Guanine Nucleotide Exchange Factors for Rho GTPases in endothelial - - PowerPoint PPT Presentation

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Guanine Nucleotide Exchange Factors for Rho GTPases in endothelial cells: Critical transducers of angiogenic signals JOSÉ VÁZQUEZ-PRADO, Ph.D.

Pharmacology Department CINVESTAV-IPN jvazquez@cinvestav.mx

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Endothelial RhoGEFs are potential targets to interfere with tumor induced angiogenesis

VEGF SDF-1

Tumor induced angiogenesis requires endothelial cell migration

β γ

α-GDP

β γ

α-GTP Rho GEF Rac GEF

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Rho GTPases: Promote actin polimerization and cell migration Rac Cdc42 Rho

Science 1998, V

• l. 279,

61 human RhoGEFs

(http://smart.embl-heidelberg.de/).

> 40 are expressed in endothelial cells. One of them is P-Rex1

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P-Rex1

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DEP PDZ DH PH Rac GTP Rac GDP Rac GTP

β γ

DEP PDZ PI3K AKT

P-Rex1

Rac GTP S1P1, CXCR4

• RhoGEFs
• RhoGEF-interacting

proteins

• and their partners ?

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GRp58 BAG5 PRKAR1A DEP PDZ DH PH Rac GTP Rac GDP DEP PDZ

mTorC2 PKA β γ

PI3K AKT S1P , SDF1 HGF

P-Rex1

PhLP-1-149 shRNA mTor shRNA P-Rex1 ∆-DH P-Rex1 Y2H

Rac GTP S1P1, CXCR4

?

Experimental approach

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mTor prey p53

• LT
• HLT

Xα-Gal

• AHLT

Xα-Gal pTD-11 P-Rex1 DEP-DEP p53 P-Rex1 DEP-DEP p53 P-Rex1 DEP-DEP

Hernandez-Negrete et al, 2007. J Biol Chem. 282. 23708

Rac GTP S1P1, CXCR4

P-Rex1- interacting proteins?

Y2H mTORC1 mTORC2

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Hernandez-Negrete et al, 2007.J Biol Chem. 282. 23708

Rac GTP S1P1, CXCR4

P-Rex1-mTOR complex ?

mTORC1 mTORC2

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GRp58

VEGF SDF-1

More than 40 RhoGEFs are expressed in endothelial cells, Their multidomain architecture suggest a potential role as signal integrators and therapeutic targets. P-Rex1 plays a role in the signaling pathway downstream of SDF1/CXCR4 leading to Rac activation, cell migration and in vitro angiogenesis. This GEFs is activated in response to Gβγ→PI3Kγ and AKT→mTor→

Conclusions:

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Future studies: Future studies:

Relevant protein

Relevant protein-

• protein interactions in live

protein interactions in live cells. cells.

RhoGEFs in endothelial cells from cancer

RhoGEFs in endothelial cells from cancer patients? patients?

RhoGEFs in mice models of cancer

RhoGEFs in mice models of cancer

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Group, 2009

Lab35F Pharmacology, Cinvestav

José Vázquez-Prado

jvazquez@cinvestav.mx

Posdoctoral fellows, Aleida Vázquez-Macias María Luisa Guzmán María Luisa Guzmán-

• Hernández.

Hernández. Ph.D. students, Alejandro Ledezma Alejandro Ledezma-

• Sánchez.

Sánchez. Jorge Carretero Jorge Carretero-

• Ortega.

Ortega. Iliana Ramirez Iliana Ramirez-

• Rangel.

Rangel. Ismael Bracho Ismael Bracho-

• Valdés

Valdés Ricardo Hernández Ricardo Hernández-

• Garc

García. a. Lydia Chavez Lydia Chavez-

• Vargas

Vargas Evelyn Robles Evelyn Robles-

• Molina

Molina Israel Valencia Israel Valencia-

• Martinez

Martinez Technician: Estanislao Escobar Estanislao Escobar-

• Islas

Islas Lab assistant: David Pérez David Pérez-

• Rangel

Rangel Collaborators, Guadalupe Reyes Guadalupe Reyes-

• Cruz,

Cruz, Cinvestav

Cinvestav

Alejandro Garcia Alejandro Garcia-

• Regalado

Regalado Claudia Gonzalez Claudia Gonzalez-

• Espinosa,

Espinosa, Cinvestav

Cinvestav

Marina Macias Marina Macias-

• Silva

Silva, IFC

, IFC-

• UNAM

UNAM

Silvio Gutkind, Silvio Gutkind, NIDCR

NIDCR-

• NIH

NIH

Joan Heller Brown Joan Heller Brown, UCSD

, UCSD

Gelasio Zarco Gelasio Zarco-

• Espinosa,

Espinosa, IMSS

IMSS-

• CMN SXXI

CMN SXXI

Gabriela Hernandez Gabriela Hernandez-

• Rivera,

Rivera, INNSZ

INNSZ

Grants: CONACyT, CONACyT, National Institutes of Health, USA National Institutes of Health, USA UC UC-

• Mexus,

Mexus, Former Ph.D students: Ivette Hernández Ivette Hernández-

• Negrete

Negrete Former M.S. students: Sandra Sanchez Sandra Sanchez-

• Barbosa

Barbosa Jonathan García Jonathan García-

• Román

Román Ana Luisa Esparza Ana Luisa Esparza-

• Silva

Silva Liseth Aldaba Liseth Aldaba-

• Muruato

Muruato

Acknowledgments: