NR1D1 controls skeletal muscle calcium homeostasis through myoregulin repression
Alexis Boulinguiez, Christian Duhem, Alicia Mayeuf-Louchart, Benoit Pourcet, Yasmine Sebti, Kateryna Kondratska, Valérie Montel, Stéphane Delhaye, Quentin Thorel, Justine Beauchamp, Aurore Hebras, Marion Gimenez, Marie Couvelaere, Mathilde Zecchin, Lise Ferri, Natalia Prevarskaya, Anne Forand, Christel Gentil, Jessica Ohana, France Piétri-Rouxel, Bruno Bastide, Bart Staels, Helene Duez, Steve Lancel
Alexis Boulinguiez, Christian Duhem, Alicia Mayeuf-Louchart, Benoit Pourcet, Yasmine Sebti, Kateryna Kondratska, Valérie Montel, Stéphane Delhaye, Quentin Thorel, Justine Beauchamp, Aurore Hebras, Marion Gimenez, Marie Couvelaere, Mathilde Zecchin, Lise Ferri, Natalia Prevarskaya, Anne Forand, Christel Gentil, Jessica Ohana, France Piétri-Rouxel, Bruno Bastide, Bart Staels, Helene Duez, Steve Lancel
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Research Article Cell biology Muscle biology

NR1D1 controls skeletal muscle calcium homeostasis through myoregulin repression

Abstract

The sarcoplasmic reticulum (SR) plays an important role in calcium homeostasis. SR calcium mishandling is described in pathological conditions, such as myopathies. Here, we investigated whether the nuclear receptor subfamily 1 group D member (NR1D1, also called REV-ERBα) regulates skeletal muscle SR calcium homeostasis. Our data demonstrate that NR1D1 deficiency in mice impaired sarco/endoplasmic reticulum calcium ATPase–dependent (SERCA-dependent) SR calcium uptake. NR1D1 acts on calcium homeostasis by repressing the SERCA inhibitor myoregulin through direct binding to its promoter. Restoration of myoregulin counteracted the effects of NR1D1 overexpression on SR calcium content. Interestingly, myoblasts from patients with Duchenne muscular dystrophy displayed lower NR1D1 expression, whereas pharmacological NR1D1 activation ameliorated SR calcium homeostasis and improved muscle structure and function in dystrophic mdx/Utr+/– mice. Our findings demonstrate that NR1D1 regulates muscle SR calcium homeostasis, pointing to its therapeutic potential for mitigating myopathy.

Authors

Alexis Boulinguiez, Christian Duhem, Alicia Mayeuf-Louchart, Benoit Pourcet, Yasmine Sebti, Kateryna Kondratska, Valérie Montel, Stéphane Delhaye, Quentin Thorel, Justine Beauchamp, Aurore Hebras, Marion Gimenez, Marie Couvelaere, Mathilde Zecchin, Lise Ferri, Natalia Prevarskaya, Anne Forand, Christel Gentil, Jessica Ohana, France Piétri-Rouxel, Bruno Bastide, Bart Staels, Helene Duez, Steve Lancel

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Figure 3

NR1D1 activation alleviates Duchenne muscular dystrophy features both in mice and human myoblasts.

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NR1D1 activation alleviates Duchenne muscular dystrophy features both in...
(A) NR1D1 expression in muscle biopsies from controls (n = 14) and patients with Duchenne muscular dystrophy (DMD, n = 23). Data are from GEO data set GSE6011. **P = 0.0092, unpaired t test. (B) NR1D1 expression in control or DMD primary myotubes. *P = 0.0404 vs. control cells, unpaired t test; n = 5–7. (C) Representative curves and (D) peak fluorescence intensity of thapsigargin-induced (TG-induced) sarcoplasmic reticulum (SR) Ca2+ release in myoblasts from controls or patients with DMD treated with SR9009 (10 μM) or vehicle. Cells were loaded with Fluo4-AM and SR Ca2+ release was induced by the addition of 1 μM TG. Results are expressed as (mean ± SEM) the Delta F/F0 ratio. **P = 0.0049 vs. control cells, unpaired t test; n = 3 controls, n = 7 in both DMD groups. (E) H&E and Sirius red staining of tibialis anterior muscles obtained from vehicle- and SR9009-injected mdx/Utr+/– mice. Scale bars: 100 μm. (F) Myofiber cross-sectional area distribution. *P < 0.05 vs. vehicle-treated mdx/Utr+/– mice by 2-way ANOVA; n = 7–9. (G) Circulating creatine phosphokinase (CPK) activity. *P = 0.0329 vs. vehicle-treated mdx/Utr+/– animals; n = 8–9. (H) Muscular hydroxyproline. *P = 0.0172; n = 8–10. (I) Col1a2 (*P = 0.0314), (J) Pdgfra (*P = 0.0164), and (K) Mln (*P = 0.0402) gene expression; n = 8–12. (L) SERCA activity (n = 8–10) in muscular microsomes from mdx/Utr+/– mice treated for 20 days with SR9009 (100 mg/kg) or vehicle. *P = 0.0266. (M) In situ measurement of gastrocnemius-developed force. *P = 0.0301; n = 4–9. (G–M) Unpaired t test.