The Antimuscarinic Agent Tolterodine Regulates Bladder Extracellular Matrix in Partial Bladder Outlet Obstruction in Rats

Recently we demonstrated the utility of a nerve-sparing mid-urethra model of partial outlet obstruction (NeMO) that has high consistency and minimal mortalities, unlike the traditional model proximal to the bladder neck. Our goal was to uncover potential therapeutic targets by investigating the genome wide transcriptional changes and pathways altered in NeMO to compare with published human bladder obstruction data. We performed RNAseq and analysed the differentially upregulated and downregulated genes for associated pathways, transcription factor binding site analysis (TFBS), upstream regulators and Gene Set Enrichment Analysis (GSEA). NeMO increased bladder mass, relative bladder mass and hyperactivity, and decreased voiding efficiency. In NeMO vs. sham, 831 genes were differentially expressed (adjusted p<0.05) and correlated significantly with at least one physiologic parameter. Gene ontology revealed an enrichment for matrix pathways in the upregulated genes, and for cardiac contraction, oxidative phosphorylation and pyruvate metabolism in downregulated genes. TFBS analysis revealed a differential regulation of up vs downregulated genes, with KLF4 strongly associated with the downregulated genes. Downregulated genes of Human bladder obstruction were also associated with the TFBS of KLF4. GSEA of the NeMO gene set confirmed the DAVID results, but also showed a cluster of cytokine activation genes. In human bladder underactive obstruction, cytokines were also highly upregulated. The common cytokine pathway upregulation provided an example of the use of RNAseq for uncovering potential new therapeutic targets. As TNF and the innate immune pathways were strongly implicated in both human and mouse, and TNF is produced by macrophages, we depletion macrophages with clodronate (CL) during NeMO. Although CL did not block hypertrophy, it significantly decreased NeMO-induced hyperactive voiding (p<0.01) and increased voiding efficiency (p<0.05). The expression of several cytokines/chemokines correlated significantly with bladder functional parameters such as residual volumes, and hyperactivity. Conclusions: Gene expression signatures of NeMO were consistent with human bladder obstruction, supporting the use of the nerve-sparing mouse obstruction model for therapeutic exploration.

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“…Previous work by us (22,29), and others (10,11,(31)(32)(33)(34)(35)(36)(37)(38)(39)(40)(41) have shown the crucial role of ECM in PBO pathology.…”

Section: Discussionmentioning

confidence: 85%

Human - murine concordance of molecular signatures in nerve-sparing murine partial bladder outlet obstruction (NeMO)

Sidler

Ahmed

Jiang

et al. 2021

Preprint

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“…After MSC treatment of PBOO rats, the bladder had physiological benefits, and Tgfb, Hif1a, Hif3a, Mtor, Col1, and Col3 were downregulated. Yang et al demonstrated that tolterodine, a drug used to delay PBOO progression, can reduce collagen volume in the bladder wall of PBOO [10]. Moreover, tolterodine also induced MMP7, ITGA4, ITGB2, TIMP1, and FN1, which may be a mechanism of tolterodine for PBOO treatment.…”

Section: Discussionmentioning

confidence: 99%

“…There are rare microarray and NGS studies on PBOO-induced gene expression. Yang et al tried to investigate the gene expressions of ECM proteins, receptors, and metabolism regulators in a rat PBOO model [10]. However, only few differentially expressed genes (DEGs) were significantly decreased in the PBOO group compared with the sham group.…”

Section: Introductionmentioning

confidence: 99%

Differentially Expressed Genes Correlated with Fibrosis in a Rat Model of Chronic Partial Bladder Outlet Obstruction

Hsueh

Chang

et al. 2021

Processes

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Chronic partial bladder outlet obstruction (PBOO) is a prevalent clinical problem that may result from multiple etiologies. PBOO may be a secondary condition to various anatomical and functional abnormalities. Bladder fibrosis is the worst outcome of PBOO. However, gene alterations and the mechanism of fibrosis development after PBOO onset are not clear. Therefore, we aimed to investigate gene expression alterations during chronic PBOO. A rat model of PBOO was established and validated by a significant increase in rat bladder weight. The bladder samples were further analyzed by microarray, and differentially expressed genes (DEGs) that are more related to PBOO compared with the control genes were selected. The data showed that 16 significantly upregulated mRNAs and 3 significantly downregulated mRNAs are involved in fibrosis. Moreover, 13 significantly upregulated mRNAs and 12 significantly downregulated mRNAs are related to TGFB signaling. Twenty-two significantly upregulated mRNAs and nine significantly downregulated mRNAs are related to the extracellular matrix. The genes with differential expressions greater than four-fold included Grem1, Thbs1, Col8a1, Itga5, Tnc, Lox, Timp1, Col4a1, Col4a2, Bhlhe40, Itga1, Tgfb3, and Gadd45b. The gene with a differential expression less than a quarter-fold was Thbs2. These findings show the potential roles of these genes in the physiology of PBOO.

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“…The animal experiments were done in adherence with the National Institutes of Health Guidelines and approved by the West China Hospital Committee on Animal Care. Based on our previous study, female rats (220–250 g) were subdivided into sham‐operated and PBOO groups, with five rats in each group 20 . Briefly, after anesthesia, a catheter with an outer diameter of 1 mm was placed into the urethra of the rat, and the bladder neck was ligated using 4‐0 wires to establish PBOO.…”

Section: Methodsmentioning

confidence: 99%

“…Based on our previous study, female rats (220-250 g) were subdivided into sham-operated and PBOO groups, with five rats in each group. 20 Briefly, after anesthesia, a catheter with an outer diameter of 1 mm was placed into the urethra of the rat, and the bladder neck was ligated using 4-0 wires to establish PBOO. The sham-operated group underwent the same procedure but without urethral ligation.…”

Section: Microarray Data Of the Obstructed Bladdermentioning

confidence: 99%

β‐Adrenoceptor regulates contraction and inflammatory cytokine expression of human bladder smooth muscle cells via autophagy under pathological hydrostatic pressure

Chen

Jin

Luo³

et al. 2020

Neurourology and Urodynamics

Self Cite

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Aims: Abnormal intravesical pressure created by partial bladder outlet obstruction (PBOO) triggered the progression from chronic inflammation to fibrosis, initiating structural and functional alterations of bladder. To elucidate the underlying mechanisms of contraction and inflammatory response, we investigated the isolated human bladder smooth muscle cells (hBSMC) under pathological hydrostatic pressure (HP) mimicking the in vivo PBOO condition. Methods: hBSMCs were subjected to HP of 200 cm H 2 O to explore the contraction and inflammatory cytokine expression of hBSMC treated with β-adrenoceptors (ADRBs) and/or autophagy signaling pathway agonists and/ or antagonists. Results: We showed that pathological HP induced the release of the proinflammatory cytokines, including monocyte chemotactic protein-1, regulated upon activation normal T cell expressed and secreted factor, and interleukin-6. HP downregulated ADRB2 and ADRB3 expression, which was consistent with the results of the PBOO rat model. ADRB2 or autophagy activation repressed pathological HP-induced proinflammatory cytokine production. ADRB2, ADRB3 or autophagy activation ameliorated the HP-enhanced contraction. The increased contraction and autophagy activity by ADRB2 agonist under HP conditions were reversed by pretreatment with antagonists of adenosine monophosphate-activated protein kinase (AMPK). Conclusion: The present study provides evidence that the ADRB3 agonist suppresses hBSMC contraction under pathological HP conditions. Moreover, the ADRB2 agonist negatively regulates the contraction and inflammatory response of hBSMCs through AMPK/mTOR-mediated autophagy under pathological HP. These findings provide a theoretical basis for potential therapeutic strategies for patients with PBOO.

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The Antimuscarinic Agent Tolterodine Regulates Bladder Extracellular Matrix in Partial Bladder Outlet Obstruction in Rats

Cited by 11 publications

References 26 publications

Human - murine concordance of molecular signatures in nerve-sparing murine partial bladder outlet obstruction (NeMO)

Human - murine concordance of molecular signatures in nerve-sparing murine partial bladder outlet obstruction (NeMO)

Differentially Expressed Genes Correlated with Fibrosis in a Rat Model of Chronic Partial Bladder Outlet Obstruction

β‐Adrenoceptor regulates contraction and inflammatory cytokine expression of human bladder smooth muscle cells via autophagy under pathological hydrostatic pressure

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