Targeted Gq-GPCR activation drives ER-dependent calcium oscillations in chondrocytes

McDonough, Ryan C.; Gilbert, Rachel M.; Gleghorn, Jason P.; Price, Christopher

doi:10.1016/j.ceca.2021.102363

Cited by 8 publications

(7 citation statements)

References 94 publications

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“…This was evidenced by ectodomain shedding of EGF and BTC, substrates of ADAM10, which was triggered only by the downstream signal of TRP channels (Fig 2E -2I). The increase in intracellular Ca 2+ concentration induced by TRP channel activation is high and long-lasting, whereas that induced by GPCRs is oscillatory and transient [34][35][36]. Previous reports showed that Ca 2+ -induced ADAM10 activation is totally dependent on anoctamin 6 (ANO6), a Ca 2+ -sensitive phosphatidylserine scramblase [37,38].…”

Section: Discussionmentioning

confidence: 99%

Ectodomain shedding of EGFR ligands serves as an activation readout for TRP channels

et al. 2023

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Transient receptor potential (TRP) channels are activated by various extracellular and intracellular stimuli and are involved in many physiological events. Because compounds that act on TRP channels are potential candidates for therapeutic agents, a simple method for evaluating TRP channel activation is needed. In this study, we demonstrated that a transforming growth factor alpha (TGFα) shedding assay, previously developed for detecting G-protein–coupled receptor (GPCR) activation, can also detect TRP channel activation. This assay is a low-cost, easily accessible method that requires only an absorbance microplate reader. Mechanistically, TRP-channel-triggered TGFα shedding is achieved by both of a disintegrin and metalloproteinase domain-containing protein 10 (ADAM10) and 17 (ADAM17), whereas the GPCR-induced TGFα shedding response depends solely on ADAM17. This difference may be the result of qualitative or quantitative differences in intracellular Ca2+ kinetics between TRP channels and GPCRs. Use of epidermal growth factor (EGF) and betacellulin (BTC), substrates of ADAM10, improved the specificity of the shedding assay by reducing background responses mediated by endogenously expressed GPCRs. This assay for TRP channel measurement will not only facilitate the high-throughput screening of TRP channel ligands but also contribute to understanding the roles played by TRP channels as regulators of membrane protein ectodomain shedding.

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Section: Discussionmentioning

confidence: 99%

Ectodomain shedding of EGFR ligands serves as an activation readout for TRP channels

et al. 2023

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“…We further tested if the Ca 2+ response was from mobilization of intracellular Ca 2+ stores or extracellular Ca 2+ influx from outside of the plasma membrane, GPCRs, including T2Rs, induce intracellular Ca 2+ responses, primarily from the ER [67][68][69][70].…”

Section: Lidocaine Stimulation Causes An Intracellular Ca 2+ Responsementioning

confidence: 99%

Lidocaine Induces Apoptosis in Head and Neck Squamous Cell Carcinoma Through Activation of Bitter Taste Receptor T2R14

Miller

Mueller

Holivert

et al. 2022

Preprint

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Head and Neck Squamous Cell Carcinomas (HNSCCs) have high mortality due to late stage diagnosis, high metastasis rates, and poor treatment options. Current therapies are invasive and aggressive, leading to a severe decline in patient quality of life (QoL). It is vital to create new therapies that are both potent and localized and/or targeted to prolong survival while maintaining QoL. Lidocaine is a local anesthetic used in HNSCC surgery settings. Lidocaine also activates bitter taste (taste family 2) receptor 14 (T2R14). T2Rs are G-protein coupled receptors (GPCRs) that increase intracellular Ca2+ when activated. T2Rs are expressed in mucosal epithelia, including internal regions of the head and neck, and are accessible drug targets. Here, we show that lidocaine increases intracellular Ca2+ and decreases cAMP in several HNSCC cell lines. Ca2+ release from the ER results in Ca2+ uptake into the mitochondria. Ca2+ mobilization is blocked with GPCR inhibitors and T2R14 antagonist, 6-methoxyflavanone. Lidocaine activation of T2R14 depolarizes the mitochondrial membrane, inhibits cell proliferation, and induces apoptosis. Lidocaine activates caspase-3 and -7 cleavage and also increases total caspase protein levels despite no changes in mRNA production. Both total and cleaved caspase products were upregulated even in the presence of cycloheximide, an inhibitor of protein synthesis. This suggests inhibition of the ubiquitin-proteasome system. It is vital to understand Lidocaine-induced apoptosis in HSNCCs to utilize its chemotherapeutic effects as a treatment option. In addition, future studies on T2R14 expression in HNSCC patients could provide insight for implementing lidocaine as a targeted topical therapy.

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“…This multicellular interaction is an example of sender-receiver type signaling, where morphogen secretion from one cell type induces a response from another cell type. Often deviations in the strength or positioning of morphogen concentrations and gradients regulate this intercellular interaction and underpin developmental diseases and congenital birth defects [11][12][13][14][15][16]. Since these interactions function at the multicellular level (tissue scale) with spatial heterogeneity, many traditional methods of inferring the gene regulatory networks that comprise these interactions are insufficient.…”

Section: Introductionmentioning

confidence: 99%

Methodology for inference of intercellular gene interactions

Modi

Zurakowski

Gleghorn

2023

Preprint

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To govern organ size, shape, and function, cell-secreted diffusible molecules called morphogens spatially pattern cell differentiation, gene expression, and proliferation. Local morphogen concentration governs cell differentiation through gene regulatory networks (GRN). Previous inference methodologies tackle intercellular GRN inference between cells of one type. This is insufficient, as many developmental systems consist of cells of different types interacting with each other. Inference methodologies of GRNs between different cell types assume knowledge of diffusible morphogen identity and concentration. This makes their applicability limited in real biological systems. Here, we develop a computational methodology to infer the intercellular GRN derived from experiments that use fluorescence from reporter proteins for gene expression measurements. For validation, we demonstrate the methodologyin silicousing three case studies based on developmental and synthetic biology. The results show that, barring practical identifiability limitations, the methodology successfully infers the intercellular GRNs.

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Targeted Gq-GPCR activation drives ER-dependent calcium oscillations in chondrocytes

Cited by 8 publications

References 94 publications

Ectodomain shedding of EGFR ligands serves as an activation readout for TRP channels

Ectodomain shedding of EGFR ligands serves as an activation readout for TRP channels

Lidocaine Induces Apoptosis in Head and Neck Squamous Cell Carcinoma Through Activation of Bitter Taste Receptor T2R14

Methodology for inference of intercellular gene interactions

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