Metabolic Regulation of Antifungal Immunity through the Mevalonate Pathway

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Genetic variation in PFKFB3 impairs antifungal immunometabolic responses and predisposes to Invasive Pulmonary Aspergillosis

Publication . Gonçalves, Samuel M.; Antunes, Daniela; Leite, Luis; Mercier, Toine; Horst, Rob Ter; Vieira, Joana; Espada, Eduardo; Pinho Vaz, Carlos; Branca, Rosa; Campilho, Fernando; Freitas, Fátima; Ligeiro, Dário; Marques, António; van de Veerdonk, Frank L.; Joosten, Leo A. B.; Lagrou, Katrien; Maertens, Johan; Netea, Mihai G.; Lacerda, João; Campos, António; Cunha, Cristina; Carvalho, Agostinho

Activation of immune cells in response to fungal infection involves the reprogramming of their cellular metabolism to support antimicrobial effector functions. Although metabolic pathways such as glycolysis are known to represent critical regulatory nodes in antifungal immunity, it remains undetermined whether these are differentially regulated at the interindividual level. In this study, we identify a key role for 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) in the immunometabolic responses to Aspergillus fumigatus. A genetic association study performed in 439 recipients of allogeneic hematopoietic stem cell transplantation (HSCT) and corresponding donors revealed that the donor, but not recipient, rs646564 variant in the PFKFB3 gene increased the risk of invasive pulmonary aspergillosis (IPA) after transplantation. The risk genotype impaired the expression of PFKFB3 by human macrophages in response to fungal infection, which was correlated with a defective activation of glycolysis and the ensuing antifungal effector functions. In patients with IPA, the risk genotype was associated with lower concentrations of cytokines in the bronchoalveolar lavage fluid samples. Collectively, these findings demonstrate the important contribution of genetic variation in PFKFB3 to the risk of IPA in patients undergoing HSCT and support its inclusion in prognostic tools to predict the risk of fungal infection in this clinical setting. IMPORTANCE The fungal pathogen Aspergillus fumigatus can cause severe and life-threatening forms of infection in immunocompromised patients. Activation of glycolysis is essential for innate immune cells to mount effective antifungal responses. In this study, we report the contribution of genetic variation in the key glycolytic activator 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) to the risk of invasive pulmonary aspergillosis (IPA) after allogeneic hematopoietic stem cell transplantation. The PFKFB3 genotype associated with increased risk of infection was correlated with an impairment of the antifungal effector functions of macrophages in vitro and in patients with IPA. This work highlights the clinical relevance of genetic variation in PFKFB3 to the risk of IPA and supports its integration in risk stratification and preemptive measures for patients at high risk of IPA.

2021-05-28Journal article

Open access

Genetic determinants of fungi-induced ROS production are associated with the risk of invasive pulmonary aspergillosis

Publication . Matzaraki, Vasiliki; Beno, Alexandra; Jaeger, Martin; Gresnigt, Mark S.; Keur, Nick; Boahen, Collins; Cunha, Cristina; Gonçalves, Samuel M.; Leite, Luis; Lacerda, João; Campos, António; van de Veerdonk, Frank L.; Joosten, Leo; Netea, Mihai G.; Carvalho, Agostinho; Kumar, Vinod

Reactive oxygen species (ROS) are an essential component of the host defense against fungal infections. However, little is known about how common genetic variation affects ROS-mediated antifungal host defense. In the present study, we investigated the genetic factors that regulate ROS production capacity in response to the two human fungal pathogens: Candida albicans and Aspergillus fumigatus. We investigated fungal-stimulated ROS production by immune cells isolated from a population-based cohort of approximately 200 healthy individuals (200FG cohort), and mapped ROS-quantitative trait loci (QTLs). We identified several genetic loci that regulate ROS levels (P < 9.99 × 10-6), with some of these loci being pathogen-specific, and others shared between the two fungi. These ROS-QTLs were investigated for their influence on the risk of invasive pulmonary aspergillosis (IPA) in a disease relevant context. We stratified hematopoietic stem-cell transplant (HSCT) recipients based on the donor's SNP genotype and tested their impact on the risk of IPA. We identified rs4685368 as a ROS-QTL locus that was significantly associated with an increased risk of IPA after controlling for patient age and sex, hematological malignancy, type of transplantation, conditioning regimen, acute graft-versus-host-disease grades III-IV, and antifungal prophylaxis. Collectively, this data provides evidence that common genetic variation can influence ROS production capacity, and, importantly, the risk of developing IPA among HSCT recipients. This evidence warrants further research for patient stratification based on the genetic profiling that would allow the identifications of patients at high-risk for an invasive fungal infection, and who would benefit the most from a preventive strategy.

2022Journal article

Open access

Phagosomal removal of fungal melanin reprograms macrophage metabolism to promote antifungal immunity

Publication . Gonçalves, Samuel M.; Duarte-Oliveira, Cláudio; Campos, Cláudia F.; Aimanianda, Vishukumar; ter Horst, Rob; Leite, Luis; Mercier, Toine; Pereira, Paulo N G; Fernández-García, Miguel; Antunes, Daniela; Rodrigues, Cláudia S.; Barbosa-Matos, Catarina; Gaifem, Joana; Mesquita, Inês; Marques, António; Osório, Nuno S.; Torrado, Egídio; Rodrigues, Fernando; Costa, Sandra; Joosten, Leo AB.; Lagrou, Katrien; Maertens, Johan; Lacerda, João; Campos, António; Brown, Gordon D.; Brakhage, Axel A.; Barbas, Coral; Silvestre, Ricardo; van de Veerdonk, Frank L.; Chamilos, Georgios; Netea, Mihai G.; Latgé, Jean-Paul; Cunha, Cristina; Carvalho, Agostinho

In response to infection, macrophages adapt their metabolism rapidly to enhance glycolysis and fuel specialized antimicrobial effector functions. Here we show that fungal melanin is an essential molecule required for the metabolic rewiring of macrophages during infection with the fungal pathogen Aspergillus fumigatus. Using pharmacological and genetic tools, we reveal a molecular link between calcium sequestration by melanin inside the phagosome and induction of glycolysis required for efficient innate immune responses. By remodeling the intracellular calcium machinery and impairing signaling via calmodulin, melanin drives an immunometabolic signaling axis towards glycolysis with activation of hypoxia-inducible factor 1 subunit alpha (HIF-1α) and phagosomal recruitment of mammalian target of rapamycin (mTOR). These data demonstrate a pivotal mechanism in the immunometabolic regulation of macrophages during fungal infection and highlight the metabolic repurposing of immune cells as a potential therapeutic strategy.

2020Journal article

Open access