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B. S. Vishwanath - One of the best experts on this subject based on the ideXlab platform.
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Echis carinatus snake venom metalloprotease-induced toxicities in mice: Therapeutic intervention by a repurposed drug, Tetraethyl thiuram disulfide (Disulfiram).
'Public Library of Science (PLoS)', 2021Co-Authors: Gotravalli V. Rudresha, Amog P. Urs, Vaddarahally N. Manjuprasanna, Krishnegowda Jayachandra, Rajesh Rajaiah, Mallanayakanakatte Milan D Gowda, B. S. VishwanathAbstract:Echis carinatus (EC) is known as saw-scaled viper and it is endemic to the Indian subcontinent. Envenoming by EC represents a major cause of snakebite mortality and morbidity in the Indian subcontinent. Zinc (Zn++) dependent snake venom metalloproteases (SVMPs) present in Echis carinatus venom (ECV) is well known to cause systemic hemorrhage and coagulopathy in experimental animals. An earlier report has shown that ECV activates neutrophils and releases neutrophil extracellular traps (NETs) that blocks blood vessels leading to severe Tissue Necrosis. However, the direct involvement of SVMPs in the release of NETs is not clear. Here, we investigated the direct involvement of EC SVMPs in observed pathological symptoms in a preclinical setup using specific Zn++ metal chelator, Tetraethyl thiuram disulfide (TTD)/disulfiram. TTD potently antagonizes the activity of SVMPs-mediated ECM protein degradation in vitro and skin hemorrhage in mice. In addition, TTD protected mice from ECV-induced footpad Tissue Necrosis by reduced expression of citrullinated H3 (citH3) and myeloperoxidase (MPO) in footpad Tissue. TTD also neutralized ECV-induced systemic hemorrhage and conferred protection against lethality in mice. Moreover, TTD inhibited ECV-induced NETosis in human neutrophils and decreased the expression of peptidyl arginine deiminase (PAD) 4, citH3, MPO, and p-ERK. Further, we demonstrated that ECV-induced NETosis and Tissue Necrosis are mediated via PAR-1-ERK axis. Overall, our results provide an insight into SVMPs-induced toxicities and the promising protective efficacy of TTD can be extrapolated to treat severe Tissue Necrosis complementing anti-snake venom (ASV)
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Echis carinatus snake venom metalloprotease-induced toxicities in mice: therapeutic intervention by a repurposed drug, tetraethylthiuram disulfide (disulfiram)
2020Co-Authors: Gotravalli V. Rudresha, Amog P. Urs, Vaddarahally N. Manjuprasanna, Mallanayakanakatte D. Milan Gowda, Krishnegowda Jayachandra, Rajesh Rajaiah, B. S. VishwanathAbstract:Echis carinatus (EC) is known as saw-scaled viper and it is endemic to the Indian subcontinent. Envenoming by EC represents a major cause of snakebite mortality and morbidity in the Indian subcontinent. Zinc (Zn++)-dependent snake venom metalloproteases (SVMPs) present in Echis carinatus venom is well known to cause systemic hemorrhage and coagulopathy in experimental animals. An earlier report has shown that ECV activates neutrophils and releases neutrophil extracellular traps (NETs) that blocks blood vessels leading to severe Tissue Necrosis. However, the direct involvement of SVMPs in the release of NETs is not clear. Here, we investigated the direct involvement of EC SVMPs in observed pathological symptoms in a preclinical setup using zinc (Zn++) metal chelator, Tetraethylthiuram disulfide (TTD)/disulfiram. TTD potently antagonizes the activity of SVMPs-mediated ECM protein degradation in vitro and skin hemorrhage in mice. In addition, TTD protected mice from ECV-induced footpad Tissue Necrosis by reduced expression of citrullinated H3 (citH3) and myeloperoxidase (MPO) in footpad Tissue. TTD also neutralized ECV-induced systemic hemorrhage and conferred protection against lethality in mice. Moreover, TTD inhibited ECV-induced NETosis in human neutrophils and decreased the expression of peptidylarginine deiminase (PAD) 4, citH3, MPO and pERK. Further, we demonstrated that ECV-induced NETosis and Tissue Necrosis is mediated via PAR-1-ERK axis. Overall, our results provide an insight into SVMPs-induced toxicities and the promising protective efficacy of TTD can be extrapolated to treat severe Tissue Necrosis complementing ASV.
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Serine protease from Tricosanthus tricuspidata accelerates healing of Echis carinatus venom-induced necrotic wound
Toxicon : official journal of the International Society on Toxinology, 2020Co-Authors: Gotravalli V. Rudresha, Manisha Choudhury, Amog P. Urs, Vaddarahally N. Manjuprasanna, Rajesh Rajaiah, B. S. VishwanathAbstract:Abstract Echis carinatus (EC) envenomation causes severe immune response by the accumulation of Tissue debris in the form of DAMPs resulting in chronic inflammation and progressive Tissue Necrosis at the bitten site. Clearing of Tissue debris is a prerequisite to enhance the healing of venom-induced necrotic wounds. Tricosanthus tricuspidata is a medicinal plant used extensively for the treatment of snake bite-induced toxicities. The active component responsible for the observed pharmacological action is a serine protease, tricuspidin. The topical application of tricuspidin was able to neutralize ECV-induced mouse footpad Tissue Necrosis and open wound in rabbits. Tricuspidin exerted its healing action via proteolytic activity as a consequence of upregulation of MMP-8 and down regulation of MMP-9. Further, tricuspidin reduced ECV-induced inflammation by decreasing the expression of TNF-α, IL-6 and MPO, and by increasing the level of VEGF-A and TGF-β1. The modulation of ECV induced immune/inflammatory mediators by tricuspidin was found to be more effective than trypsin. Moreover, tricuspidin and trypsin activated MAPKs via protease activated receptors-2 (PAR-2). These data indicate that the proteolytic activity of tricuspidin directly involved in the healing of ECV-induced chronic wound.
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plant dnases are potent therapeutic agents against echis carinatus venom induced Tissue Necrosis in mice
Journal of Cellular Biochemistry, 2019Co-Authors: Gotravalli V. Rudresha, Amog P. Urs, Vaddarahally N. Manjuprasanna, Kanve Nagaraj Suvilesh, Puttappa Sharanappa, B. S. VishwanathAbstract:Echis carinatus envenomation leads to severe Tissue Necrosis at the bitten site by releasing DNA from immune cells that blocks the blood flow. An earlier report has shown that exogenous DNase 1 offers protection against such severe local Tissue Necrosis. Tricosanthus tricuspidata is a medicinal plant and the paste prepared from its leaves has been used extensively for the treatment of snakebite-induced Tissue Necrosis. Most studies including reports from our laboratory focused on plant secondary metabolite as therapeutic molecules against snakebite envenomation. However, the involvement of hydrolytic enzymes including DNase in treating snake venom-induced Tissue Necrosis has not been addressed. Several folk medicinal plants used against snakebite treatment showed the presence of DNase activity and found to be rich in T. tricuspidata. Further, purified T. tricuspidata DNase showed a single sharp peak in reversed-phase high-performance liquid chromatography (RP-HPLC) with an apparent molecular mass of 17 kDa. T. tricuspidata DNase exhibited potent DNA degrading activity performed using agarose gel electrophoresis, spectrophotometric assay, and DNA zymography. In addition, purified DNase from T. tricuspidata was able to neutralize E. carinatus venom-induced mouse tail Tissue Necrosis and normalized elevated serum creatine kinase (CK) and lactate dehydrogenase (LDH) levels 30 minutes post venom injection. T. tricuspidata DNase was also able to reverse E. carinatus venom-induced histopathological changes and collagen depletion in mice tail Tissue. All these observed pharmacological actions of T. tricuspidata DNase were inhibited by sodium fluoride (NaF). This study provides scientific validation of the traditional use of T. tricuspidata leaf paste in the healing of snakebite-induced Tissue Necrosis and might be exploited to treat snake venom-induced local toxicity.
Christopher D Lascola - One of the best experts on this subject based on the ideXlab platform.
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bag3 bcl 2 associated athanogene 3 coding variant in mice determines susceptibility to ischemic limb muscle myopathy by directing autophagyclinical perspective
Circulation, 2017Co-Authors: Joseph M Mcclung, Timothy J Mccord, Terence E Ryan, Cameron A Schmidt, Thomas D Green, Jessica L Reinardy, Sarah B Mueller, Talaignair N Venkatraman, Kevin W. Southerland, Christopher D LascolaAbstract:Background: Critical limb ischemia is a manifestation of peripheral artery disease that carries significant mortality and morbidity risk in humans, although its genetic determinants remain largely unknown. We previously discovered 2 overlapping quantitative trait loci in mice, Lsq-1 and Civq-1 , that affected limb muscle survival and stroke volume after femoral artery or middle cerebral artery ligation, respectively. Here, we report that a Bag3 variant (Ile81Met) segregates with Tissue protection from hind-limb ischemia. Methods: We treated mice with either adeno-associated viruses encoding a control (green fluorescent protein) or 2 BAG3 (Bcl-2–associated athanogene-3) variants, namely Met81 or Ile81, and subjected the mice to hind-limb ischemia. Results: We found that the BAG3 Ile81Met variant in the C57BL/6 (BL6) mouse background segregates with protection from Tissue Necrosis in a shorter congenic fragment of Lsq-1 (C.B6– Lsq1-3 ). BALB/c mice treated with adeno-associated virus encoding the BL6 BAG3 variant (Ile81; n=25) displayed reduced limb-Tissue Necrosis and increased limb Tissue perfusion compared with Met81- (n=25) or green fluorescent protein– (n=29) expressing animals. BAG3 Ile81 , but not BAG3 Met81 , improved ischemic muscle myopathy and muscle precursor cell differentiation and improved muscle regeneration in a separate, toxin-induced model of injury. Systemic injection of adeno-associated virus–BAG3 Ile81 (n=9), but not BAG3 Met81 (n=10) or green fluorescent protein (n=5), improved ischemic limb blood flow and limb muscle histology and restored muscle function (force production). Compared with BAG3 Met81 , BAG3 Ile81 displayed improved binding to the small heat shock protein (HspB8) in ischemic skeletal muscle cells and enhanced ischemic muscle autophagic flux. Conclusions: Taken together, our data demonstrate that genetic variation in BAG3 plays an important role in the prevention of ischemic Tissue Necrosis. These results highlight a pathway that preserves Tissue survival and muscle function in the setting of ischemia.
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a bag3 coding variant in mice determines susceptibility to ischemic limb muscle myopathy by directing autophagy
Circulation, 2017Co-Authors: Joseph M Mcclung, Timothy J Mccord, Terence E Ryan, Cameron A Schmidt, Thomas D Green, Jessica L Reinardy, Sarah B Mueller, Talaignair N Venkatraman, Kevin W. Southerland, Christopher D LascolaAbstract:Background —Critical limb ischemia (CLI) is a manifestation of peripheral artery disease (PAD) that carries significant mortality and morbidity risk in humans, although its genetic determinants remain largely unknown. We previously discovered two overlapping quantitative trait loci (QTL) in mice, Lsq-1 and Civq-1 , that affected limb muscle survival and stroke volume following femoral artery or middle cerebral artery ligation, respectively. Here we report that a Bag3 variant (Ile81Met) segregates with Tissue protection from hindlimb ischemia (HLI). Methods —We treated mice with either adeno-associated viruses (AAV) encoding a control (GFP), or two BAG3 variants, namely Met81 or Ile81, and subjected the mice to hindlimb ischemia. Results —We found that the BAG3 Ile81Met variant in the C57BL/6 (BL6) mouse background segregates with protection from Tissue Necrosis in a shorter congenic fragment of Lsq-1 (C.B6- Lsq1-3 ). Treating BALB/c mice with AAV encoding the BL6 BAG3 variant (Ile81) ( n =25) displayed reduced limb Tissue Necrosis and increased limb Tissue perfusion compared to Met81- ( n =25) or GFP- ( n =29) expressing animals. BAG3Ile81, but not BAG3Met81, improved ischemic muscle myopathy and muscle precursor cell differentiation and improved muscle regeneration in a separate, toxin-induced model of injury. Systemic injection of AAV-BAG3Ile81 ( n =9), but not BAG3Met81 ( n =10) or GFP ( n =5), improved ischemic limb blood flow, limb muscle histology, and restored muscle function (force production). Compared to BAG3Met81, BAG3Ile81 displayed improved binding to the small heat shock protein (HspB8) in ischemic skeletal muscle cells and enhanced ischemic muscle autophagic flux. Conclusions —Taken together, our data demonstrate that genetic variation in BAG3 plays an important role in the prevention of ischemic Tissue Necrosis. These results highlight a pathway that preserves Tissue survival and muscle function in the setting of ischemia.
Gotravalli V. Rudresha - One of the best experts on this subject based on the ideXlab platform.
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Echis carinatus snake venom metalloprotease-induced toxicities in mice: Therapeutic intervention by a repurposed drug, Tetraethyl thiuram disulfide (Disulfiram).
'Public Library of Science (PLoS)', 2021Co-Authors: Gotravalli V. Rudresha, Amog P. Urs, Vaddarahally N. Manjuprasanna, Krishnegowda Jayachandra, Rajesh Rajaiah, Mallanayakanakatte Milan D Gowda, B. S. VishwanathAbstract:Echis carinatus (EC) is known as saw-scaled viper and it is endemic to the Indian subcontinent. Envenoming by EC represents a major cause of snakebite mortality and morbidity in the Indian subcontinent. Zinc (Zn++) dependent snake venom metalloproteases (SVMPs) present in Echis carinatus venom (ECV) is well known to cause systemic hemorrhage and coagulopathy in experimental animals. An earlier report has shown that ECV activates neutrophils and releases neutrophil extracellular traps (NETs) that blocks blood vessels leading to severe Tissue Necrosis. However, the direct involvement of SVMPs in the release of NETs is not clear. Here, we investigated the direct involvement of EC SVMPs in observed pathological symptoms in a preclinical setup using specific Zn++ metal chelator, Tetraethyl thiuram disulfide (TTD)/disulfiram. TTD potently antagonizes the activity of SVMPs-mediated ECM protein degradation in vitro and skin hemorrhage in mice. In addition, TTD protected mice from ECV-induced footpad Tissue Necrosis by reduced expression of citrullinated H3 (citH3) and myeloperoxidase (MPO) in footpad Tissue. TTD also neutralized ECV-induced systemic hemorrhage and conferred protection against lethality in mice. Moreover, TTD inhibited ECV-induced NETosis in human neutrophils and decreased the expression of peptidyl arginine deiminase (PAD) 4, citH3, MPO, and p-ERK. Further, we demonstrated that ECV-induced NETosis and Tissue Necrosis are mediated via PAR-1-ERK axis. Overall, our results provide an insight into SVMPs-induced toxicities and the promising protective efficacy of TTD can be extrapolated to treat severe Tissue Necrosis complementing anti-snake venom (ASV)
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Echis carinatus snake venom metalloprotease-induced toxicities in mice: therapeutic intervention by a repurposed drug, tetraethylthiuram disulfide (disulfiram)
2020Co-Authors: Gotravalli V. Rudresha, Amog P. Urs, Vaddarahally N. Manjuprasanna, Mallanayakanakatte D. Milan Gowda, Krishnegowda Jayachandra, Rajesh Rajaiah, B. S. VishwanathAbstract:Echis carinatus (EC) is known as saw-scaled viper and it is endemic to the Indian subcontinent. Envenoming by EC represents a major cause of snakebite mortality and morbidity in the Indian subcontinent. Zinc (Zn++)-dependent snake venom metalloproteases (SVMPs) present in Echis carinatus venom is well known to cause systemic hemorrhage and coagulopathy in experimental animals. An earlier report has shown that ECV activates neutrophils and releases neutrophil extracellular traps (NETs) that blocks blood vessels leading to severe Tissue Necrosis. However, the direct involvement of SVMPs in the release of NETs is not clear. Here, we investigated the direct involvement of EC SVMPs in observed pathological symptoms in a preclinical setup using zinc (Zn++) metal chelator, Tetraethylthiuram disulfide (TTD)/disulfiram. TTD potently antagonizes the activity of SVMPs-mediated ECM protein degradation in vitro and skin hemorrhage in mice. In addition, TTD protected mice from ECV-induced footpad Tissue Necrosis by reduced expression of citrullinated H3 (citH3) and myeloperoxidase (MPO) in footpad Tissue. TTD also neutralized ECV-induced systemic hemorrhage and conferred protection against lethality in mice. Moreover, TTD inhibited ECV-induced NETosis in human neutrophils and decreased the expression of peptidylarginine deiminase (PAD) 4, citH3, MPO and pERK. Further, we demonstrated that ECV-induced NETosis and Tissue Necrosis is mediated via PAR-1-ERK axis. Overall, our results provide an insight into SVMPs-induced toxicities and the promising protective efficacy of TTD can be extrapolated to treat severe Tissue Necrosis complementing ASV.
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Serine protease from Tricosanthus tricuspidata accelerates healing of Echis carinatus venom-induced necrotic wound
Toxicon : official journal of the International Society on Toxinology, 2020Co-Authors: Gotravalli V. Rudresha, Manisha Choudhury, Amog P. Urs, Vaddarahally N. Manjuprasanna, Rajesh Rajaiah, B. S. VishwanathAbstract:Abstract Echis carinatus (EC) envenomation causes severe immune response by the accumulation of Tissue debris in the form of DAMPs resulting in chronic inflammation and progressive Tissue Necrosis at the bitten site. Clearing of Tissue debris is a prerequisite to enhance the healing of venom-induced necrotic wounds. Tricosanthus tricuspidata is a medicinal plant used extensively for the treatment of snake bite-induced toxicities. The active component responsible for the observed pharmacological action is a serine protease, tricuspidin. The topical application of tricuspidin was able to neutralize ECV-induced mouse footpad Tissue Necrosis and open wound in rabbits. Tricuspidin exerted its healing action via proteolytic activity as a consequence of upregulation of MMP-8 and down regulation of MMP-9. Further, tricuspidin reduced ECV-induced inflammation by decreasing the expression of TNF-α, IL-6 and MPO, and by increasing the level of VEGF-A and TGF-β1. The modulation of ECV induced immune/inflammatory mediators by tricuspidin was found to be more effective than trypsin. Moreover, tricuspidin and trypsin activated MAPKs via protease activated receptors-2 (PAR-2). These data indicate that the proteolytic activity of tricuspidin directly involved in the healing of ECV-induced chronic wound.
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plant dnases are potent therapeutic agents against echis carinatus venom induced Tissue Necrosis in mice
Journal of Cellular Biochemistry, 2019Co-Authors: Gotravalli V. Rudresha, Amog P. Urs, Vaddarahally N. Manjuprasanna, Kanve Nagaraj Suvilesh, Puttappa Sharanappa, B. S. VishwanathAbstract:Echis carinatus envenomation leads to severe Tissue Necrosis at the bitten site by releasing DNA from immune cells that blocks the blood flow. An earlier report has shown that exogenous DNase 1 offers protection against such severe local Tissue Necrosis. Tricosanthus tricuspidata is a medicinal plant and the paste prepared from its leaves has been used extensively for the treatment of snakebite-induced Tissue Necrosis. Most studies including reports from our laboratory focused on plant secondary metabolite as therapeutic molecules against snakebite envenomation. However, the involvement of hydrolytic enzymes including DNase in treating snake venom-induced Tissue Necrosis has not been addressed. Several folk medicinal plants used against snakebite treatment showed the presence of DNase activity and found to be rich in T. tricuspidata. Further, purified T. tricuspidata DNase showed a single sharp peak in reversed-phase high-performance liquid chromatography (RP-HPLC) with an apparent molecular mass of 17 kDa. T. tricuspidata DNase exhibited potent DNA degrading activity performed using agarose gel electrophoresis, spectrophotometric assay, and DNA zymography. In addition, purified DNase from T. tricuspidata was able to neutralize E. carinatus venom-induced mouse tail Tissue Necrosis and normalized elevated serum creatine kinase (CK) and lactate dehydrogenase (LDH) levels 30 minutes post venom injection. T. tricuspidata DNase was also able to reverse E. carinatus venom-induced histopathological changes and collagen depletion in mice tail Tissue. All these observed pharmacological actions of T. tricuspidata DNase were inhibited by sodium fluoride (NaF). This study provides scientific validation of the traditional use of T. tricuspidata leaf paste in the healing of snakebite-induced Tissue Necrosis and might be exploited to treat snake venom-induced local toxicity.
Gregory G. Neely - One of the best experts on this subject based on the ideXlab platform.
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Molecular dissection of box jellyfish venom cytotoxicity highlights an effective venom antidote
Nature Communications, 2019Co-Authors: Man-tat Lau, Jamie Seymour, John Manion, Jamie B. Littleboy, Lisa Oyston, Thang M. Khuong, Qiao-ping Wang, David T. Nguyen, Daniel Hesselson, Gregory G. NeelyAbstract:The box jellyfish Chironex fleckeri is extremely venomous, and envenoming causes Tissue Necrosis, extreme pain and death within minutes after severe exposure. Despite rapid and potent venom action, basic mechanistic insight is lacking. Here we perform molecular dissection of a jellyfish venom-induced cell death pathway by screening for host components required for venom exposure-induced cell death using genome-scale lenti-CRISPR mutagenesis. We identify the peripheral membrane protein ATP2B1, a calcium transporting ATPase, as one host factor required for venom cytotoxicity. Targeting ATP2B1 prevents venom action and confers long lasting protection. Informatics analysis of host genes required for venom cytotoxicity reveal pathways not previously implicated in cell death. We also discover a venom antidote that functions up to 15 minutes after exposure and suppresses Tissue Necrosis and pain in mice. These results highlight the power of whole genome CRISPR screening to investigate venom mechanisms of action and to rapidly identify new medicines. Box jellyfish venom causes Tissue damage, pain, and death through unknown molecular mechanisms. Here, Lau et al. perform a CRISPR screen to identify genes required for venom action and use this information to develop an antidote that blocks venom-induced pain and Tissue damage in vivo.
Joseph M Mcclung - One of the best experts on this subject based on the ideXlab platform.
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bag3 bcl 2 associated athanogene 3 coding variant in mice determines susceptibility to ischemic limb muscle myopathy by directing autophagyclinical perspective
Circulation, 2017Co-Authors: Joseph M Mcclung, Timothy J Mccord, Terence E Ryan, Cameron A Schmidt, Thomas D Green, Jessica L Reinardy, Sarah B Mueller, Talaignair N Venkatraman, Kevin W. Southerland, Christopher D LascolaAbstract:Background: Critical limb ischemia is a manifestation of peripheral artery disease that carries significant mortality and morbidity risk in humans, although its genetic determinants remain largely unknown. We previously discovered 2 overlapping quantitative trait loci in mice, Lsq-1 and Civq-1 , that affected limb muscle survival and stroke volume after femoral artery or middle cerebral artery ligation, respectively. Here, we report that a Bag3 variant (Ile81Met) segregates with Tissue protection from hind-limb ischemia. Methods: We treated mice with either adeno-associated viruses encoding a control (green fluorescent protein) or 2 BAG3 (Bcl-2–associated athanogene-3) variants, namely Met81 or Ile81, and subjected the mice to hind-limb ischemia. Results: We found that the BAG3 Ile81Met variant in the C57BL/6 (BL6) mouse background segregates with protection from Tissue Necrosis in a shorter congenic fragment of Lsq-1 (C.B6– Lsq1-3 ). BALB/c mice treated with adeno-associated virus encoding the BL6 BAG3 variant (Ile81; n=25) displayed reduced limb-Tissue Necrosis and increased limb Tissue perfusion compared with Met81- (n=25) or green fluorescent protein– (n=29) expressing animals. BAG3 Ile81 , but not BAG3 Met81 , improved ischemic muscle myopathy and muscle precursor cell differentiation and improved muscle regeneration in a separate, toxin-induced model of injury. Systemic injection of adeno-associated virus–BAG3 Ile81 (n=9), but not BAG3 Met81 (n=10) or green fluorescent protein (n=5), improved ischemic limb blood flow and limb muscle histology and restored muscle function (force production). Compared with BAG3 Met81 , BAG3 Ile81 displayed improved binding to the small heat shock protein (HspB8) in ischemic skeletal muscle cells and enhanced ischemic muscle autophagic flux. Conclusions: Taken together, our data demonstrate that genetic variation in BAG3 plays an important role in the prevention of ischemic Tissue Necrosis. These results highlight a pathway that preserves Tissue survival and muscle function in the setting of ischemia.
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a bag3 coding variant in mice determines susceptibility to ischemic limb muscle myopathy by directing autophagy
Circulation, 2017Co-Authors: Joseph M Mcclung, Timothy J Mccord, Terence E Ryan, Cameron A Schmidt, Thomas D Green, Jessica L Reinardy, Sarah B Mueller, Talaignair N Venkatraman, Kevin W. Southerland, Christopher D LascolaAbstract:Background —Critical limb ischemia (CLI) is a manifestation of peripheral artery disease (PAD) that carries significant mortality and morbidity risk in humans, although its genetic determinants remain largely unknown. We previously discovered two overlapping quantitative trait loci (QTL) in mice, Lsq-1 and Civq-1 , that affected limb muscle survival and stroke volume following femoral artery or middle cerebral artery ligation, respectively. Here we report that a Bag3 variant (Ile81Met) segregates with Tissue protection from hindlimb ischemia (HLI). Methods —We treated mice with either adeno-associated viruses (AAV) encoding a control (GFP), or two BAG3 variants, namely Met81 or Ile81, and subjected the mice to hindlimb ischemia. Results —We found that the BAG3 Ile81Met variant in the C57BL/6 (BL6) mouse background segregates with protection from Tissue Necrosis in a shorter congenic fragment of Lsq-1 (C.B6- Lsq1-3 ). Treating BALB/c mice with AAV encoding the BL6 BAG3 variant (Ile81) ( n =25) displayed reduced limb Tissue Necrosis and increased limb Tissue perfusion compared to Met81- ( n =25) or GFP- ( n =29) expressing animals. BAG3Ile81, but not BAG3Met81, improved ischemic muscle myopathy and muscle precursor cell differentiation and improved muscle regeneration in a separate, toxin-induced model of injury. Systemic injection of AAV-BAG3Ile81 ( n =9), but not BAG3Met81 ( n =10) or GFP ( n =5), improved ischemic limb blood flow, limb muscle histology, and restored muscle function (force production). Compared to BAG3Met81, BAG3Ile81 displayed improved binding to the small heat shock protein (HspB8) in ischemic skeletal muscle cells and enhanced ischemic muscle autophagic flux. Conclusions —Taken together, our data demonstrate that genetic variation in BAG3 plays an important role in the prevention of ischemic Tissue Necrosis. These results highlight a pathway that preserves Tissue survival and muscle function in the setting of ischemia.
