Liver-Specific siRNA-Mediated Stat3 or C3 Knockdown Improves the Outcome of Experimental Autoimmune Myocarditis

Lidia Avalle; Francesca Marino; Annalisa Camporeale; Chiara Guglielmi; Daniele Viavattene; Silvio Bandini; Laura Conti; James Cimino; Marco Forni; Cristina Zanini; Alessandra Ghigo; Roman L Bogorad; Federica Cavallo; Paolo Provero; Victor Koteliansky; Valeria Poli

doi:10.1016/j.omtm.2020.05.023

Liver-Specific siRNA-Mediated Stat3 or C3 Knockdown Improves the Outcome of Experimental Autoimmune Myocarditis

Mol Ther Methods Clin Dev. 2020 May 22:18:62-72. doi: 10.1016/j.omtm.2020.05.023. eCollection 2020 Sep 11.

Authors

Lidia Avalle¹, Francesca Marino¹, Annalisa Camporeale¹, Chiara Guglielmi¹, Daniele Viavattene¹, Silvio Bandini¹, Laura Conti¹, James Cimino¹, Marco Forni², Cristina Zanini², Alessandra Ghigo¹, Roman L Bogorad³, Federica Cavallo¹, Paolo Provero^{1

4}, Victor Koteliansky^{5

6}, Valeria Poli¹

Affiliations

¹ Department of Molecular Biotechnology and Health Science, University of Torino, Via Nizza 52, Torino 10126, Italy.
² EuroClone S.p.A Research Laboratory, Molecular Biotechnology Center, University of Turin, Torino 10126, Italy.
³ David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02142, USA.
⁴ Center for Translational Genomics and Bioinformatics, IRCCS San Raffaele Scientific Institute, 20132 Milan, Italy.
⁵ Skolkovo Institute of Science and Technology, Skolkovo, Moscow 121205, Russia.
⁶ Department of Chemistry, MV Lomonosov Moscow State University, Moscow 119991, Russia.

Abstract

Myocarditis can lead to autoimmune disease, dilated cardiomyopathy, and heart failure, which is modeled in the mouse by cardiac myosin immunization (experimental autoimmune myocarditis [EAM]). Signal transducer and activator of transcription 3 (STAT3) systemic inhibition exerts both preventive and therapeutic effects in EAM, and STAT3 constitutive activation elicits immune-mediated myocarditis dependent on complement C3 and correlating with activation of the STAT3-interleukin 6 (IL-6) axis in the liver. Thus, liver-specific STAT3 inhibition may represent a therapeutic option, allowing to bypass the heart toxicity, predicted by systemic STAT3 inhibition. We therefore decided to explore the effectiveness of silencing liver Stat3 and C3 in preventing EAM onset and/or the recovery of cardiac functions. We first show that complement C3 and C5 genetic depletion significantly prevents the onset of spontaneous myocarditis, supporting the complement cascade as a viable target. In order to interfere with complement production and STAT3 activity specifically in the liver, we took advantage of liver-specific Stat3 or C3 small interfering (si)RNA nanoparticles, demonstrating that both siRNAs can significantly prevent myocarditis onset and improve the recovery of heart functions in EAM. Our data demonstrate that liver-specific Stat3/C3 siRNAs may represent a therapeutic option for autoimmune myocarditis and suggest that complement levels and activation might be predictive of progression to dilated cardiomyopathy.

Keywords: C3/C5 null mice; STAT3; acute-phase response (APR); complement; constitutively active STAT3 mice; experimental autoimmune myocarditis (EAM); hepatotropic siRNA therapy.