The DWORF micropeptide enhances contractility and prevents heart failure in a mouse model of dilated cardiomyopathy

Makarewich, Catherine A.; Munir, Amir Z.; Schiattarella, Gabriele Giacomo; Bezprozvannaya, Svetlana; Raguimova, Olga N.; Cho, Ellen E.; Vidal, Alexander; Robia, Seth L.; Bassel‐Duby, Rhonda; Olson, Eric N.

doi:10.7554/elife.38319

Cited by 96 publications

(126 citation statements)

References 50 publications

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“…2). This is consistent with previous in vivo observations (18) and fluorescence-based measurements showing that SERCA has a higher apparent affinity for DWORF as compared to PLB (21). DWORF is reported to interact with SERCA2a to form a heterodimer (22).…”

Section: Discussionsupporting

confidence: 92%

The transmembrane domain of DWORF activates SERCA directly; P15 and W22 residues are essential

Stroik

Yuen

et al. 2020

Preprint

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The Ca-ATPase isoform 2a (SERCA2a) re-sequesters cytosolic Ca2+ into the sarcoplasmic reticulum (SR) of cardiac myocytes, enabling muscle relaxation during diastole. A central factor in heart failure is abnormally high cytosolic [Ca2+], resulting in pathophysiology and decreased cardiac performance. Therefore, augmentation of the SERCA2a Ca2+ transport activity is a promising therapeutic approach. A novel transmembrane peptide, dwarf open reading frame (DWORF), is proposed to enhance SR Ca2+ uptake and myocyte contractility by displacing the protein phospholamban (PLB) from its inhibitory site on SERCA2a. In the present study, we have developed several cell-based FRET biosensor systems for time-resolved FRET (TR-FRET) measurements of the protein-protein interactions and structural changes in SERCA2a complexes with PLB and/or DWORF. To test the hypothesis that DWORF competes with PLB to occupy the putative SERCA2a binding site, we transiently transfected DWORF into a stable cell line expressing SERCA2a labeled with green fluorescent protein (GFP, the FRET donor) and PLB labeled with red fluorescent protein (RFP, the FRET acceptor). We observed a significant decrease in FRET efficiency, consistent with a decrease in the fraction of SERCA2a bound to PLB. Functional analysis demonstrates that DWORF activates SERCA in both the presence and absence of PLB. Furthermore, using site-directed mutagenesis, we generated DWORF variants that do not activate SERCA, thus identifying residues that are necessary for functional SERCA2a-DWORF interactions. This work advances our mechanistic understanding of the regulation of SERCA2a by small transmembrane proteins and sets the stage for future therapeutic development in heart failure research.

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Section: Discussionsupporting

confidence: 92%

The transmembrane domain of DWORF activates SERCA directly; P15 and W22 residues are essential

Stroik

Yuen

et al. 2020

Preprint

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“…Overexpression of PLN and increases in the ratio of PLN relative to SERCA has been implicated in dilated cardiomyopathy, heart failure, and skeletal muscle myopathies. 11,[47][48][49] Thus, finding ways to reduce the amount of PLN relative to SERCA is an active field of research. 50,51 Long considered to be the functional homolog of PLN, sarcolipin (SLN) is a small 31 amino acid protein that binds to and regulates the SERCA pump.…”

Section: Serca Uncoupling: a Role For Nnat?mentioning

confidence: 99%

Neuronatin regulates whole‐body metabolism: is thermogenesis involved?

Braun¹,

Geromella²,

Hamstra³

et al. 2020

FASEB BioAdvances

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The neuronatin (NNAT, 3.9 kb) gene is paternally inherited with the maternal allele silenced via DNA methylation. 1,2 Genomic imprinting such as this results in monoallelic expression-a characteristic of several other genes critical for growth, development, and metabolism. 3 Through alternative splicing, NNAT mRNA produces two isoforms, NNATα (81 amino acids) and NNATβ (54 amino acids), 4,5 though the extent with which these isoforms are functionally redundant or divergent remain to be uncovered. NNAT/Nnat mRNA is enriched in the developing brain, and while downregulated

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“…Thus, changes in PLN expression and phosphorylation may disrupt Ca 2+ homeostasis and cause the dilatation and weakening of the left ventricle. Eventually, the reduced rate of muscle relaxation and poor cardiac output lead to heart failure in DCM patients and, as such, SERCA2a and PLN have emerged as important therapeutic targets (MacLennan & Kranias, 2003; Makarewich et al., 2018).…”

Section: Introductionmentioning

confidence: 99%

“…Furthermore, recent evidence suggests that the inhibitory effects of PLN can be alleviated by displacing it from SERCA via a novel micropeptide, DWORF (dwarf open reading frame). When DWORF is overexpressed in cardiac muscle, SERCA function improves and DCM pathology is mitigated in mice (Makarewich et al., 2018). Collectively, these studies demonstrate the efficacy of targeting SERCA2a and/or PLN for DCM.…”

Section: Introductionmentioning

confidence: 99%

Low‐dose lithium feeding increases the SERCA2a‐to‐phospholamban ratio, improving SERCA function in murine left ventricles

Hamstra

Kurgan

Baranowski

et al. 2020

Experimental Physiology

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The sarco(endo)plasmic reticulum Ca 2+ -ATPase (SERCA) pump is responsible for regulating calcium (Ca 2+ ) within myocytes, with SERCA2a being the dominant isoform in cardiomyocytes.Its inhibitor, phospholamban (PLN), acts by decreasing the affinity of SERCA for Ca 2+ . Changes in the SERCA2a:PLN ratio can cause Ca 2+ dysregulation often seen in patients with dilated cardiomyopathy and heart failure. The enzyme glycogen synthase kinase-3 (GSK3) is known to downregulate SERCA function by decreasing the SERCA2a:PLN ratio. In this study, we sought to determine whether feeding mice low-dose lithium, a natural GSK3 inhibitor, would improve left ventricular SERCA function by altering the SERCA2a:PLN ratio. To this end, male wildtype C57BL/6J mice were fed low-dose lithium via drinking water (10 mg kg −1 day −1 LiCl for 6 weeks) and left ventricles were harvested. GSK3 activity was significantly reduced in LiClfed versus control-fed mice. The apparent affinity of SERCA for Ca 2+ was also increased (pCa 50 ; control, 6.09 ± 0.03 versus LiCl, 6.26 ± 0.04, P < 0.0001) along with a 2.0-fold increase in SERCA2a:PLN ratio in LiCl-fed versus control-fed mice. These findings suggest that low-dose lithium supplementation can improve SERCA function by increasing the SERCA2a:PLN ratio.Future studies in murine preclinical models will determine whether GSK3 inhibition via low-dose lithium could be a potential therapeutic strategy for dilated cardiomyopathy and heart failure. K E Y W O R D Scalcium, cardiac muscle, GSK3, lithium supplementation, phospholamban, SERCA 1 wileyonlinelibrary.com/journal/eph Experimental Physiology. 2020;105:666-675.

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The DWORF micropeptide enhances contractility and prevents heart failure in a mouse model of dilated cardiomyopathy

Cited by 96 publications

References 50 publications

The transmembrane domain of DWORF activates SERCA directly; P15 and W22 residues are essential

The transmembrane domain of DWORF activates SERCA directly; P15 and W22 residues are essential

Neuronatin regulates whole‐body metabolism: is thermogenesis involved?

Low‐dose lithium feeding increases the SERCA2a‐to‐phospholamban ratio, improving SERCA function in murine left ventricles

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