Treatment With Methotrexate Associated With Lipid Core Nanoparticles Prevents Aortic Dilation in a Murine Model of Marfan Syndrome

Maria Carolina Guido; Natalia de Menezes Lopes; Camila Inagaki Albuquerque; Elaine Rufo Tavares; Leonardo Jensen; Priscila de Oliveira Carvalho; Thauany Martins Tavoni; Ricardo Ribeiro Dias; Lygia da Veiga Pereira; Francisco Rafael Martins Laurindo; Raul Cavalcante Maranhão

doi:10.3389/fcvm.2022.893774

Treatment With Methotrexate Associated With Lipid Core Nanoparticles Prevents Aortic Dilation in a Murine Model of Marfan Syndrome

Front Cardiovasc Med. 2022 Jun 10:9:893774. doi: 10.3389/fcvm.2022.893774. eCollection 2022.

Affiliations

¹ Laboratory of Metabolism and Lipids, Heart Institute (InCor) of the Medical School Hospital, University of São Paulo, São Paulo, Brazil.
² Laboratory of Hypertension, Heart Institute (InCor) of the Medical School Hospital, University of São Paulo, São Paulo, Brazil.
³ Department of Cardiovascular Surgery, Heart Institute (InCor), Medical School Hospital, University of São Paulo, São Paulo, Brazil.
⁴ Department of Genetics and Evolutionary Biology, Institute of Biosciences, University of São Paulo, São Paulo, Brazil.
⁵ Laboratory of Vascular Biology, Heart Institute (InCor), Medical School Hospital, University of São Paulo, São Paulo, Brazil.

Abstract

In Marfan syndrome (MFS), dilation, dissection, and rupture of the aorta occur. Inflammation can be involved in the pathogenicity of aortic defects and can thus be a therapeutic target for MFS. Previously, we showed that the formulation of methotrexate (MTX) associated with lipid nanoparticles (LDE) has potent anti-inflammatory effects without toxicity. To investigate whether LDEMTX treatment can prevent the development of aortic lesions in the MFS murine model. MgΔloxPneo MFS (n = 40) and wild-type (WT, n = 60) mice were allocated to 6 groups weekly injected with IP solutions of: (1) only LDE; (2) commercial MTX; (3) LDEMTX (dose = 1mg/kg) between 3rd and 6th months of life. After 12 weeks of treatments, animals were examined by echocardiography and euthanatized for morphometric and molecular studies. MFS mice treated with LDEMTX showed narrower lumens in the aortic arch, as well as in the ascending and descending aorta. LDEMTX reduced fibrosis and the number of dissections in MFS but not the number of elastic fiber disruptions. In MFS mice, LDEMTX treatment lowered protein expression of pro-inflammatory factors macrophages (CD68), T-lymphocytes (CD3), tumor necrosis factor-α (TNF-α), apoptotic factor cleaved-caspase 3, and type 1 collagen and lowered the protein expression of the transforming growth factor-β (TGF-β), extracellular signal-regulated kinases ½ (ERK1/2), and SMAD3. Protein expression of CD68 and CD3 had a positive correlation with an area of aortic lumen (r ² = 0.36; p < 0.001), suggesting the importance of inflammation in the causative mechanisms of aortic dilation. Enhanced adenosine availability by LDEMTX was suggested by higher aortic expression of an anti-adenosine A2a receptor (A2a) and lower adenosine deaminase expression. Commercial MTX had negligible effects. LDEMTX prevented the development of MFS-associated aortic defects and can thus be a candidate for testing in clinical studies.

Keywords: Marfan syndrome; adenosine; aortic dilation; inflammation; lipid nanoparticle.