Final Report Summary - ATHERO-B-CELL (Targeting and exploiting B cell functions for treatment in cardiovascular disease)
Executive Summary:
The Athero-B-Cell Consortium enabled the first investigation into the role of B cell heterogeneity in cardiovascular disease (CVD) revealing new roles for B cells in atherosclerosis, its main driver. As the inflammatory component of atherosclerosis is increasingly proven to be pathogenic, the higher becomes the need to target it without compromising host defense. Hence targeting pathogenic immune cell subsets while sparing beneficial ones is an important therapeutic strategy.
The Athero-B-Cell Consortium mapped B cell subsets in large biobanks in the peripheral circulation of patients with CVD and in selected patients within the plaque, identifying both pathogenic and protective subsets variably associated with CV outcomes. Moreover, we identified a new function through which B cells can increase atherogenesis, the production of pro-inflammatory and pathogenic antibodies directed against ApoA1, the main component of the high-density lipoprotein particles. We also defined master regulators of B regulatory cells, a subset of B cells with anti-atherogenic potential. The Athero-B-Cell Consortium was also one the first group world-wide to apply nascent single cell technology, such as mass cytometry, to vascular tissue defining its immune cell heterogeneity in health and disease to unprecedented detail with new SME-developed analytical tools based on machine-learning algorithms.
The key driver of the discovery engine of Athero-B-Cell has been the application of bioinformatic approaches by the SME to the new and existing omics and single cell data from the Academic partners. This Academia-SME partnership led to the identification of targets for which specific locked-nucleic acid-based treatment candidates have been generated and studied in vitro and in vivo in models of CVD. New mouse models were also designed and generated to achieve a modeling of atherosclerosis that more accurately reflects the human phenotypes and the role of B cells in the pathogenesis of CVD. This level of innovation will feed our research in the years to come.
B cells are an important immune cellular component of the pathogenesis of atherosclerosis and mitigation of the proatherogenic or autoreactive B cells and exploitation of the protective ability of certain B cell subsets offer therapeutic promise.
Project Context and Objectives:
Cardiovascular disease (CVD) causes nearly half of all deaths in Europe (45%), costing a dramatic €210 billion a year (European Heart Network,www.ehnheart.org). Indeed, CVD is still the world’s leading cause of death, claiming 17.1 million lives each year worldwide, of which almost 2 million are in the EU (European Heart Network, www.ehnheart.org). The European Commission noted that by 2050, the number of people over 50 will rise by 35% and the number of those over 85 will triple (Communication on the Flagship Initiative Innovation Union, 2010). The WHO has estimated that rising life expectancy coupled with adverse trends in major cardiovascular risk factors including obesity and type II diabetes could lead to a doubling in the absolute incidence of CVD by 2050. (EU Report: cardiovascular research in the European Union, 2009.) This is already noticeable, with a decrease in mortality but an increase in prevalence of CVD in the last 10 years (Davies, 2007). In addition, in countries with less medical care, the incidence of CVD is rising and will increase dramatically (Anderson, 2007;Fuster, 2007). Thus, new and innovative treatments will be essential for maintaining quality of life whilst containing costs.
Despite the reduction in mortality that has been achieved over the past decades, 70% of CV events cannot be prevented with the treatment of known risk factors (Baigent, 2005). Aggressive glucose metabolism control in Type 2 Diabetes did not automatically result in CVD prevention (Mazzone, 2010) and lipid-lowering agents beyond statins have not yet delivered the expected reduction in CV events (Gotto, 2012) highlighting a complex relationship between inflammation and hyperlipidemia. The treatment of risk factors, albeit beneficial, has reached its full potential in CVD and new treatments should be targeted to novel disease factors.
The once dominant lipid insudation theory on the origin of atherosclerosis has been now superseded to accommodate more dynamic models of pathogenesis of atherosclerosis (Libby, 2012). The “response to injury” paradigm – proposed by Russell Ross (Ross, 1999) – explains the changes in the intima upon lipid accumulation characterised by the influx of smooth muscle cells and formation of a two-compartment plaque: the mature fibro-atheroma. Rupture of the fibrous cap allows the prothrombotic tissue factor-rich necrotic core to contact blood and platelets causing thrombosis. Thrombogenic factors in the blood then promote rapid thrombus propagation and coronary thrombosis (Naghavi, 2003). Plaque rupture and subsequent thrombosis is often asymptomatic with its healing leading to discontinuous plaque progression (Mann, 1999). However, if plaque rupture leads to acute or subacute occlusion of blood flow, acute myocardial damage ensues (Falk, 1995). Work over the past 35 years has proven that inflammation is integral to the pathogenesis of atherosclerosis. Activation of both innate and adaptive immunity occurs within the atherosclerotic plaque (Hansson, 2002), indicating a coordinated immune response rather than a non-specific inflammatory infiltrate. These paradigms, identified in experimental animal studies were validated against clinical data. The role of inflammation in atherosclerosis started with the finding of Goran Hansson that atherosclerotic plaques overexpress HLA-DR, an indirect sign of cytokine production and antigen presentation (Jonasson, 1985). It was subsequently demonstrated that macrophage recruitment into the arterial intima is required for the development of lesions and is associated with plaque activity, fibrous cap rupture and plaque erosion (Lessner, 2002; Swirski, 2006;Yan, 2007). Macrophage derived matrix metalloproteinases are now held responsible for the thinning of the fibrous cap and the risk of rupture (Falk, 1995). Toll-like receptors, innate immune pattern recognition receptors (PRRs) that detect pathogen-associated molecular patterns (PAMPs), are important mediators of the inflammatory response in atherosclerosis (Cole, 2010; Cole, 2011; Monaco, 2009). Besides innate immune responses that are mounted by macrophages and dendritic cells, adaptive immune responses with clonally expanded T cell populations contribute to this process. T-lymphocytes eventually reduce smooth muscle cell proliferation and repair mechanisms through the production of IFNγ leading to thinning of the fibrous cap (Hansson, 1989).
The recent Canakinumab Antiinflammatory Thrombosis Outcomes Study (CANTOS) has provided the first proof-of-concept that that targeting inflammation in cardiovascular disease is clinically relevant. In this study, in patients on statin therapy but with elevated C-reactive protein (CRP) levels, a significant reduction of CV events by the inhibition of interleukin-1β (IL-1β) without further influencing lipid levels was observed (Ridker, 2017). However, anti-IL-1β antibodies increases the infection risk. Thus, precision drugs targeting the immune response in atherosclerosis with limited side effects are required for the treatment of CVD.
Significant bottlenecks are preventing translation to the clinic including Host defense impairment with broad acting anti-inflammatories, the premature testing and failure of biologics (Chung, 2003;Mann, 2004) that discouraged Pharma from conducting clinical trials in CVD (Couzin-Frankel, 2012), a lack of knowledge of the best therapeutic targets due to a lack of preclinical data from human models of CVD and the fact that the immune system has opposing roles in CVD (Hansson, 2011).
B cells are a classic example of the contrasting role of immune cell subsets in CVD as they have been shown to carry out both pro- and anti-atherogenic functions and these dual roles are executed by different B cell subsets in CVD. B cells have been described in murine aortic root atherosclerotic lesions both at early and advanced stages of disease. CD22+ B cells were chiefly in clusters at the base of the plaques (Zhou, 1999). B cell depletion via an anti CD20 antibody reduced the development of atherosclerosis and the activation of dendritic cells and T cells, consistent with a pivotal role of B cells in the induction and perpetuation of the disease (Ait-Oufella, 2010). Transfer of splenic B cells to lymphocyte- depleted ApoE-/- mice or to B cell-deficient ApoE-/- mice increased atherosclerosis (Kyaw, 2010). In contrast to these results, suggesting a pathogenic involvement of B cells in atherosclerosis, there are studies demonstrating that B cells also play a protective role. Splenectomy in ApoE mice leads to moderately increased atherosclerotic lesions (Caligiuri, 2002). Adoptive transfer of total splenic B cells from atherosclerotic mice rescues splenectomised mice from increased atherogenesis (Caligiuri, 2002). The anti-atherogenic function of B cells has been assigned to “Innate” B1 cells, a lineage of B cells that has a peritoneal origin in mouse, and is distinct from bone marrow derived B cells - hereafter called B2 (Bouaziz, 2008). B1a B cells mediate the protective effect of splenic B cells via production of natural IgM antibodies (Kyaw, 2011) that recognise a phosphocholine motif contained within oxidised (ox) LDL and apoptotic cells (Binder, 2004; Chou, 2009 ; Horkko, 1999). Nevertheless, transfer of bone marrow from B cell deficient mice (mMT) to LDLR-/- mice, yields a 30% increase in lesion area (Major, 2002), suggesting that bone marrow – derived B cell subsets may also be protective.
B cell biology is a fertile new ground for identifying therapeutic targets in human disease, but limited knowledge is hampering their discovery. The success of therapeutic B cell depletion (rituximab) in autoimmune disease is very heterogeneous (Edwards, 2006) highlighting the complexity of B cell function, and our limited our understanding of their function in the context of human disease. B-cells are indispensable for antibody (Ab) generation. However, B-cells also produce cytokines, present antigens and restrain excessive immune responses (Harris, 2000; Mauri, 2012; Shlomchik, 2001). New data reveals that B cells participate in directing the magnitude and quality of T cell responses to foreign and self-antigens (Lund, 2010).
Phenotypic and functional heterogeneity have been clearly defined for effector and memory T cells. However B cell functional effector subsets and their role in disease are less well understood. Mature B cells can be further subdivided into marginal zone, follicular, B1, memory, regulatory and germinal centre (GC) subtypes (Fairfax, 2008) which complicates matters. Thus, understanding the distinction between suppressive and pathogenic B-cells and the “pathways” controlling their differentiation/maturation in CVD will improve targeting of pathogenic B-cell-lineages and will help to identify those patients most likely to benefit from such therapies.
The Athero-B-Cell project aimed to identify differences in B cell effector subsets relevant to atherosclerosis development and investigate how these differences could be used to develop new CVD therapeutics through combining the multidisciplinary knowledge and expertise of its Partners on B cells in inflammation and atherosclerosis with the data mining and therapeutic targeting platforms of world leading SMEs.
Athero-B-Cell aimed to bridge the gap between the laboratory and the clinic by bringing together basic and clinical scientists with internationally recognised expertise in: the biology of B cell subsets (Mauri-UCL, Malin-KI, Patrone-UniGenova), omics of leukocytes in large-scale clinical studies (Hoefer, UMCU), humoral immunity in CVD (Nordin Fredrickson-ULund, Mach-UniGeneva) and inflammatory signalling in atherosclerosis (Monaco-UOXF). Furthermore, the project sought to create an SME-Academic collaboration to generate a new pipeline of therapeutic targets whose potential will be realised by the SMEs.
The predominant aim of Athero-B-Cell was to identify, characterise and validate new approaches for manipulating B cell subsets in CVD using available and emerging -omics data from large-scale clinical studies.
Objectives:
1) To define targets to reduce activation of pathogenic B cells and antibodies
2) To identify enhancers of atheroprotective B regulatory cell populations and antibodies
3) To validate molecular and miRNA targets to modulate B cell behaviour in CVD with innovative locked nucleic acid (LNA) based antisense platforms (that have reached Phase 2 clinical trials in human).
Athero-B-Cell aims to mine available and new omics data from large-scale clinical studies to identify a specific B cell signature that is associated with cardiovascular disease and specific B cell effector functions including antibody and cytokine production as well as regulatory ability. The aim is to generate novel candidate therapeutic targets to modulate B cell behaviour in CVD.
Project Results:
Over the 60 months of the Athero-B-Cell project, the Consortium Partners have developed novel research tools (including a panel of antibodies to enable analysis and phenotyping of human B cell subsets in clinical cohorts and the generation of new bioinformatics research tools) to generate further interesting results and publications. The main work and accomplishments of the project are summarised below.
Work Package 1: Discovery of B cell specific therapeutic targets for CVD
The key efforts of the WP1 Beneficiaries revolved around the execution of B cell specific –omics, phenotypic studies, and antibody production in blood and serum samples from CVD cohorts. B cells were isolated from 295 samples from the Athero-Express study and high-throughput transcriptomics analysis was performed to map B cell-related changes specifically associated with CVD and CVD outcomes. Bioinformatics was applied to the data and identified differentially expressed genes in patients with versus without a follow-up cardiovascular event. Functional network and pathway analysis was also performed. A further study on PMBC used for the ‘omics’ study revealed that high numbers of unswitched and switched memory B cells are associated with a lower risk of secondary cardiovascular events in a population with severe cardiovascular disease.
In addition, a cytometry by time of flight (CyTOF) workflow was established. Mass cytometry (CyTOF) allows allows the simultaneous analysis of multiple parameters on single cells. The technique employs tagging of antibodies with isotopically pure rare earth metals, mostly lanthanides, which are then used to stain cells of interest. Due to the pure and distinct isotopic masses of the metals, the mass cytometry detection system overcomes the problem of channel overlap experienced in flow cytometry caused by flurochrome overlap. As a result, the introduction of CyTOF has greatly expanded the number of parameters that can be measured per single cell in a sample. More than 40 parameters at a time have been successfully measured per sample and as little as 10,000 cells have been used as starting cells for CyTOF analysis. This makes CyTOF one of the most robust techniques for identification of a whole repertoire of immune cells when tissues/cells for analysis are limited. CyTOF represents a step-change in our ability to phenotype cells in tissues in a broad and high-dimensional manner.
CyTOF was used to determine the immune cell landscape in aortas of ApoE-/- mice and revealed differences in the immune cell census of aortas from mice fed a chow and a high fat diet. This analysis revealed at least 13 leucocyte populations in the aortas of ApoE-/- mice, including major myeloid and lymphoid cell subsets. Feeding ApoE-/- mice a high fat diet led to increased monocytes and pDC and decreased cDC2 in the aortas. Although statistical significance was not reached, a trend towards increased neutrophils and reduced B cells was also observed in aortas of high fat compared to chow fed ApoE-/- mice. Myeloid cells (neutrophils, eosinophils, monocytes, macrophages, dendritic cells) are known to play important roles in atherosclerosis development. Myeloid cells (gated as Lin-CD11blo-hi) accounted for around 75% of the CD45+ leucocytes in the ApoE-/- aortas in our study. At least 13 myeloid cell subsets were found in the aortas of ApoE-/- mice. Differences in aortic myeloid populations were observed between ApoE-/- mice fed a chow and a high fat diet.
Studies in this work package also evaluated the relationship between anti-ApoA-1 antibody levels and clinical outcomes.
Anti-apoA-1 IgG were found to be independent predictors of nonfatal incident CAD in the general population. In a prospectively studied, population-based cohort of 5220 subjects (mean age 52.6±10.7 years, 47.4% males), followed over a median period of 5.6 years, subjects positive versus negative for anti-apoA-1 IgG presented a total CAD rate of 3.9% versus 2.8% (P=0.077) and a nonfatal CAD rate of 3.6% versus 2.3% (P=0.018) respectively. After multivariate adjustment for established cardiovascular risk factors, the hazard ratios of anti-apoA-1 IgG for total and nonfatal CAD were: hazard ratio=1.36 (95% confidence interval, 0.94-1.97; P=0.105) and hazard ratio=1.53 (95% confidence interval, 1.03-2.26; P=0.034) respectively. The strength of this association was dependent on a functional polymorphism of the CD14 receptor gene (Arterioscler Thromb Vasc Biol. 2017;37:2342-2349).
Furthermore, studies examined whether anti-apoA-1 IgG: (a) predict all-cause mortality in the general population and (b) are associated with single-nucleotide polymorphisms (SNPs) in a genome-wide association study (GWAS). Anti-apoA-1 IgG positivity independently predicted all-cause mortality: hazard ratio (HR) = 1.54 95% confidence interval (95% CI): 1.11-2.13 P = 0.01 indicating that preclinical autoimmunity to anti-apoA-1 IgG may represent a novel mortality risk factor (Front Immunol. 2017;8:437).
Work Package 2: Target Selection
Work package 2 led to the development of tools to aid target selection. SME-based in silico platforms were needed to integrate the –omics data for selection of the pathways forming a specific B cell signature associated with CVD outcomes and B cell effector functions. A fully automated solution that can perform effective data collection, quality assurance, processing and normalisation, as well as comprehensive statistical and functional analysis has been developed and utilised on datasets of the consortium partners.
Secondly, an ad-hoc panel of antibodies was described and used to interrogate the B cell compartment in cardiovascular disease. B cell content of 804 PBMC samples, collected as part of the prospective Malmo Diet and Cancer cardiovascular case control cohort, were phenotyped by flow cytometry. Initial analysis of the data has revealed interesting associations between specific B cell subsets.
Work Package 3: Validation of novel targets for B cell responses with pro-atherogenic outcomes
In WP3, novel locked nucleic acids (LNAs) and animal models against targets involved in B cell activity have been generated.
In recent years, RNA has become an attractive target for therapeutic intervention since RNA mediates most cellular gene expression. Synthetic oligonucleotides are the rational way to target RNA as using a sequence complementary to a specific mRNA, can inhibit its expression and therefore reduce target protein levels. Over the past decades, considerable work has attempted to develop RNA binding oligonucleotides with improved nuclease stability and binding affinity). The discovery of Locked Nucleic Acid (LNA) chemistry opened new avenues for antisense oligonucleotide therapeutics. LNA is an RNA modification with an oxy-methylene bridge between the 2’ and 4’-positions of the ribose. The methylene bridge creates a bi-cyclic structure ‘locking’ the ribose into a conformation ideal for Watson-Crick binding. When incorporated into a DNA or RNA oligonucleotide, LNA makes the pairing with a complementary nucleotide more rapid and the stability of the resulting duplex is increased. In addition, affinity to complementary RNA sequences and resistance to nuclease digestion is greatly enhanced by LNA.
LNA based antisense oligonucleotides possess the following features: 1) Excellent specificity, providing optimal targeting, 2) Increased affinity to targets providing improved potency in-vivo, 3) Strong pharmacology, cell permeability upon systemic delivery without complicated delivery vehicles, 4) Scalable and cost-effective manufacturing, 5) Well-tolerated with predictable toxicology and 6) Potential for oral delivery. The additional advantage of using LNA platforms is that it allows us to target intracellular molecules not easily targeted with small molecule inhibitors and/or antibodies. LNA technology provides an exciting opportunity to develop novel therapies to specifically target sub-populations of B cells.
Based on input from the Scientific Partners in the Consortium and from data generated as part of WP1 and WP2, several targets were chosen for RICC to develop LNAs against. LNAs against these selected targets have been generated and have been found to display powerful knockdown of the target in vitro and in vivo and had safe pharmacology in mouse. These compounds have been applied to in vitro experiments and to murine models of cardiovascular disease including a collar-induced injury model and a hypercholesterolaemic model of atherosclerosis.
Autoantibodies have been shown to play a critical role in predicting major adverse cardiovascular events in atherosclerotic patients. In this work package, we also aimed to assess the role of IgG auto-antibodies against ApoA-1 in mouse CV models. The role of anti-apoA-1 IgG on tissue factor (TF) expression and activation, a key coagulation regulator underlying atherothrombosis was examined. ApoE-/-, ApoE-/-TLR2-/- and ApoE-/-TLR4-/- mice were passively immunised with anti-apoA-1 or control IgG. intraplaque TF expression was significantly increased in ApoE-/- Mice that received anti-apoA-1 IgG passive immunisation. This was attenuated in ApoE-/- mice lacking either TLR2 or TLR4. Together with data from human samples, our results support a possible causal relationship between anti-apoA-1 IgG and atherothrombosis (Pagone et al., 2016 Thromb Haemost 116(3) 554-564).
Work Package 4: Validation of targets related to the enhancement of Bregs as a novel therapeutic strategy
AtheroBCell partners involved in this work package made significant progress towards the definition of the role of B cells in experimental models of atherosclerosis and also the generation of mouse models that specifically target B cells in hyperlipidemic mice.
As part of this project, the existence and role of regulatory B cells in vascular disease was examined. A well-established model of neointima formation based on the placement of a perivascular collar around the murine carotid artery in ApoE-/- mice was used to examine the role of Bcells and regulatory B cells in atherosclerosis. In this model, carotid injury leads to the development of fibro-atheromatous lesions with a macrophage-rich core. B cells were purified from the spleen and draining lymph nodes of ApoE-/- mice and transferred to syngeneic mice prior to collar placement. Adoptive transfer of B cells isolated from ApoE-/- LN significantly protected recipient mice from lesion development compared to control-treated mice or mice receiving splenic B cells. Moreover, treatment with LN-derived B cells reduced macrophage infiltration. The protective effect of lymph node B cells was found to be partially due to the cytokine interleukin (IL)-10. A significant increase in the number of total B cells was observed in the lymph nodes, but not in the spleens, of ApoE-/- compared to wild-type mice. Follicular B cells (CD21intCD23intCD24int) were increased in the LN of ApoE-/- compared to WT mice and in addition, an unexpected significant increase of CD21hiCD23hiCD24hi transitional two-marginal-zone precursor B cells, which were previously shown to have immunosuppressive Breg cell function, was observed in the LN of ApoE-/- compared to WT mice. Concomitantly, ApoE-/- mice displayed reduced levels of CD21hiCD23hiCD24hi B cells in the spleen compared to WT mice. We speculated that the CD21hiCD23hiCD24hi Breg subset, previously demonstrated to suppress immune-mediated experimental disease, might be responsible for the protective effect of the LN-derived B cells in our model. To prove the direct involvement of this subset in protection, we performed adoptive transfer experiments using purified CD21hiCD23hiCD24hi cells or follicular B cells isolated by FACS sorting from the LNs of ApoE-/- mice. We observed a significant reduction in lesion development after adoptive transfer of CD21hiCD23hiCD24hi B cells but not after transfer of the same number of follicular B cells. Similarly, neointimal macrophage content was reduced in the CD21hiCD23hiCD24hi B cell treated group compared to control and the follicular B cell treated group. The protective effect of CD21hiCD23hiCD24hi B cells was associated with an increase in levels of IL-10 in the serum. In conclusion, the AtheroBCell consortium described for the first time that a B2 Breg subset protects from neointima formation. We also revealed that B cells can provide vascular protection by means other than antibody generation, e.g. IL-10 production and the induction of Tr1 cells. (Strom, Cross, Cole et al., 2015. Thromb Haemost 114(4): 835-847).
Based on the expert knowledge of the AtheroBcell Partners and the data emerging from ‘-omics’ analysis in the project, several novel mouse models were designed and generated. One model will allow the specific knockdown of Bregs in atherosclerosis as well as fate-mapping experiments that will provide insight into the atherosclerotic ontogeny of the Breg lineage. Other models will allow tracking and deletion of molecules involved in Bcell biology and atherogenesis. Initial experiments in several of these mouse models are ongoing.
Moreover in this work package, we aimed to to investigate if B cells pulsed with p210-CTB can protect against atherosclerosis. Apolipoprotein B-100 is the major protein in LDL and we have previously identified certain peptide sequences as targets for autoimmune responses in humans and shown that prototype vaccines based on particularly peptide number 210 can inhibit the development of atherosclerosis in mice. The protective response induced by p210 immunization strategies has been shown to be associated with an increase in regulatory T cells (Tregs). Furthermore, p210 has been used together with other ApoB100 peptides in low dose continuous subcutaneous administration resulting in reduced lesion development and an increased Treg population. This work has led to results that are due to be published in the near future.
Potential Impact:
The Canakinumab Antiinflammatory Thrombosis Outcomes Study (CANTOS) has provided the first proof-of-concept that that targeting inflammation in cardiovascular disease is clinically relevant. (Ridker, NEJM 2017). However, anti-IL-1β antibodies increases the infection risk. Thus, precision drugs targeting the immune response in atherosclerosis with limited side effects are required for the treatment of CVD.
B cell targeting is an important therapeutic opportunity that has not being thoroughly explored in CVD. By mapping B cell heterogeneity in CVD, the Athero-B-Cell consortium has enabled the discovery and characterization of on/off switches of B cell phenotypes that can either contribute or protect form atherosclerosis and other vascular events, as well as novel effector pathways that B cell use to worsen disease such as autoreactive antibodies. The first impact is in the generation of a target portfolio that can be further pushed down the translational path by Academia and SME.
The second large impact is the SME success. The SMEs involved in Athero-B-Cell have gone from strength to strength during the course of the Consortium, furthering their target portfolio, skill development, knowledge and base of operation. Albeit Athero-B-Cell was not the only factor contributing their growth, this level of research activity led to a spur of further opportunities.
The third large impact of Athero-B-Cell has been its collaborative and cross-fertilising model of research commitment mainly based on data and model sharing, which has supported the research and the interactions of young scientists between all the Partners, cementing group and institutional interaction and data and knowledge exchange. This has led to stronger partnership between the participating Institutions and durable relationships being established, leading to at least 2 further collaborative applications in the near future.
The fourth impact are is the dissemination and training activities of the young scientists of Athero-B-Cell. The Investigators of Athero-B-Cell, many of which are females, albeit established, are relatively young themselves and these FP7 funds supported their ascending trajectory of them and also of their young post-docs and PhD students they harbour in their labs.
Besides the generation of novel scientific knowledge that can be exploited by the wider scientific
community, the project has also led to several discoveries that can be directly exploited
commercially. The most directly exploitable results are represented by Patent applications citing Athero-B-Cell as a founding source that have been listed in Template B2.
List of Websites:
The project website has the address: http://www.atherobcell.eu/.
Contractors involved:
- The Chancellor, Master and Scholars of University of Oxford (UK): Claudia Monaco
- Roche Innovation Center Copenhagen A/S (Denmark): Steffen Schmidt
- Université de Genève (Switzerland): François Mach
- Università di Genova (Italy): Alessio Nencioni
- Lunds Universitet (Sweden): Gunilla Nordin Fredrikson
- University Medical Center Utrecht (The Netherlands): Imo Hoefer
- Karolinska Institutet (Sweden): Stephen Malin
- University College London (UK): Claudia Mauri
- Bering Limited (UK): Ignat Drodzov
- genOway SA (France): Vincent Dubus
- ALTA Ricerca e Svluppo in Biotecnologie S.r.l.u. (Italy): Riccardo Bertini
Coordinator contact details:
The Chancellor, Master and Scholars of University of Oxford
Prof. Claudia Monaco
Kennedy Institute of Rheumatology
Roosevelt Drive, Headington, Oxford, OX3 7FY, UK.
The Athero-B-Cell Consortium enabled the first investigation into the role of B cell heterogeneity in cardiovascular disease (CVD) revealing new roles for B cells in atherosclerosis, its main driver. As the inflammatory component of atherosclerosis is increasingly proven to be pathogenic, the higher becomes the need to target it without compromising host defense. Hence targeting pathogenic immune cell subsets while sparing beneficial ones is an important therapeutic strategy.
The Athero-B-Cell Consortium mapped B cell subsets in large biobanks in the peripheral circulation of patients with CVD and in selected patients within the plaque, identifying both pathogenic and protective subsets variably associated with CV outcomes. Moreover, we identified a new function through which B cells can increase atherogenesis, the production of pro-inflammatory and pathogenic antibodies directed against ApoA1, the main component of the high-density lipoprotein particles. We also defined master regulators of B regulatory cells, a subset of B cells with anti-atherogenic potential. The Athero-B-Cell Consortium was also one the first group world-wide to apply nascent single cell technology, such as mass cytometry, to vascular tissue defining its immune cell heterogeneity in health and disease to unprecedented detail with new SME-developed analytical tools based on machine-learning algorithms.
The key driver of the discovery engine of Athero-B-Cell has been the application of bioinformatic approaches by the SME to the new and existing omics and single cell data from the Academic partners. This Academia-SME partnership led to the identification of targets for which specific locked-nucleic acid-based treatment candidates have been generated and studied in vitro and in vivo in models of CVD. New mouse models were also designed and generated to achieve a modeling of atherosclerosis that more accurately reflects the human phenotypes and the role of B cells in the pathogenesis of CVD. This level of innovation will feed our research in the years to come.
B cells are an important immune cellular component of the pathogenesis of atherosclerosis and mitigation of the proatherogenic or autoreactive B cells and exploitation of the protective ability of certain B cell subsets offer therapeutic promise.
Project Context and Objectives:
Cardiovascular disease (CVD) causes nearly half of all deaths in Europe (45%), costing a dramatic €210 billion a year (European Heart Network,www.ehnheart.org). Indeed, CVD is still the world’s leading cause of death, claiming 17.1 million lives each year worldwide, of which almost 2 million are in the EU (European Heart Network, www.ehnheart.org). The European Commission noted that by 2050, the number of people over 50 will rise by 35% and the number of those over 85 will triple (Communication on the Flagship Initiative Innovation Union, 2010). The WHO has estimated that rising life expectancy coupled with adverse trends in major cardiovascular risk factors including obesity and type II diabetes could lead to a doubling in the absolute incidence of CVD by 2050. (EU Report: cardiovascular research in the European Union, 2009.) This is already noticeable, with a decrease in mortality but an increase in prevalence of CVD in the last 10 years (Davies, 2007). In addition, in countries with less medical care, the incidence of CVD is rising and will increase dramatically (Anderson, 2007;Fuster, 2007). Thus, new and innovative treatments will be essential for maintaining quality of life whilst containing costs.
Despite the reduction in mortality that has been achieved over the past decades, 70% of CV events cannot be prevented with the treatment of known risk factors (Baigent, 2005). Aggressive glucose metabolism control in Type 2 Diabetes did not automatically result in CVD prevention (Mazzone, 2010) and lipid-lowering agents beyond statins have not yet delivered the expected reduction in CV events (Gotto, 2012) highlighting a complex relationship between inflammation and hyperlipidemia. The treatment of risk factors, albeit beneficial, has reached its full potential in CVD and new treatments should be targeted to novel disease factors.
The once dominant lipid insudation theory on the origin of atherosclerosis has been now superseded to accommodate more dynamic models of pathogenesis of atherosclerosis (Libby, 2012). The “response to injury” paradigm – proposed by Russell Ross (Ross, 1999) – explains the changes in the intima upon lipid accumulation characterised by the influx of smooth muscle cells and formation of a two-compartment plaque: the mature fibro-atheroma. Rupture of the fibrous cap allows the prothrombotic tissue factor-rich necrotic core to contact blood and platelets causing thrombosis. Thrombogenic factors in the blood then promote rapid thrombus propagation and coronary thrombosis (Naghavi, 2003). Plaque rupture and subsequent thrombosis is often asymptomatic with its healing leading to discontinuous plaque progression (Mann, 1999). However, if plaque rupture leads to acute or subacute occlusion of blood flow, acute myocardial damage ensues (Falk, 1995). Work over the past 35 years has proven that inflammation is integral to the pathogenesis of atherosclerosis. Activation of both innate and adaptive immunity occurs within the atherosclerotic plaque (Hansson, 2002), indicating a coordinated immune response rather than a non-specific inflammatory infiltrate. These paradigms, identified in experimental animal studies were validated against clinical data. The role of inflammation in atherosclerosis started with the finding of Goran Hansson that atherosclerotic plaques overexpress HLA-DR, an indirect sign of cytokine production and antigen presentation (Jonasson, 1985). It was subsequently demonstrated that macrophage recruitment into the arterial intima is required for the development of lesions and is associated with plaque activity, fibrous cap rupture and plaque erosion (Lessner, 2002; Swirski, 2006;Yan, 2007). Macrophage derived matrix metalloproteinases are now held responsible for the thinning of the fibrous cap and the risk of rupture (Falk, 1995). Toll-like receptors, innate immune pattern recognition receptors (PRRs) that detect pathogen-associated molecular patterns (PAMPs), are important mediators of the inflammatory response in atherosclerosis (Cole, 2010; Cole, 2011; Monaco, 2009). Besides innate immune responses that are mounted by macrophages and dendritic cells, adaptive immune responses with clonally expanded T cell populations contribute to this process. T-lymphocytes eventually reduce smooth muscle cell proliferation and repair mechanisms through the production of IFNγ leading to thinning of the fibrous cap (Hansson, 1989).
The recent Canakinumab Antiinflammatory Thrombosis Outcomes Study (CANTOS) has provided the first proof-of-concept that that targeting inflammation in cardiovascular disease is clinically relevant. In this study, in patients on statin therapy but with elevated C-reactive protein (CRP) levels, a significant reduction of CV events by the inhibition of interleukin-1β (IL-1β) without further influencing lipid levels was observed (Ridker, 2017). However, anti-IL-1β antibodies increases the infection risk. Thus, precision drugs targeting the immune response in atherosclerosis with limited side effects are required for the treatment of CVD.
Significant bottlenecks are preventing translation to the clinic including Host defense impairment with broad acting anti-inflammatories, the premature testing and failure of biologics (Chung, 2003;Mann, 2004) that discouraged Pharma from conducting clinical trials in CVD (Couzin-Frankel, 2012), a lack of knowledge of the best therapeutic targets due to a lack of preclinical data from human models of CVD and the fact that the immune system has opposing roles in CVD (Hansson, 2011).
B cells are a classic example of the contrasting role of immune cell subsets in CVD as they have been shown to carry out both pro- and anti-atherogenic functions and these dual roles are executed by different B cell subsets in CVD. B cells have been described in murine aortic root atherosclerotic lesions both at early and advanced stages of disease. CD22+ B cells were chiefly in clusters at the base of the plaques (Zhou, 1999). B cell depletion via an anti CD20 antibody reduced the development of atherosclerosis and the activation of dendritic cells and T cells, consistent with a pivotal role of B cells in the induction and perpetuation of the disease (Ait-Oufella, 2010). Transfer of splenic B cells to lymphocyte- depleted ApoE-/- mice or to B cell-deficient ApoE-/- mice increased atherosclerosis (Kyaw, 2010). In contrast to these results, suggesting a pathogenic involvement of B cells in atherosclerosis, there are studies demonstrating that B cells also play a protective role. Splenectomy in ApoE mice leads to moderately increased atherosclerotic lesions (Caligiuri, 2002). Adoptive transfer of total splenic B cells from atherosclerotic mice rescues splenectomised mice from increased atherogenesis (Caligiuri, 2002). The anti-atherogenic function of B cells has been assigned to “Innate” B1 cells, a lineage of B cells that has a peritoneal origin in mouse, and is distinct from bone marrow derived B cells - hereafter called B2 (Bouaziz, 2008). B1a B cells mediate the protective effect of splenic B cells via production of natural IgM antibodies (Kyaw, 2011) that recognise a phosphocholine motif contained within oxidised (ox) LDL and apoptotic cells (Binder, 2004; Chou, 2009 ; Horkko, 1999). Nevertheless, transfer of bone marrow from B cell deficient mice (mMT) to LDLR-/- mice, yields a 30% increase in lesion area (Major, 2002), suggesting that bone marrow – derived B cell subsets may also be protective.
B cell biology is a fertile new ground for identifying therapeutic targets in human disease, but limited knowledge is hampering their discovery. The success of therapeutic B cell depletion (rituximab) in autoimmune disease is very heterogeneous (Edwards, 2006) highlighting the complexity of B cell function, and our limited our understanding of their function in the context of human disease. B-cells are indispensable for antibody (Ab) generation. However, B-cells also produce cytokines, present antigens and restrain excessive immune responses (Harris, 2000; Mauri, 2012; Shlomchik, 2001). New data reveals that B cells participate in directing the magnitude and quality of T cell responses to foreign and self-antigens (Lund, 2010).
Phenotypic and functional heterogeneity have been clearly defined for effector and memory T cells. However B cell functional effector subsets and their role in disease are less well understood. Mature B cells can be further subdivided into marginal zone, follicular, B1, memory, regulatory and germinal centre (GC) subtypes (Fairfax, 2008) which complicates matters. Thus, understanding the distinction between suppressive and pathogenic B-cells and the “pathways” controlling their differentiation/maturation in CVD will improve targeting of pathogenic B-cell-lineages and will help to identify those patients most likely to benefit from such therapies.
The Athero-B-Cell project aimed to identify differences in B cell effector subsets relevant to atherosclerosis development and investigate how these differences could be used to develop new CVD therapeutics through combining the multidisciplinary knowledge and expertise of its Partners on B cells in inflammation and atherosclerosis with the data mining and therapeutic targeting platforms of world leading SMEs.
Athero-B-Cell aimed to bridge the gap between the laboratory and the clinic by bringing together basic and clinical scientists with internationally recognised expertise in: the biology of B cell subsets (Mauri-UCL, Malin-KI, Patrone-UniGenova), omics of leukocytes in large-scale clinical studies (Hoefer, UMCU), humoral immunity in CVD (Nordin Fredrickson-ULund, Mach-UniGeneva) and inflammatory signalling in atherosclerosis (Monaco-UOXF). Furthermore, the project sought to create an SME-Academic collaboration to generate a new pipeline of therapeutic targets whose potential will be realised by the SMEs.
The predominant aim of Athero-B-Cell was to identify, characterise and validate new approaches for manipulating B cell subsets in CVD using available and emerging -omics data from large-scale clinical studies.
Objectives:
1) To define targets to reduce activation of pathogenic B cells and antibodies
2) To identify enhancers of atheroprotective B regulatory cell populations and antibodies
3) To validate molecular and miRNA targets to modulate B cell behaviour in CVD with innovative locked nucleic acid (LNA) based antisense platforms (that have reached Phase 2 clinical trials in human).
Athero-B-Cell aims to mine available and new omics data from large-scale clinical studies to identify a specific B cell signature that is associated with cardiovascular disease and specific B cell effector functions including antibody and cytokine production as well as regulatory ability. The aim is to generate novel candidate therapeutic targets to modulate B cell behaviour in CVD.
Project Results:
Over the 60 months of the Athero-B-Cell project, the Consortium Partners have developed novel research tools (including a panel of antibodies to enable analysis and phenotyping of human B cell subsets in clinical cohorts and the generation of new bioinformatics research tools) to generate further interesting results and publications. The main work and accomplishments of the project are summarised below.
Work Package 1: Discovery of B cell specific therapeutic targets for CVD
The key efforts of the WP1 Beneficiaries revolved around the execution of B cell specific –omics, phenotypic studies, and antibody production in blood and serum samples from CVD cohorts. B cells were isolated from 295 samples from the Athero-Express study and high-throughput transcriptomics analysis was performed to map B cell-related changes specifically associated with CVD and CVD outcomes. Bioinformatics was applied to the data and identified differentially expressed genes in patients with versus without a follow-up cardiovascular event. Functional network and pathway analysis was also performed. A further study on PMBC used for the ‘omics’ study revealed that high numbers of unswitched and switched memory B cells are associated with a lower risk of secondary cardiovascular events in a population with severe cardiovascular disease.
In addition, a cytometry by time of flight (CyTOF) workflow was established. Mass cytometry (CyTOF) allows allows the simultaneous analysis of multiple parameters on single cells. The technique employs tagging of antibodies with isotopically pure rare earth metals, mostly lanthanides, which are then used to stain cells of interest. Due to the pure and distinct isotopic masses of the metals, the mass cytometry detection system overcomes the problem of channel overlap experienced in flow cytometry caused by flurochrome overlap. As a result, the introduction of CyTOF has greatly expanded the number of parameters that can be measured per single cell in a sample. More than 40 parameters at a time have been successfully measured per sample and as little as 10,000 cells have been used as starting cells for CyTOF analysis. This makes CyTOF one of the most robust techniques for identification of a whole repertoire of immune cells when tissues/cells for analysis are limited. CyTOF represents a step-change in our ability to phenotype cells in tissues in a broad and high-dimensional manner.
CyTOF was used to determine the immune cell landscape in aortas of ApoE-/- mice and revealed differences in the immune cell census of aortas from mice fed a chow and a high fat diet. This analysis revealed at least 13 leucocyte populations in the aortas of ApoE-/- mice, including major myeloid and lymphoid cell subsets. Feeding ApoE-/- mice a high fat diet led to increased monocytes and pDC and decreased cDC2 in the aortas. Although statistical significance was not reached, a trend towards increased neutrophils and reduced B cells was also observed in aortas of high fat compared to chow fed ApoE-/- mice. Myeloid cells (neutrophils, eosinophils, monocytes, macrophages, dendritic cells) are known to play important roles in atherosclerosis development. Myeloid cells (gated as Lin-CD11blo-hi) accounted for around 75% of the CD45+ leucocytes in the ApoE-/- aortas in our study. At least 13 myeloid cell subsets were found in the aortas of ApoE-/- mice. Differences in aortic myeloid populations were observed between ApoE-/- mice fed a chow and a high fat diet.
Studies in this work package also evaluated the relationship between anti-ApoA-1 antibody levels and clinical outcomes.
Anti-apoA-1 IgG were found to be independent predictors of nonfatal incident CAD in the general population. In a prospectively studied, population-based cohort of 5220 subjects (mean age 52.6±10.7 years, 47.4% males), followed over a median period of 5.6 years, subjects positive versus negative for anti-apoA-1 IgG presented a total CAD rate of 3.9% versus 2.8% (P=0.077) and a nonfatal CAD rate of 3.6% versus 2.3% (P=0.018) respectively. After multivariate adjustment for established cardiovascular risk factors, the hazard ratios of anti-apoA-1 IgG for total and nonfatal CAD were: hazard ratio=1.36 (95% confidence interval, 0.94-1.97; P=0.105) and hazard ratio=1.53 (95% confidence interval, 1.03-2.26; P=0.034) respectively. The strength of this association was dependent on a functional polymorphism of the CD14 receptor gene (Arterioscler Thromb Vasc Biol. 2017;37:2342-2349).
Furthermore, studies examined whether anti-apoA-1 IgG: (a) predict all-cause mortality in the general population and (b) are associated with single-nucleotide polymorphisms (SNPs) in a genome-wide association study (GWAS). Anti-apoA-1 IgG positivity independently predicted all-cause mortality: hazard ratio (HR) = 1.54 95% confidence interval (95% CI): 1.11-2.13 P = 0.01 indicating that preclinical autoimmunity to anti-apoA-1 IgG may represent a novel mortality risk factor (Front Immunol. 2017;8:437).
Work Package 2: Target Selection
Work package 2 led to the development of tools to aid target selection. SME-based in silico platforms were needed to integrate the –omics data for selection of the pathways forming a specific B cell signature associated with CVD outcomes and B cell effector functions. A fully automated solution that can perform effective data collection, quality assurance, processing and normalisation, as well as comprehensive statistical and functional analysis has been developed and utilised on datasets of the consortium partners.
Secondly, an ad-hoc panel of antibodies was described and used to interrogate the B cell compartment in cardiovascular disease. B cell content of 804 PBMC samples, collected as part of the prospective Malmo Diet and Cancer cardiovascular case control cohort, were phenotyped by flow cytometry. Initial analysis of the data has revealed interesting associations between specific B cell subsets.
Work Package 3: Validation of novel targets for B cell responses with pro-atherogenic outcomes
In WP3, novel locked nucleic acids (LNAs) and animal models against targets involved in B cell activity have been generated.
In recent years, RNA has become an attractive target for therapeutic intervention since RNA mediates most cellular gene expression. Synthetic oligonucleotides are the rational way to target RNA as using a sequence complementary to a specific mRNA, can inhibit its expression and therefore reduce target protein levels. Over the past decades, considerable work has attempted to develop RNA binding oligonucleotides with improved nuclease stability and binding affinity). The discovery of Locked Nucleic Acid (LNA) chemistry opened new avenues for antisense oligonucleotide therapeutics. LNA is an RNA modification with an oxy-methylene bridge between the 2’ and 4’-positions of the ribose. The methylene bridge creates a bi-cyclic structure ‘locking’ the ribose into a conformation ideal for Watson-Crick binding. When incorporated into a DNA or RNA oligonucleotide, LNA makes the pairing with a complementary nucleotide more rapid and the stability of the resulting duplex is increased. In addition, affinity to complementary RNA sequences and resistance to nuclease digestion is greatly enhanced by LNA.
LNA based antisense oligonucleotides possess the following features: 1) Excellent specificity, providing optimal targeting, 2) Increased affinity to targets providing improved potency in-vivo, 3) Strong pharmacology, cell permeability upon systemic delivery without complicated delivery vehicles, 4) Scalable and cost-effective manufacturing, 5) Well-tolerated with predictable toxicology and 6) Potential for oral delivery. The additional advantage of using LNA platforms is that it allows us to target intracellular molecules not easily targeted with small molecule inhibitors and/or antibodies. LNA technology provides an exciting opportunity to develop novel therapies to specifically target sub-populations of B cells.
Based on input from the Scientific Partners in the Consortium and from data generated as part of WP1 and WP2, several targets were chosen for RICC to develop LNAs against. LNAs against these selected targets have been generated and have been found to display powerful knockdown of the target in vitro and in vivo and had safe pharmacology in mouse. These compounds have been applied to in vitro experiments and to murine models of cardiovascular disease including a collar-induced injury model and a hypercholesterolaemic model of atherosclerosis.
Autoantibodies have been shown to play a critical role in predicting major adverse cardiovascular events in atherosclerotic patients. In this work package, we also aimed to assess the role of IgG auto-antibodies against ApoA-1 in mouse CV models. The role of anti-apoA-1 IgG on tissue factor (TF) expression and activation, a key coagulation regulator underlying atherothrombosis was examined. ApoE-/-, ApoE-/-TLR2-/- and ApoE-/-TLR4-/- mice were passively immunised with anti-apoA-1 or control IgG. intraplaque TF expression was significantly increased in ApoE-/- Mice that received anti-apoA-1 IgG passive immunisation. This was attenuated in ApoE-/- mice lacking either TLR2 or TLR4. Together with data from human samples, our results support a possible causal relationship between anti-apoA-1 IgG and atherothrombosis (Pagone et al., 2016 Thromb Haemost 116(3) 554-564).
Work Package 4: Validation of targets related to the enhancement of Bregs as a novel therapeutic strategy
AtheroBCell partners involved in this work package made significant progress towards the definition of the role of B cells in experimental models of atherosclerosis and also the generation of mouse models that specifically target B cells in hyperlipidemic mice.
As part of this project, the existence and role of regulatory B cells in vascular disease was examined. A well-established model of neointima formation based on the placement of a perivascular collar around the murine carotid artery in ApoE-/- mice was used to examine the role of Bcells and regulatory B cells in atherosclerosis. In this model, carotid injury leads to the development of fibro-atheromatous lesions with a macrophage-rich core. B cells were purified from the spleen and draining lymph nodes of ApoE-/- mice and transferred to syngeneic mice prior to collar placement. Adoptive transfer of B cells isolated from ApoE-/- LN significantly protected recipient mice from lesion development compared to control-treated mice or mice receiving splenic B cells. Moreover, treatment with LN-derived B cells reduced macrophage infiltration. The protective effect of lymph node B cells was found to be partially due to the cytokine interleukin (IL)-10. A significant increase in the number of total B cells was observed in the lymph nodes, but not in the spleens, of ApoE-/- compared to wild-type mice. Follicular B cells (CD21intCD23intCD24int) were increased in the LN of ApoE-/- compared to WT mice and in addition, an unexpected significant increase of CD21hiCD23hiCD24hi transitional two-marginal-zone precursor B cells, which were previously shown to have immunosuppressive Breg cell function, was observed in the LN of ApoE-/- compared to WT mice. Concomitantly, ApoE-/- mice displayed reduced levels of CD21hiCD23hiCD24hi B cells in the spleen compared to WT mice. We speculated that the CD21hiCD23hiCD24hi Breg subset, previously demonstrated to suppress immune-mediated experimental disease, might be responsible for the protective effect of the LN-derived B cells in our model. To prove the direct involvement of this subset in protection, we performed adoptive transfer experiments using purified CD21hiCD23hiCD24hi cells or follicular B cells isolated by FACS sorting from the LNs of ApoE-/- mice. We observed a significant reduction in lesion development after adoptive transfer of CD21hiCD23hiCD24hi B cells but not after transfer of the same number of follicular B cells. Similarly, neointimal macrophage content was reduced in the CD21hiCD23hiCD24hi B cell treated group compared to control and the follicular B cell treated group. The protective effect of CD21hiCD23hiCD24hi B cells was associated with an increase in levels of IL-10 in the serum. In conclusion, the AtheroBCell consortium described for the first time that a B2 Breg subset protects from neointima formation. We also revealed that B cells can provide vascular protection by means other than antibody generation, e.g. IL-10 production and the induction of Tr1 cells. (Strom, Cross, Cole et al., 2015. Thromb Haemost 114(4): 835-847).
Based on the expert knowledge of the AtheroBcell Partners and the data emerging from ‘-omics’ analysis in the project, several novel mouse models were designed and generated. One model will allow the specific knockdown of Bregs in atherosclerosis as well as fate-mapping experiments that will provide insight into the atherosclerotic ontogeny of the Breg lineage. Other models will allow tracking and deletion of molecules involved in Bcell biology and atherogenesis. Initial experiments in several of these mouse models are ongoing.
Moreover in this work package, we aimed to to investigate if B cells pulsed with p210-CTB can protect against atherosclerosis. Apolipoprotein B-100 is the major protein in LDL and we have previously identified certain peptide sequences as targets for autoimmune responses in humans and shown that prototype vaccines based on particularly peptide number 210 can inhibit the development of atherosclerosis in mice. The protective response induced by p210 immunization strategies has been shown to be associated with an increase in regulatory T cells (Tregs). Furthermore, p210 has been used together with other ApoB100 peptides in low dose continuous subcutaneous administration resulting in reduced lesion development and an increased Treg population. This work has led to results that are due to be published in the near future.
Potential Impact:
The Canakinumab Antiinflammatory Thrombosis Outcomes Study (CANTOS) has provided the first proof-of-concept that that targeting inflammation in cardiovascular disease is clinically relevant. (Ridker, NEJM 2017). However, anti-IL-1β antibodies increases the infection risk. Thus, precision drugs targeting the immune response in atherosclerosis with limited side effects are required for the treatment of CVD.
B cell targeting is an important therapeutic opportunity that has not being thoroughly explored in CVD. By mapping B cell heterogeneity in CVD, the Athero-B-Cell consortium has enabled the discovery and characterization of on/off switches of B cell phenotypes that can either contribute or protect form atherosclerosis and other vascular events, as well as novel effector pathways that B cell use to worsen disease such as autoreactive antibodies. The first impact is in the generation of a target portfolio that can be further pushed down the translational path by Academia and SME.
The second large impact is the SME success. The SMEs involved in Athero-B-Cell have gone from strength to strength during the course of the Consortium, furthering their target portfolio, skill development, knowledge and base of operation. Albeit Athero-B-Cell was not the only factor contributing their growth, this level of research activity led to a spur of further opportunities.
The third large impact of Athero-B-Cell has been its collaborative and cross-fertilising model of research commitment mainly based on data and model sharing, which has supported the research and the interactions of young scientists between all the Partners, cementing group and institutional interaction and data and knowledge exchange. This has led to stronger partnership between the participating Institutions and durable relationships being established, leading to at least 2 further collaborative applications in the near future.
The fourth impact are is the dissemination and training activities of the young scientists of Athero-B-Cell. The Investigators of Athero-B-Cell, many of which are females, albeit established, are relatively young themselves and these FP7 funds supported their ascending trajectory of them and also of their young post-docs and PhD students they harbour in their labs.
Besides the generation of novel scientific knowledge that can be exploited by the wider scientific
community, the project has also led to several discoveries that can be directly exploited
commercially. The most directly exploitable results are represented by Patent applications citing Athero-B-Cell as a founding source that have been listed in Template B2.
List of Websites:
The project website has the address: http://www.atherobcell.eu/.
Contractors involved:
- The Chancellor, Master and Scholars of University of Oxford (UK): Claudia Monaco
- Roche Innovation Center Copenhagen A/S (Denmark): Steffen Schmidt
- Université de Genève (Switzerland): François Mach
- Università di Genova (Italy): Alessio Nencioni
- Lunds Universitet (Sweden): Gunilla Nordin Fredrikson
- University Medical Center Utrecht (The Netherlands): Imo Hoefer
- Karolinska Institutet (Sweden): Stephen Malin
- University College London (UK): Claudia Mauri
- Bering Limited (UK): Ignat Drodzov
- genOway SA (France): Vincent Dubus
- ALTA Ricerca e Svluppo in Biotecnologie S.r.l.u. (Italy): Riccardo Bertini
Coordinator contact details:
The Chancellor, Master and Scholars of University of Oxford
Prof. Claudia Monaco
Kennedy Institute of Rheumatology
Roosevelt Drive, Headington, Oxford, OX3 7FY, UK.