Epidermal Growth Factor Receptor(EGFR) signaling pathway

Samantha 03/09/11

Outline

• EGFR signaling pathway• EGFR endocytosis• Crosstalk• Clinic therapy

The significant of EGFR signaling pathway

http://www.informedicalcme.com/egfr/tyrosine-kinase-inhibitors-sensitivity/

EGFR signaling pathway

• EGF Receptors• Downstream of EGFR• A new role for Egfr in cancer

The discovery of epidermal growth factor

http://nobelprize.org/nobel_prizes/medicine/laureates/1986/press.html

http://nobelprize.org/nobel_prizes/medicine/laureates/1986/press.html

The discovery of epidermal growth factor

EGF Receptor Members

Table 15-4 Molecular Biology of the Cell (© Garland Science 2008)

EGF Receptor Members

EGF receptor members:Vertibrates: EGFR; ErbB2; ErbB3; ErbB4C. elegans: Let-23Drosophila: DER

C. elegans and Drosophila have served as useful model systems for studying the signaling processes triggered by the EGF receptor.

RTKs have 3 functional regions:

1. N-terminal extracellular region w/ 1 or more ligand-binding sites

2. Single transmembrane -helix domain

3. C-terminal cytoplasmic region w/ catalytic domain + phosphorylation sites

Lodish (2004) Fig 14-5

Recall:

EGF receptor structure

ErbB3--has impaired kinase activity, and only signals when heterodimerizing with another receptor type.

Bogdan et al. 2000

EGF receptor structure

Eigenbrot et al;

Ligands:

Activation of the EGF receptor

Roepstorff et al.2008

Spices Ligands

C. elegans Lin-3, a soluble TGFa-like ligand.

Drosophila Spitz, Vein, Grken, Keren

Human More than dozen ligands

ErbB2 doesn’t bind any EGF-like ligands on its own, but can heterodimerize with other EGF-bound receptors.

ErbB3 has impaired kinase activity, and only signals when heterodimerizing with another receptor type.

cis- &/or trans-phosphorylation

Receptor tyrosine kinase activation by dimerization

Lodish (2004) Fig 14-5

only trans-phosphorylation

Recall:

Hubbard et al. (2009)

Activation of the EGF receptor

In the cytoplasm, the two tyrosine kinase domains form an asymmetric dimer, with the c-terminal lobe.

Juxtamembrane region of EGFR stabilizes formation of the asymmetric kinase dimer.

EGFR is not activated by autophosphorylation of the activation loop.

Downstream of EGFR

Bogdan et al. 2000

Downstream of EGFR

SIGMA-ALDRICH

Negative regulator of EGFR signaling

Bogdan et al. 2000

Models of Signaling in Membrane Rafts

Model 1a: - raft-associated receptors activated

by dimerization.

Model 1b: - dimerization increases affinity for

raft.

Model 2: - clustering of multiple rafts triggers

signaling

Simons & Toomre 2000

Recall:

Lipid rafts and EGFR

methyl-B-cyclodextrin cholesterol depletion

inhibition of EGF-stimulated PI turnover.BUT, led to an enhancement of EGF-stimulated MAP kinase activity.

Lipid rafts disruption appears to have both positive and negative effects on receptor tyrosine kinase-mediated signaling

Lipid rafts and EGFR

methyl-B-cyclodextrin cholesterol depletion

EGF receptor function is suppressed when the receptor is localized to lipid rafts.

The enhancement of EGF binding and receptor autophosphorylation An increase in intrinsic receptor kinase activity .

A new role of Egfr as a transcription factor

Wen, 2010

A new role of Egfr in cancer

Engelman etal. 2008

Outline

• EGFR signaling pathway• EGFR endocytosis• Crosstalk• Clinic therapy

EGFR endocytosis

• Ligand-induced endocytosis of EGFR• Novel way of EGFR endocytosis

Negative regulator of EGFR signaling

Bogdan et al. 2000

Grb-dependent pathway: Grb2-Cb1 complex is recruited to C-terminal of EGFR.Adds mono-or polyubiquitins to EGFR.Activated EGFR is transported to clathrin coated pits.

Roepstorff et al. 2008

Clathrin-mediated EGFR endocytosis

Clathrin-mediated EGFR endocytosisGrb-independent pathwayCb1 can mediate ubiquination of “X” internalization of EGFR.EGFR can directly interact with AP-2

Sorkin et al.2008

Clathrin-mediated EGFR endocytosis

A, EGFR is recycled back to the plasma membrane.B, EGFR will be degraded in lysosomes.

Clathrin-mediated EGFR endocytosis & signaling

Signal transduction molecules affect membrane trafficking.Membrane trafficking can regulate signal transduction events.

Signal transduction Membrane trafficking

AKT and ERK require CME, EGFR-activated DNA synthesis depends on functional CME.

The initial phase of EGFR signaling is endocyotosis-independent, but Later events require clathrin-mediated EGFR Endocytosis(CME).

Clathrin-independent EGFR endocytosisUnder conditions of receptor overexpression and/or high ligand concentrations, clathrin-independent internalization determines the overall rate of the EGFR uptake into the cell

A-431 cells COS cells Several types of cells

Formation of micro-and macropinocytic vesiles

Large vesicular structures

Vesicular-tubular endocytic compartment originated from the plasma membrane dorsal ruffles

The novel function of EGFR dimerization in internalization

Inhibition of EGFR kinase activation did not block EGF-inducedEGFR internalization. Absence of EGFR dimerization, EGF binding can not stimulate EGFR endocytosis.

EGFR dimerization can mediate the EGF-induced cellular process independently of its role in activating EGFR tyrosien kinase.

Outline

• EGFR signaling pathway• EGFR endocytosis• Crosstalk• Clinic therapy

Cross-talk

• EGFR signaling and Notch pathway• EGFR signaling and GPCR• EGFR and Wnt signaling • EGF and insulin cell signaling pathway

EGFR signaling and Notch pathway

Hasson and Paroush, 2006

EGFR signaling and GPCR

Prenzel et al .2000

EGFR and Wnt signaling

EGF and insulin cell signaling pathway

Outline

• EGFR signaling pathway• EGFR endocytosis• Crosstalk• Clinic therapy

Clinic therapy

• EGFR inhibitor• New possible therapies

Clinic Therapy

• Monoclonal antibodies (mAbs)directed against the EGFR extracellular domain

• Small molecule tyrosine kinase inhibitors (TKIs)directed against the tyrosine kinase domain.

EGFR inhibitor

https://www. medscape.com

Natural inhibitors include potato carboxypeptidase inhibitor(PCI), which contains a small cysteine-rich module, called a T-knot scaffold, that is shared by several different protein families, including the EGF family.

New possible therapies

• Nuclear EGFR and EGFR-targeted Therapy• Therapy based on EGFR kinase-indepenent activity

References• 1, Robert N.Jorissen, Francesca Walker, Normand Pouliot , Thomas P.J.Garrett, Colin W. Ward, and Antony W.Burgess. Epidermal growth factor receptor: mechanisms of activation and signaling. Experimental Cell Research 284(2003)31-53.• 2, Sven Bogdan and Christian Klambt. Epidermal growth factor receptor signaling. Current Biology (2000) Vol10 No8.• 3, Shiaw-Yih Lin, Keishi Makino, Weiya Xia, Angabin Matin, Yong Wen, Ka Yin Kwong, Lilly Bourguignon and Mien-Chie Hung. Nuclear localization of EGF receptor and its potential new role as a transcription factor. Nature Cell Biology Vol3(2001).• 4, Zhang Weihua, Rachel Tsan, Wei-Chien Huang, Qiuyu Wu, Chao-Hua Chiu, Isaiah J. Fidler, and Mien-Chie Huang. Survival of Cancer Cells is Maintained by EGFR independent of Its Kinase Activity. Cancer Cell 13, 385-393, (2008).• 5, Jeffrey A. Engelman and Lewis C. Cantley. A sweet new role for EGFR in cancer. Cancer Cell13(2008).• 6, Kirstine Roepstorff, Lene Grovdal, Michael Grandal, Mads Lerdrup, BoVanDeurs. Endocytic downregulation of ErbB receptors: mechanisms and relevance in cancer. Histochem Cell Biol(2008)129:563-578.• 7, Amandio V Vieira, Christophe Lamaze, Sandra L. Schmid. Control of EGF Receptor Signaling By Clathrin- Mediated Endocytosis. Science Vol274(1996)• 8, Alexander Sorkin, Lai Kuan Goh. Endocytosis and Intracellular trafficking of ErbBs. Experimental Cell Research 315(2009)683-696.

References• 9.Norbert Prezel, Esther Zwick, Michael Leserer and Axel Ullrich. Tyrosine Kinase signaling in breast cancer Epidermal growth factor receptor: Convergence point for signal integration and diversification. Breast Cancer Research(2000), 2:184-190.• 10,Qian Wang, Greg Villeneuve and Zhixiang Wang. Control of epidermal growth factor receptor endocytosis by receptor dimerization, rater than receptor kinase activation. 2005 EuROPEAN MOLECULAR BIOLOGY ORGANIZATION.• 11, Rafal Zielinski, Pawel F Przytycki, Jie Zheng, David Zhang, Teresa M Przytycka and Jacek Capala. The crosstalk between EGF, IGF, and Insuin cell signaling pathways-computational and experimental analysis. BMC Systems Biology 2009, 3:88• 12, Tianhui Hu, Cunxi Li. Convergence between Wnt-B-catenin and EGFR signaling in cancer. Molecular cancer 2010, 9:236.• 13, P Hasson and Z Paroush. Crosstalk between the EGFR and other signaling pathways at the level of the global transcriptional corepressor Groucho/TLE. British Journal of Cancer(2006)94,771-775.• 14, Elizabeth A Musgrove, Wnt signallin gvia the epidermal growth factor receptor: a role in breast cancer? Breast Cancer Res 2004, 6:65-68.• 15, Stevan R. Hubbard. The Juxtamembrane Region of EGFR Takes Center Stage. Cell137, June 26 2009.