Jeffrey E Cohen1, Brendan P Purcell1, John W MacArthur1, Anbin Mu1, Yasuhiro Shudo1, Jay B Patel1, Christopher M Brusalis1, Alen Trubelja1, Alexander S Fairman1, Bryan B Edwards1, Mollie S Davis1, George Hung1, William Hiesinger1, Pavan Atluri1, Kenneth B Margulies1, Jason A Burdick1, Y Joseph Woo2. 1. From the Department of Cardiothoracic Surgery, Stanford University, CA (J.E.C., J.W.M., Y.S., J.B.P., B.B.E., Y.J.W.); and Departments of Surgery, Division of Cardiovascular Surgery (J.E.C., J.W.M., C.M.B., A.T., A.S.F., G.H., W.H., P.A.), Bioengineering (B.P.P., M.S.D., J.A.B.), and Cardiology (A.M., K.B.M.), University of Pennsylvania, Philadelphia. 2. From the Department of Cardiothoracic Surgery, Stanford University, CA (J.E.C., J.W.M., Y.S., J.B.P., B.B.E., Y.J.W.); and Departments of Surgery, Division of Cardiovascular Surgery (J.E.C., J.W.M., C.M.B., A.T., A.S.F., G.H., W.H., P.A.), Bioengineering (B.P.P., M.S.D., J.A.B.), and Cardiology (A.M., K.B.M.), University of Pennsylvania, Philadelphia. joswoo@stanford.edu.
Abstract
BACKGROUND: Neuregulin-1β (NRG) is a member of the epidermal growth factor family possessing a critical role in cardiomyocyte development and proliferation. Systemic administration of NRG demonstrated efficacy in cardiomyopathy animal models, leading to clinical trials using daily NRG infusions. This approach is hindered by requiring daily infusions and off-target exposure. Therefore, this study aimed to encapsulate NRG in a hydrogel to be directly delivered to the myocardium, accomplishing sustained localized NRG delivery. METHODS AND RESULTS: NRG was encapsulated in hydrogel, and release over 14 days was confirmed by ELISA in vitro. Sprague-Dawley rats were used for cardiomyocyte isolation. Cells were stimulated by PBS, NRG, hydrogel, or NRG-hydrogel (NRG-HG) and evaluated for proliferation. Cardiomyocytes demonstrated EdU (5-ethynyl-2'-deoxyuridine) and phosphorylated histone H3 positivity in the NRG-HG group only. For in vivo studies, 2-month-old mice (n=60) underwent left anterior descending coronary artery ligation and were randomized to the 4 treatment groups mentioned. Only NRG-HG-treated mice demonstrated phosphorylated histone H3 and Ki67 positivity along with decreased caspase-3 activity compared with all controls. NRG was detected in myocardium 6 days after injection without evidence of off-target exposure in NRG-HG animals. At 2 weeks, the NRG-HG group exhibited enhanced left ventricular ejection fraction, decreased left ventricular area, and augmented borderzone thickness. CONCLUSIONS: Targeted and sustained delivery of NRG directly to the myocardial borderzone augments cardiomyocyte mitotic activity, decreases apoptosis, and greatly enhances left ventricular function in a model of ischemic cardiomyopathy. This novel approach to NRG administration avoids off-target exposure and represents a clinically translatable strategy in myocardial regenerative therapeutics.
BACKGROUND:Neuregulin-1β (NRG) is a member of the epidermal growth factor family possessing a critical role in cardiomyocyte development and proliferation. Systemic administration of NRG demonstrated efficacy in cardiomyopathy animal models, leading to clinical trials using daily NRG infusions. This approach is hindered by requiring daily infusions and off-target exposure. Therefore, this study aimed to encapsulate NRG in a hydrogel to be directly delivered to the myocardium, accomplishing sustained localized NRG delivery. METHODS AND RESULTS: NRG was encapsulated in hydrogel, and release over 14 days was confirmed by ELISA in vitro. Sprague-Dawley rats were used for cardiomyocyte isolation. Cells were stimulated by PBS, NRG, hydrogel, or NRG-hydrogel (NRG-HG) and evaluated for proliferation. Cardiomyocytes demonstrated EdU (5-ethynyl-2'-deoxyuridine) and phosphorylated histone H3 positivity in the NRG-HG group only. For in vivo studies, 2-month-old mice (n=60) underwent left anterior descending coronary artery ligation and were randomized to the 4 treatment groups mentioned. Only NRG-HG-treated mice demonstrated phosphorylated histone H3 and Ki67 positivity along with decreased caspase-3 activity compared with all controls. NRG was detected in myocardium 6 days after injection without evidence of off-target exposure in NRG-HG animals. At 2 weeks, the NRG-HG group exhibited enhanced left ventricular ejection fraction, decreased left ventricular area, and augmented borderzone thickness. CONCLUSIONS: Targeted and sustained delivery of NRG directly to the myocardial borderzone augments cardiomyocyte mitotic activity, decreases apoptosis, and greatly enhances left ventricular function in a model of ischemic cardiomyopathy. This novel approach to NRG administration avoids off-target exposure and represents a clinically translatable strategy in myocardial regenerative therapeutics.
Authors: Juli R Bagó; Carolina Soler-Botija; Laura Casaní; Elisabeth Aguilar; Maria Alieva; Núria Rubio; Antoni Bayes-Genis; Jerónimo Blanco Journal: Int J Cardiol Date: 2013-10-06 Impact factor: 4.164
Authors: Katrien Lemmens; Paul Fransen; Stanislas U Sys; Dirk L Brutsaert; Gilles W De Keulenaer Journal: Circulation Date: 2004-01-19 Impact factor: 29.690
Authors: Katashi Okoshi; Masaharu Nakayama; Xinhua Yan; Marina P Okoshi; Adam J T Schuldt; Mark A Marchionni; Beverly H Lorell Journal: Circulation Date: 2004-08-02 Impact factor: 29.690
Authors: Elena Tous Kichula; Hua Wang; Shauna M Dorsey; Spencer E Szczesny; Dawn M Elliott; Jason A Burdick; Jonathan F Wenk Journal: Ann Biomed Eng Date: 2013-11-23 Impact factor: 3.934
Authors: Brian D Polizzotti; Balakrishnan Ganapathy; Stuart Walsh; Sangita Choudhury; Niyatie Ammanamanchi; David G Bennett; Cristobal G dos Remedios; Bernhard J Haubner; Josef M Penninger; Bernhard Kühn Journal: Sci Transl Med Date: 2015-04-01 Impact factor: 17.956
Authors: John W MacArthur; Amanda N Steele; Andrew B Goldstone; Jeffrey E Cohen; William Hiesinger; Y Joseph Woo Journal: Curr Treat Options Cardiovasc Med Date: 2017-04
Authors: Lei Cai; Ruby E Dewi; Andrew B Goldstone; Jeffrey E Cohen; Amanda N Steele; Y Joseph Woo; Sarah C Heilshorn Journal: Adv Healthc Mater Date: 2016-10-06 Impact factor: 9.933
Authors: Yasuhiro Shudo; Andrew B Goldstone; Jeffrey E Cohen; Jay B Patel; Michael S Hopkins; Amanda N Steele; Bryan B Edwards; Masashi Kawamura; Shigeru Miyagawa; Yoshiki Sawa; Y Joseph Woo Journal: J Thorac Cardiovasc Surg Date: 2017-05-24 Impact factor: 5.209
Authors: Yasuhiro Shudo; Jeffrey E Cohen; Andrew B Goldstone; John W MacArthur; Jay Patel; Bryan B Edwards; Michael S Hopkins; Amanda N Steele; Lydia-Marie Joubert; Shigeru Miyagawa; Yoshiki Sawa; Y Joseph Woo Journal: Cytotherapy Date: 2016-04 Impact factor: 5.414