Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf ·...
Transcript of Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf ·...
![Page 1: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/1.jpg)
Lecture V
Other angiogenic mediators VEGF family
![Page 2: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/2.jpg)
Vascular endothelial growth factorVascular endothelial growth factor--A (VEGF)A (VEGF)
Yancopoulos, Science 2000
Receptors on endothelial
cells
VEGF-A is a major angiogenic growth factor. It acts on endothelial cells, being produced by numerous cell types, including vascular smooth muscle cells (VSMC), fibroblasts or tumor cells.
![Page 3: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/3.jpg)
VEGFs growth factors activate the class of receptorsVEGFs growth factors activate the class of receptors::VEGFRs and neuropilinsVEGFRs and neuropilins
![Page 4: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/4.jpg)
Gene Sequence homology Splice variants
Main activity
VEGF-A
121,145, 165,183, 189, 206
regulation of normal and
pathological angiogenesis, survival
VEGF-B
45% homology with VEGF-A
167, 186
embryonic myocardial
vascularization
VEGF-C
30% homology with VEGF-A 165 27% homology with VEGF-B 167
388
promotion of angiogenesis and
lymphangiogenesis
VEGF-D
61% homology with VEGF-C
31% with VEGF-A 165
333
regulator of lymphangiogenesis, proliferation of endothelial cells
PlGF
42% homology with VEGF-A
131, 152
modulation of pathological angiogenesis, migration of
endothelial cells and monocytes
![Page 5: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/5.jpg)
Placenta growth factor
- expressed primarily in the placenta, especially late stages of gestation
- expression up-regulated during pathological conditions: heart ischemiamyocardial infarction, wound healing
- binds to VEGFR-1
![Page 6: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/6.jpg)
Placental growth factor
Trends Cardiovasc Med. 2002
131 aa
152 aa
203 aa
Signal peptide
![Page 7: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/7.jpg)
Molecular characterization of PlGF
* Biologicaly active glicoprotein acting as homodimer 46 kDa, as well as heterodimer with VEGF
* PlGF 1 – 131 a.a. secretory protein, undergoes N glycosylation
* PlGF 2 – 152 a.a. highly cationic 21 a.a. carboxyterminal (encoded in the 6th exon) enables it’s association with membrane by binding to polyanionic heparan sulphate
* PlGF 3 – the longest of all three isoforms, extra 72 a.a.(216 b.p.) insert between 4th and the 5th exon, N glycosylation
![Page 8: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/8.jpg)
Receptor for PlGF
* Distinct pathophysiological roles of PlGF are mediated by binding to the fms-like tyrosine kinase receptor 1 Flt 1/ VEGFR1
* PlGF binds to Flt 1/ VEGFR1 with high affinity leading to its trans phosphorylation, thus activating signal transduction cascade
* PlGF does not cause trans phosphorylation and further signal transduction cascade through the KDR/ VEGFR2
![Page 9: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/9.jpg)
Role Role of PlGF of PlGF –– effect of PlGF gene knockouteffect of PlGF gene knockout
1. Does not affect embryonic angiogenesis 2. Impaired inflammatory angiogenesis 3. Bone-marrow transplant restored inflammatory angiogenesis in
adult PlGF-ko mice 4. Synergism with VEGF, not bFGF 5. Amplify the angiogenic response to VEGF 6. Effect of PlGF in inflammation is mediated by VEGFR1, but it is not
angiogenic
![Page 10: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/10.jpg)
Role Role of PlGF in embryo and in the adult of PlGF in embryo and in the adult
- amplification of VEGF-driven angiogenesis by PlGF
Carmeliet et al., 2003
![Page 11: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/11.jpg)
Intramolecular crossIntramolecular cross--talk talk through flkthrough flk--1/1/fltflt--1 1 dimerization dimerization
Carmeliet et al., 2003
![Page 12: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/12.jpg)
Synergism between Synergism between VEGF VEGF and PlGF and PlGF
1. By activating VEGFR-1, PlGF induces an intermolecular cross-talk between VEGFR-1 and VEGFR-2, what activates VEGFR-2 and enhances the response to VEGF
2. PlGF, as a subunit of the PlGF/VEGF heterodimer, stimulatesangiogenesis by inducing the formation of VEGFR-1/VEGFR-2 heterodimers, which transphosphorylate each other in intramolecular reaction
3. PlGF upregulates VEGF, thereby amplifying the synergism
![Page 13: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/13.jpg)
Embryonic stem cells derived tumors Embryonic stem cells derived tumors
Wild type
PlGF-/-
VEGF-/-
Carmeliet et al., 2002
![Page 14: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/14.jpg)
Impaired wound healing in PlGFImpaired wound healing in PlGF--//-- mice mice
Carmeliet et al., 2002
![Page 15: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/15.jpg)
Vascular response Vascular response to to PlGF adenoviral gene PlGF adenoviral gene transfer transfer in the in the ear of nude ear of nude mice mice
0 days 14 days 28 days
Luttuin et al., Nature Med. 2002
![Page 16: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/16.jpg)
Nature Med. 2001
Role Role of PlGF in recruiment of progenitor cells of PlGF in recruiment of progenitor cells
![Page 17: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/17.jpg)
Different cells express Different cells express VEGF VEGF receptorsreceptors
VEGFR-1
ENDOTHELIAL CELLS; osteoblasts, monocyte/macrophages,pericytes, placental trophoblasts, renal mesangial cells, some hHematopoietic cells
VEGFR-2
ENDOTHELIAL CELLS; neuronal cells, osteoblasts,pancreatic ductal cells, retinal progenitor cells, megakaryocytes, some hematopoietic cells
VEGFR-3
Present on all endothelia during development, but in the adult becomes restricted to lymphatic EC and certain fenestratedblood vascular EC. Up-regulated on blood vascular EC in tumors
![Page 18: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/18.jpg)
Inhibition of PlGF signaling results in:
- reduced mobilization of bone-marrow-derived myeloid progenitors into the peripheral blood
- impaired infiltration of Flt-1 expressing leukocytes in inflammed tissues
- defective activation of myeloid cells
-Inhibition of the atherosclerotic plaque growth
AntiAnti--inflammatory effect of antiinflammatory effect of anti--Flt1 (VEGFR1)Flt1 (VEGFR1)
![Page 19: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/19.jpg)
AntiAnti--FltFlt--1 1 inhibits vascularisation inhibits vascularisation ((Matrigel plugMatrigel plug) )
Luttuin et al., Nature Med. 2002
![Page 20: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/20.jpg)
Luttuin et al., Nature Med. 2002
![Page 21: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/21.jpg)
VEGF/VEGFR-2 axis – important for physiological (developmental) and pathological angiogenesis
PlGF/VEGFR-1axis – specifically crucial for pathological angiogenesis
![Page 22: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/22.jpg)
VEGFVEGF--B B
-a ligand for VEGFR-1
-Two isoforms: VEGFB-167 (binds to heparan sulphate) and VEGFB-186
- may contribute to the embryonic myocardial vascularization
- may play a role in pathological angiogenesis (eg synovial inflammation was reduced in VEGFb knockout mice )
![Page 23: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/23.jpg)
Li &Erriskon, 2001
Heparin –binding domainVEGF homology domain
unique VEGFB186 domain
Silk domain
![Page 24: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/24.jpg)
Functions and activity of Functions and activity of VEGFVEGF--BB
- binds to VEGFR-1 - knockouts are not lethal - activates Akt kinase (like VEGF-A) - increases eNOS expression - induced angiogenesis in subcutaneously injected Matrigel –
- angiogenesis was blocked by L-NAME and ant-VEGFR1
Seems to be required for coronary artery development
![Page 25: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/25.jpg)
Lymphatic Lymphatic system system Gasparo Assellius – 1627
Functions:
1. Collection of fluids extravasated from the vessels 2. Fat uptake in the gut 3. Immune functions
4. Role in pathology: lymphaedema (insuficient) tumor growth and metastatis (overgrowth of
lymphatic vessels)
![Page 26: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/26.jpg)
Vascular Vascular system system and lymphatic and lymphatic systemsystem
Jones N et al., Nature Rev Mol cell Biol. April 2001
![Page 27: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/27.jpg)
Karkkainen et al., Nature Cell Biol 2002
Development of lymphatic Development of lymphatic system system
- from embryonic vein - from lymphangioblasticprecursors in the tissues
![Page 28: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/28.jpg)
Rafii S & Skobe M, Nature Med. February 2003
Segregation of blood and lymphatic vessels Segregation of blood and lymphatic vessels
![Page 29: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/29.jpg)
Development of the lymphatic Development of the lymphatic system system
Blood vessels lymph vessels
VEGFR-3LYVE-1
VEGFR-1VEGF-2Tie-2Nrp-1CD31CD31
RK Jain & TP Padera, Science 2003: 299: 209-210.
VEGFR-3Prox-1SLC
Prox-1LYVE-1
![Page 30: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/30.jpg)
ProxProx--1 1 –– a master regulator a master regulator of the lymphatic vasculature phenotype of the lymphatic vasculature phenotype Prospero-like protein
![Page 31: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/31.jpg)
Prox-1 up-regulates lymphangiogenic genes
Knockouts of Prox-1:
+/- - appear normal at birth, but die within 2-3 days after birth
-/- - die at E14.5-15.5 due to combination of several phenotypic alterations, including lack of lymphatic vessel formation
![Page 32: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/32.jpg)
Rafii S & Skobe M, Nature Med. February 2003
Segregation of blood and lymphatic vessels Segregation of blood and lymphatic vessels
![Page 33: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/33.jpg)
Markers for the lymphatic vessels
![Page 34: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/34.jpg)
Gale & Yancopoulos, 1999
![Page 35: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/35.jpg)
VEGF-C - like VEGF, stimulates the migration of EC and increase vascular permeability and EC proliferation (but at higherconcentrations than VEGF)
- binds to VEGFR-3 (FLT4) and VEGFR-2; binds also to Nrp-2
- its expression does not seem to be regulated by hypoxia, but is by pro-inflammatory cytokines
- regulate physiological and pathological-blood vessel growth in vivo
- regulate growth of lymphatic vessels
- overexpression of VEGF-C in skin keratinocytes leads to dermal-lymphatic vessel hyperplasia
- knockouts – results in defects in intestinal and cutaneous lymphatic vessels
![Page 36: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/36.jpg)
VEGFRVEGFR--3 3 and lymphangiogenesis and lymphangiogenesis
VEGFR-3 is initially expressed in all venous endothelial cells
Only Prox-1-expressing cells start the sprouting process and simultaneously upregulate other lymphatic markers, such as podoplanin and LYVE-1
Signaling by VEGFR-3 alone is sufficient to induce lymphangiogenesis
Knockout of VEGFR-3 Lethal at E 9.5 - enlarged vessel lumens, cardiovascular failure
![Page 37: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/37.jpg)
Proteolytic activations of VEGFsProteolytic activations of VEGFs
McColl et al., APMIS 2004
Solubilisation Release from ECM
Binding to Neuropilin-1
Binding to VEGFR-2 and increased affinity to VEGFR-3 and Neuropilin-1
Binding to VEGFR-2 and increased affinity to VEGFR-3 and neuropilin-1
VEGF-A VEGF-B186 VEGF-C VEGF-D
![Page 38: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/38.jpg)
Processing of VEGF-C
![Page 39: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/39.jpg)
VEGF-C acquires the capacity to bind VEGFR-2
- binding activity of processed VEGF-D to VEGR-2 increasesby 290-fold
Processing signifcantly enhances activity ofProcessing signifcantly enhances activity of VEGFVEGF--CC andand DD
![Page 40: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/40.jpg)
1. In Vegfc-/- mice, endothelial cells commit to the lymphatic lineage but do not sprout to form lymph vessels.
2. Sprouting was rescued by VEGF-C and VEGF-D but not by VEGF, indicating VEGF receptor 3 specificity.
3. The lack of lymphatic vessels resulted in prenatal death due to fluid accumulation in tissues, and Vegfc+/- mice developed cutaneous lymphatic hypoplasia and lymphedema.
VEGF-C is the paracrine factor essential for lymphangiogenesis, and show that both Vegfc
alleles are required fornormal lymphatic development.
Effect of VEGF-C gene knockout
Karkkainen MJ, Nature Immunol 2004
![Page 41: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/41.jpg)
VEGFVEGF--D D c-FOS induced growth factor (FIGF)
- 61% of aa identity to VEGF-C
- binds VEGFR-3 and VEGFR-2 (but mice VEGF-D does not bind to VEGFR2)
- also proteolytically processed in a similar way like VEGF-C
- stimulates EC proliferation
- is lymphangiogenic when overexpressed in skin keratinocytes
- little is known about VEGF-D expression in physiological conditions
![Page 42: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/42.jpg)
VEGFVEGF--D D knockoutknockout
?
![Page 43: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/43.jpg)
Karkkainen et al., Nature Cell Biol 2002
VEGFVEGF--C C and and VEGFVEGF--D D signalingsignaling
![Page 44: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/44.jpg)
VEGFR-3
Present on all endothelia during development, but in the adult becomes restricted to lymphatic EC and certain fenestratedblood vascular EC. Up-regulated on blood vascular EC in tumors
![Page 45: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/45.jpg)
Functions of Functions of VEGFVEGF--C C and and VEGFVEGF--D D in tumors in tumors
![Page 46: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/46.jpg)
Functions of Functions of VEGFVEGF--C C and and VEGFVEGF--D D in tumors in tumors
- produced by tumor and stromal cells
- signal through VEGFR-2 – leads to angiogenesis and tumor growth
- signal through VEGFR3- on lymphatic endothelial cells – promotes metastasis
- modulate also immune functions: VEGF-C – is chemotactic for macrophages – express VEGFR-3macrophages also express and secrete VEGF-C and VEGF-D
![Page 47: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/47.jpg)
VEGF-A and VEGF-D induces strong capillary enlargement in skeletal muscle
Rissanen et al., Circ Res 2003
![Page 48: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/48.jpg)
Regulation of VEGFs expression Regulation of VEGFs expression
McColl et al., APMIS 2004
![Page 49: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/49.jpg)
![Page 50: Other angiogenic mediators VEGF familybiotka.mol.uj.edu.pl/zbm/handouts/2006_05_angiogeneza.pdf · 2009-02-19 · VEGF-A 121,145, 165,183, 189, 206 regulation of normal and pathological](https://reader034.fdocuments.in/reader034/viewer/2022042310/5ed86b83b7394006ce60297f/html5/thumbnails/50.jpg)