Cholesterol absorption blocker ezetimibe prevents muscle wasting in severe dysferlin‐deficient and mdx mice
Abstract Background Muscular dystrophy (MD) causes muscle wasting and is often lethal in patients due to a lack of proven therapies. In contrast, mouse models of MD are notoriously mild. We have previously shown severe human‐like muscle pathology in mdx [Duchenne MD (DMD)] and dysferlin‐deficient li...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2022-02-01
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| Series: | Journal of Cachexia, Sarcopenia and Muscle |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/jcsm.12879 |
| _version_ | 1827267262105518080 |
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| author | Zoe White Marine Theret Nadia Milad Lin Wei Tung William Wei‐Han Chen Martin G. Sirois Fabio Rossi Pascal Bernatchez |
| author_facet | Zoe White Marine Theret Nadia Milad Lin Wei Tung William Wei‐Han Chen Martin G. Sirois Fabio Rossi Pascal Bernatchez |
| author_sort | Zoe White |
| collection | DOAJ |
| description | Abstract Background Muscular dystrophy (MD) causes muscle wasting and is often lethal in patients due to a lack of proven therapies. In contrast, mouse models of MD are notoriously mild. We have previously shown severe human‐like muscle pathology in mdx [Duchenne MD (DMD)] and dysferlin‐deficient limb‐girdle MD type 2B (LGMD2B) mice by inactivating the gene encoding for apolipoprotein E (ApoE), a lipid transporter synthesized by the liver, brain and adipocytes to regulate lipid and fat metabolism. Having recently established that human DMD is a novel type of primary genetic dyslipidaemia with elevated cholesterol, we sought to determine whether cholesterol could exacerbate the muscle wasting process observed in severe rodent MD. Methods Severe mdx and dysferlin knock‐out mice lacking ApoE were treated with ezetimibe (15 mg/kg/day), a clinically approved drug exhibiting few pleiotropic effects. In separate studies, dietary cholesterol was raised (from 0.2% to 2% cholesterol) in combination with experimental micro‐injury and direct cholesterol injection assays. Muscles were assessed histologically for changes in collagen and adipocyte infiltration and both transcriptomic and cellular changes by RNA‐seq and fluorescence‐activated cell sorting analysis. Results Treatment of severe DMD and LGMD2B mice with ezetimibe completely prevented clinical signs of ambulatory dysfunction (0% incidence vs. 33% for vehicle treatment; P < 0.05). Histological analyses revealed that ezetimibe‐reduced fibro‐fatty infiltration up to 84% and 63% in severely affected triceps (P ≤ 0.0001) and gastrocnemius (P ≤ 0.003) muscles, resulting in a respective 1.9‐fold and 2.2‐fold retention of healthy myofibre area (P ≤ 0.0001). Additionally, raising dietary cholesterol and thus concentrations of plasma low‐density lipoprotein‐associated cholesterol (by 250%; P < 0.0001) reduced overall survivability (by 100%; P < 0.001) and worsened muscle damage in the LGMD2B triceps by 767% (P < 0.03). Micro‐pin‐induced mechanical injury in LGMD2B mice fed a high cholesterol diet exacerbated muscle damage by 425% (P < 0.03) and increased macrophage recruitment (by 98%; P = 0.03) compared with those injured on a chow diet. Parallel RNA‐seq analyses revealed that injury in cholesterol‐fed mice also modulated the expression of 3671 transcripts (1953 up‐regulated), with fibrogenic, inflammatory and programmed cell death‐associated pathways among the most enriched. Mice lacking dysferlin also displayed heightened muscle necrosis (by 123%; P < 0.0001) following a direct intramuscular injection of cholesterol compared with control mice. Conclusions Cholesterol exacerbates rodent MD. Specific inhibition of cholesterol absorption with ezetimibe may safely attenuate human MD severity and delay death. |
| first_indexed | 2024-04-09T12:37:12Z |
| format | Article |
| id | doaj.art-1734f6d867e74b1188de0ce7c5e14a22 |
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| language | English |
| last_indexed | 2025-03-22T04:30:57Z |
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| publisher | Wiley |
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| series | Journal of Cachexia, Sarcopenia and Muscle |
| spelling | doaj.art-1734f6d867e74b1188de0ce7c5e14a222024-04-28T03:10:19ZengWileyJournal of Cachexia, Sarcopenia and Muscle2190-59912190-60092022-02-0113154456010.1002/jcsm.12879Cholesterol absorption blocker ezetimibe prevents muscle wasting in severe dysferlin‐deficient and mdx miceZoe White0Marine Theret1Nadia Milad2Lin Wei Tung3William Wei‐Han Chen4Martin G. Sirois5Fabio Rossi6Pascal Bernatchez7Department of Anesthesiology, Pharmacology & Therapeutics University of British Columbia (UBC) Vancouver BC CanadaSchool of Biomedical Engineering, Department of Medical Genetics University of British Columbia (UBC) Vancouver BC CanadaDepartment of Anesthesiology, Pharmacology & Therapeutics University of British Columbia (UBC) Vancouver BC CanadaSchool of Biomedical Engineering, Department of Medical Genetics University of British Columbia (UBC) Vancouver BC CanadaDepartment of Anesthesiology, Pharmacology & Therapeutics University of British Columbia (UBC) Vancouver BC CanadaMontreal Heart Institute, Department of Pharmacology and Physiology Université de Montréal Montreal QC CanadaSchool of Biomedical Engineering, Department of Medical Genetics University of British Columbia (UBC) Vancouver BC CanadaDepartment of Anesthesiology, Pharmacology & Therapeutics University of British Columbia (UBC) Vancouver BC CanadaAbstract Background Muscular dystrophy (MD) causes muscle wasting and is often lethal in patients due to a lack of proven therapies. In contrast, mouse models of MD are notoriously mild. We have previously shown severe human‐like muscle pathology in mdx [Duchenne MD (DMD)] and dysferlin‐deficient limb‐girdle MD type 2B (LGMD2B) mice by inactivating the gene encoding for apolipoprotein E (ApoE), a lipid transporter synthesized by the liver, brain and adipocytes to regulate lipid and fat metabolism. Having recently established that human DMD is a novel type of primary genetic dyslipidaemia with elevated cholesterol, we sought to determine whether cholesterol could exacerbate the muscle wasting process observed in severe rodent MD. Methods Severe mdx and dysferlin knock‐out mice lacking ApoE were treated with ezetimibe (15 mg/kg/day), a clinically approved drug exhibiting few pleiotropic effects. In separate studies, dietary cholesterol was raised (from 0.2% to 2% cholesterol) in combination with experimental micro‐injury and direct cholesterol injection assays. Muscles were assessed histologically for changes in collagen and adipocyte infiltration and both transcriptomic and cellular changes by RNA‐seq and fluorescence‐activated cell sorting analysis. Results Treatment of severe DMD and LGMD2B mice with ezetimibe completely prevented clinical signs of ambulatory dysfunction (0% incidence vs. 33% for vehicle treatment; P < 0.05). Histological analyses revealed that ezetimibe‐reduced fibro‐fatty infiltration up to 84% and 63% in severely affected triceps (P ≤ 0.0001) and gastrocnemius (P ≤ 0.003) muscles, resulting in a respective 1.9‐fold and 2.2‐fold retention of healthy myofibre area (P ≤ 0.0001). Additionally, raising dietary cholesterol and thus concentrations of plasma low‐density lipoprotein‐associated cholesterol (by 250%; P < 0.0001) reduced overall survivability (by 100%; P < 0.001) and worsened muscle damage in the LGMD2B triceps by 767% (P < 0.03). Micro‐pin‐induced mechanical injury in LGMD2B mice fed a high cholesterol diet exacerbated muscle damage by 425% (P < 0.03) and increased macrophage recruitment (by 98%; P = 0.03) compared with those injured on a chow diet. Parallel RNA‐seq analyses revealed that injury in cholesterol‐fed mice also modulated the expression of 3671 transcripts (1953 up‐regulated), with fibrogenic, inflammatory and programmed cell death‐associated pathways among the most enriched. Mice lacking dysferlin also displayed heightened muscle necrosis (by 123%; P < 0.0001) following a direct intramuscular injection of cholesterol compared with control mice. Conclusions Cholesterol exacerbates rodent MD. Specific inhibition of cholesterol absorption with ezetimibe may safely attenuate human MD severity and delay death.https://doi.org/10.1002/jcsm.12879DysferlinDystrophinCholesterolTriglyceridesPlasma lipids |
| spellingShingle | Zoe White Marine Theret Nadia Milad Lin Wei Tung William Wei‐Han Chen Martin G. Sirois Fabio Rossi Pascal Bernatchez Cholesterol absorption blocker ezetimibe prevents muscle wasting in severe dysferlin‐deficient and mdx mice Journal of Cachexia, Sarcopenia and Muscle Dysferlin Dystrophin Cholesterol Triglycerides Plasma lipids |
| title | Cholesterol absorption blocker ezetimibe prevents muscle wasting in severe dysferlin‐deficient and mdx mice |
| title_full | Cholesterol absorption blocker ezetimibe prevents muscle wasting in severe dysferlin‐deficient and mdx mice |
| title_fullStr | Cholesterol absorption blocker ezetimibe prevents muscle wasting in severe dysferlin‐deficient and mdx mice |
| title_full_unstemmed | Cholesterol absorption blocker ezetimibe prevents muscle wasting in severe dysferlin‐deficient and mdx mice |
| title_short | Cholesterol absorption blocker ezetimibe prevents muscle wasting in severe dysferlin‐deficient and mdx mice |
| title_sort | cholesterol absorption blocker ezetimibe prevents muscle wasting in severe dysferlin deficient and mdx mice |
| topic | Dysferlin Dystrophin Cholesterol Triglycerides Plasma lipids |
| url | https://doi.org/10.1002/jcsm.12879 |
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