Project acronym CYTRIX
Project Engineering Cytokines for Super-Affinity Binding to Matrix in Regenerative Medicine
Researcher (PI) Jeffrey Alan Hubbell
Host Institution (HI) ECOLE POLYTECHNIQUE FEDERALE DE LAUSANNE
Country Switzerland
Call Details Advanced Grant (AdG), LS7, ERC-2013-ADG
Summary In physiological situations, the extracellular matrix (ECM) sequesters cytokines, localizes them, and modulates their signaling. Thus, physiological signaling from cytokines occurs primarily when the cytokines are interacting with the ECM. In therapeutic use of cytokines, however, this interaction and balance have not been respected; rather the growth factors are merely injected or applied as soluble molecules, perhaps in controlled release forms. This has led to modest efficacy and substantial concerns on safety. Here, we will develop a protein engineering design for second-generation cytokines to lead to their super-affinity binding to ECM molecules in the targeted tissues; this would allow application to a tissue site to yield a tight association with ECM molecules there, turning the tissue itself into a reservoir for cytokine sequestration and presentation. To accomplish this, we have undertaken preliminary work screening a library of cytokines for extraordinarily high affinity binding to a library of ECM molecules. We have thereby identified a small peptide domain within placental growth factor-2 (PlGF-2), namely PlGF-2123-144, that displays super-affinity for a number of ECM proteins. Also in preliminary work, we have demonstrated that recombinant fusion of this domain to low-affinity binding cytokines, namely VEGF-A, PDGF-BB and BMP-2, confers super-affinity binding to ECM molecules and accentuates their functionality in vivo in regenerative medicine models. In the proposed project, based on this preliminary data, we will push forward this protein engineering design, pursuing super-affinity variants of VEGF-A and PDGF-BB in chronic wounds, TGF-beta3 and CXCL11 in skin scar reduction, FGF-18 in osteoarthritic cartilage repair and CXCL12 in stem cell recruitment to ischemic cardiac muscle. Thus, we seek to demonstrate a fundamentally new concept and platform for second-generation growth factor protein engineering.
Summary
In physiological situations, the extracellular matrix (ECM) sequesters cytokines, localizes them, and modulates their signaling. Thus, physiological signaling from cytokines occurs primarily when the cytokines are interacting with the ECM. In therapeutic use of cytokines, however, this interaction and balance have not been respected; rather the growth factors are merely injected or applied as soluble molecules, perhaps in controlled release forms. This has led to modest efficacy and substantial concerns on safety. Here, we will develop a protein engineering design for second-generation cytokines to lead to their super-affinity binding to ECM molecules in the targeted tissues; this would allow application to a tissue site to yield a tight association with ECM molecules there, turning the tissue itself into a reservoir for cytokine sequestration and presentation. To accomplish this, we have undertaken preliminary work screening a library of cytokines for extraordinarily high affinity binding to a library of ECM molecules. We have thereby identified a small peptide domain within placental growth factor-2 (PlGF-2), namely PlGF-2123-144, that displays super-affinity for a number of ECM proteins. Also in preliminary work, we have demonstrated that recombinant fusion of this domain to low-affinity binding cytokines, namely VEGF-A, PDGF-BB and BMP-2, confers super-affinity binding to ECM molecules and accentuates their functionality in vivo in regenerative medicine models. In the proposed project, based on this preliminary data, we will push forward this protein engineering design, pursuing super-affinity variants of VEGF-A and PDGF-BB in chronic wounds, TGF-beta3 and CXCL11 in skin scar reduction, FGF-18 in osteoarthritic cartilage repair and CXCL12 in stem cell recruitment to ischemic cardiac muscle. Thus, we seek to demonstrate a fundamentally new concept and platform for second-generation growth factor protein engineering.
Max ERC Funding
2 368 170 €
Duration
Start date: 2014-05-01, End date: 2019-04-30
Project acronym GLIOMA
Project Molecular Mechanisms of Glioma Genesis and Progression
Researcher (PI) Joan Seoane
Host Institution (HI) FUNDACIO PRIVADA INSTITUT D'INVESTIGACIO ONCOLOGICA DE VALL-HEBRON (VHIO)
Country Spain
Call Details Starting Grant (StG), LS6, ERC-2007-StG
Summary Glioma is the most common and aggressive tumour of the brain and its most malignant form, glioblastoma multiforme, is nowadays virtually not curable. Very little is known about glioma genesis and progression at the molecular level and not much progress has been achieved in the treatment of this disease during the last years. The understanding of the molecular mechanisms involved in the biology of glioma is essential for the development of successful and rational therapeutic strategies. Our project aims to: 1- Study the role of the TGF-beta, Shh, Notch, and Wnt signal transduction pathways in glioma. These pathways have been implicated in glioma but still not much is known about their specific mechanisms of action. 2- Study of a cell population within the tumour mass that has stem-cell-like characteristics, the glioma stem cells, and how the four mentioned pathways regulate their biology. 3- Study the role of a transcription factor, FoxG1, that has an important oncogenic role in some gliomas and that it is regulated by the four mentioned pathways interconnecting some of them. Our approach will be based on a tight collaboration with clinical researchers of our hospital and the study of patient-derived tumours. We will analyse human biopsies, generate primary cultures of human tumour cells, isolate the stem-cell-like population of patient-derived gliomas and generate mouse models for glioma based on the orthotopical inoculation of human glioma stem cells in the mouse brain to generate tumours with the same characteristics as the original human tumour. In addition, we will also study genetically modified mouse models and established cell lines. We expect that our results will help understand the biology of glioma and cancer, and we aspire to translate our discoveries to a more clinical ambit identifying molecular markers of diagnosis and prognosis, markers of response to therapies, and unveil new therapeutic targets against this deadly disease.
Summary
Glioma is the most common and aggressive tumour of the brain and its most malignant form, glioblastoma multiforme, is nowadays virtually not curable. Very little is known about glioma genesis and progression at the molecular level and not much progress has been achieved in the treatment of this disease during the last years. The understanding of the molecular mechanisms involved in the biology of glioma is essential for the development of successful and rational therapeutic strategies. Our project aims to: 1- Study the role of the TGF-beta, Shh, Notch, and Wnt signal transduction pathways in glioma. These pathways have been implicated in glioma but still not much is known about their specific mechanisms of action. 2- Study of a cell population within the tumour mass that has stem-cell-like characteristics, the glioma stem cells, and how the four mentioned pathways regulate their biology. 3- Study the role of a transcription factor, FoxG1, that has an important oncogenic role in some gliomas and that it is regulated by the four mentioned pathways interconnecting some of them. Our approach will be based on a tight collaboration with clinical researchers of our hospital and the study of patient-derived tumours. We will analyse human biopsies, generate primary cultures of human tumour cells, isolate the stem-cell-like population of patient-derived gliomas and generate mouse models for glioma based on the orthotopical inoculation of human glioma stem cells in the mouse brain to generate tumours with the same characteristics as the original human tumour. In addition, we will also study genetically modified mouse models and established cell lines. We expect that our results will help understand the biology of glioma and cancer, and we aspire to translate our discoveries to a more clinical ambit identifying molecular markers of diagnosis and prognosis, markers of response to therapies, and unveil new therapeutic targets against this deadly disease.
Max ERC Funding
1 566 000 €
Duration
Start date: 2008-08-01, End date: 2014-07-31
Project acronym RESCARF
Project Renal stem cells: possible role in kidney pathologies and as new theraputic tools
Researcher (PI) Paola Romagnani
Host Institution (HI) UNIVERSITA DEGLI STUDI DI FIRENZE
Country Italy
Call Details Starting Grant (StG), LS6, ERC-2007-StG
Summary Chronic Kidney Disease (CKD) affects 11% of the adult population and is considered by the WHO as one of the health emergencies of the 21st century. Although cell therapy might be beneficial for CKD, human stem cells that might be used to improve kidney function were so far unknown. Recently, we demonstrated the existence of resident stem cells in the urinary pole of the Bowman’s capsule of adult human kidney and therefore named as adult parietal epithelial multipotent progenitors (APEMP). Injection of APEMP in SCID mice affected by acute renal failure, induced regeneration of tubular structures and reduced morphological and functional kidney damage. More recently, we found that APEMP are highly represented in embryonic kidneys and constitute the common progenitor of tubular cells and podocytes. The first aim of this project is to assess the regenerative properties of APEMP in in vivo models of glomerular injury and their potential use as a novel therapeutic tool to prevent the deterioration of kidney function in chronic renal failure. Second, we will try to identify the mechanisms that regulate the growth, survival, differentiation, and migration of APEMP, which is critical to set up cell therapies of renal injury which should be effective and safe. To this end, the role of different molecular pathways such as Sonic hedgehog, Wnt/beta-catenin, Notch, TGF-beta/BMP and of CXCR4, CXCR7 or CXCR3-B chemokine receptors in the regenerative activity of APEMP will be investigated. Third, to assess whether APEMP directly contribute to kidney regeneration after glomerular or tubular damage, transgenic animals in which APEMP are genetically tagged will be generated. Fourth, by using transgenic animals we will try to understand if an alteration of APEMP growth and/or differentiation is implicated in the pathogenesis of some renal disorders that frequently progress towards end stage renal disease.
Summary
Chronic Kidney Disease (CKD) affects 11% of the adult population and is considered by the WHO as one of the health emergencies of the 21st century. Although cell therapy might be beneficial for CKD, human stem cells that might be used to improve kidney function were so far unknown. Recently, we demonstrated the existence of resident stem cells in the urinary pole of the Bowman’s capsule of adult human kidney and therefore named as adult parietal epithelial multipotent progenitors (APEMP). Injection of APEMP in SCID mice affected by acute renal failure, induced regeneration of tubular structures and reduced morphological and functional kidney damage. More recently, we found that APEMP are highly represented in embryonic kidneys and constitute the common progenitor of tubular cells and podocytes. The first aim of this project is to assess the regenerative properties of APEMP in in vivo models of glomerular injury and their potential use as a novel therapeutic tool to prevent the deterioration of kidney function in chronic renal failure. Second, we will try to identify the mechanisms that regulate the growth, survival, differentiation, and migration of APEMP, which is critical to set up cell therapies of renal injury which should be effective and safe. To this end, the role of different molecular pathways such as Sonic hedgehog, Wnt/beta-catenin, Notch, TGF-beta/BMP and of CXCR4, CXCR7 or CXCR3-B chemokine receptors in the regenerative activity of APEMP will be investigated. Third, to assess whether APEMP directly contribute to kidney regeneration after glomerular or tubular damage, transgenic animals in which APEMP are genetically tagged will be generated. Fourth, by using transgenic animals we will try to understand if an alteration of APEMP growth and/or differentiation is implicated in the pathogenesis of some renal disorders that frequently progress towards end stage renal disease.
Max ERC Funding
820 200 €
Duration
Start date: 2008-10-01, End date: 2012-09-30
Project acronym TARG-SUP
Project Targeting TGF-β activation, likely the core mechanism of immunosuppression by human regulatory T cells.
Researcher (PI) Sophie Elizabeth J. Lucas
Host Institution (HI) UNIVERSITE CATHOLIQUE DE LOUVAIN
Country Belgium
Call Details Consolidator Grant (CoG), LS6, ERC-2015-CoG
Summary Regulatory T lymphocytes (Tregs) inhibit immune responses and are required to maintain immune tolerance. Tregs express membrane protein GARP, which displays latent TGF-β1 on the cell surface. Immunosuppression by human Tregs appears to require GARP-mediated activation of TGF-β1.
My objectives are to unravel the molecular aspects of TGF-β1 activation by GARP and determine the functional importance of this process in physiological and pathological conditions where Tregs or other GARP-expressing cells are present. As this implies the development of new tools to modulate GARP-dependent TGF-β1 activation and Treg immunosuppression, we will also explore their potential for the treatment of immune-related human diseases, and notably cancer.
More specifically, I will:
- Derive antibodies that modulate GARP-mediated TGF-β1 production by human Tregs and perform structural analyses in the presence of these antibodies to identify tri-dimensional changes in GARP/TGF-β1 complexes that lead to the release of active TGF-β1.
- Identify and characterize additional proteins implicated in TGF-β1 activation by human Tregs, as GARP is required but not sufficient for TGF-β1 activation by Tregs.
- Determine the immunological and clinical impact of inhibitory anti-GARP mAbs on cancer in mice. We will derive anti-murine GARP mAbs. As an alternative, we will generate mutant mice expressing a chimeric mouse/human GARP that is recognized by anti-human GARP mAbs. The antibodies will be tested in tumour-bearing mice treated or not with other immunotherapies including vaccines or immunostimulatory antibodies.
- Determine whether blocking anti-GARP mAbs improve immune responses to microbial vaccines or to chronic infections, as these represent important applications for transient inhibition of Treg activity in humans.
- Analyse the expression and roles of GARP in non-Treg cells to better understand GARP functions, which remain largely unknown, and predict potential toxicities of anti-GARP mAbs.
Summary
Regulatory T lymphocytes (Tregs) inhibit immune responses and are required to maintain immune tolerance. Tregs express membrane protein GARP, which displays latent TGF-β1 on the cell surface. Immunosuppression by human Tregs appears to require GARP-mediated activation of TGF-β1.
My objectives are to unravel the molecular aspects of TGF-β1 activation by GARP and determine the functional importance of this process in physiological and pathological conditions where Tregs or other GARP-expressing cells are present. As this implies the development of new tools to modulate GARP-dependent TGF-β1 activation and Treg immunosuppression, we will also explore their potential for the treatment of immune-related human diseases, and notably cancer.
More specifically, I will:
- Derive antibodies that modulate GARP-mediated TGF-β1 production by human Tregs and perform structural analyses in the presence of these antibodies to identify tri-dimensional changes in GARP/TGF-β1 complexes that lead to the release of active TGF-β1.
- Identify and characterize additional proteins implicated in TGF-β1 activation by human Tregs, as GARP is required but not sufficient for TGF-β1 activation by Tregs.
- Determine the immunological and clinical impact of inhibitory anti-GARP mAbs on cancer in mice. We will derive anti-murine GARP mAbs. As an alternative, we will generate mutant mice expressing a chimeric mouse/human GARP that is recognized by anti-human GARP mAbs. The antibodies will be tested in tumour-bearing mice treated or not with other immunotherapies including vaccines or immunostimulatory antibodies.
- Determine whether blocking anti-GARP mAbs improve immune responses to microbial vaccines or to chronic infections, as these represent important applications for transient inhibition of Treg activity in humans.
- Analyse the expression and roles of GARP in non-Treg cells to better understand GARP functions, which remain largely unknown, and predict potential toxicities of anti-GARP mAbs.
Max ERC Funding
1 993 125 €
Duration
Start date: 2016-04-01, End date: 2021-03-31
Project acronym TGF-MEPPA
Project Terrestrial Gamma Flashes-the Most Energetic Photon Phenomenon in our Atmosphere
Researcher (PI) Nikolai oestgaard
Host Institution (HI) UNIVERSITETET I BERGEN
Country Norway
Call Details Advanced Grant (AdG), PE10, ERC-2012-ADG_20120216
Summary "Only 20 years after the discovery of Cosmic Gamma-ray Bursts from the universe another completely unknown phenomenon involving gamma-rays was discovered by coincidence the BATSE instrument on the Compton Gamma-Ray Observatory. Short-lived (~1 ms) and very energetic photon emissions (>1 MeV and later: >40 MeV) were found to originate from the Earth’s atmosphere and were named Terrestrial Gamma Flashes (TGFs). These flashes are the most energetic natural photon phenomenon that is known to exist on Earth, in which also anti-matter is produced. Based on the few datasets available to date we believe that TGFs are related to electric discharges in thunderstorm systems and that electrons accelerated to relativistic energies are involved to produce bremsstrahlung of such high energies. However, it is not known how frequent TGFs are, the altitude range and the spatial extent of their source region, to what kind of thunderstorms and lightning they are related or the implications of relativistic electrons and positrons ejected into space. There is no consensus on how TGFs are produced. All these questions need to be answered before we understand how important they are and how they may affect the Earth’s electrical circuit and atmosphere.
The goal of the TGF-MEPPA project is to attack these questions by combining modelling of electron acceleration in thunderstorm electric fields, X- and gamma-ray production and propagation, lightning development with unprecedented measurements of TGFs from three different altitudes: 350 km, 30 km and 20 km to obtain the most comprehensive and detailed dataset needed to make significant advances in the TGF research. I will also perform electric discharge experiments in the laboratory. The goal is to establish a consistent model for the TGF-production and answer the question ‘How common are TGFs?’ to determine their implications for the Earth’s electrical circuit, atmosphere and outer space."
Summary
"Only 20 years after the discovery of Cosmic Gamma-ray Bursts from the universe another completely unknown phenomenon involving gamma-rays was discovered by coincidence the BATSE instrument on the Compton Gamma-Ray Observatory. Short-lived (~1 ms) and very energetic photon emissions (>1 MeV and later: >40 MeV) were found to originate from the Earth’s atmosphere and were named Terrestrial Gamma Flashes (TGFs). These flashes are the most energetic natural photon phenomenon that is known to exist on Earth, in which also anti-matter is produced. Based on the few datasets available to date we believe that TGFs are related to electric discharges in thunderstorm systems and that electrons accelerated to relativistic energies are involved to produce bremsstrahlung of such high energies. However, it is not known how frequent TGFs are, the altitude range and the spatial extent of their source region, to what kind of thunderstorms and lightning they are related or the implications of relativistic electrons and positrons ejected into space. There is no consensus on how TGFs are produced. All these questions need to be answered before we understand how important they are and how they may affect the Earth’s electrical circuit and atmosphere.
The goal of the TGF-MEPPA project is to attack these questions by combining modelling of electron acceleration in thunderstorm electric fields, X- and gamma-ray production and propagation, lightning development with unprecedented measurements of TGFs from three different altitudes: 350 km, 30 km and 20 km to obtain the most comprehensive and detailed dataset needed to make significant advances in the TGF research. I will also perform electric discharge experiments in the laboratory. The goal is to establish a consistent model for the TGF-production and answer the question ‘How common are TGFs?’ to determine their implications for the Earth’s electrical circuit, atmosphere and outer space."
Max ERC Funding
2 492 811 €
Duration
Start date: 2013-03-01, End date: 2018-02-28
