Pub Date : 2026-01-13DOI: 10.1126/scisignal.adw3709
Vera-Marie E. Dunlock, Sergi Regot
T cells achieve precise antigen discrimination by relying on the temporal stability of T cell receptor (TCR) interactions with antigens. Given that time is central to antigen discrimination, we used real-time, single-cell imaging in a controlled TCR-antigen system to characterize extracellular signal–regulated kinase (ERK) signaling dynamics as a function of antigen affinity to better understand the temporal patterns of signaling downstream of the TCR. We found that intermediate-affinity antigens elicited pulsatile ERK activity at different frequencies and that T cell activation correlated with the cumulative amount of ERK activity. Mechanistically, we found that the ERK pulsing frequency depended on the rate of activity of the Src family kinase LCK at the plasma membrane, whereas mitogen-activated protein kinase (MAPK) kinase (MEK) modulated the amplitude of ERK signaling. Moreover, we showed that ERK activity dynamics in T cells depended on members of two upstream MAP3K groups: mixed lineage kinases (MLKs) and RAFs, which played distinct roles promoting or sustaining the formation of upstream signaling condensates containing the transmembrane adaptor molecule LAT. Together, our findings reveal insights into the spatiotemporal organization of TCR signaling activities and their roles in T cell activation.
{"title":"Antigen affinity modulates ERK pulsing frequency during T cell activation","authors":"Vera-Marie E. Dunlock, Sergi Regot","doi":"10.1126/scisignal.adw3709","DOIUrl":"10.1126/scisignal.adw3709","url":null,"abstract":"<div >T cells achieve precise antigen discrimination by relying on the temporal stability of T cell receptor (TCR) interactions with antigens. Given that time is central to antigen discrimination, we used real-time, single-cell imaging in a controlled TCR-antigen system to characterize extracellular signal–regulated kinase (ERK) signaling dynamics as a function of antigen affinity to better understand the temporal patterns of signaling downstream of the TCR. We found that intermediate-affinity antigens elicited pulsatile ERK activity at different frequencies and that T cell activation correlated with the cumulative amount of ERK activity. Mechanistically, we found that the ERK pulsing frequency depended on the rate of activity of the Src family kinase LCK at the plasma membrane, whereas mitogen-activated protein kinase (MAPK) kinase (MEK) modulated the amplitude of ERK signaling. Moreover, we showed that ERK activity dynamics in T cells depended on members of two upstream MAP3K groups: mixed lineage kinases (MLKs) and RAFs, which played distinct roles promoting or sustaining the formation of upstream signaling condensates containing the transmembrane adaptor molecule LAT. Together, our findings reveal insights into the spatiotemporal organization of TCR signaling activities and their roles in T cell activation.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":"19 920","pages":""},"PeriodicalIF":6.6,"publicationDate":"2026-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145958201","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-13DOI: 10.1126/scisignal.adp4326
Kai J. Winterberg, Vanessa Schwentner, Feng Gu, Franziska Möckl, Gaoyang Li, Andreas Bauche, Stefanie Etzold, Anette Rosche, Mariella Weiß, Nikolaus Thuille, Fritz Förster, Lena Woelk, René Werner, Dejan Kovacevic, Boris Fehse, Roberta Kurelic, Mikolaj Nawrocki, Samuel Huber, Hans-Willi Mittrücker, Chris Meier, Christa E. Müller, Gottfried Baier, Bjørn S. Skålhegg, Xavier de Deken, Christian Wahl-Schott, Thomas Mair, Bente Siebels, Roger Cugota Canals, Francesca Odoardi, Dmitri Lodygin, Alexander Flügel, Viacheslav O. Nikolaev, Björn-Philipp Diercks, Andreas H. Guse
T cell activation critically depends on the calcium ion (Ca2+)–mobilizing second messenger NAADP (nicotinic acid adenine dinucleotide phosphate), which induces the formation of Ca2+ microdomains that initiate global Ca2+ signals. NAADP is produced in immune synapses in T cells by dual NADPH oxidase 2 (DUOX2). Here, we investigated the mechanisms that stimulate DUOX2 activity in T cells. DUOX2 activity was enhanced by a modest increase in intracellular Ca2+ concentration, similar to that induced by Ca2+ microdomains that arise in resting T cells through different T cell receptor (TCR)–independent mechanisms. In addition, DUOX2 was activated in vitro by phosphorylation of threonine-789 mediated by PKA Cβ or PKCθ, and genetic deficiency of PKA Cβ2 or PKCθ decreased NAADP-dependent Ca2+ microdomain formation in T cells. PKA Cβ2 was activated downstream of adenosine A2A receptors, independently of the TCR. In contrast, PKCθ was activated by the tyrosine kinase LCK downstream of TCR stimulation. Inhibition of A2A receptors or PKCθ to prevent full DUOX2 activation decreased the production of the proinflammatory cytokine IL-17 by effector T cells. Thus, full stimulation of NAADP signaling that is critical for T cell activation requires integration of multiple TCR-independent and -dependent signals with different spatiotemporal characteristics by DUOX2, a fine-tuning mechanism that could be relevant for inflammation.
T细胞的激活主要依赖于钙离子（Ca2+）动员第二信使NAADP（烟酸腺嘌呤二核苷酸磷酸），它诱导Ca2+微域的形成，启动全局Ca2+信号。NAADP通过双NADPH氧化酶2 （DUOX2）在T细胞的免疫突触中产生。在这里，我们研究了刺激T细胞中DUOX2活性的机制。DUOX2活性通过细胞内Ca2+浓度的适度增加而增强，类似于通过不同的T细胞受体（TCR）独立机制在静息T细胞中产生的Ca2+微域诱导。此外，在体外，PKA Cβ或PKCθ介导的苏氨酸-789磷酸化激活了DUOX2， PKA Cβ2或PKCθ的遗传缺陷减少了T细胞中naadp依赖性Ca2+微结构域的形成。PKA c - β2在腺苷A2A受体下游被激活，独立于TCR。相反，PKCθ在TCR刺激的下游被酪氨酸激酶LCK激活。抑制A2A受体或PKCθ以防止DUOX2完全激活，可减少效应T细胞产生的促炎细胞因子IL-17。因此，充分刺激对T细胞活化至关重要的NAADP信号需要通过DUOX2整合具有不同时空特征的多个tcr独立和依赖信号，这是一种可能与炎症相关的微调机制。
{"title":"Multiple signaling events are required for NAADP synthesis by DUOX2 and formation of Ca2+ microdomains to initiate T cell activation","authors":"Kai J. Winterberg, Vanessa Schwentner, Feng Gu, Franziska Möckl, Gaoyang Li, Andreas Bauche, Stefanie Etzold, Anette Rosche, Mariella Weiß, Nikolaus Thuille, Fritz Förster, Lena Woelk, René Werner, Dejan Kovacevic, Boris Fehse, Roberta Kurelic, Mikolaj Nawrocki, Samuel Huber, Hans-Willi Mittrücker, Chris Meier, Christa E. Müller, Gottfried Baier, Bjørn S. Skålhegg, Xavier de Deken, Christian Wahl-Schott, Thomas Mair, Bente Siebels, Roger Cugota Canals, Francesca Odoardi, Dmitri Lodygin, Alexander Flügel, Viacheslav O. Nikolaev, Björn-Philipp Diercks, Andreas H. Guse","doi":"10.1126/scisignal.adp4326","DOIUrl":"10.1126/scisignal.adp4326","url":null,"abstract":"<div >T cell activation critically depends on the calcium ion (Ca<sup>2+</sup>)–mobilizing second messenger NAADP (nicotinic acid adenine dinucleotide phosphate), which induces the formation of Ca<sup>2+</sup> microdomains that initiate global Ca<sup>2+</sup> signals. NAADP is produced in immune synapses in T cells by dual NADPH oxidase 2 (DUOX2). Here, we investigated the mechanisms that stimulate DUOX2 activity in T cells. DUOX2 activity was enhanced by a modest increase in intracellular Ca<sup>2+</sup> concentration, similar to that induced by Ca<sup>2+</sup> microdomains that arise in resting T cells through different T cell receptor (TCR)–independent mechanisms. In addition, DUOX2 was activated in vitro by phosphorylation of threonine-789 mediated by PKA Cβ or PKCθ, and genetic deficiency of PKA Cβ2 or PKCθ decreased NAADP-dependent Ca<sup>2+</sup> microdomain formation in T cells. PKA Cβ2 was activated downstream of adenosine A<sub>2A</sub> receptors, independently of the TCR. In contrast, PKCθ was activated by the tyrosine kinase LCK downstream of TCR stimulation. Inhibition of A<sub>2A</sub> receptors or PKCθ to prevent full DUOX2 activation decreased the production of the proinflammatory cytokine IL-17 by effector T cells. Thus, full stimulation of NAADP signaling that is critical for T cell activation requires integration of multiple TCR-independent and -dependent signals with different spatiotemporal characteristics by DUOX2, a fine-tuning mechanism that could be relevant for inflammation.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":"19 920","pages":""},"PeriodicalIF":6.6,"publicationDate":"2026-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145958202","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-06DOI: 10.1126/scisignal.adw1483
Mengru Gao, ShaoKun Zang, Yanqing Zhu, Kun Xi, Yage Du, Shizhuo Cheng, Luwei Miao, Yanhui Lu, Chunyou Mao, Yan Zhang, Xin Ma
Hydroxycarboxylic acid receptor 1 (HCAR1) is a class A G protein–coupled receptor (GPCR) that is activated by the endogenous metabolite l-lactate and that plays an important role in various metabolic and inflammatory disorders. HCAR1 uses distinct ligand recognition and self-activation mechanisms to mediate specific pathophysiological functions through Gαi/o and β-arrestin signaling pathways. To support effective drug development targeting HCAR1, we investigated ligand recognition and activation mechanisms through cryo–electron microscopy (cryo-EM) structures of the HCAR1-Gαi1 complex in the apo state or with l-lactate or with the synthetic agonist CHBA. Compared with other HCARs, HCAR1 has a more compact binding pocket, which is stabilized by three unique disulfide bonds. l-lactate exhibited a flexible binding mode and relatively weak intermolecular interactions, thus requiring millimolar concentrations for receptor activation. In contrast, the binding of CHBA was more stable because of its chlorinated benzene ring, thus resulting in improved agonist potency. Structural comparisons with HCAR2 identified critical residues that restrict the size of the binding pocket of HCAR1 and influence ligand selectivity. Self-activation of HCAR1 is driven by conformational rearrangements within extracellular loop 2, with Phe168ECL2 playing a pivotal role as the key agonist. Together, these results clarify the mechanisms underlying HCAR1 activation, self-activation, and ligand selectivity, providing a structural framework for the design of high-affinity, selective agonists and inverse agonists with minimized off-target effects.
{"title":"Structural insights into the activation mechanism of the human metabolite receptor HCAR1","authors":"Mengru Gao, ShaoKun Zang, Yanqing Zhu, Kun Xi, Yage Du, Shizhuo Cheng, Luwei Miao, Yanhui Lu, Chunyou Mao, Yan Zhang, Xin Ma","doi":"10.1126/scisignal.adw1483","DOIUrl":"10.1126/scisignal.adw1483","url":null,"abstract":"<div >Hydroxycarboxylic acid receptor 1 (HCAR1) is a class A G protein–coupled receptor (GPCR) that is activated by the endogenous metabolite <span>l</span>-lactate and that plays an important role in various metabolic and inflammatory disorders. HCAR1 uses distinct ligand recognition and self-activation mechanisms to mediate specific pathophysiological functions through Gα<sub>i/o</sub> and β-arrestin signaling pathways. To support effective drug development targeting HCAR1, we investigated ligand recognition and activation mechanisms through cryo–electron microscopy (cryo-EM) structures of the HCAR1-Gα<sub>i1</sub> complex in the apo state or with <span>l</span>-lactate or with the synthetic agonist CHBA. Compared with other HCARs, HCAR1 has a more compact binding pocket, which is stabilized by three unique disulfide bonds. <span>l</span>-lactate exhibited a flexible binding mode and relatively weak intermolecular interactions, thus requiring millimolar concentrations for receptor activation. In contrast, the binding of CHBA was more stable because of its chlorinated benzene ring, thus resulting in improved agonist potency. Structural comparisons with HCAR2 identified critical residues that restrict the size of the binding pocket of HCAR1 and influence ligand selectivity. Self-activation of HCAR1 is driven by conformational rearrangements within extracellular loop 2, with Phe168<sup>ECL2</sup> playing a pivotal role as the key agonist. Together, these results clarify the mechanisms underlying HCAR1 activation, self-activation, and ligand selectivity, providing a structural framework for the design of high-affinity, selective agonists and inverse agonists with minimized off-target effects.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":"19 919","pages":""},"PeriodicalIF":6.6,"publicationDate":"2026-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145905282","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-06DOI: 10.1126/scisignal.adx8300
Hyunbae Kim, Pattaraporn Thepsuwan, Juncheng Wei, Donghong Ju, Qi Chen, Xiaohong Zhang, Li Li, Jie Xu, Xin Tong, Shengyi Sun, Chuan He, Lei Yin, Deyu Fang, Kezhong Zhang
Hepatic lipid metabolism is regulated by circadian rhythms and dynamically responds to nutrient availability, such that lipid synthesis, oxidation, and storage are temporally coordinated. We demonstrated that the endoplasmic reticulum (ER)–localized E3 ubiquitin ligase HRD1 stimulated lipid accumulation in the liver by decreasing the N6-methyladenosine (m6A) methylation and expression of mRNAs encoding factors involved in lipid metabolism. In mouse livers, m6A RNA modification and the expression of mRNAs encoding the m6A writer METTL14 and the m6A reader YTHDF3 were under circadian control and inversely correlated with the abundance of HRD1. m6A RNA sequencing analyses revealed that HRD1 and the m6A writer METTL14 had opposing roles in the m6A modification and expression of mRNAs encoding factors involved in fatty acid metabolism. In vivo, hepatic lipid accumulation and triglyceride amounts were decreased in mice with hepatic HRD1 deficiency fed a high-fat diet but increased in mice with hepatic METTL14 or YTHDF deficiency fed normal chow. Mechanistically, HRD1 mediated the polyubiquitination and degradation of PPARα, which transcriptionally activated METTL14 and YTHDF3 expression in the liver. Our work identifies a pathway regulated by circadian rhythms or nutrients in which HRD1 promotes the degradation of PPARα to decrease the m6A modification and expression of hepatic mRNAs encoding factors involved in lipid metabolism.
{"title":"The ubiquitin E3 ligase HRD1 restricts hepatic lipid metabolism by suppressing PPARα-driven m6A RNA modification","authors":"Hyunbae Kim, Pattaraporn Thepsuwan, Juncheng Wei, Donghong Ju, Qi Chen, Xiaohong Zhang, Li Li, Jie Xu, Xin Tong, Shengyi Sun, Chuan He, Lei Yin, Deyu Fang, Kezhong Zhang","doi":"10.1126/scisignal.adx8300","DOIUrl":"10.1126/scisignal.adx8300","url":null,"abstract":"<div >Hepatic lipid metabolism is regulated by circadian rhythms and dynamically responds to nutrient availability, such that lipid synthesis, oxidation, and storage are temporally coordinated. We demonstrated that the endoplasmic reticulum (ER)–localized E3 ubiquitin ligase HRD1 stimulated lipid accumulation in the liver by decreasing the <i>N</i><sup>6</sup>-methyladenosine (m6A) methylation and expression of mRNAs encoding factors involved in lipid metabolism. In mouse livers, m6A RNA modification and the expression of mRNAs encoding the m6A writer METTL14 and the m6A reader YTHDF3 were under circadian control and inversely correlated with the abundance of HRD1. m6A RNA sequencing analyses revealed that HRD1 and the m6A writer METTL14 had opposing roles in the m6A modification and expression of mRNAs encoding factors involved in fatty acid metabolism. In vivo, hepatic lipid accumulation and triglyceride amounts were decreased in mice with hepatic HRD1 deficiency fed a high-fat diet but increased in mice with hepatic METTL14 or YTHDF deficiency fed normal chow. Mechanistically, HRD1 mediated the polyubiquitination and degradation of PPARα, which transcriptionally activated <i>METTL14</i> and <i>YTHDF3</i> expression in the liver. Our work identifies a pathway regulated by circadian rhythms or nutrients in which HRD1 promotes the degradation of PPARα to decrease the m6A modification and expression of hepatic mRNAs encoding factors involved in lipid metabolism.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":"19 919","pages":""},"PeriodicalIF":6.6,"publicationDate":"2026-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145905283","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-23DOI: 10.1126/scisignal.adt1539
Adam M. Boulton, Megan E. Grund, Yuxin Wang, Erica M. Orsini, Yan Liu, Susamma Abraham, Lisa M. Grove, Ryan Musich, Caitlin M. Snyder, Haley Ricci, Amber Cardani-Boulton, Vidula Vachharajani, Mitchell A. Olman, Rachel G. Scheraga
The nuclear factor κB (NF-κB) signaling pathway plays a critical role in activating macrophages in the pathogenesis of many inflammatory diseases. Tissue mechanical properties are important in modulating key cellular proinflammatory responses. Here, we investigated how the mechanosensitive membrane cation channel TRPV4 (transient receptor potential vanilloid 4) limits macrophage proinflammatory responses in bacterial pneumonia. We found that TRPV4 suppressed proinflammatory gene expression in alveolar macrophages in response to Pseudomonas aeruginosa pneumonia in mice and in response to agonists of various Toll-like receptors (TLRs) in vitro. TRPV4 suppressed proinflammatory gene expression in macrophages by decreasing the activity of the NF-κB subunit p65. Upon stimulation of macrophages with bacterial lipopolysaccharide, a fraction of TRPV4 translocated from the endoplasmic reticulum to the plasma membrane, releasing p65 for nuclear translocation. TRPV4 interacted with p65 through an N-terminal cytoplasmic ankyrin repeat domain (ANKRD) that shares sequence homology with the p65-binding ANKRD of the NF-κB inhibitor IκBα. Given the diverse roles of TRPV4 and NF-κB in various cell types, our identification of cross-talk between a mechanosensitive channel and p65 in macrophages suggests application to many NF-κB–dependent diseases, such as cancer and atherosclerosis.
{"title":"The mechanosensitive channel TRPV4 inhibits pulmonary inflammation by limiting NF-κB signaling in alveolar macrophages","authors":"Adam M. Boulton, Megan E. Grund, Yuxin Wang, Erica M. Orsini, Yan Liu, Susamma Abraham, Lisa M. Grove, Ryan Musich, Caitlin M. Snyder, Haley Ricci, Amber Cardani-Boulton, Vidula Vachharajani, Mitchell A. Olman, Rachel G. Scheraga","doi":"10.1126/scisignal.adt1539","DOIUrl":"10.1126/scisignal.adt1539","url":null,"abstract":"<div >The nuclear factor κB (NF-κB) signaling pathway plays a critical role in activating macrophages in the pathogenesis of many inflammatory diseases. Tissue mechanical properties are important in modulating key cellular proinflammatory responses. Here, we investigated how the mechanosensitive membrane cation channel TRPV4 (transient receptor potential vanilloid 4) limits macrophage proinflammatory responses in bacterial pneumonia. We found that TRPV4 suppressed proinflammatory gene expression in alveolar macrophages in response to <i>Pseudomonas aeruginosa</i> pneumonia in mice and in response to agonists of various Toll-like receptors (TLRs) in vitro. TRPV4 suppressed proinflammatory gene expression in macrophages by decreasing the activity of the NF-κB subunit p65. Upon stimulation of macrophages with bacterial lipopolysaccharide, a fraction of TRPV4 translocated from the endoplasmic reticulum to the plasma membrane, releasing p65 for nuclear translocation. TRPV4 interacted with p65 through an N-terminal cytoplasmic ankyrin repeat domain (ANKRD) that shares sequence homology with the p65-binding ANKRD of the NF-κB inhibitor IκBα. Given the diverse roles of TRPV4 and NF-κB in various cell types, our identification of cross-talk between a mechanosensitive channel and p65 in macrophages suggests application to many NF-κB–dependent diseases, such as cancer and atherosclerosis.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":"18 918","pages":""},"PeriodicalIF":6.6,"publicationDate":"2025-12-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145808792","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-23DOI: 10.1126/scisignal.ads7662
Qianqian Hu, Lily L. Remsing Rix, Bina Desai, Daria Miroshnychenko, Xueli Li, Eric A. Welsh, Bin Fang, Gabriela M. Wright, Neelkamal Chaudhary, Jodi L. Kroeger, Robert C. Doebele, John M. Koomen, Eric B. Haura, Andriy Marusyk, Uwe Rix
Cancer-associated fibroblasts (CAFs) are associated with tumor progression and drug resistance. Here, we investigated the mechanisms underlying the cross-talk between CAFs and tumor cells in non–small cell lung cancer (NSCLC). In NSCLC cell lines with EML4-ALK fusions, we observed substantial CAF-mediated drug resistance to clinically used inhibitors of the tyrosine kinase ALK. Array-based cytokine profiling of CAF-derived conditioned medium indicated that a major contributor to the phenomenon was the secreted growth factor HGF, and blocking its receptor MET overcame paracrine resistance to ALK inhibition. However, cell-selective labeling of the proteome in cocultures also revealed an equally important contribution by the fibronectin-integrin pathway, specifically integrin β1, which was confirmed through pharmacological inhibition and cell-specific silencing or knockout. Concurrent targeting of MET and integrin signaling effectively abrogated ALK inhibitor resistance in EML4-ALK+ NSCLC cells cocultured with CAFs. Moreover, the combination of the ALK inhibitor alectinib with the MET inhibitor capmatinib and/or the integrin inhibitor cilengitide was more effective than single-agent treatment in suppressing tumor growth in allografted mice. The findings illustrate a previously unappreciated complex nature of concurrent paracrine and juxtacrine mechanisms of CAF-driven resistance that may inform the development of more effective therapeutic approaches.
{"title":"Cancer-associated fibroblasts confer ALK inhibitor resistance in EML4-ALK–driven lung cancer by concurrent integrin and MET signaling","authors":"Qianqian Hu, Lily L. Remsing Rix, Bina Desai, Daria Miroshnychenko, Xueli Li, Eric A. Welsh, Bin Fang, Gabriela M. Wright, Neelkamal Chaudhary, Jodi L. Kroeger, Robert C. Doebele, John M. Koomen, Eric B. Haura, Andriy Marusyk, Uwe Rix","doi":"10.1126/scisignal.ads7662","DOIUrl":"10.1126/scisignal.ads7662","url":null,"abstract":"<div >Cancer-associated fibroblasts (CAFs) are associated with tumor progression and drug resistance. Here, we investigated the mechanisms underlying the cross-talk between CAFs and tumor cells in non–small cell lung cancer (NSCLC). In NSCLC cell lines with <i>EML4-ALK</i> fusions, we observed substantial CAF-mediated drug resistance to clinically used inhibitors of the tyrosine kinase ALK. Array-based cytokine profiling of CAF-derived conditioned medium indicated that a major contributor to the phenomenon was the secreted growth factor HGF, and blocking its receptor MET overcame paracrine resistance to ALK inhibition. However, cell-selective labeling of the proteome in cocultures also revealed an equally important contribution by the fibronectin-integrin pathway, specifically integrin β<sub>1</sub>, which was confirmed through pharmacological inhibition and cell-specific silencing or knockout. Concurrent targeting of MET and integrin signaling effectively abrogated ALK inhibitor resistance in <i>EML4-ALK<sup>+</sup></i> NSCLC cells cocultured with CAFs. Moreover, the combination of the ALK inhibitor alectinib with the MET inhibitor capmatinib and/or the integrin inhibitor cilengitide was more effective than single-agent treatment in suppressing tumor growth in allografted mice. The findings illustrate a previously unappreciated complex nature of concurrent paracrine and juxtacrine mechanisms of CAF-driven resistance that may inform the development of more effective therapeutic approaches.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":"18 918","pages":""},"PeriodicalIF":6.6,"publicationDate":"2025-12-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145808794","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-23DOI: 10.1126/scisignal.aee7080
Annalisa M. VanHook
A bacterial virulence factor subverts host cell apoptosis by acting as a protein recombinase.
细菌毒力因子通过作为蛋白质重组酶破坏宿主细胞凋亡。
{"title":"Shigella mixes and matches host proteins","authors":"Annalisa M. VanHook","doi":"10.1126/scisignal.aee7080","DOIUrl":"10.1126/scisignal.aee7080","url":null,"abstract":"<div >A bacterial virulence factor subverts host cell apoptosis by acting as a protein recombinase.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":"18 918","pages":""},"PeriodicalIF":6.6,"publicationDate":"2025-12-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145808793","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-23DOI: 10.1126/scisignal.adv0660
Nicholas M. Venetos, Colin T. Stomberski, Hua-Lin Zhou, Zhaoxia Qian, Precious J. McLaughlin, Puneet K. Bansal, John Feczko, Ilya Bederman, Hoa Nguyen, Alfred Hausladen, Joseph C. Schindler, Zachary W. Grimmett, Henri Brunengraber, Richard T. Premont, Jonathan S. Stamler
Lipid homeostasis is subject to control by posttranslational modification machinery, such as sirtuin deacetylases that reverse coenzyme A (CoA)–dependent acetylation. Here, we showed that a mammalian denitrosylase (SCoR2), which counteracts CoA-dependent S-nitrosylation, promoted both fat storage and lipogenesis to impair metabolic health. In mice, SCoR2 protein abundance correlated with body mass, and deleting or pharmacologically inhibiting SCoR2 prevented both diet-induced obesity and metabolic dysfunction–associated steatotic liver disease (MASLD). Loss of SCoR2 in adipocytes promoted the S-nitrosylation of the actin cytoskeletal regulator myosin 9, which inhibited the activity of the lipogenesis-promoting transcription factors PPARγ, SREBP1, and CEBPα to prevent fat storage. In hepatocytes, inhibition of SCoR2-mediated denitrosylation of lipogenic enzymes reduced fat synthesis and induced fat oxidation. In humans, an obesity-linked polymorphism was associated with increased SCoR2 mRNA expression, and in patient adipose and liver tissues, SCoR2 protein or mRNA abundance directly correlated with adipocyte size or MASLD. These results indicate that SCoR2 regulates nutrient metabolism, similar to sirtuins, and is a potential drug target for obesity and MASLD.
{"title":"The protein denitrosylase SCoR2 regulates lipogenesis and fat storage","authors":"Nicholas M. Venetos, Colin T. Stomberski, Hua-Lin Zhou, Zhaoxia Qian, Precious J. McLaughlin, Puneet K. Bansal, John Feczko, Ilya Bederman, Hoa Nguyen, Alfred Hausladen, Joseph C. Schindler, Zachary W. Grimmett, Henri Brunengraber, Richard T. Premont, Jonathan S. Stamler","doi":"10.1126/scisignal.adv0660","DOIUrl":"10.1126/scisignal.adv0660","url":null,"abstract":"<div >Lipid homeostasis is subject to control by posttranslational modification machinery, such as sirtuin deacetylases that reverse coenzyme A (CoA)–dependent acetylation. Here, we showed that a mammalian denitrosylase (SCoR2), which counteracts CoA-dependent S-nitrosylation, promoted both fat storage and lipogenesis to impair metabolic health. In mice, SCoR2 protein abundance correlated with body mass, and deleting or pharmacologically inhibiting SCoR2 prevented both diet-induced obesity and metabolic dysfunction–associated steatotic liver disease (MASLD). Loss of <i>SCoR2</i> in adipocytes promoted the S-nitrosylation of the actin cytoskeletal regulator myosin 9, which inhibited the activity of the lipogenesis-promoting transcription factors PPARγ, SREBP1, and CEBPα to prevent fat storage. In hepatocytes, inhibition of SCoR2-mediated denitrosylation of lipogenic enzymes reduced fat synthesis and induced fat oxidation. In humans, an obesity-linked polymorphism was associated with increased <i>SCoR2</i> mRNA expression, and in patient adipose and liver tissues, SCoR2 protein or mRNA abundance directly correlated with adipocyte size or MASLD. These results indicate that SCoR2 regulates nutrient metabolism, similar to sirtuins, and is a potential drug target for obesity and MASLD.</div>","PeriodicalId":21658,"journal":{"name":"Science Signaling","volume":"18 918","pages":""},"PeriodicalIF":6.6,"publicationDate":"2025-12-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145808796","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
京公网安备 11010802042870号
京ICP备2023020795号-1
扫码关注我们
求助内容：
应助结果提醒方式：