Neuregulin-1 Induces Cardiac Hypertrophy and Impairs Cardiac Performance in Post–Myocardial Infarction Rats

HomeCirculationVol. 142, No. 13Neuregulin-1 Induces Cardiac Hypertrophy and Impairs Cardiac Performance in Post–Myocardial Infarction Rats Free AccessLetterPDF/EPUBAboutView PDFView EPUBSections ToolsAdd to favoritesDownload citationsTrack citationsPermissions ShareShare onFacebookTwitterLinked InMendeleyRedditDiggEmail Jump toFree AccessLetterPDF/EPUBNeuregulin-1 Induces Cardiac Hypertrophy and Impairs Cardiac Performance in Post–Myocardial Infarction Rats Magdalena Zurek, PhD Edvin Johansson, PhD Malin Palmer, MSc Tamsin Albery, MSc Karin Johansson, MSc Katarina Rydén-Markinhutha, MSc Qing-Dong WangMD, PhD Magdalena ZurekMagdalena Zurek Magdalena Zurek, PhD, Cardiovascular, Renal and Metabolism, BioPharmaceuticals R&D, AstraZeneca Gothenburg, Pepparedsleden 1, SE-431 83 Mölndal, Sweden. Email E-mail Address: [email protected] https://orcid.org/0000-0003-2471-9346 Imaging and AI, Clinical Pharmacology and Safety Sciences, BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden (M.Z., E.J.). *Drs Zurek and Wang contributed equally. Search for more papers by this author , Edvin JohanssonEdvin Johansson Imaging and AI, Clinical Pharmacology and Safety Sciences, BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden (M.Z., E.J.). Antaros Medical, Mölndal, Sweden (E.J.). Search for more papers by this author , Malin PalmerMalin Palmer Bioscience Cardiovascular, Research and Early Development, Cardiovascular, Renal and Metabolism (CVRM), BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden (M.P., T.A., K.J., K.R.-M., Q.-D.W.). Search for more papers by this author , Tamsin AlberyTamsin Albery Bioscience Cardiovascular, Research and Early Development, Cardiovascular, Renal and Metabolism (CVRM), BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden (M.P., T.A., K.J., K.R.-M., Q.-D.W.). Search for more papers by this author , Karin JohanssonKarin Johansson Bioscience Cardiovascular, Research and Early Development, Cardiovascular, Renal and Metabolism (CVRM), BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden (M.P., T.A., K.J., K.R.-M., Q.-D.W.). Search for more papers by this author , Katarina Rydén-MarkinhuthaKatarina Rydén-Markinhutha Bioscience Cardiovascular, Research and Early Development, Cardiovascular, Renal and Metabolism (CVRM), BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden (M.P., T.A., K.J., K.R.-M., Q.-D.W.). Search for more papers by this author , Qing-Dong WangQing-Dong Wang Qing-Dong Wang, MD, PhD, Cardiovascular, Renal and Metabolism, BioPharmaceuticals R&D, AstraZeneca Gothenburg, Pepparedsleden 1, SE-431 83 Mölndal, Sweden. Email E-mail Address: [email protected] Bioscience Cardiovascular, Research and Early Development, Cardiovascular, Renal and Metabolism (CVRM), BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden (M.P., T.A., K.J., K.R.-M., Q.-D.W.). *Drs Zurek and Wang contributed equally. Search for more papers by this author Originally published28 Sep 2020https://doi.org/10.1161/CIRCULATIONAHA.119.044313Circulation. 2020;142:1308–1311All experimental work was compliant with European Union Directive 2010/63/EU and approved by the local ethics committee.NRG-1 (neuregulin-1) has an indispensable role in cardiac development and may be important for maintaining adult cardiac function. In multiple animal studies, NRG-1, or its analogue GGF2 (human glial growth factor), has been shown to increase the left ventricular ejection fraction (LVEF), suggesting that NRG-1 could be a promising heart failure treatment. Recently, recombinant human NRG-1 and GGF2 proteins have been moved into clinical testing for chronic heart failure treatment. According to ClinicalTrials.gov, rhNRG-1 (recombinant human NRG-1; Neucardin) is currently being evaluated in a phase III trial and recombinant human GGF2 (cimaglermin alfa) has completed two phase I trials. The mechanisms underlying the effect of NRG-1 on LVEF are not fully understood, but improved cardiomyocyte structure, contractility, and proliferation are proposed explanations.1 It has also been suggested that NRG-1 induces cardiomyocyte hypertrophy1; however, the associated potentially detrimental impact on left ventricular (LV) function has been largely neglected.The enthusiasm for NRG-1 in heart failure is to a large extent based on consistently reported LVEF improvement and reductions of LV end-systolic volume and end-diastolic volume, but studies seldom report left ventricular mass (LVM), cardiac output or stroke volume, or other functional parameters such as strain. This makes it difficult to interpret the merit of the LVEF improvements, because the presence of hypertrophy in itself leads to LVEF increase.2 The 1 exception to report findings beyond LVEF, LV end-systolic volume, and LV end-diastolic volume was from a porcine post–myocardial infarction (MI) study,3 where interventricular septal wall thickness and heart weight were reported to increase after GGF2 treatment, but no discussion was held about the potential impact on LV dimensions.Here, we report a cardiac magnetic resonance study with multiple readouts in post-MI rats treated with rhNRG-1 and demonstrate that rhNRG-1 induces hypertrophy and worsens cardiac performance.To assess LV function and remodeling, we performed cardiac magnetic resonance before and after 7 days of rhNRG-1 treatment with multiple doses (0.6, 2.5, and 5 µg·kg–1·h–1) or albumin (vehicle) in rats post-MI, with the treatment starting 1 week after permanent coronary artery ligation. rhNRG-1 increased LVEF and reduced LV end-systolic volume and LV end-diastolic volume dose-dependently, consistent with earlier reports (Figure A through C).1 The LVEF improvement was accompanied by an LVM/body weight increase (Figure D). An increased LV wall thickness after rhNRG-1 treatment was also apparent (Figure E).Download figureDownload PowerPointFigure. The impact of neuregulin-1 on cardiac hypertrophy and performance demonstrated in 2 myocardial infarction (MI) rat models.In A through I, results obtained from a subacute post-MI model (n=12 vehicle group, n=7–9 treatment groups) with 1 week of vehicle or rhNRG-1 treatment (0.6, 2.5, and 5 µg·kg–1·h–1) initiated 7 days post-MI are shown. In J through M, data from a chronic ischemia/reperfusion (I/R) model (n=7 per group) with 1 week of vehicle or rhNRG-1 treatment (1 µg·kg–1·h–1), initiated 4 weeks after reperfusion, followed by 2 weeks of washout, are given together with data from sham-operated animals (n=3). The rhNRG-1 treatment was associated with increased left ventricular ejection fraction (LVEF; A and K) and reduced left ventricular end-systolic (LVESV) and end-diastolic volumes (LVEDV; B, C, and L) in both models at all investigated dose levels. Furthermore, hypertrophy as assessed via left ventricular mass-to-body weight ratio (LVM/BW) was detected in the chronic model (M) and for 1 dose level in the subacute model (D). After 2 weeks of washout, none of the effects on ventricular volumes or mass was sustained (K through M). Global longitudinal strain (GLS) as determined via feature-tracking was either unchanged or worsened across the dose groups (F). For the study population, a correlation between a worsening in GLS and a reduction in cardiac output was found (G). In E, short-axis and 4-chamber view images at end-diastole (ED) and end-systole (ES) are given, depicting induced hypertrophy and systolic LV obliteration after 1 week of treatment; Arrows point to the thickened LV wall. A dose-dependent increase in NT-proBNP concentration during treatment was present (H), and at the final day of treatment, the NT-proBNP concentration correlated negatively with cardiac output (I). No difference in the number of Ki67-positive cells, determined by immunohistochemical staining, in the peri-infarct area was found after 2 weeks of washout between the vehicle and rhNRG-1 (1.0 µg·kg–1·h–1) groups (J). Group means are given as mean±SEM and statistical tests are performed on change from baseline values (A through D, F, K through M). P values are calculated by 1-way ANOVA with the Dunnett multiple comparisons test (A through D, F), 2-way repeated-measures ANOVA (H), and unpaired 1-tailed t test (J through M). Pearson correlations coefficients were determined for G and I. Significance levels are given as ****P<0.0001, ***P<0.001, **P<0.01, *P<0.05 and nonsignificance (ns; P>0.05). NT-proBNP indicates N-terminal pro-B-type natriuretic peptide; and rhNRG-1, recombinant human neuregulin-1.The global longitudinal strain has emerged as a prognostic end point enabling detection of subclinical myocardial dysfunction, serving as a more sensitive marker of LV function than LVEF in patients with preserved and reduced ejection fraction.4 Our findings demonstrate that, despite large LVEF improvement, no improvement or even a worsening of myocardial global longitudinal strain is present (Figure F). The worsening correlated with a dose-dependent reduction in cardiac output (Figure G), in turn, a product of dose-dependent decreases in stroke volume and heart rate (data not shown). rhNRG-1 elevated plasma levels of N-terminal pro-B-type natriuretic peptide dose-dependently (Figure H), further supporting a prohypertrophic effect of NRG-1 or heart failure worsening. Increased N-terminal pro-B-type natriuretic peptide levels were reported earlier, for example, in the phase I study (Single Ascending Doses of GGF2 in Patients With Left Ventricular Dysfunction and Symptomatic Heart Failure; NCT01258387), a 2-fold increase in N-terminal pro-B-type natriuretic peptide was detected after 7 days of cimaglermin treatment, but the implications of these results were not discussed. In our study, N-terminal pro-B-type natriuretic peptide correlated inversely with cardiac output (Figure I), further undermining the proposed beneficial impact of rhNRG-1 on cardiac performance.In addition to the subacute post-MI heart failure model, we investigated rats subjected to 35 minutes of coronary artery ligation followed by reperfusion. rhNRG-1 treatment (1 µg·kg–1·h–1) was initiated 4 weeks after reperfusion, and continued for 1 week, followed by 2 weeks of washout to test if effects were sustained. Similar to our findings in the post-MI model, rhNRG-1 increased LVEF (Figure K), reduced LV end-systolic volume and LV end-diastolic volume (Figure L), increased LVM (Figure M), and reduced stroke volume and cardiac output (data not shown). However, the effects were not sustained after 2 weeks washout. No impact on global longitudinal strain was observed in the reperfusion model, consistent with results in the post-MI model at lower doses (data not shown).NRG-1 has been shown to induce cardiomyocyte proliferation, but effects were only evident in cardiomyocytes isolated from young individuals.5 Because the effect of NRG-1 was lost 14 days after treatment, the LVM increase is most likely attributable to cardiomyocyte hypertrophy rather than proliferation. Staining of the proliferation marker Ki67 revealed no difference in the number of Ki67-positive cells between the vehicle and rhNRG1 groups (Figure J).In summary, our study used a range of complementary end points to LV geometry to assess the effects of NRG-1, enabling us to unmask a negative impact on cardiac performance. Our study cautions the enthusiasm on the therapeutic value of NRG-1 in chronic heart failure. We recommend reporting LVM, stroke volume/cardiac output, and global longitudinal strain along with LVEF and LV volumes in future studies to enable more comprehensive discussions on the effects of NRG-1 on cardiac function.AcknowledgmentsThe authors thank Animal Sciences and Technologies, AstraZeneca, Gothenburg, Sweden, for providing excellent technical assistance.Sources of FundingThe study was supported by AstraZeneca.DisclosuresNone.Footnotes*Drs Zurek and Wang contributed equally.The data that support the findings of this study, and the experimental procedures and protocols, as well, are available from the corresponding author on reasonable request.https://www.ahajournals.org/journal/circQing-Dong Wang, MD, PhD, Cardiovascular, Renal and Metabolism, BioPharmaceuticals R&D, AstraZeneca Gothenburg, Pepparedsleden 1, SE-431 83 Mölndal, Sweden. Email qing-dong.[email protected]comMagdalena Zurek, PhD, Cardiovascular, Renal and Metabolism, BioPharmaceuticals R&D, AstraZeneca Gothenburg, Pepparedsleden 1, SE-431 83 Mölndal, Sweden. Email [email protected]comReferences1. De Keulenaer GW, Feyen E, Dugaucquier L, Shakeri H, Shchendrygina A, Belenkov YN, Brink M, Vermeulen Z, Segers VFM. Mechanisms of the multitasking endothelial protein NRG-1 as a compensatory factor during chronic heart failure.Circ Heart Fail. 2019; 12:e006288. doi: 10.1161/CIRCHEARTFAILURE.119.006288LinkGoogle Scholar2. MacIver DH. A new method for quantification of left ventricular systolic function using a corrected ejection fraction.Eur J Echocardiogr. 2011; 12:228–234. doi: 10.1093/ejechocard/jeq185CrossrefMedlineGoogle Scholar3. Galindo CL, Kasasbeh E, Murphy A, Ryzhov S, Lenihan S, Ahmad FA, Williams P, Nunnally A, Adcock J, Song Y, et al.. Anti-remodeling and anti-fibrotic effects of the neuregulin-1β glial growth factor 2 in a large animal model of heart failure.J Am Heart Assoc. 2014; 3:e000773. doi: 10.1161/JAHA.113.000773LinkGoogle Scholar4. Park JJ, Park JB, Park JH, Cho GY. Global longitudinal strain to predict mortality in patients with acute heart failure.J Am Coll Cardiol. 2018; 71:1947–1957. doi: 10.1016/j.jacc.2018.02.064CrossrefMedlineGoogle Scholar5. Polizzotti BD, Ganapathy B, Walsh S, Choudhury S, Ammanamanchi N, Bennett DG, dos Remedios CG, Haubner BJ, Penninger JM, Kühn B. Neuregulin stimulation of cardiomyocyte regeneration in mice and human myocardium reveals a therapeutic window.Sci Transl Med. 2015; 7:281ra45. doi: 10.1126/scitranslmed.aaa5171CrossrefMedlineGoogle Scholar Previous Back to top Next FiguresReferencesRelatedDetails September 29, 2020Vol 142, Issue 13Article InformationMetrics Download: 1,274 © 2020 American Heart Association, Inc.https://doi.org/10.1161/CIRCULATIONAHA.119.044313PMID: 32986480 Originally publishedSeptember 28, 2020 Keywordsneuregulinsventricular dysfunction, lefthypertrophyPDF download SubjectsTranslational StudiesMyocardial RegenerationHeart FailureHypertrophy