Expression of Inflammatory Genes in Murine Lungs in a Model of Experimental Pulmonary Hypertension: Effects of an Antibody-Based Targeted Delivery of Interleukin-9

Highlights What are the main findings? F8IL9 therapy leads to a downregulation of certain inflammatory genes in PH-induced mice and displays beneficial effects on disease severity. What is the implication of the main finding? Targeted delivery of IL9 via F8 constitutes a novel treatment strategy in PH which likely has anti-inflammatory and therefore beneficial effects in PH. The results of this study offer inspiration to further investigate inflammation and remodeling in PH as a target for future specific therapies.Highlights What are the main findings? F8IL9 therapy leads to a downregulation of certain inflammatory genes in PH-induced mice and displays beneficial effects on disease severity. What is the implication of the main finding? Targeted delivery of IL9 via F8 constitutes a novel treatment strategy in PH which likely has anti-inflammatory and therefore beneficial effects in PH. The results of this study offer inspiration to further investigate inflammation and remodeling in PH as a target for future specific therapies.Abstract Background: Pathogenesis of pulmonary hypertension (PH) is a multifactorial process driven by inflammation and pulmonary vascular remodeling. To target these two aspects of PH, we recently tested a novel treatment: Interleukin-9 (IL9) fused to F8, an antibody that binds to the extra-domain A of fibronectin (EDA+ Fn). As EDA+ Fn is not found in healthy adult tissue but is expressed during PH, IL9 is delivered specifically to the tissue affected by PH. We found that F8IL9 reduced pulmonary vascular remodeling and attenuated PH compared with sham-treated mice. Purpose: To evaluate possible F8IL9 effects on PH-associated inflammatory processes, we analysed the expression of genes involved in pulmonary immune responses. Methods: We applied the monocrotaline (MCT) model of PH in mice (n = 44). Animals were divided into five experimental groups: sham-induced animals without PH (control, n = 4), MCT-induced PH without treatment (PH, n = 8), dual endothelin receptor antagonist treatment (dual ERA, n = 8), F8IL9 treatment (n = 12, 2 formats with n = 6 each), or with KSFIL9 treatment (KSFIL9, n = 12, 2 formats with n = 6 each, KSF: control antibody with irrelevant antigen specificity). After 28 days, a RT-PCR gene expression analysis of inflammatory response (84 genes) was performed in the lung. Results: Compared with the controls, 19 genes exhibited relevant (+2.5-fold) upregulation in the PH group without treatment. Gene expression levels in F8IL9-treated lung tissue were reduced compared to the PH group without treatment. This was the case especially for CCL20, CXCL5, C-reactive protein, pentraxin related (CRPPR), and Kininogen-1 (KNG1). Conclusion: In accordance with the hypothesis stated above, F8IL9 treatment diminished the upregulation of some genes associated with inflammation in a PH animal model. Therefore, we hypothesize that IL9-based immunocytokine treatment will likely modulate various inflammatory pathways.