Proyectos - Internacionales - Programas de Investigación e Innovación de la UE

We are investigating how tissue damage and microbial signals lead to mitochondrial reprogramming in dendritic cells (DCs) and macrophages, and how manipulation of mitochondrial metabolism regulates myeloid cell function. This project aims to identify new targets to exploit DCs and macrophages for improved immunotherapy. So far we have: 1. Characterized the metabolic reprogramming after DC stimulation with different stimuli both in mouse and human DCs. 2. We are performing the analysis of how innate sensing connects with mitochondrial adaptations in DCs; 3. We have addressed the effect of drugs targeting mitochondrial biology and now we are analyzing the effect of genetic manipulation of mitochondrial biology on DC function in vitro. 4. We are assessing the functional in vivo effects of targeting mitochondrial biology in DCs in homeostasis and disease.

The project aims 1. the analysis of how sensing of external stimuli by DCs leads to mitochondrial adaptations; 2. The investigation in how mitochondrial metabolism may impact in DC function.

The main results obtained in these aims are:
1. In the current reporting period we have found how macrophage polarization is regulated by Fgr kinase-mediated phosphorylation of mitochondrial CII, which impacts on proinflammatory macrophages in the adipose tissue. Fgr deficiency leads to improved glucose metabolism and leaner mice with reduced liver steatosis (Acín-Pérez et al. Nat Metabolism 2020). In addition, we found that a polybacterial preparation induces metabolic reprogramming and trained immunity, protects against viral infection and enhances vaccine immunogenicity (Brandi et al. 2022. Cell Reports; del Fresno et al. Front. Immunol. 2021).
2. Our novel genetic approaches based on CD11c-selective deletion of key proteins affecting OXPHOS complexes have revealed diverse results depending on the targeted complex. We performed the analysis of CD11c-expressing macrophages. We found that distinct physiological functions direct the metabolic requirements of tissue-resident macrophages. Using the CD11c-Cre driver line, we observed a profound phenotype in some CD11c-expressing macrophage (MF) populations. Using our CD11c∆Tfam mice and Lysozyme M-Cre Tfamf/f (LysM∆Tfam) mice, we found that mitochondrial impairment differentially affects presence and identity of MFs correlating with their expression levels of OXPHOS-related genes. Alveolar macrophages are drastically affected and the oxphos is needed for macrophage lipid handling activity. Macrophages in tissues exposed to lipids in homeostasis are more dependent on oxphos for their metabolism. In addition, proinflammatory macrophages in the adipose tissue are also oxphos-dependent. Due to the lack of this proinflammatory macrophages, LysM∆Tfam mice are leaner and show better glucose metabolism and less liver steatosis than control mice (Wculek et al. submitted).

In relation with the previous mentioned objectives, the progress and expected potential are:
1. We have characterized how innate stimuli lead to distinct metabolic signatures and have described modulatory mechanisms that affect metabolic reprogramming and trained immunity (Saz-Leal et al. Cell Reports. 2018). We found that a polybacterial preparation induces metabolic reprogramming and trained immunity, protects against viral infection and enhances vaccine immunogenicity (Brandi et al. 2022. Cell Reports; del Fresno et al. Front. Immunol. 2021).We are now working on new regulators of trained immunity with particular focus on the role of mitochondrial metabolism.
2. In our biased approach we have established the involvement of the HIF-1 pathway in alveolar macrophage function and the importance of sensing oxygen for terminal differentiation (Izquierdo et al. Cell Reports. 2018). We are also analyzing other sensing pathways that affect DC and macrophage mitochondrial metabolism.
3. We have found a new function in inflammation of DCs (Del Fresno et al. Science. 2018) and a new pathway of sensing microbiota that affects immunity in the gut (Martínez-López et al. Immunity. 2019). This has potential impact as new functional targets that can be affected by the manipulation of mitochondrial metabolism, and we are currently exploring this avenue. In fact, we have recently described that Fgr kinase can modulate mitochondrial complex II activity and macrophage polarization, which affects host metabolism. Fgr promotes proinflammatory macrophage polarization, obesity and metabolic syndrome (Acín-Pérez et al. Nat Metab. 2020).
4. Our work has established that cDC1s can be effectively used for cancer immunotherapy (Wculek et al. JITC. 2019). The potential is to manipulate metabolism in cDC1s to improve cancer immunotherapy.

Atherosclerosis, a leading cause of heart disease, poses a challenge due to its complex and often asymptomatic progression. In this context, the EIC-funded ABCardionostics project will develop a multi-marker PET/MRI system. Specifically, ABCardionostics aims to enhance the staging and personalised treatment of patients with vulnerable atherosclerosis. The project focuses on creating human antibodies (HuAbs) that target specific monocyte/macrophage populations involved in plaque development. These antibodies will provide detailed insights into plaque composition and progression. Using advanced biotechnologies, including phage display and multispectral mapping, ABCardionostics will pioneer in vivo imaging techniques and novel immunotherapy strategies, potentially transforming diagnostic and therapeutic practices for atherosclerosis and beyond.

Ischemic heart disease is the main cause of death in the EU, straining patients and economies. Regenerative Medicine has failed at delivering a definitive solution, and even the breakthrough of cell reprogramming, biomaterials or 3D printing, have not been able to find a curative solution. Generating a muscle with efficient pumping requires a careful recapitulation of the myocardial architecture. BRAV∃ is born with the ambition of shaping this quantum leap in the field. The overall concept is to provide a lasting functional support to injured hearts through the fabrication of regenerative personalized advanced tissue engineering-based biological ventricular assist devices (BioVADs). To do so, we will apply multimodal deep cardiac phenotyping, coupled to advanced Computational Modelling and biomechanical analysis in a large animal model of disease, to create a personalised 3D printable design. We will for the first time create a fibre-reinforced human heart-sized cardiac tissue able to recapitulate the low Young´s Modulus of the myocardium while withstanding pressures generated during the cardiac circle. Using the latest human induced pluripotent stem cell (hiPSC) technology and industrial-scale growth and differentiation, we will cellularize this novel human heart-sized constructs, creating a highly efficiently aligned cardiac tissue (including vasculature). BioVADs will be matured in in-Consortium built electromechanical stimulation bioreactors before transplantation in a porcine model of disease. We anticipate our BioVADs will constitute a one-shot regenerative treatment of IHD, decreasing the burden on healthcare providers and improving the quality of life of patients. Crucially, we will for the first time generate a wealth of information on heart development at a human scale. Delivering this novel application whilst developing the technological environment (bioreactor, chamber, pacemaker) will boost the capacity of the EU to grow economically and lead the field.

This project aims to investigate the early associations between cardiovascular health and Alzheimer’s disease (AD) pathology, with a focus on subclinical atherosclerosis (SCAth) and cardiovascular risk factors (CVRFs), in a large cohort of asymptomatic, middle-aged individuals. The overarching objective is to determine how cardiovascular health in subclinical stages influences both cerebrovascular integrity and early AD biomarkers, and to examine how these factors contribute to cognitive decline, with particular emphasis on sex-specific differences.

The proposed research leverages data from the PESA-Brain study, a deeply phenotyped cohort integrating advanced neuroimaging and cognitive testing, with longitudinal cardiovascular and blood-based biomarkers assessments. We will first test the relationships between cardiovascular health markers and cerebrovascular health, focusing on brain perfusion, microstructure, and microinfarcts. We will then explore the links between cardiovascular health and AD pathology, using plasma and brain amyloid and tau biomarkers, as well as cognitive outcomes, including memory and executive function.

The project will employ advanced statistical approaches, such as linear mixed models and mediation analysis, to handle the complexity of multimodal data and examine causal relationships between cardiovascular health, brain pathology, and cognition. By including sex-stratified analyses, the research will provide critical insights into gender-specific susceptibilities, which have been underexplored in previous studies.

The relevance of this project to the work programme lies in its potential to shift the focus of AD prevention to midlife, offering novel insights into the neurobiological pathways that link cardiovascular dysfunction and neurodegeneration. These findings could pave the way for earlier interventions targeting modifiable CVRFs, significantly reducing the incidence and societal burden of dementia in aging populations.

Cardiovascular disease remains the leading cause of mortality worldwide, with atherosclerosis (AT) as the primary underlying factor. Ageing is the strongest risk factor for both AT and CVD. Emerging evidence indicates that the B cell-mediated immune response plays a pivotal role in the development and progression of AT, although the nature of this response and its triggering antigens are poorly understood. In our prior research, we identified ALDH4A1 as novel AT antigen and showed that anti-ALDH4A1 A12 antibody is atheroprotective in mice. Our recent preliminary data show increased titers of anti-ALDH4A1 antibodies in aged mice as well as a rise in age-associated B cells in AT mice. These findings suggest a strong link between ageing, the B cell immune response, and AT. ANTI-ATHERO aims to functionally dissect the AT antibody response with the ultimate goal of identifying age-associated antigens that can serve as targets for specific immunotherapies. We will pursue the following aims: 1) characterize the early and aged B cell immune response in AT with time-stamping mouse models, 2) generate a single cell clonal atlas of time-stamped AT B cells and identify individual antibodies and antigens associated with the early and the aged AT antibody response, 3) translate our findings to human disease by mapping the B cell response in clinical AT and testing the identified antigens as markers of disease progression and, 4) evaluate the therapeutic potential of the identified antibodies and antigens through passive and active immunization strategies. We anticipate that the ANTI-ATHERO project will generate transformative insights into the interplay between ageing, the immune system, and AT. Our findings have the potential to significantly advance not only cardiovascular research but also contribute to a broader understanding of other age-related diseases and autoimmune disorders.