Download Angiogenesis

PhD program
Molecular Signal Tranduction
TUMOR ANGIOGENESIS
Erhard Hofer
Department of Vascular Biology and Thrombosis Research
Center for Biomolecular Medicine and Pharmacology
Medical University of Vienna
Part I: Overview of vessel formation
1- Angiogenesis and vasculogenesis
2- Important factors and receptors
3- VEGF receptor signaling
4- Tumor angiogenesis
5- Anti-angiogenesis therapies
Literature:
Books:
B. Alberts et al., Molecular Biology of the Cell,
5th Edition, Taylor and Francis Inc., 2007
Pg. 1279-1283
R.A. Weinberg, The Biology of Cancer,
Garland Science, 2007
Pg. 556-585
Tumor Angiogenesis - Basic mechanisms and Cancer Therapy,
D. Marme, N. Fusenig, ed.
Springer Verlag 2008
Angiogenesis - From basic science to clinical application
N. Ferrara, ed.
CRC Press, Taylor&Francis Group, 2007
Literature:
Reviews:
Nature Insight Angiogenesis
G.D. Yancopoulos et al. (2000). Vascular-specific growth factors and blood vessel
formation. Nature 407, 242-248.
Angiogenesis Focus, Nature Med 9, June 2003
Peter Carmeliet, Angiogenesis in Health and Disease
Napoleone Ferrara et al., The biology of VEGF and its receptors
Rakesh K. Jain, Molecular regulation of vessel maturation
Shanin Rafii and David Lyden, Therapeutic stem and progenitor cell
transplantation for organ vascularization and regeneration
Christopher W. Pugh and Peter J. Ratcliffe, Regulation of angiogenesis by hypoxia:
role of the HIF system
Angiogenesis, Nature Reviews Cancer 3, June 2003
Gabriele Bergers and Laura E. Benjamin, Tumorigenesis and the
angiogenic switch
C.J. Schofield and P.J. Ratcliffe. Oxygen sensing by HIF hydroxylases.
Nature Rev. Mol. Cell Biol. 5, 343-354 (2004)
Nature Insight Angiogenesis, Vol. 438, pg. 931-974, December 2005
Carmeliet, Angiogenesis in life, disease and medicine
Coultas, Endothelial cells and VEGF in vascular development
Alitalo, Lymphangiogenesis in development and human disease
Greenberg, From angiogenesis to neuropathology
Gariano, Retinal angiogenesis in development and disease
Ferrara, Angiogenesis as a therapeutic target
P. Carmeliet and M. Tessier-Lavigne, Common mechanisms of nerve and blood
vessel wiring, Nature 436, 195-200 (2005)
J. Folkman, Angiogenesis: an organizing principle for drug discovery ?
Nature Reviews Drug Discovery 6, 273-286 (2007)
Role of notch:
Adams and Alitalo, Molecular regulation of angiogenesis and lymphangiogenesis,
Nature Rev Mol Cell Biol 8, 464-478 (2007)
Germain et al., Hypoxia-driven angiogenesis, Curr Opinion in Hematol 17 (2010)
Guidance cues:
Larrivee et al., Guidance of vascular development: Lessons from the nervous system,
Circulation Research 104, 428-441 (2009)
Gaur et al., Role of class 3 semaphorins and their receptors in tumor growth
and angiogenesis, Clin Cancer Res 15, 6763-70 (2009)
Unterlagen:
http://mailbox.univie.ac.at/erhard.hofer
Student point, Vorlesungsunterlagen
[email protected]
Structure of vessels and capillaries
Small artery: Monocellular layer of endothelial cells
Capillary: endothelial cell,
basal lamina, pericytes
Angiogenesis:
Sprouting of cells from mature endothelial cells of the vessel wall
(secretion of proteases, resolution of
Basal lamina, migration towards
Chemotactic gradient, proliferation,
Tube formation)
VEGF is factor largely specific for
endothelial cells,
bFGF can also induce,
not specific for EC)
Mouse cornea:
wounding induces
angiogenesis,
chemotactic
response to
angiogenic factors
Sprouting towards chemotactic gradient: VEGF
Hypoxia - HIF - VEGF
every cell must be within 50 to 100 mm of a capillary
HIF: hypoxia inducible factor
VEGF: vascular endothelial growth factor
Von Hippel-Lindau Tumor Suppressor, HIF and VEGF
VEGF-gene:
Regulated by HIF,
HIF is continously produced,
ubiquitinylated,
degraded in proteasome,
therefore low concentration;
Ubiquitinylation dependent on
Hippel-Lindau tumor
suppressor
(part of an E3 ubiquitin-ligase
complex)
HIF1a is modified by a
prolyl hydroxylase,
then better interaction with
vHL protein, high turnover;
Hydroxylase is regulated by O2
capillaries sprouting in
the retina of an embryonic mouse
capillary lumen opening up
behind the tip cell
(red dye injected)
Vasculogenesis
Formation of vessels by
differentiation of cells from
angioblasts in the yolk sac
of the embryo:
Is differentiation and proliferation
of endothelial cells
in a non-vascularized tissue
Leads to formation of a primitive
tubular network
Has to undergo angiogenic
remodeling to stable vascular
system
Postnatal vasculogenesis
Hemangioblast
Angioblast
EC
Factors and receptors
Endothelium-specific factors:
VEGF family: 5 factors
Angiopoietin family : 4 factors
Ephrin family : at least 1 factor
Non EC-specific factors :
bFGF
PDGF
TGF-b
VEGF/VEGFR family
VEGF/VEGFR:
VEGF-A: initiation of vasculogenesis
and sprouting angiogenesis,
Immature vessels,
Vascular permeability factor,
Haploid insufficiency in k.o. mice,
PlGF: remodeling of adult vessels
VEGF-B: heart vascularization ?
VEGF-C: lymphatic vessels
VEGF-D: lymphatic vessels ?
VEGFR-2: growth and permeability
VEGFR-1: negative role ?, decoy receptor,
synergism with VEGFR-2 in
tumor angiogenesis
VEGFR-3: lymphatic vessels
Network of lymphatic vessels (red) and capillaries (green):
Lymphatic vessels are larger, not supported by underlying mural cells
Figure 13.31 The Biology of Cancer (© Garland Science 2007)
Differential signaling by tyr kinase receptors
EC “specific” factors/receptors:
VEGFR2
VEGFR1
VEGF-A, PLGF
VEGFR2
VEGF-A
VEGFR3
TIE1
TIE2
VEGF-C
?
ANG1,2
Y799
Y820
P38, src (vascular leakage?)
Y925
Y936
Y951
TSAd (migration)
Y994
Y1006
Y1052
Y1057
PI-3 kinase (survival)
Y1080
Y1104
gene regulation
Y1128
Y1134
Y1175
Y1212
Y1221
Y1303
Y1307
Y1317
PLC-g
proliferation
vasculogenesis
angiogenesis
Sakurai et al.
PNAS 2005
82 of the most strongly VEGFregulated genes (over 5-fold) compared
to EGF and IL-1 induction
VEGF + IL-1 cluster
VEGF + EGF + IL-1 cluster
VEGF + EGF cluster
VEGF cluster
Overlapping and specific gene repertoires
of VEGF, EGF and IL-1
VEGF
20%
EGF
20%
IL-1
60%
About 60 genes reproducibly induced by VEGF over 3-fold
VEGF-induced genes overlap to a large degree
with IL1-induced genes (50-60 %)
20 % of genes are preferentially induced by VEGF
Signaling by receptors of endothelial cells
IL-1
VEGF-A
IL-1R
EGF
VEGFR-2
EGFR
p38
P
PLC-g
MyD88
951
TsAd
P
P
1175
1175
PI3K
IKKa/IKKb/IKKg
Ca++
P
Grb2
Actin
cytoskeleton
PKC
Ras
Akt
IB
NFB
NFB
Calcineurin
NFAT
EGR-1
EGR-1
gene regulation
proliferation
inflammation
Raf
MEK/ERK
Raf
MEK/ERK
survival
migration
permeability
angiogenesis
Hofer E., Schweighofer B. Signaling transduction induced in endothelial cells by growth factor receptors involved in
angiogenesis. Thrombosis ang haemostasis 2007
Guidance molecules in endothelial tip cell
attraction and repulsion
Carmeliet P, Nature. 2005
Eichmann A, Curr Opin
Neurobiol. 2005
Angiopoietins und Tie Receptors:
Ang1: remodeling and maturation
Quiescence and stability
Resistance to permeability,
Supports interaction with other cells and matrix,
Vessel size (VEGF number of vessels),
Repair of damaged vessels
Ang2: natural antagonist,
Overexpression similar Ang-1 k.o. oder Tie-2 k.o.,
Destabilization signal for initiation of vascular remodeling
Either regression or increased VEGF sensitivity
Ang2 is induced in tumors
Ang3: ?
Ang4: ?
Tie2: binds Ang1-4
Tie1: ?
Ephrins und Eph-Receptors:
Largest family of growth factor receptors,
Relevant for vascular system:
Ephrin B2/ Eph B4 : remodeling and maturation
Different for early arterial (Ephrin B2)
and venous vessels (EphB4),
Hypothesis: role for fusion of arterial/
venous vessels
Growth of tumor vessels
3-incorporation of
BM-derived precursors
2-Intussusceptive
growth
1-Sprouting
4-Cooption of existing vessels
5-Lymphangiogenesis
Role of VEGF and Ang2 for tumor angiogenesis,
VEGF-blockade is promising for anti-ngiogenesis therapy
Concept 1: non-vascularized Tumor
Concept 2: many tumors “home in” onto vessels, occupate existing vessels,
Vessel produces Ang2, first tumor regression, then VEGF production by tumor
Recruitment of capillaries by an implanted tumor
Figure 13.32a The Biology of Cancer (© Garland Science 2007)
Chaotic organization of tumor-associated vasculature
Figure 13.34a The Biology of Cancer (© Garland Science 2007)
Structure and function of tumor vessels:
Chaotic architecture and blood flow
Therefore hypoxic and acidic regions in tumor
Permeability strongly increased
fenestrae
enlarged Junctions
No functional lymphatics inside the tumor
enlarged in surrounding,
increases metastasis
Mosaic vessels
Abnormale endothelium
Tumor vessel is only partially overlaid by pericytes and SMC
Figure 13.33 The Biology of Cancer (© Garland Science 2007)
The Rip-Tag model of islet tumor cell progression
Transgene: SV40 large and small T transcription driven by insulin promoter
Transcription in b-cells of islets of Langerhans
Figure 13.37 The Biology of Cancer (© Garland Science 2007)
The angiogenic switch and recruitment of inflammatory cells
Figure 13.38b The Biology of Cancer (© Garland Science 2007)
Heterotypic interactions as targets for therapeutic intervention
Figure 13.49 The Biology of Cancer (© Garland Science 2007)
Inhibition of tumor angiogenesis
(Combination with
5-fluorouracil for
colorectal cancer)
1-Bevacizumab
2-VEGF-trap
3-Pegaptinib
(Macular degeneration)
4
6- downstream
Signals ?
5- SU11248
Bay43-9006
Bevacizumab
Colorectal cancer
Phase III
Combination therapy
Hurwitz et al. 2004
Mass et al. 2004
IFL:
Irinotecan
5-fluorouracil
Leucovorin
Median survival benefit
of two trials (2004):
3.7-4.7 months
Gentherapien:
rAdenoviren
rRetroviren
Targeting of viruses to tumors, tumor endothelium
Targeting of liposomes to tumors, tumor endothelium
Oncolytic viruses
BM progenitor cells home to tumor vasculature
Next meeting in Zürich, June 15-18, 2011
organized by Michael Detmar, ETH
Ralf Adams, Max-Planck-Institute, Müns ter, Ger many
Kari Alitalo, University of Helsinki, Finland
Hirofumi Arakawa, National Cancer Center Research In stitute, Tokyo, Japan
Hellmut Augustin, German Cancer Research Center, Heidelberg, Germany
Roy Bicknell, University of Birmingha m, UK
Georg Breier, Technical University Dresden, Germany
Peter Carmeliet, Catholic University of Leuven, Belgium
Michael Detmar, Swiss Federal Inst itute of Techno logy Zur ich, Switzerland
Anna Dimberg, Uppsala University, Sweden
Anne Eichmann, INSERM U833, College de France, Paris, France
Britta Engelhardt, University of Bern, Switze rland
Napoleone Ferrara, Genentech Inc ., San Francisco, USA
Holger Gerhardt, London Research Institute, Cancer Research UK
Dontscho Kerjaschki, Medical University of V ienna, Austria
Alexander Koch, Gene ntech In c., San Francisco, USA
Donald McDonald, University of California, San Francisco, USA
Gera Neufeld, Israel Inst itute of Technology, Ha ifa, Israe l
Jaques Pouyssegur, Inst itute of Developmental Biology and Cancer, Nice,
France
Masabumi Shibuya, University of Tokyo, Japan
Dietmar Vestweber, Max-Planck-Institute, Münster, Germany