TY - JOUR
T1 - Stimulating the autophagic-lysosomal axis enhances host defense against fungal infection in a zebrafish model of invasive Aspergillosis
AU - Forn-Cuní, G.
AU - Welvaarts, L.
AU - Stel, F. M.
AU - van den Hondel, C. J.
AU - Arentshorst, M.
AU - Ram, A. F. J.
AU - Meijer, A. H.
N1 - Funding Information: We thank Michiel van der Vaart and Monica Varela for confocal microscopy advice and critical proofreading of the manuscript, and all members of the fish facility team for zebrafish care at the Institute of Biology Leiden. The A. fumigatus melanin-deficient strain ∆pksP was a kind gift from Dr. Jean-Paul Latgé (Institut Pasteur, Paris, France). The fluorescent zebrafish lines labelling Lc3, macrophages or neutrophils were kind gifts from Dan Klionsky (University of Michigan, USA), Georges Lutfalla (University of Montpellier, France), and Stephen Renshaw (University of Sheffield). This work and G.F.-C. were supported by a European Marie Curie fellowship (H2020-COFUND-2015-FP707404). Funding Information: This work was supported by the H2020 Marie Skłodowska-Curie Actions [H2020-COFUND-2015-FP707404]. We thank Michiel van der Vaart and Monica Varela for confocal microscopy advice and critical proofreading of the manuscript, and all members of the fish facility team for zebrafish care at the Institute of Biology Leiden. The A. fumigatus melanin-deficient strain ∆pksP was a kind gift from Dr. Jean-Paul Latgé (Institut Pasteur, Paris, France). The fluorescent zebrafish lines labelling Lc3, macrophages or neutrophils were kind gifts from Dan Klionsky (University of Michigan, USA), Georges Lutfalla (University of Montpellier, France), and Stephen Renshaw (University of Sheffield). This work and G.F.-C. were supported by a European Marie Curie fellowship (H2020-COFUND-2015-FP707404). Publisher Copyright: © 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.
PY - 2023
Y1 - 2023
N2 - The increasing prevalence of antifungal-resistant human pathogenic fungi, particularly azole-resistant Aspergillus fumigatus, is a life-threatening challenge to the immunocompromised population. Autophagy-related processes such as LC3-associated phagocytosis have been shown to be activated in the host response against fungal infection, but their overall effect on host resistance remains uncertain. To analyze the relevance of these processes in vivo, we used a zebrafish animal model of invasive Aspergillosis. To confirm the validity of this model to test potential treatments for this disease, we confirmed that immunosuppressive treatments or neutropenia rendered zebrafish embryos more susceptible to A. fumigatus. We used GFP-Lc3 transgenic zebrafish to visualize the autophagy-related processes in innate immune phagocytes shortly after phagocytosis of A. fumigatus conidia, and found that both wild-type and melanin-deficient conidia elicited Lc3 recruitment. In macrophages, we observed GFP-Lc3 accumulation in puncta after phagocytosis, as well as short, rapid events of GFP-Lc3 decoration of single and multiple conidia-containing vesicles, while neutrophils covered single conidia-containing vesicles with bright and long-lasting GFP-Lc3 signal. Next, using genetic and pharmacological stimulation of three independent autophagy-inducing pathways, we showed that the antifungal autophagy response improves the host survival against A. fumigatus infection, but only in the presence of phagocytes. Therefore, we provide proof-of-concept that stimulating the (auto)phagolysosomal pathways is a promising approach to develop host-directed therapies against invasive Aspergillosis, and should be explored further either as adjunctive or stand-alone therapy for drug-resistant Aspergillus infections. Abbreviations: DMSO: dimethyl sulfoxide; HR: hazard ratio; HDT: host-directed therapy; Hpf: hours post fertilization; IA: invasive Aspergillosis; LAP: LC3-associated phagocytosis; MTZ: metronidazole; PTU: N-phenylthiourea; ROS: reactive oxygen species.
AB - The increasing prevalence of antifungal-resistant human pathogenic fungi, particularly azole-resistant Aspergillus fumigatus, is a life-threatening challenge to the immunocompromised population. Autophagy-related processes such as LC3-associated phagocytosis have been shown to be activated in the host response against fungal infection, but their overall effect on host resistance remains uncertain. To analyze the relevance of these processes in vivo, we used a zebrafish animal model of invasive Aspergillosis. To confirm the validity of this model to test potential treatments for this disease, we confirmed that immunosuppressive treatments or neutropenia rendered zebrafish embryos more susceptible to A. fumigatus. We used GFP-Lc3 transgenic zebrafish to visualize the autophagy-related processes in innate immune phagocytes shortly after phagocytosis of A. fumigatus conidia, and found that both wild-type and melanin-deficient conidia elicited Lc3 recruitment. In macrophages, we observed GFP-Lc3 accumulation in puncta after phagocytosis, as well as short, rapid events of GFP-Lc3 decoration of single and multiple conidia-containing vesicles, while neutrophils covered single conidia-containing vesicles with bright and long-lasting GFP-Lc3 signal. Next, using genetic and pharmacological stimulation of three independent autophagy-inducing pathways, we showed that the antifungal autophagy response improves the host survival against A. fumigatus infection, but only in the presence of phagocytes. Therefore, we provide proof-of-concept that stimulating the (auto)phagolysosomal pathways is a promising approach to develop host-directed therapies against invasive Aspergillosis, and should be explored further either as adjunctive or stand-alone therapy for drug-resistant Aspergillus infections. Abbreviations: DMSO: dimethyl sulfoxide; HR: hazard ratio; HDT: host-directed therapy; Hpf: hours post fertilization; IA: invasive Aspergillosis; LAP: LC3-associated phagocytosis; MTZ: metronidazole; PTU: N-phenylthiourea; ROS: reactive oxygen species.
KW - Aspergillus
KW - autophagic defense
KW - fungal infection
KW - host-pathogen interaction
KW - immunomodulation
KW - innate immunity
KW - phagocytes
KW - zebrafish
UR - http://www.scopus.com/inward/record.url?scp=85133258648&partnerID=8YFLogxK
U2 - https://doi.org/10.1080/15548627.2022.2090727
DO - https://doi.org/10.1080/15548627.2022.2090727
M3 - Article
C2 - 35775203
SN - 1554-8627
VL - 19
SP - 324
EP - 337
JO - Autophagy
JF - Autophagy
IS - 1
ER -