 Oct;26(5):1017-35.– http://www.sciencedirect.com/science/article/pii/S0889858812001281

Contact: [email protected]