The Therapeutic Vaccine For Pulmonary Artery Hypertension Targeting IL-11RA

Objective: Pulmonary Artery Hypertension(PAH)is a cardiopulmonary disease characterized by pulmonary vascular remodeling.Although current treatment methods can effectively reduce pulmonary artery pressure,the mortality rate of PAH remains high.New therapeutic strategies of PAH should be established.This project intends to develop a therapeutic vaccine against IL-11 RA to improve pulmonary vascular fibrosis,delay or reverse pulmonary vascular remodeling,and provide a new method for the treatment of PAH.Method: Using weighted gene coexpression network analysis(WGCNA)method,the coexpression networks were constructed based on GSE117261,which contained transcriptome data from the PAH lung tissues.To find the gene module related to PAH,the functional enrichment analysis was performed by Metascape.The specific peptides to IL11 RA were screened out based on structure and functional sites of IL-11RA’s molecular,and the immunoreactivity,immunogenicity of peptides.The IL-11 RA vaccine was prepared by Sulfo-SMCC coupling peptides and Qβ virus-like particles(VLP).Immunized the mice by IL-11 RA vaccine and the antibody titers were measured by ELISA.The experimental PAH in mice induced by hypoxia exposure for four weeks combined with subcutaneous injection of Sugen5416.Right ventricular systolic pressure,the weight of right ventricle,septum,and left ventricle were measured.The expression of the gene IL-11,IL-11 RA,TGF-β,MMP1,TIMP1,COLⅠ,and COLⅢ was measured by RT-q PCR.Protein expression and phosphorylation were detected by Western Blots.The concentration of IL-11 in serum was estimated by ELISA.Hematoxylin-eosin(H&E)staining was used to evaluate the pulmonary artery remodeling and cardiomyocyte size,Masson staining for exhibiting fibrosis,and renal PAS staining was used to observe the structure of the kidney.The expression and localization of IL-11 RA were detected by immunofluorescence.Results: A total of 13 gene coexpression modules were identified through WGCNA,among which the pink module has the most significant correlation ship with PAH.Functional enrichment analysis showed that genes in the pink module mainly involved in the extracellular matrix and immune function.Three peptides from IL-11 RA were screened out and conjugated with Qβ VLP to prepare the vaccines,which induced high titer of antibodies in mice.In mice experimental pulmonary artery hypertension,the IL-11 RA vaccine reduced right ventricular systolic pressure,right ventricular remodeling,pulmonary artery media thickening,and cardiomyocyte hypertrophy.The IL-11 RA vaccine can alleviate the deposition of collagen around lung vessels and decrease the expression of gene TGF-β,COLⅠ,COLⅢ,MMP1,and TIMP1 at the transcription level.In lung tissue,IL-11 RA is mainly expressed in alveolar epithelial cells,bronchial epithelial cells,pulmonary vascular endothelial cells,and fibroblasts,which concluded immunofluorescence.The phosphorylation level of Erk could reduce by the IL-11 RA vaccine.The PAS staining of the kidneys showed that the IL-11 RA vaccine did not cause significant damage,which preliminarily proved the vaccine’s safety.Conclusion: Extracellular matrix remodeling played a pivotal role in the pathogenesis of PAH.IL-11 RA vaccine can inhibit TGF-β,reduce the expression of gene COLⅠ,COLⅢ,MMP,and TIMP,alleviate the collagen deposition around PAH lung vessels,and decrease the remodeling of pulmonary vessels.Our results exhibit a prospect for the treatment of PAH.