Muscle regenerative capacity and aging
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1 Muscle regenerative capacity and aging International Conference on Frailty & Sarcopenia Research Charlotte A. Peterson Miami, Florida March 1-3, 2018
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3 Satellite cells: Resident muscle stem cells Relaix & Zammit, 2012
4 Plasmalemma Basal lamina Myofibril Mitochondria Satellite cell nucleus Myonucleus
5 Pax7 IHC to identify satellite cells Supplement Figure 2F: Dystrophin staining to demonstrate how CSA was determined
6 Myofiber repair and regeneration is mediated by satellite cells Adapted from Abmayr & Pavlath Development 2012
7 Normal compared to regenerated single fibers
8 Pax7-DTA mouse for satellite cell depletion Pax7-Cre ER V Rosa26- fl stop V fl -DTA Pax7-DTA Mauro, 1961 >4 months of age, ~ 25g Tamoxifen: 2mg/day for 5 days IP, 2 week washout Only mice with >90% satellite cell depletion are used
9 Tamoxifen Vehicle Satellite cell-depleted muscle does not regenerate PBS BaCl µm
10 Other roles for satellite cells? Pax7+ Satellite Cell Adapted from Brack & Rando, Cell Stem Cell, 2012
11 Dogma: The myonuclear domain remains constant during adult muscle growth via satellite cell fusion Resistance training Satellite cell proliferation Fusion to the myofiber Myofiber hypertrophy Satellite cell Myonucleus
12 muscle weight (mg/g) Model of muscle hypertrophy 1,8 1,4 0,9 0,5 0,0 CONT SA1 SA2 SA4 Synergist ablation (SA) Imposes functional overload of plantaris muscle Substitute for resistance exercise training
13 Early growth response following synergist ablation is unaffected by satellite cell depletion McCarthy et al., Development, 2011
14 Hypertrophy occurs in the absence of myonuclear accretion in satellite cell-depleted muscle nucleus inside the fiber (myonucleus) nucleus outside fiber
15 Muscle growth plateaus in the absence of satellite cell participation Fry et al., FASEB J, 2014
16 Vehicle Tamoxifen Vehicle Tamoxifen Sham SA-8 Collagen Glycoaminoglycans ECM accumulation is increased in satellite cell-depleted muscle following 8 weeks of overload WGA Area Fiber Vehicle Tamoxifen * * 0 Sham SA-8
17 In vitro analysis of the myogenic progenitor cell (MPC)-fibroblast interaction Well (Fibroblasts) Insert (MPCs) Well (Fibroblasts) Insert (Fibroblasts)
18 Relative mrna expression Activated satellite cell/mpc secretory products inhibit ECM gene expression in fibroblasts 3 2,5 + Fibroblasts + MPCs 2 1,5 * * * 1 0,5 0 Col1a2 Col3a1 Col6a2 Fbnct Fry et al., Cell Stem Cell 2017
19 Do satellite cells play a role in normal muscle maintenance? Pax7- DTA Pax7+ Satellite Cell Early Brack & Rando, Cell Stem Cell 2012
20 Dogma: Loss of satellite cell activity causes sarcopenia
21 Satellite cell depletion as a premature muscle aging model? Tamoxifen/Vehicle Pax7-DTA mouse Muscle collected from: 5-month olds - 1 month of satellite cell depletion (young) 24-month olds - 20 months of satellite cell depletion (old)
22 Mean Fiber CSA (µm 2 ) Pax7+ Cells / Fiber Tam Vehicle Long term satellite cell depletion does not exacerbate sarcopenia Young Old 0,12 0,08 0,04 0 * * * * * * Plantaris Gastroc TA/EDL Soleus Plantaris Gastroc TA/EDL Soleus * * 0 Plantaris Gastroc TA/EDL Soleus
23 Tamoxifen Vehicle Satellite cell-depleted aged muscle does not regenerate PBS BaCl 2 Fry et al., Nature Medicine, 2016
24 #Nuclei/100um Fiber Length Myonuclear number is not altered in satellite celldepleted muscle Vehicle Tamoxifen Young Old
25 % WGA Increased fibrosis in lifelong satellite cell-depleted muscle Vehicle Tamoxifen # Vehicle Tamoxifen
26 Growth in response to overload is impaired in aged mice regardless of satellite cell content Lee et al., J. Gerontol., 2016
27 Myonuclear accretion does not drive myofiber growth in aged muscle
28 Conclusions Muscle regeneration, maintenance and hypertrophy have different satellite cell requirements. Satellite cell-depleted muscle becomes fibrotic over time which may limit long term growth. Satellite cells repress ECM production by fibroblasts which may facilitate muscle remodeling. Depletion of satellite cells in adult mice impairs muscle regenerative capacity without affecting sarcopenia. Satellite cell loss with age may contribute to muscle fibrosis, limiting muscle adaptability. Satellite cell fusion and myonuclear accretion are insufficient to drive myofiber growth in aged muscle.
29 Take Home Messages Compensatory mechanisms exist in the adult mouse that enable muscle maintenance and adaptation independent of satellite cells. In addition to therapeutic potential of satellite cells in treating degenerative muscle diseases by promoting regeneration, satellite cells may be useful in reducing fibrosis associated with aging, independent of their ability to contribute nuclei to myofibers.
30 Acknowledgements Christopher Fry Undergrads: Tyler Kirby Angel Ho Alex Bugg Jonah Lee Honglu Liu Abigail Troske Jyothi Mula Amreen Farooqui Sam Crow Janna Jackson Drake Hyman Bailee Taylor Sarah White Tyler Kmiec Kaitlyn Frick Jake Beggs Ashley Bayes Grace Walton Kelcye Garrison Kate Kosmac Clyde Dean Kevin Murach Zakkary Hardyniec Cory Dungan Rod Erfani Ivan Vechetti Vandre Casagrande-Figueiredo Bailey Peck Davis Englund Yuan Wen John McCarthy Esther Dupont-Versteegden
Myogenic Progenitor Cells Control Extracellular Matrix Production by Fibroblasts during Skeletal Muscle Hypertrophy
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Supplementary Figure 1. Satellite cell contribution to myofibers in whole. gastrocnemius/plantaris/soleus, diaphragm, and EOM of 12 or 20 month
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