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Pro-inflammatory cytokines drive functional deregulation of potassium channels in primary synovial fibroblasts

ABSTRACT: Pro-inflammatory cytokines drive functional deregulation of potassium channels in primary synovial fibroblasts

PROVIDER: PRJEB31797 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR3237330_1.fastq.gz	Fastqsanger.gz
	ERR3237330_2.fastq.gz	Fastqsanger.gz
	ERR3237331_1.fastq.gz	Fastqsanger.gz
	ERR3237331_2.fastq.gz	Fastqsanger.gz
	ERR3237332_1.fastq.gz	Fastqsanger.gz

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Pro-inflammatory cytokines drive functional deregulation of potassium channels in primary synovial fibroblasts

Project description:The synovium secretes synovial fluid but is also richly innervated with nociceptors and acts as a gateway between avascular joint tissues and the circulatory system. Resident fibroblast-like synoviocytes’ (FLS) calcium-activated potassium channels (KCa) change in activity in arthritis models and this correlates with FLS activation. To deepen our understanding of the synovium in the context of synovial joint health and disease, the electrophysiological profile of FLS cells needs to be characterised, along with the ion channels that are present. Ion channels are an essential component of any cell membrane that controls ion movement in and out of the cell and play an important role in a multitude of cell regulating processes, typically by modulating the membrane potential To this end, we chose to perform an UNBIASED screen to identify/screen the ion channels that are transcribed in these cells and determine if this "channelome" changes with cytokine exposure. PCR/Westerns naturally bias toward the known proteins and transcripts and so we used Next Generation sequencing to scan the entire transcriptome. Objective: To determine the mechanism of this activation in an in vitro model of inflammatory arthritis; 72hr treatment with the cytokines TNFa and IL1b.

2020-03-06 | E-MTAB-7798 | biostudies-arrayexpress

KCNQ POTASSIUM CHANNELS MODULATE WNT ACTIVITY IN GASTRO-OESOPHAGEAL ADENOCARCINOMAS

Project description:Voltage sensitive potassium channels play an important role in controlling membrane potential and ionic homeostasis in the gut and have been implicated in gastrointestinal (GI) cancers. Through large scale analysis of 897 patients with Gastro-oesophageal adenocarcincomas (GOA) coupled with in vitro models, we find KCNQ family genes are mutated in ~30% of patients, and play therapeutically targetable roles in GOA cancer growth. KCNQ1 and KCNQ3 mediate the WNT pathway and MYC to increase proliferation through resultant effects on cadherins junctions. This also highlights novel roles for KCNQ3 in non-excitable tissues. We additionally discover that activity of KCNQ3 sensitises cancer cells to existing potassium channel inhibitors, and that inhibition of KCNQ activity reduces proliferation of GOA cancer cells. These findings reveal a novel and exploitable role for potassium channels in the advancement of human cancer, and highlight that supplemental treatments for GOAs may exist through KCNQ inhibitors.

2023-09-13 | GSE242782 | GEO

E-MTAB-7798

Project description:Pro-inflammatory cytokines drive functional deregulation of potassium channels in primary synovial fibroblasts

2023-06-16 | E-MTAB-7798 | ExpressionAtlas

Identification of proteins associated with Kv1.2 potassium channels.

Project description:Kv1.2 is a prominent voltage-gated potassium channel that influences action potential generation and propagation in the central nervous system. We explored multi-protein complexes containing Kv1.2 using mass spectrometry of immunoprecipitated samples. Proteins identified as candidate interactors were prioritized based on abundance, and were screened for functional effects on Kv1.2 channels using electrophysiology.

2018-11-04 | PXD011010 | Pride

Mechanism of pharmacochaperoning in ATP-sensitive potassium channels revealed by cryo-EM

Project description:ATP-sensitive potassium (K-ATP) channels composed of a pore-forming Kir6.2 potassium channel and a regulatory ABC transporter sulfonylurea receptor 1 (SUR1) regulate insulin secretion in pancreatic beta-cells to maintain glucose homeostasis. Mutations that impair channel folding or assembly prevent cell surface expression and cause congenital hyperinsulinism. Structurally diverse K-ATP inhibitors have been shown to act as pharmacochaperones to correct mutant channel expression, but the mechanism is unknown. Here, we compare cryoEM structures of K-ATP channels bound to pharmacochaperones glibenclamide, repaglinide, and carbamazepine. DSP cross-linking mass spectrometry was used to partially confirm cryoEM structures.

2019-11-12 | PXD013873 | Pride

Mechanism of pharmacochaperoning in ATP-sensitive potassium channels revealed by cryo-EM

Project description:ATP-sensitive potassium (K-ATP) channels composed of a pore-forming Kir6.2 potassium channel and a regulatory ABC transporter sulfonylurea receptor 1 (SUR1) regulate insulin secretion in pancreatic beta-cells to maintain glucose homeostasis. Mutations that impair channel folding or assembly prevent cell surface expression and cause congenital hyperinsulinism. Structurally diverse K-ATP inhibitors have been shown to act as pharmacochaperones to correct mutant channel expression, but the mechanism is unknown. Here, we compare cryoEM structures of K-ATP channels bound to pharmacochaperones glibenclamide, repaglinide, and carbamazepine. CyanurBiotinDimercaptoPropionylSuccinimide (CBDPS) cross-linking mass spectrometry was used to partially confirm cryoEM structures.

2019-08-12 | PXD014498 | Pride

Co-expression of calcium channels and delayed rectifier potassium channels protects the heart from proarrhythmic events

Project description:Co-expression of calcium channels and delayed rectifier potassium channels protects the heart from proarrhythmic events

| PRJNA611588 | ENA

Pro-inflammatory Cytokines Drive Deregulation of Potassium Channel Expression in Primary Synovial Fibroblasts.

Project description:The synovium secretes synovial fluid, but is also richly innervated with nociceptors and acts as a gateway between avascular joint tissues and the circulatory system. Resident fibroblast-like synoviocytes' (FLS) calcium-activated potassium channels (K Ca) change in activity in arthritis models and this correlates with FLS activation.ObjectiveTo investigate this activation in an in vitro model of inflammatory arthritis; 72 h treatment with cytokines TNFα and IL1β.MethodsFLS cells were isolated from rat synovial membranes. We analyzed global changes in FLS mRNA by RNA-sequencing, then focused on FLS ion channel genes and the corresponding FLS electrophysiological phenotype and finally modeling data with ingenuity pathway analysis (IPA) and MATLAB.ResultsIPA showed significant activation of inflammatory, osteoarthritic and calcium signaling canonical pathways by cytokines, and we identified ∼200 channel gene transcripts. The large K Ca (BK) channel consists of the pore forming Kcnma1 together with β-subunits. Following cytokine treatment, a significant increase in Kcnma1 RNA abundance was detected by qPCR and changes in several ion channels were detected by RNA-sequencing, including a loss of BK channel β-subunit expression Kcnmb1/2 and an increase in Kcnmb3. In electrophysiological experiments, there was a decrease in over-all current density at 20 mV without change in chord conductance at this potential.ConclusionTNFα and IL1β treatment of FLS in vitro recapitulated several common features of inflammatory arthritis at the transcriptomic level, including increase in Kcnma1 and Kcnmb3 gene expression.

| S-EPMC7105747 | biostudies-literature

Identification of potassium and chloride channels in eccrine sweat glands

Project description:K and Cl channels play critical role for sweat secretion, however, individual K or Cl channels and their functions are largely unknown. By comparing expreession profilings between wild-type and Eda mutant Tabby footpads we identified 4 K and 2 Cl channels highly expressed in mouse eccrine sweat glands.

2015-12-09 | GSE73715 | GEO

Co-expression of calcium channels and delayed rectifier potassium channels protects the heart from proarrhythmic events [Figure 2 mRNA data]

Project description:Co-expression of calcium channels and delayed rectifier potassium channels protects the heart from proarrhythmic events [Figure 2 mRNA data]

| PRJNA611596 | ENA

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