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. 2022 Sep 5;14(17):7052-7064.
doi: 10.18632/aging.204271. Epub 2022 Sep 5.

Bruton tyrosine kinase (BTK) may be a potential therapeutic target for interstitial cystitis/bladder pain syndrome

Guang Wang  1 , Tong-Xin Yang  2 , Jiong-Ming Li  2 , Zi-Ye Huang  2 , Wen-Bo Yang  1 , Pei Li  2 , Da-Lin He  1
Affiliations

Bruton tyrosine kinase (BTK) may be a potential therapeutic target for interstitial cystitis/bladder pain syndrome

Guang Wang et al. Aging (Albany NY). .

Abstract

Aims: To determine the potential diagnostic and therapeutic targets of Interstitial Cystitis/Bladder Pain Syndrome (IC/BPS).

Methods: We selected the GSE11783, GSE57560 and GSE621 datasets from the GEO database and merged them. R software was used to screen differentially expressed genes (DEGs) between IC/BPS and normal bladder tissues. The "String" online tool is used to analyze DEGs interaction and functional protein enrichment. CIBERSORT online tool was used to analyze the infiltration of immune cells. In addition, we verified the function of BTK in IC/BPS at the clinical samples and cells level.

Results: Bioinformatics analysis revealed that 5 genes were significantly overexpressed in IC/BPS, and the protein-protein interaction diagram showed that BTK was a critical link between these five proteins. At the same time, functional enrichment showed that they were significantly related to innate immunity. Immunoinfiltration showed that mast cell resting in IC/BPS was significantly higher. IHC staining of clinical samples showed that the mast cell markers Tryptase and BTK were highly expressed in IC/BPS tissues. At the cell level, knockdown of BTK inhibited proliferation, migration, invasion, and degranulation of mast cells.

Conclusions: This study provides a new perspective for understanding the molecular mechanisms involved in IC/BPS and suggests that BTK may be a target for treating IC/BPS.

Keywords: BTK; bioinformatics; biomarker; interstitial cystitis; mast cells.

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Conflict of interest statement

CONFLICTS OF INTEREST: The authors declare that they have no conflicts of interest.

Figures

Figure 1
Figure 1
Identification of DEGs in GEO dataset of IC/BPS patients. (A, B) The ggplot2 package performed principal component analysis (PCA) on the (A) unnormalized and (B) normalized GSE11783, GSE57560 and GSE621 datasets. (C) Volcano maps of the merged datasets. (D) Heatmaps of cluster analysis by DEGs expression in GSE11783, GSE57560 and GSE621 datasets.
Figure 2
Figure 2
PPI and functional enrichment analysis.
Figure 3
Figure 3
Immune-infiltration analysis of GEO datasets of IC/BPS patients by the CIBERSORTx online tool. Violin Plot and were used to compare the immune cell score difference between IC/BPS and normal bladder tissues (Blue indicates normal bladder tissues; red indicates IC/BPS bladder tissues. Wilcoxon signed rank test was used to compare and calculate the statistical p-value).
Figure 4
Figure 4
Differential expression of BTK in IC/BPS clinical samples. (A) HE staining shows the pathological state of IC/BPS samples. Scale bar: 100 and 50 μm. (B) Masson staining is used to distinguish collagen fibers from muscle fibers. Collagen fibers are blue and smooth muscle fibers are red. The staining results show that compared with NC group, collagen fibers in IC/BPS group are significantly reduced. Scale bar: 100 and 50 μm. (C, D) IHC staining showed the difference of (C) Tryptase and (D) BTK expression in normal and IC/BPS tissues. Scale bar: 20 μm. (E) Western blotting exhibited the expression level of BTK in normal and IC/BPS tissues. In all cases, Values are mean ± SD (n=10 for each group; *P<0.05, **P<0.01).
Figure 5
Figure 5
In vitro verification of the effect of BTK on the proliferation and metastasis of HMC-1 cells. (A) Western blotting displayed the knockdown efficiency of sh-BTK. (B) HMC-1 cell proliferation was detected by CCK-8 kit. (C) Wound-Healing assay showed the migration ability of HMC-1 cells. Scale bar: 100 μm. (D) Transwell assay was used to detect HCC-1 cell invasion. Scale bar: 100 μm. (E–G) Quantifications of mast cell mediators, including (E) Tryptase, (F) β-hexosaminidase and (G) Histamine. In all cases, Values are mean ± SD (n=3 for each group; *P<0.05, **P<0.01).
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References

    1. Akiyama Y, Homma Y, Maeda D. Pathology and terminology of interstitial cystitis/bladder pain syndrome: A review. Histol Histopathol. 2019; 34:25–32. 10.14670/HH-18-028 - DOI - PubMed
    1. Hanno PM, Erickson D, Moldwin R, Faraday MM, and American Urological Association. Diagnosis and treatment of interstitial cystitis/bladder pain syndrome: AUA guideline amendment. J Urol. 2015; 193:1545–53. 10.1016/j.juro.2015.01.086 - DOI - PubMed
    1. Fall M, Nordling J, Cervigni M, Dinis Oliveira P, Fariello J, Hanno P, Kåbjörn-Gustafsson C, Logadottir Y, Meijlink J, Mishra N, Moldwin R, Nasta L, Quaghebeur J, et al.. Hunner lesion disease differs in diagnosis, treatment and outcome from bladder pain syndrome: an ESSIC working group report. Scand J Urol. 2020; 54:91–8. 10.1080/21681805.2020.1730948 - DOI - PubMed
    1. Chelimsky G, Heller E, Buffington CA, Rackley R, Zhang D, Chelimsky T. Co-morbidities of interstitial cystitis. Front Neurosci. 2012; 6:114. 10.3389/fnins.2012.00114 - DOI - PMC - PubMed
    1. van de Merwe JP, Nordling J, Bouchelouche P, Bouchelouche K, Cervigni M, Daha LK, Elneil S, Fall M, Hohlbrugger G, Irwin P, Mortensen S, van Ophoven A, Osborne JL, et al.. Diagnostic criteria, classification, and nomenclature for painful bladder syndrome/interstitial cystitis: an ESSIC proposal. Eur Urol. 2008; 53:60–7. 10.1016/j.eururo.2007.09.019 - DOI - PubMed

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