Sc-RNA sequencing of skeletal muscle macrophages during T. gondii infection and injury
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PROVIDER: PRJNA450034 | ENA |
REPOSITORIES: ENA
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Project description:The proper coordination of macrophages is essential for skeletal muscle repair. However, the full heterogenity of macrophages responding to injury has yet to be elucidated on the single-cell level. Furthermore, chronic inflammation may shift macrophage heterogeneity at steady-state and affect the generation of heterogeneity necessary for tissue repair. We use scRNA-sequencing to determine the heterogeneity of skeletal muscle macrophages during infection and wound repair (uninfected and infected). We identify multiple transcriptionally distinct macrophage subsets during injury in uninfected mice as well as during infection at steady-state. Interestingly, in response to injury, infected skeletal muscle macrophages fail to generate certain subsets of reparative macrophages in a timely fashion.
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Project description:Toxoplasma gondii is an obligatory intracellular parasite that causes persistent infections in birds and mammals including ~30% of the world’s human population. Differentiation from proliferative and metabolically active tachyzoites to largely dormant bradyzoites initiates the chronic phase of infection and occurs predominantly in brain and muscle tissues. Here we used murine skeletal muscle cells (SkMCs) to decipher host cellular factors that favor T. gondii bradyzoite formation in terminally differentiated and syncytial myotubes, but not in proliferating myoblast precursors. Genome-wide transcriptome analyses of T. gondii-infected SkMCs and non-infected controls identified ~6,500 genes which were differentially expressed (DEGs) in myotubes compared to myoblasts, largely irrespective of infection. On the other hand, genes related to central carbohydrate metabolism, to redox homeostasis, and to the Nrf2-dependent stress response pathway were enriched in both infected myoblast precursors and myotubes. Stable isotope-resolved metabolite profiling indicated increased fluxes into the oxidative branch of the pentose phosphate pathway (OxPPP) in infected myoblasts and into the TCA cycle in infected myotubes. High OxPPP activity in infected myoblasts was associated with increased NADPH/NADP+ ratio while myotubes exhibited higher ROS levels and lower expression of anti-oxidants and detoxification enzymes. Pharmacological reduction of ROS levels in SkMCs inhibited bradyzoite differentiation, while increased ROS induced bradyzoite formation. Thus, we identified a novel host cell-dependent mechanism that triggers stage conversion of T. gondii into persistent tissue cysts in its natural host cell type.
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Project description:Skeletal muscle injury-responsive macrophage subsets in larval zebrafish
| PRJNA684129 | ENA
Project description:Single-cell RNA-sequencing (scRNA-seq) was applied to identify and characterise macrophage subsets that responded to larval zebrafish muscle injury. The Tg(mpeg1:mCherry) transgenic zebrafish line was utilised to isolate mCherry-expressing macrophages by FACS. Following needle-stab muscle injury of a 4 days post fertilisation (dpf) larvae, the wound site was dissected out at 1, 2, and 3 days post injury (dpi) for macrophage isolation. Macrophages isolated from 4 dpf-uninjured larvae were also included. This analysis led to the identification of 8 discrete clusters of macrophages, one of which corresponded to uninjured macrophages. The 7 wound-present macrophage subsets highlighted greater macrophage heterogeneity than previously described in an in vivo skeletal muscle injury context.
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Project description:The oxidative phosphorylation (OXPHOS) system is dynamic and the respiratory complexes (RCs) coexist with super-assembled quaternary structures called supercomplexes (SCs). How assembly occurs and the physiological role of supercomplex assembly is still under intensive investigation. The Cox7a family member Cox7a2l, also known as Scaf1, promotes CIII-CIV super assembly and energetic efficiency in zebrafish, mice, and humans. Here we studied the role of a second member of the Cox7a family, Cox7a1 in SC assembly and striated muscle physiology. We found that this protein drives CIV homodimer formation, which increases CIV activity. The substantial reduction in CIV2 formation led to a profound metabolic rearrangement with a consequent non-pathological loss of skeletal muscle performance. Ablation of Cox7a1 also rewired heart metabolism. Already in homeostatic conditions, cox7a1-/- hearts revealed a pro-regenerative metabolic profile. While overall cardiac function was not affected, the absence of Cox7a1 altered the cardiac regenerative response. The effects of cox7a1 and cox7a2l loss of function on skeletal muscle and myocardium physiology and injury response were not identical, revealing that there is a high specificity of Cox7a isoform in controlling OXPHOS assembly and striated muscle metabolism. While in both cases overall OXPHOS activity is modified, the loss of CIII-CIV heterodimer formation or CIV homodimer formation have very distinct metabolic and physiological consequences, highlighting the complexity of OXPHOS function and the importance of cox7a1 in striated muscle maturation.
2024-03-20 | GSE254466 | GEO
Project description:Toxoplasmosis is a zoonotic disease caused by a ubiquitous protozoan parasite called Toxoplasma gondii, which can
infect all mammal and bird species throughout the world. seroprevalence varies widely between countries. Studies
have estimated that between 7-34% of people in the UK have been infected with T. gondii. The vast majority of these
people will not have noticed any symptoms, however about 10% of people develop a mild to moderate self limiting
flu-like illness. Following the acute active stage of the infection the parasite persists in the body in the form of cysts,
particularly in heart and skeletal muscle and nervous system tissues, for many years, and usually for life. In
immunocompetent persons these cysts do not pose a health risk. We will use RNA-seq to quantify the transcriptional
response of macrophages to T gondii infection.
This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/
| EGAS00001001708 | EGA