Influence of pole‐like kinase 1 on controlling contractility of the urinary tract and gene activated patterns of human bladder smooth muscle cells

Abstract

BackgroundPole like kinase (PLK)‐mediated promotion of proliferation and smooth muscle contraction in the lower urinary tract has been recently suggested and may offer putative targets and novel compounds for treatments of lower urinary tract symptoms. However, PLK function for contractility of smooth muscle and proliferation of bladder smooth muscle cells are unknown and the specificity of PLK inhibitors has been questioned.MethodsHere, human bladder smooth muscle cells (hBSMCs) inhibited with five PLK inhibitors (TAK960, MLN0905, Wortmannin, GW843682X, Centrinone‐B) and hBSMCs cultured in smooth muscle medium were compared. hBSMCs characteristics according to criteria set by the International Society of Cellular Therapy were confirmed by flow cytometry. Proliferation and viability were assessed by CCK‐8 assay. Contractions of human ureterovesical junction smooth muscle from donation after circulatory death renal transplantation were studied in an organ bath, where they were induced by the α1‐adrenoceptor agonists noradrenaline, phenylephrine, and electric field stimulation. Transcriptome sequencing of hBSMCs were obtained using NEBNext®UltraTM RNA Library Prep Kit for Illumina®. Gene set enrichment analysis was performed using the Kyoto Encyclopedia of Genes and Genomes and gene ontology terms.ResultsViability and proliferation of hBSMCs were reduced in a concentration‐ and time‐dependent manner. TAK960 inhibits adrenergic contractions of human ureterovesical junction smooth muscle, and proliferation of hBSMCs. No significant differences in other PLK inhibitors combined with control were found. Compared to hBSMCs in control medium, 323 upregulated genes and 160 downregulated genes were significantly altered by TAK960 24hours 500nM, respectively (FDR=0.05, FC=1.5). Genes most prominently and significantly upregulated by PLK inhibiting condition were IFI30, GBP4, VTN, SCG2, OAS2, RRAD, TNFSF10, KRT18, SFN, meanwhile, downregulated was CCL11 (log2FC ≥1.5 or ≤ ‐1.5). The most significantly relative signaling pathway included cytokine‐cytokine receptor interaction, systemic lupus erythematosus and transcriptional misregulation cancer (enrichment factor ≥0.05). Accession to cite for these sequences read archive data is PRJNA714246.ConclusionOur findings point to a certain role of PLK‐1 in of proliferation in hBSMCs and contractility in ureterovesical junction smooth muscle, which may be of relevance in lower urinary tract symptom. PLK‐1 inhibitors block these effects and show shared and divergent effects. PLK‐1 regulation of bladder smooth muscle growth and contraction may include genomic and nongenomic mechanisms.