Vascular growth factors and their inhibitors have a great potential for the treatment of human diseases. We focus on their preclinical and translational use in studies to restore homeostasis and to improve tissue functions in cardiovascular disease and several other human diseases.
Kari Alitalo
GROUP LEADER
MD, PHD, ACADEMY PROFESSOR
CONTACT
KARI.ALITALO@HELSINKI.FI
+358 50 500 3572
The aim of our research is to establish novel therapy modalities based on knowledge of the biology of vascular endothelial growth factors (VEGFs), angiopoietins (Ang), and the growth of blood vessels (angiogenesis) and lymphatic vessels (lymphangiogenesis). Current cardiovascular and cancer therapies are often insufficient, unsuccessful or not suitable for all patients, thus novel therapies are urgently needed. Inhibition of vessel growth is already used in the clinics, but with limited success. On the other hand, stimulation of the growth of blood vessels, angiogenesis, and of arteriogenesis, the growth of (collateral) arteries, has been unsuccessfully tried for the treatment of tissue ischemia.
We explore the functions and the translational potential of the vascular growth factors and receptors that we have discovered. Our results have suggested therapeutic utility in a number of human diseases. In our studies, we use molecular genetic models and functional genomics, single-cell transcriptomics, proteomics and metabolomics, viral vectors for gene delivery and biologicals that block growth factor-receptor interactions. Using thes technology platforms, we are well suited and networked for new discoveries with the aim to advance current therapy of cardiovascular diseases and several other human diseases. Three of our previous concepts have already been translated to clinical development and we aim to provide additional therapeutic modalities.
Our work on endothelial growth factors and our new discoveries should provide the possibility of exploring additional treatments for human diseases in which the vasculature is an essential participant.
Members of the VEGF family, comprising five mammalian proteins, are major regulators of blood and lymphatic vessel development and growth. VEGFs stimulate angiogenesis and lymphangiogenesis by activating VEGF receptor (VEGFR) tyrosine kinases in endothelial cells. In physiological and pathological angiogenesis, lack of sufficient oxygen induces the production of VEGF, which stimulates vessel growth mainly via VEGFR-2, whereas VEGFR-1 serves mostly as a decoy receptor without effective signal transduction. Although antibodies that block VEGF have been successfully used in tumor therapy, attempts to use VEGF for proangiogenic therapy in tissue ischemia have been hampered by VEGF-induced vascular leakage and growth of abnormal vessels.
VEGF-B has remained rather enigmatic since its discovery more than 20 years ago. However, we have shown that VEGF-B can provide protection from myocardial infarction by expanding the coronary vasculature and cardiac muscle mass. Furthermore, VEGF-B expression is decreased in failing hearts. It seems that VEGF-B administration provides a unique possibility to remodel the myocardial vasculature to boost blood flow in the heart and in adipose tissue. Our experiments indicate that VEGF-B can also improve glucose and insulin tolerance in obese mice.
In adults, the VEGF-C and VEGF-D receptor, VEGFR-3, is expressed primarily in the lymphatic vasculature, where VEGF-C acts as a regulator of lymphatic vessel growth. VEGFR-3 is also expressed in growing blood vessels. In 2015, we discovered a meningeal lymphatic vessel system around the brain. The meningeal lymphatics could be involved in a number of neurodegenerative and neuroinflammatory diseases of considerable socioeconomic burden. We currently study possible functions of VEGF-C/VEGFR-3 in cerebrovascular dynamics, fluid drainage and cellular trafficking in mouse models of Alzheimer’s disease and of multiple sclerosis.
Angiopoietins (Ang1, Ang2, Ang4) are critical for vascular stabilization after angiogenesis. We have recently shown how the angiopoietin receptor complex Tie1/Tie2 regulates vessel leakage in inflammation, that blocking of Ang2 attenuates neuroinflammation, that Ang2 mutations are associated with human lymphedema and that the full lymphangiogenic activity of VEGF-C requires autocrine Ang2 signaling by the lymphatic endothelium. The unique role of the Ang-Tie signaling pathway in vascular stability suggests that it could serve as a target of therapeutic intervention in diseases in which the vascular integrity is compromised.
Leppäpuska IM, Hartiala P, Suominen S, Suominen E, Kaartinen I, Mäki M, Seppänen M, Kiiski J, Viitanen T, Lahdenperä O, Vuolanto A, Alitalo K, Saarikko AM. Phase 1 Lymfactin® study: 24-month efficacy and safety results of combined adenoviral VEGF-C and lymph node transfer treatment for upper extremity lymphedema. J Plast Reconstr Aesthet Surg. 75:3938-3945, 2022.
Results of phase I clinical trial on adenoviral VEGF-C in lymphedema.
Korhonen EA, Murtomäki A, Jha SK, Anisimov A, Pink A, Zhang Y, Stritt S, Liaqat I, Stanczuk L, Alderfer L, Sun Z, Kapiainen E, Singh A, Sultan I, Lantta A, Leppänen VM, Eklund L, He Y, Augustin HG, Vaahtomeri K, Saharinen P, Mäkinen T, Alitalo K. Lymphangiogenesis requires Ang2/Tie/PI3K signaling for VEGFR3 cell-surface expression. J Clin Invest. 132(15):e155478, 2022
This study explains the mechanism by which angiopoietin2 and Tie1 mutations cause lymphedema.
Leppänen VM, Brouillard P, Korhonen EA, Sipilä T, Jha SK, Revencu N, Labarque V, Fastré E, Schlögel M, Ravoet M, Singer A, Luzzatto C, Angelone D, Crichiutti G, D’Elia A, Kuurne J, Elamaa H, Koh GY, Saharinen P, Vikkula M, Alitalo K. Characterization of ANGPT2 mutations associated with primary lymphedema. Sci Transl Med 12: eaax8013, 2020.
Discovery of mutations in angiopoietin 2 growth factor gene in primary lymphedema patients
Antila S, Karaman S, Nurmi H, Airavaara M, Voutilainen MH, Mathivet T, Chilov D, Li Z, Koppinen T, Park J-H, Fang S, Aspelund A, Saarma M, Eichmann A, Thomas J-L, Alitalo K. Development and plasticity of meningeal lymphatic vessels. J Exp Med 214: 3645-3667, 2017.
Meningeal lymphatics develop postnatally and require continuous VEGF-C stimulation for maintenance.
Aspelund A, Antila S, Proulx ST, Karlsen TV, Karaman S, Detmar M, Wiig H, Alitalo K. A dural lymphatic vascular system that drains brain interstitial fluid and macromolecules. J Exp Med 212: 991-9, 2015.
Listed amongst the 10 Breakthroughs of 2015 by Science and as one of 10 Notable advances 2015 by Nature Medicine.
Aspelund A, Tammela T, Antila S Nurmi H, Leppänen VM, Zarkada G, Stanczuk L, Francois M, Mäkinen T, Saharinen P, Immonen I, Alitalo K. The Schlemm’s canal is a VEGF-C/VEGFR-3-responsive lymphatic-like vessel. J Clin Invest 124: 3975-3986, 2014.
Schlemm’s canal that regulates intraocular pressure is a target of the lymphangiogenic growth factor VEGF-C.
Jeltsch M, Jha SK, Tvorogov D, Anisimov A, Leppänen V-M, Holopainen T, Kivelä R, Ortega S, Kärpänen T, Alitalo K. CCBE1 enhances lymphangiogenesis via a disintegrin and metalloprotease with thrombospondin motifs-3-mediated vascular endothelial growth factor-C activation. Circulation 129: 1962-1971, 2014.
Discovery of the first protease that activates VEGF-C and the mechanism of its regulation by CCBE1.
Leppänen V-M, Tvorogov D, Kisko K, Prota AE, Jeltsch M, Anisimov A, Markovic-Mueller S, Stuttfeld E, Goldie KN, Ballmer-Hofer K, Alitalo K. Structural and mechanistic insights into VEGF receptor 3 ligand binding and activation. Proc Natl Acad Sci USA 110: 12960-12965, 2013.
Atomic resolution structure of the VEGF-C/VEGFR3 complex reveals a homologous receptor-receptor interaction site that is compatible with blocking antibody function (Tvorogov et al., 2010).
Tammela T, Zarkada G, Nurmi H, Jakobsson L, Heinolainen K, Tvorogov D, Zheng W, Franco CA, Murtomäki A, Aranda E, Miura N, Ylä-Herttuala S, Fruttiger M, Mäkinen T, Eichmann A, Pollard JW, Gerhardt H, Alitalo K. VEGFR-3 controls tip to stalk conversion at vessel fusion sites by reinforcing Notch signalling. Nature Cell Biol 13: 1202-1213, 2011.
The mechanism of ligand-independent VEGFR-3 function in blood vessel fusions.
Tammela T, Saaristo A, Holopainen T, Ylä-Herttuala S, Andersson LC, Virolainen S, Immonen I, Alitalo K. Photodynamic ablation of lymphatic vessels and intralymphatic cancer cells prevents metastasis. Sci Transl Med 3: 69ra11, 2011.
A new method to inhibit in transit/satellite metastasis.
Tvorogov D, Anisimov A, Zheng W, Leppänen V-M, Tammela T, Laurinavicius S, Holnthoner W, Heloterä H, Holopainen T, Jeltsch M, Kalkkinen N, Lankinen H, Ojala P, Alitalo K. Effective Suppression of Vascular Network Formation by Combination of Antibodies Blocking VEGFR Ligand Binding and Receptor Dimerization. Cancer Cell 18: 630-40, 2010.
These results indicate that VEGF receptor directed antibodies that inhibit ligand binding and receptor dimerization have additive or even synergistic effects.
D’Amico G, Korhonen EA, Waltari M, Saharinen P, Laakkonen P, Alitalo K. Loss of endothelial Tie1 receptor impairs lymphatic vessel development-brief report. Arterioscler Thromb Vasc Biol. 30: 207-9, 2010
First indication that the Tie1 receptor is involved in lymphangiogenesis.
V-M Leppänen, A Prota, M Jeltsch, A Anisimov, N Kalkkinen, T Strandin, H Lankinen, A Goldman, K Ballmer-Hofer and K Alitalo: Structural determinants of growth factor binding and specificity by VEGF-Receptor 2. Proc Natl Acad Sci, USA, 107: 2425-2430, 2010.
The structure of VEGF-C, and its complex with the main angiogenic receptor VEGFR-2.
Tammela T, Zarkada G, Wallgard E, Murtomäki A, Suchting S, Wirzenius M, Waltari M, Hellström M, Schomber T, Peltonen R, Freitas C, Duarte A, Isoniemi H, Laakkonen P, Christofori G, Ylä-Herttuala S, Shibuya M, Pytowski B, Eichmann A, Betsholtz C, Alitalo K. Blocking VEGFR-3 suppresses angiogenic sprouting and vascular network formation. Nature 454: 656-60, 2008.
This paper reveals a new mechanism involved in blood vessel sprouting and provides an additional target for angiogenesis inhibition.
Tammela T, Saaristo A, Holopainen T, Lyytikkä J, Kotronen A, Pitkonen M, Abo-Ramadan U, Ylä-Herttuala S, Petrova TV, Alitalo K. Therapeutic differentiation and maturation of lymphatic vessels after lymph node dissection and transplantation. Nature Medicine 13: 1458-66, 2007.
The first evidence that collecting lymphatic vessels can differentiate from lymphatic capillaries in adults and that VEGF-C gene therapy induces this process.
He Y, Rajantie I, Pajusola K, Jeltsch M, Holopainen T, Ylä-Herttuala S, Harding T, Jooss K, Takahashi T, Alitalo K. Vascular endothelial cell growth factor receptor 3-mediated activation of lymphatic endothelium is crucial for tumor cell entry and spread via lymphatic vessels. Cancer Res 65: 4739-46, 2005.
The mechanism of VEGF-C induced lymphatic sprouting towards as well as dilation of the draining lymphatic vessels, both contributing to lymphatic metastasis. These processes were blocked dose-dependently by inhibition of VEGFR-3.
Petrova TV, Kärpänen T, Norrmen C, Mellor R, Tamakoshi T, Finegold D, Ferrell R, Kerjaschki D, Mortimer P, Ylä-Herttuala S, Miura N, Alitalo K. Defective valves and abnormal mural cell recruitment underlie lymphatic vascular failure in lymphedema distichiasis. Nature Medicine 10: 974-81, 2004.
Mechanism of development of Lymphedema distichiasis.
Kärkkäinen MJ, Haiko P, Sainio K, Partanen J, Taipale J, Petrova TV, Jeltsch M, Jackson DG, Talikka M, Rauvala H, Betsholtz C, Alitalo K. Vascular endothelial growth factor C is required for sprouting of the first lymphatic vessels from embryonic veins. Nature Immunology 5: 74-80, 2004.
The results of this paper indicate that VEGF-C is the paracrine factor essential for lymphangiogenesis, and that both Vegfc alleles are required for normal lymphatic development.
He Y, Kozaki K, Kärpänen T, Koshikawa K, Ylä-Herttuala S, Takahashi T, Alitalo K. Suppression of tumor lymphangiogenesis and lymph node metastasis by blocking vascular endothelial growth factor receptor 3 signaling. J Natl Cancer Inst 94: 819-25, 2002.
Development of VEGFR-3 signaling inhibitor for suppression of tumor lymphangiogenesis and metastasis to regional lymph nodes.
Mäkinen T, Jussila L, Veikkola T, Kärpänen T, Kettunen MI, Pulkkanen KJ, Kauppinen R, Jackson DG, Kubo H, Nishikawa S-I, Ylä-Herttuala S, Alitalo K. Inhibition of lymphangiogenesis with resulting lymphedema in transgenic mice expressing soluble VEGF receptor-3. Nature Medicine 7: 199-205, 2001.
Demonstration that a soluble form of VEGFR-3 is a potent inhibitor of VEGF-C/VEGF-D signaling and lymphangiogenesis.
Mandriota SJ, Jussila L, Jeltsch M, Compagni A, Baetens D, Prevo R, Banerji S, Huarte J, Montesano R, Jackson DG, Orci L, Alitalo K, Christofori G, Pepper MS. Vascular endothelial growth factor-C-mediated lymphangiogenesis promotes tumour metastasis. EMBO J 20: 672-82, 2001.
Demonstration that VEGF-C-induced lymphangiogenesis mediates tumour cell dissemination and the formation of lymph node metastases. Acknowledgements:” The project presented in this manuscript was conceived and started in Helsinki, and the work is the result of an equal contribution from the laboratories in Helsinki, Vienna and Geneva, together with a major contribution from the Oxford group.”
Kärpänen T, Egeblad M, Kärkkäinen MJ, Kubo H, Ylä-Herttuala S, Jaattela M, Alitalo K. Vascular endothelial growth factor C promotes tumor lymphangiogenesis and intralymphatic tumor growth. Cancer Res 61: 1786-90, 2001.
These data show that VEGF-C facilitates tumor metastasis to the lymphatic vessels and that tumor spread is inhibited by blocking the interaction between VEGF-C and its receptor.
Kärkkäinen MJ, Saaristo A, Jussila L, Karila KA, Lawrence EC, Pajusola K, Bueler H, Eichmann A, Kauppinen R, Kettunen MI, Ylä-Herttuala S, Finegold DN, Ferrell RE, Alitalo K. A model for gene therapy of human hereditary lymphedema. Proc Natl Acad Sci USA 98: 12677-82, 2001.
First demonstration that growth factor gene therapy would be applicable to human lymphedema and provide a paradigm for other diseases associated with mutant receptors.
Kärkkäinen MJ, Ferrell RE, Lawrence EC, Kimak MA, Levinson KL, McTigue MA, Alitalo K, Finegold DN: Missense mutations interfere with VEGFR-3 signalling in primary lymphoedema. Nature Genetics 25: 153-159, 2000.
Heterozygous missense mutations of VEGFR-3 were shown to inactivate the tyrosine kinase and downstream gene activation in primary lymphedema, indicating that mutations interfering with VEGFR-3 signal transduction are a cause of primary lymphoedema.
Dumont, DJ, Jussila L, Taipale J, Lymboussaki A, Mustonen T, Pajusola K, Breitman M, Alitalo K. Cardiovascular failure in mouse embryos deficient in VEGF receptor-3. Science 282: 946-949, 1998.
This paper shows that VEGFR-3 has an essential role in the development of the embryonic cardiovascular system before the emergence of the lymphatic vessels.
Jeltsch M, Kaipainen A, Joukov V, Meng X, Lakso M, Rauvala H, Swartz M, Fukumura D, Rakesh KJ, Alitalo K. Hyperplasia of lymphatic vessels in VEGF-C transgenic mice. Science 276: 1423-1425, 1997.
First evidence that VEGF-C induces lymphangiogenesis.
Joukov V, Pajusola K, Kaipainen, Chilov DA, Lahtinen I, Kukk E, Saksela O, Kalkkinen N, Alitalo K. A novel vascular endothelial growth factor, VEGF-C, is a ligand for the Flt4 (VEGFR-3) and KDR (VEGFR-2) receptor tyrosine kinases. EMBO J 15: 290-298, 1996.
Isolation of the first ligand for VEGFR3.
Joukov V, Pajusola K, Kaipainen, Chilov DA, Lahtinen I, Kukk E, Saksela O, Kalkkinen N, Alitalo K. A novel vascular endothelial growth factor, VEGF-C, is a ligand for the Flt4 (VEGFR-3) and KDR (VEGFR-2) receptor tyrosine kinases. EMBO J 15: 290-298, 1996.
Isolation of VEGF-C, the first ligand for VEGFR3.
Kaipainen A, Korhonen J, Mustonen T, van Hinsbergh VWM, Fang G-H, Dumont D, Breitman M, Alitalo K. Expression of the fms-like tyrosine kinase 4 gene becomes restricted to lymphatic endothelium during development. Proc Natl Acad Sci 92: 3566-3570, 1995.
First evidence that VEGFR3 mediated signals are selective for lymphatic vessels in adults.
Partanen J, Armstrong E, Mäkelä TP, Korhonen J, Sandberg M, Renkonen R, Knuutila S, Huebner K, Alitalo K. A novel endothelial cell surface receptor tyrosine kinase with extracellular epidermal growth factor homology domains. Mol Cell Biol 12: 1698-1707, 1992.
Discovery of the endothelial Tie receptor (later renamed Tie1)
Andrey Anisimov, Ph.D., Adjunct Professor, Senior Research Scientist
Salli Antila, M.D., Ph.D. Student
Iina Häkkänen, M.Sc., Research Assistant
Marko Hyytiäinen, Ph.D., Senior Research Scientist
Pauliina Kallio, M.D., Ph.D. Student
Zhilin Li, Ph.D., Postdoctoral Researcher
Ibrahim Sultan, M.Sc., Ph.D. Student
Aino Tedeton, Ph.D., Postdoctoral Researcher