Transcriptomic Analysis Reveals a Link Between Hippo Signaling Pathway and Macrophages in Lungs of Mice with OVA-Induced Allergic Asthma

Abstract

Purpose The Hippo signaling pathway participates in the restriction of cell proliferation and organ growth. Activated macrophages have been implicated in the pathogenesis of allergic asthma. Recent studies have shown that Hippo signaling pathway may also be involved in the regulation of asthma. However, the link between Hippo signaling pathway and macrophages in the context of allergic asthma has not been investigated. The purpose of this study was to explore the link between Hippo signaling pathway and macrophages using a mice model of OVA-induced allergic asthma. Methods Mice models of asthma were established. Lung tissues were collected from mice and pooled for mRNA sequencing and bioinformatics analysis. The relative mRNA expression of Hippo signalling pathway-related proteins Yap1, Lef1 and Ctgf was also measured. Double immunofluorescence staining was performed on lung tissues to evaluate macrophage marker F4/80 expression and Yap1/Lef1/Ctgf expression. Results Results of the RNA-Seq of lung tissues demonstrated that the Hippo signaling pathway was down-regulated in OVA-induced allergic asthma. Using the cytoHubba tool kits in Cytoscape, the following top 10 hub genes of Hippo signalling pathway were identified: Yap1, Lef1, Ctgf, Ccnd1, Axin2, Smad7, Wnt4, Wnt3a, Pard6b, and Wwc1 . Using the seq-ImmuCC ( http://218.4.234.74:3200/immune/ ), a negative correlation was found between macrophages and Hippo signaling pathway activity (R 2 = 0.93). The mRNA expression levels of pulmonary Yap1, Lef1, and Ctgf were down-regulated in the mice model of OVA-induced allergic asthma. Moreover, double-stained immunofluorescence for F4/80 and Yap1, Lef1, Ctgf in mouse lung sections respectively revealed that macrophage proliferation was correlated with downregulation of the Hippo signaling pathway in the mice model of OVA-induced allergic asthma. Conclusion These results demonstrated that the Hippo signaling pathway was down-regulated in asthma mice, and the proliferation of macrophages was associated with downregulation of the Hippo signaling pathway. These findings reveal novel insights into the pathogenesis and treatment of asthma.

Integrative bioinformatics analysis to explore a robust diagnostic signature and landscape of immune cell infiltration in sarcoidosis

“…This result was consistent with previous enrichment analyses, demonstrating that IFN-γ and Th17 cells had an essential effect on the development and progression of granulomatous tissues in sarcoidosis (30,31). Lymphoid enhancer-binding factor 1 (LEF1) is one of the Hippo signaling pathway hub genes, and it has been suggested that macrophage proliferation is related to the downregulation of the Hippo signaling pathway (32). The pathology of sarcoidosis is characterized by chronic granulomas with a core infiltration of macrophages and a peripheral infiltration of lymphocytes visually (33).…”

Mycobacterium vaccae alleviates allergic airway inflammation and airway hyper‐responsiveness in asthmatic mice by altering intestinal microbiota

“…The 16S rDNA sequencing and metabolite data analysed during this study are included in its supplementary information files. The transcriptome data used in this study were retrieved from our previous study [23]. To minimize the misuse of experimental animals, the same batch of animals was used in both studies, and all other experiments in the two studies were independent and repeatable.…”

“…After the final challenge, as described previously, specific airway resistance was measured to determine airway responsiveness [23]. The mice were placed in the testing chamber for an adaptation period of 5 min, then they were separately exposed to aerosols of phosphate buffer saline (PBS) (baseline) and increasing doses of methacholine (MCh) (6.25, 12.5, and 25 mg/mL) for 3 min.…”

“…Bronchoalveolar lavage fluid (BALF) was collected as previously described [23]. The trachea was cannulated, and BALF was collected by sequentially lavaging the lungs thrice with ice-cold PBS solution (0.5 mL).…”

FKN secreted by kidney epithelial cells regulates macrophage activation in lupus nephritis via the Hippo signaling pathway

“…52 CTGF is the key downstream target of YAP. 53 Continuous activation of YAP increases the expression of CTGF in renal tubular epithelial cells. 54 We found abnormal expression in kidney tissues of LN patients.…”