The Warburg effect is necessary to promote glycosylation in the blastema during zebrafish tail regeneration

Sinclair, Jason; Hoying, David R.; Bresciani, Erica; Nogare, Damian Dalle; Needle, Carli D.; Berger, Alexandra; Wu, Weiwei; Bishop, Kevin; Elkahloun, Abdel G.; Chitnis, Ajay B.; Liu, Paul

doi:10.1038/s41536-021-00163-x

Cited by 34 publications

(40 citation statements)

References 95 publications

(117 reference statements)

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“…Increased expression of glycolysis genes has also been observed in zebrafish following larval tail amputation, with glycolysis inhibition resulting in abnormal blastema formation (Sinclair et a., 2021), and we additionally find that activity of the glycolytic enzymes hexokinase and LDH are required for larval tail regeneration. Thus, aerobic glycolysis is required for successful regeneration through the formation or output of the blastema-like notochord bead.…”

Section: A Role For Aerobic Glycolysis In Early Wound Healing and For...supporting

confidence: 68%

Aerobic glycolysis is important for zebrafish larval wound closure and tail regeneration

Carney

Amaya

2021

Preprint

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The underlying mechanisms of appendage regeneration remain largely unknown, and uncovering these mechanisms in capable organisms have far-reaching implications for potential treatments in humans. Recent studies implicate a requirement for metabolic reprogramming reminiscent of the Warburg effect during successful appendage and organ regeneration. Changes are thus predicted to be highly dynamic, methods permitting direct visualization of metabolites at the tissue and organismal level, in real time, would offer a significant advance in defining the influence of metabolism on regeneration and healing. We sought to examine whether glycolytic activity was altered during larval fin regeneration, utilising the genetically encoded biosensor, Laconic, enabling the spatiotemporal assessment of lactate levels in living zebrafish. We present evidence for a rapid increase in lactate levels within minutes following injury, with a role of aerobic glycolysis in actomyosin contraction and wound closure. We also find a second wave of lactate production, associated with overall larval tail regeneration. Chemical inhibition of glycolysis attenuates both contraction of the wound and regrowth of tissue following tail amputation, suggesting aerobic glycolysis is necessary at two distinct stages of regeneration.SUMMARY STATEMENTBy combining a genetically encoded lactate FRET sensor with chemical inhibitors, we demonstrate a critical role for the Warburg effect and metabolic reprogramming during zebrafish wound closure and tail regeneration.

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Section: A Role For Aerobic Glycolysis In Early Wound Healing and For...supporting

confidence: 68%

Aerobic glycolysis is important for zebrafish larval wound closure and tail regeneration

Carney

Amaya

2021

Preprint

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“…Nevertheless, our data suggests that a reduced intracellular redox state in macrophages supports wound healing; this is supported by the results of the metformin treatment showing that an increase in the optical redox ratio ( Figure 5D ) is associated with improved wound healing ( Figure 6H ) and the converse by Stat6 depletion ( Figure 6D and J ). Our results are in line with recent zebrafish studies that found that tail transection leads to a shift to glucose metabolism and wound healing is blocked upon 2-DG treatment ( Sinclair et al, 2021 ), and progenerative macrophages display a glycolytic phenotype in context of muscle injury as indicated by increased levels of intracellular NADH measured by two-photon autofluorescence imaging and bioluminescence-based assay ( Ratnayake et al, 2021 ).…”

Section: Discussionsupporting

confidence: 92%

In vivo fluorescence lifetime imaging of macrophage intracellular metabolism during wound responses in zebrafish

Miskolci

Tweed

Lasarev

et al. 2022

eLife

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The function of macrophages in vitro is linked to their metabolic rewiring. However, macrophage metabolism remains poorly characterized in situ. Here, we used two-photon intensity and lifetime imaging of autofluorescent metabolic coenzymes, nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) and flavin adenine dinucleotide (FAD), to assess the metabolism of macrophages in the wound microenvironment. Inhibiting glycolysis reduced NAD(P)H mean lifetime and made the intracellular redox state of macrophages more oxidized, as indicated by reduced optical redox ratio. We found that TNFα+ macrophages had lower NAD(P)H mean lifetime and were more oxidized compared to TNFα− macrophages. Both infection and thermal injury induced a macrophage population with a more oxidized redox state in wounded tissues. Kinetic analysis detected temporal changes in the optical redox ratio during tissue repair, revealing a shift toward a more reduced redox state over time. Metformin reduced TNFα+ wound macrophages, made intracellular redox state more reduced and improved tissue repair. By contrast, depletion of STAT6 increased TNFα+ wound macrophages, made redox state more oxidized and impaired regeneration. Our findings suggest that autofluorescence of NAD(P)H and FAD is sensitive to dynamic changes in intracellular metabolism in tissues and can be used to probe the temporal and spatial regulation of macrophage metabolism during tissue damage and repair.

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“…This metabolic shift implies that the migratory layer has a higher metabolic demand, which is supplied by glycolytic energy metabolism, which is fast but inefficient compared with OXPHOS [ 51 ]. In another study, zebrafish with their tails chopped off have a metabolic change that favours glucose metabolism during early regeneration [ 52 ]. Blocking glucose metabolism with 2-DG did not affect the embryo’s general health, but it prevented the tail from regrowing after amputation due to the inability to develop a functioning blastema, reinforcing that glycolysis plays a quintessential role in wound healing [ 52 ].…”

Section: Discussionmentioning

confidence: 99%

“…In another study, zebrafish with their tails chopped off have a metabolic change that favours glucose metabolism during early regeneration [ 52 ]. Blocking glucose metabolism with 2-DG did not affect the embryo’s general health, but it prevented the tail from regrowing after amputation due to the inability to develop a functioning blastema, reinforcing that glycolysis plays a quintessential role in wound healing [ 52 ]. In our study, abrogating the energy production by 2-DG drastically reduced ePRP-mediated wound healing abilities in fibroblasts ( Figure 8 A,B).…”

Section: Discussionmentioning

confidence: 99%

Enhanced Platelet-Rich Plasma (ePRP) Stimulates Wound Healing through Effects on Metabolic Reprogramming in Fibroblasts

Weng

Cheng

Lee

et al. 2021

IJMS

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As a source of growth factors for expediting wound healing and tissue regeneration, plasma-rich plasma (PRP) has been extensively applied in diverse fields including orthopaedics, ophthalmology, oral and maxillofacial surgery, dentistry, and gynaecology. However, the function of PRP in metabolic regulations remains enigmatic. A standardized method was devised herein to enrich growth factors and to lyophilize it as enhanced PRP (ePRP) powder, which could become ubiquitously available without mechanical centrifugation in clinical practice. To identify metabolic reprogramming in human dermal fibroblasts under ePRP treatment, putative metabolic targets were identified by transcriptome profiling and validated for their metabolic effects and mechanism. ePRP does not only promote wound healing but re-aligns energy metabolism by shifting to glycolysis through stimulation of glycolytic enzyme activity in fibroblasts. On the contrary, oxygen consumption rates and several mitochondrial respiration activities were attenuated in ePRP-treated fibroblasts. Furthermore, ePRP treatment drives the mitochondrial resetting by hindering the mitochondrial biogenesis-related genes and results in a dampened mitochondrial mass. Antioxidant production was further increased by ePRP treatment to prevent reactive oxygen species formation. Besides, ePRP also halts the senescence progression of fibroblasts by activating SIRT1 expression. Importantly, the glycolytic inhibitor 2-DG can completely reverse the ePRP-enhanced wound healing capacity, whereas the mitochondrial inhibitor oligomycin cannot. This is the first study to utilize PRP for comprehensively investigating its effects on the metabolic reprogramming of fibroblasts. These findings indicate that PRP’s primary metabolic regulation is to promote metabolic reprogramming toward glycolytic energy metabolism in fibroblasts, preserving redox equilibrium and allowing anabolic pathways necessary for the healing and anti-ageing process.

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The Warburg effect is necessary to promote glycosylation in the blastema during zebrafish tail regeneration

Cited by 34 publications

References 95 publications

Aerobic glycolysis is important for zebrafish larval wound closure and tail regeneration

Aerobic glycolysis is important for zebrafish larval wound closure and tail regeneration

In vivo fluorescence lifetime imaging of macrophage intracellular metabolism during wound responses in zebrafish

Enhanced Platelet-Rich Plasma (ePRP) Stimulates Wound Healing through Effects on Metabolic Reprogramming in Fibroblasts

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