Pedro J. del Rivero Morfin, Diego Scala Chavez, Srinidhi Jayaraman, Lin Yang, Stefanie M. Geisler, Audrey L. Kochiss, Petronel Tuluc, Henry M. Colecraft, Steven O. Marx, X. Shawn Liu, Anjali M. Rajadhyaksha, Manu Ben-Johny
L-type Ca2+ channels (CaV1.2/1.3) convey influx of calcium ions that orchestrate a bevy of biological responses including muscle contraction, neuronal function, and gene transcription. Deficits in CaV1 function play a vital role in cardiac and neurodevelopmental disorders. Here, we develop a genetically encoded enhancer of CaV1.2/1.3 channels (GeeCL) to manipulate Ca2+ entry in distinct physiological settings. We functionalized a nanobody that targets the CaV complex by attaching a minimal effector domain from an endogenous CaV modulator—leucine-rich repeat containing protein 10 (Lrrc10). In cardiomyocytes, GeeCL selectively increased L-type current amplitude. In neurons in vitro and in vivo, GeeCL augmented excitation-transcription (E-T) coupling. In all, GeeCL represents a powerful strategy to boost CaV1.2/1.3 function and lays the groundwork to illuminate insights on neuronal and cardiac physiology and disease.
L 型 Ca2+ 通道（CaV1.2/1.3）传递钙离子的流入，协调肌肉收缩、神经元功能和基因转录等一系列生物反应。CaV1 功能缺陷在心脏和神经发育疾病中起着至关重要的作用。在这里，我们开发了一种基因编码的 CaV1.2/1.3 通道增强子（GeeCL），以在不同的生理环境中操纵 Ca2+ 进入。我们从内源性 CaV 调节器--富含亮氨酸重复序列的蛋白 10（Lrrc10）中提取了一个最小效应结构域，使纳米抗体功能化，从而靶向 CaV 复合物。在心肌细胞中，GeeCL 可选择性地增加 L 型电流振幅。在体外和体内神经元中，GeeCL增强了兴奋-转录（E-T）耦合。总之，GeeCL代表了一种增强CaV1.2/1.3功能的强大策略，为深入了解神经元和心脏的生理学和疾病奠定了基础。
{"title":"A genetically encoded actuator boosts L-type calcium channel function in diverse physiological settings","authors":"Pedro J. del Rivero Morfin, Diego Scala Chavez, Srinidhi Jayaraman, Lin Yang, Stefanie M. Geisler, Audrey L. Kochiss, Petronel Tuluc, Henry M. Colecraft, Steven O. Marx, X. Shawn Liu, Anjali M. Rajadhyaksha, Manu Ben-Johny","doi":"10.1126/sciadv.adq3374","DOIUrl":"10.1126/sciadv.adq3374","url":null,"abstract":"<div >L-type Ca<sup>2+</sup> channels (Ca<sub>V</sub>1.2/1.3) convey influx of calcium ions that orchestrate a bevy of biological responses including muscle contraction, neuronal function, and gene transcription. Deficits in Ca<sub>V</sub>1 function play a vital role in cardiac and neurodevelopmental disorders. Here, we develop a genetically encoded enhancer of Ca<sub>V</sub>1.2/1.3 channels (GeeC<sub>L</sub>) to manipulate Ca<sup>2+</sup> entry in distinct physiological settings. We functionalized a nanobody that targets the Ca<sub>V</sub> complex by attaching a minimal effector domain from an endogenous Ca<sub>V</sub> modulator—leucine-rich repeat containing protein 10 (Lrrc10). In cardiomyocytes, GeeC<sub>L</sub> selectively increased L-type current amplitude. In neurons in vitro and in vivo, GeeC<sub>L</sub> augmented excitation-transcription (E-T) coupling. In all, GeeC<sub>L</sub> represents a powerful strategy to boost Ca<sub>V</sub>1.2/1.3 function and lays the groundwork to illuminate insights on neuronal and cardiac physiology and disease.</div>","PeriodicalId":21609,"journal":{"name":"Science Advances","volume":null,"pages":null},"PeriodicalIF":11.7,"publicationDate":"2024-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.science.org/doi/reader/10.1126/sciadv.adq3374","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142547185","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Philippa R. Kennedy, Upasana Sunil Arvindam, Shee Kwan Phung, Brianna Ettestad, Xueyang Feng, Yunmin Li, Quinlan M. Kile, Peter Hinderlie, Melissa Khaw, Rih-Sheng Huang, Marissa Kaufman, Patrycja Puchalska, Amanda Russell, Jonah Butler, Lucas Abbott, Paul McClure, Xianghua Luo, Quynhanh T. Lu, Bruce R. Blazar, Peter A. Crawford, James Lim, Jeffrey S. Miller, Martin Felices
Limited oxygen (hypoxia) in solid tumors poses a challenge to successful immunotherapy with natural killer (NK) cells. NK cells have impaired cytotoxicity when cultured in hypoxia (1% oxygen) but not physiologic (>5%) or atmospheric oxygen (20%). We found that changes to cytotoxicity were regulated at the transcriptional level and accompanied by metabolic dysregulation. Dosing with interleukin-15 (IL-15) enhanced NK cell cytotoxicity in hypoxia, but preactivation with feeder cells bearing IL-21 and 4-1BBL was even better. Preactivation resulted in less perturbed metabolism in hypoxia; greater resistance to oxidative stress; and no hypoxia-induced loss of transcription factors (T-bet and Eomes), activating receptors, adhesion molecules (CD2), and cytotoxic proteins (TRAIL and FasL). There remained a deficit in CD122/IL-2Rβ when exposed to hypoxia, which affected IL-15 signaling. However, tri-specific killer engager molecules that deliver IL-15 in the context of anti-CD16/FcγRIII were able to bypass this deficit, enhancing cytotoxicity of both fresh and preactivated NK cells in hypoxia.
实体瘤中有限的氧气（缺氧）给使用自然杀伤（NK）细胞成功进行免疫治疗带来了挑战。在缺氧（1% 氧气）条件下培养的 NK 细胞的细胞毒性会受损，而在生理氧（>5%）或大气氧（20%）条件下培养的 NK 细胞的细胞毒性则不会受损。我们发现，细胞毒性的变化受转录水平的调控，并伴随着代谢失调。使用白细胞介素-15（IL-15）可增强NK细胞在缺氧条件下的细胞毒性，但使用含有IL-21和4-1BBL的饲养细胞进行预激活效果更好。预激活后，NK细胞在缺氧条件下的新陈代谢扰动较小；对氧化应激的抵抗力较强；没有因缺氧引起的转录因子（T-bet 和 Eomes）、活化受体、粘附分子（CD2）和细胞毒性蛋白（TRAIL 和 FasL）的损失。当暴露于缺氧环境时，CD122/IL-2Rβ仍然缺失，这影响了IL-15信号的传递。然而，在抗CD16/FcγRIII背景下传递IL-15的三特异性杀手吞噬分子能够绕过这一缺陷，增强缺氧环境下新鲜和预活化NK细胞的细胞毒性。
{"title":"Metabolic programs drive function of therapeutic NK cells in hypoxic tumor environments","authors":"Philippa R. Kennedy, Upasana Sunil Arvindam, Shee Kwan Phung, Brianna Ettestad, Xueyang Feng, Yunmin Li, Quinlan M. Kile, Peter Hinderlie, Melissa Khaw, Rih-Sheng Huang, Marissa Kaufman, Patrycja Puchalska, Amanda Russell, Jonah Butler, Lucas Abbott, Paul McClure, Xianghua Luo, Quynhanh T. Lu, Bruce R. Blazar, Peter A. Crawford, James Lim, Jeffrey S. Miller, Martin Felices","doi":"10.1126/sciadv.adn1849","DOIUrl":"10.1126/sciadv.adn1849","url":null,"abstract":"<div >Limited oxygen (hypoxia) in solid tumors poses a challenge to successful immunotherapy with natural killer (NK) cells. NK cells have impaired cytotoxicity when cultured in hypoxia (1% oxygen) but not physiologic (>5%) or atmospheric oxygen (20%). We found that changes to cytotoxicity were regulated at the transcriptional level and accompanied by metabolic dysregulation. Dosing with interleukin-15 (IL-15) enhanced NK cell cytotoxicity in hypoxia, but preactivation with feeder cells bearing IL-21 and 4-1BBL was even better. Preactivation resulted in less perturbed metabolism in hypoxia; greater resistance to oxidative stress; and no hypoxia-induced loss of transcription factors (T-bet and Eomes), activating receptors, adhesion molecules (CD2), and cytotoxic proteins (TRAIL and FasL). There remained a deficit in CD122/IL-2Rβ when exposed to hypoxia, which affected IL-15 signaling. However, tri-specific killer engager molecules that deliver IL-15 in the context of anti-CD16/FcγRIII were able to bypass this deficit, enhancing cytotoxicity of both fresh and preactivated NK cells in hypoxia.</div>","PeriodicalId":21609,"journal":{"name":"Science Advances","volume":null,"pages":null},"PeriodicalIF":11.7,"publicationDate":"2024-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.science.org/doi/reader/10.1126/sciadv.adn1849","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142547199","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Min Ge, Zesong Ruan, Ya-Xuan Zhu, Wencheng Wu, Chuang Yang, Han Lin, Jianlin Shi
In the competition between the pathogen infection and the host defense, infectious microorganisms may enter the host cells by evading host defense mechanisms and use the intracellular biomolecules as replication nutrient. Among them, intracellular Staphylococcus aureus relies on the host cells to protect itself from the attacks by antibiotics or immune system to achieve long-term colonization in the host, and the consequent clinical therapeutic failures and relapses after antibiotic treatment. Here, we demonstrate that intracellular S. aureus surviving well even in the presence of vancomycin can be effectively eliminated using an emerging cell-mimicking therapeutic strategy. These cell mimics with natural killer cell–like activity (NKMs) are composed of a redox-responsive degradable carrier, and perforin and granzyme B within the carrier. NKMs perform far more effectivly than clinical antibiotics in treating intracellular bacterial infections, providing a direct evidence of the NK cell–mimicking immune mechanism in the treatment of intracellular S. aureus.
在病原体感染与宿主防御的竞争中，感染性微生物可能通过逃避宿主防御机制进入宿主细胞，并利用细胞内的生物大分子作为复制养分。其中，细胞内金黄色葡萄球菌依靠宿主细胞保护自身免受抗生素或免疫系统的攻击，从而实现在宿主体内的长期定植，并因此导致临床治疗失败和抗生素治疗后复发。在这里，我们证明了即使在万古霉素的作用下也能很好存活的金黄色葡萄球菌，可以通过一种新兴的细胞模拟治疗策略有效地消灭。这些具有类似自然杀伤细胞活性的细胞模拟物（NKMs）由氧化还原反应可降解载体以及载体内的穿孔素和颗粒酶 B 组成。在治疗细胞内细菌感染方面，NKMs 的效果远远优于临床抗生素，直接证明了 NK 细胞模拟免疫机制在治疗细胞内金黄色葡萄球菌方面的作用。
{"title":"A natural killer cell mimic against intracellular pathogen infections","authors":"Min Ge, Zesong Ruan, Ya-Xuan Zhu, Wencheng Wu, Chuang Yang, Han Lin, Jianlin Shi","doi":"10.1126/sciadv.adp3976","DOIUrl":"10.1126/sciadv.adp3976","url":null,"abstract":"<div >In the competition between the pathogen infection and the host defense, infectious microorganisms may enter the host cells by evading host defense mechanisms and use the intracellular biomolecules as replication nutrient. Among them, intracellular <i>Staphylococcus aureus</i> relies on the host cells to protect itself from the attacks by antibiotics or immune system to achieve long-term colonization in the host, and the consequent clinical therapeutic failures and relapses after antibiotic treatment. Here, we demonstrate that intracellular <i>S. aureus</i> surviving well even in the presence of vancomycin can be effectively eliminated using an emerging cell-mimicking therapeutic strategy. These cell mimics with natural killer cell–like activity (NKMs) are composed of a redox-responsive degradable carrier, and perforin and granzyme B within the carrier. NKMs perform far more effectivly than clinical antibiotics in treating intracellular bacterial infections, providing a direct evidence of the NK cell–mimicking immune mechanism in the treatment of intracellular <i>S. aureus</i>.</div>","PeriodicalId":21609,"journal":{"name":"Science Advances","volume":null,"pages":null},"PeriodicalIF":11.7,"publicationDate":"2024-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.science.org/doi/reader/10.1126/sciadv.adp3976","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142547186","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Paul Scesa, Helen Nguyen, Paige Weiss, Alejandra P. Rodriguez, Matthew Garchow, Shannon I. Ohlemacher, Evangelia Prappas, Serena A. Caplins, Carole A. Bewley, Laine Bohnert, Amanda J. Zellmer, Eric M. Wood, Eric W. Schmidt, Patrick J. Krug
Secondary metabolites often function as antipredator defenses, but when bioactive at low concentrations, their off-target effects on other organisms may be overlooked. Candidate “keystone molecules” are proposed to affect community structure and ecosystem functions, generally originating as defenses of primary producers; the broader effects of animal chemistry remain largely unexplored, however. Here, we characterize five previously unreported polyketides (alderenes A to E) biosynthesized by sea slugs reaching exceptional densities (up to 9000 slugs per square meter) in Northern Hemisphere estuaries. Alderenes comprise only 0.1% of slug wet weight, yet rendered live slugs or dead flesh unpalatable to three co-occurring consumers, making a potential food resource unavailable and redirecting energy flow in critical nursery habitat. Alderenes also displaced infauna from the upper sediment of the mudflat but attracted ovipositing snails. By altering communities, such compounds may have unexpected cascading effects on processes ranging from bioturbation to reproduction of species not obviously connected to the producing organisms, warranting greater attention by ecologists.
{"title":"Defensive polyketides produced by an abundant gastropod are candidate keystone molecules in estuarine ecology","authors":"Paul Scesa, Helen Nguyen, Paige Weiss, Alejandra P. Rodriguez, Matthew Garchow, Shannon I. Ohlemacher, Evangelia Prappas, Serena A. Caplins, Carole A. Bewley, Laine Bohnert, Amanda J. Zellmer, Eric M. Wood, Eric W. Schmidt, Patrick J. Krug","doi":"10.1126/sciadv.adp8643","DOIUrl":"10.1126/sciadv.adp8643","url":null,"abstract":"<div >Secondary metabolites often function as antipredator defenses, but when bioactive at low concentrations, their off-target effects on other organisms may be overlooked. Candidate “keystone molecules” are proposed to affect community structure and ecosystem functions, generally originating as defenses of primary producers; the broader effects of animal chemistry remain largely unexplored, however. Here, we characterize five previously unreported polyketides (alderenes A to E) biosynthesized by sea slugs reaching exceptional densities (up to 9000 slugs per square meter) in Northern Hemisphere estuaries. Alderenes comprise only 0.1% of slug wet weight, yet rendered live slugs or dead flesh unpalatable to three co-occurring consumers, making a potential food resource unavailable and redirecting energy flow in critical nursery habitat. Alderenes also displaced infauna from the upper sediment of the mudflat but attracted ovipositing snails. By altering communities, such compounds may have unexpected cascading effects on processes ranging from bioturbation to reproduction of species not obviously connected to the producing organisms, warranting greater attention by ecologists.</div>","PeriodicalId":21609,"journal":{"name":"Science Advances","volume":null,"pages":null},"PeriodicalIF":11.7,"publicationDate":"2024-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.science.org/doi/reader/10.1126/sciadv.adp8643","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142547194","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Elena Beanato, Hyuk-June Moon, Fabienne Windel, Pierre Vassiliadis, Maximillian J. Wessel, Traian Popa, Menoud Pauline, Esra Neufeld, Emanuela De Falco, Baptiste Gauthier, Melanie Steiner, Olaf Blanke, Friedhelm C. Hummel
Because of the depth of the hippocampal-entorhinal complex (HC-EC) in the brain, understanding of its role in spatial navigation via neuromodulation was limited in humans. Here, we aimed to better elucidate this relationship in healthy volunteers, using transcranial temporal interference electric stimulation (tTIS), a noninvasive technique allowing to selectively neuromodulate deep brain structures. We applied tTIS to the right HC-EC in either continuous or intermittent theta-burst stimulation patterns (cTBS or iTBS), compared to a control condition, during a virtual reality–based spatial navigation task and concomitant functional magnetic resonance imaging. iTBS improved spatial navigation performance, correlated with hippocampal activity modulation, and decreased grid cell–like activity in EC. Collectively, these data provide the evidence that human HC-EC activity can be directly and noninvasively modulated leading to changes of spatial navigation behavior. These findings suggest promising perspectives for patients suffering from cognitive impairment such as following traumatic brain injury or dementia.
{"title":"Noninvasive modulation of the hippocampal-entorhinal complex during spatial navigation in humans","authors":"Elena Beanato, Hyuk-June Moon, Fabienne Windel, Pierre Vassiliadis, Maximillian J. Wessel, Traian Popa, Menoud Pauline, Esra Neufeld, Emanuela De Falco, Baptiste Gauthier, Melanie Steiner, Olaf Blanke, Friedhelm C. Hummel","doi":"10.1126/sciadv.ado4103","DOIUrl":"10.1126/sciadv.ado4103","url":null,"abstract":"<div >Because of the depth of the hippocampal-entorhinal complex (HC-EC) in the brain, understanding of its role in spatial navigation via neuromodulation was limited in humans. Here, we aimed to better elucidate this relationship in healthy volunteers, using transcranial temporal interference electric stimulation (tTIS), a noninvasive technique allowing to selectively neuromodulate deep brain structures. We applied tTIS to the right HC-EC in either continuous or intermittent theta-burst stimulation patterns (cTBS or iTBS), compared to a control condition, during a virtual reality–based spatial navigation task and concomitant functional magnetic resonance imaging. iTBS improved spatial navigation performance, correlated with hippocampal activity modulation, and decreased grid cell–like activity in EC. Collectively, these data provide the evidence that human HC-EC activity can be directly and noninvasively modulated leading to changes of spatial navigation behavior. These findings suggest promising perspectives for patients suffering from cognitive impairment such as following traumatic brain injury or dementia.</div>","PeriodicalId":21609,"journal":{"name":"Science Advances","volume":null,"pages":null},"PeriodicalIF":11.7,"publicationDate":"2024-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.science.org/doi/reader/10.1126/sciadv.ado4103","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142547203","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Matteo Fiorenza, Johan Onslev, Carlos Henríquez-Olguín, Kaspar W. Persson, Sofie A. Hesselager, Thomas E. Jensen, Jørgen F. P. Wojtaszewski, Morten Hostrup, Jens Bangsbo
Preclinical models suggest mitochondria-derived oxidative stress as an underlying cause of insulin resistance. However, it remains unknown whether this pathophysiological mechanism is conserved in humans. Here, we used an invasive in vivo mechanistic approach to interrogate muscle insulin action while selectively manipulating the mitochondrial redox state in humans. To this end, we conducted insulin clamp studies combining intravenous infusion of a lipid overload with intake of a mitochondria-targeted antioxidant (mitoquinone). Under lipid overload, selective modulation of mitochondrial redox state by mitoquinone enhanced insulin-stimulated glucose uptake in skeletal muscle. Mechanistically, mitoquinone did not affect canonical insulin signaling but augmented insulin-stimulated glucose transporter type 4 (GLUT4) translocation while reducing the mitochondrial oxidative burden under lipid oversupply. Complementary ex vivo studies in human muscle fibers exposed to high intracellular lipid levels revealed that mitoquinone improves features of mitochondrial bioenergetics, including diminished mitochondrial H2O2 emission. These findings provide translational and mechanistic evidence implicating mitochondrial oxidants in the development of lipid-induced muscle insulin resistance in humans.
{"title":"Reducing the mitochondrial oxidative burden alleviates lipid-induced muscle insulin resistance in humans","authors":"Matteo Fiorenza, Johan Onslev, Carlos Henríquez-Olguín, Kaspar W. Persson, Sofie A. Hesselager, Thomas E. Jensen, Jørgen F. P. Wojtaszewski, Morten Hostrup, Jens Bangsbo","doi":"10.1126/sciadv.adq4461","DOIUrl":"10.1126/sciadv.adq4461","url":null,"abstract":"<div >Preclinical models suggest mitochondria-derived oxidative stress as an underlying cause of insulin resistance. However, it remains unknown whether this pathophysiological mechanism is conserved in humans. Here, we used an invasive in vivo mechanistic approach to interrogate muscle insulin action while selectively manipulating the mitochondrial redox state in humans. To this end, we conducted insulin clamp studies combining intravenous infusion of a lipid overload with intake of a mitochondria-targeted antioxidant (mitoquinone). Under lipid overload, selective modulation of mitochondrial redox state by mitoquinone enhanced insulin-stimulated glucose uptake in skeletal muscle. Mechanistically, mitoquinone did not affect canonical insulin signaling but augmented insulin-stimulated glucose transporter type 4 (GLUT4) translocation while reducing the mitochondrial oxidative burden under lipid oversupply. Complementary ex vivo studies in human muscle fibers exposed to high intracellular lipid levels revealed that mitoquinone improves features of mitochondrial bioenergetics, including diminished mitochondrial H<sub>2</sub>O<sub>2</sub> emission. These findings provide translational and mechanistic evidence implicating mitochondrial oxidants in the development of lipid-induced muscle insulin resistance in humans.</div>","PeriodicalId":21609,"journal":{"name":"Science Advances","volume":null,"pages":null},"PeriodicalIF":11.7,"publicationDate":"2024-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.science.org/doi/reader/10.1126/sciadv.adq4461","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142547215","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lakes emit substantial amounts of carbon dioxide (CO2) into the atmosphere, but why they do remains debated. The long-standing vision of lakes as solely respirators of the organic matter leaking from the soils has been challenged by evidence that inorganic carbon produced by weathering of the catchment bedrock could also support lake CO2 emissions. How inorganic carbon inputs ultimately generate lake CO2 outgassing remains a blind spot. We develop and introduce a calcite module in a coupled one-dimensional physical-biogeochemical model that we use to simulate the carbon cycle of the large Lake Geneva over the past 40 years. We mechanistically demonstrate how the so-far neglected process of calcite precipitation boosts net CO2 emissions at the annual scale. Far from being anecdotal, we show that calcite precipitation could explain CO2 outgassing across various lakes globally, including some of the largest lakes in the world.
{"title":"Calcite precipitation: The forgotten piece of lakes’ carbon cycle","authors":"Gaël Many, Nicolas Escoffier, Pascal Perolo, Fabian Bärenbold, Damien Bouffard, Marie-Elodie Perga","doi":"10.1126/sciadv.ado5924","DOIUrl":"10.1126/sciadv.ado5924","url":null,"abstract":"<div >Lakes emit substantial amounts of carbon dioxide (CO<sub>2</sub>) into the atmosphere, but why they do remains debated. The long-standing vision of lakes as solely respirators of the organic matter leaking from the soils has been challenged by evidence that inorganic carbon produced by weathering of the catchment bedrock could also support lake CO<sub>2</sub> emissions. How inorganic carbon inputs ultimately generate lake CO<sub>2</sub> outgassing remains a blind spot. We develop and introduce a calcite module in a coupled one-dimensional physical-biogeochemical model that we use to simulate the carbon cycle of the large Lake Geneva over the past 40 years. We mechanistically demonstrate how the so-far neglected process of calcite precipitation boosts net CO<sub>2</sub> emissions at the annual scale. Far from being anecdotal, we show that calcite precipitation could explain CO<sub>2</sub> outgassing across various lakes globally, including some of the largest lakes in the world.</div>","PeriodicalId":21609,"journal":{"name":"Science Advances","volume":null,"pages":null},"PeriodicalIF":11.7,"publicationDate":"2024-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.science.org/doi/reader/10.1126/sciadv.ado5924","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142547188","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Qiang Shen, Kiichi Murakami, Valentin Sotov, Marcus Butler, Pamela S. Ohashi, Michael Reedijk
Aberrant Notch, which is a defining feature of triple-negative breast cancer (TNBC) cells, regulates intercellular communication in the tumor immune microenvironment (TIME). This includes tumor-associated macrophage (TAM) recruitment through Notch-dependent cytokine secretion, contributing to an immunosuppressive TIME. Despite the low response rate of TNBC to immune checkpoint blockade (ICB), here, we report that inhibition of Notch-driven cytokine-mediated programs reduces TAMs and induces responsiveness to sequentially delivered ICB. This is characterized by the emergence of GrB+ cytotoxic T lymphocytes (CTLs) in the primary tumor. A more impressive effect of sequential treatment is observed in the lung where TAM depletion and increased CTLs are accompanied by near-complete abolition of metastases. This is due to (i) therapeutic reduction in Notch-dependent, prometastatic circulating factors released by the primary tumor, and (ii) elevated PD ligand 1 (PD-L1) in lung metastases, rendering them profoundly sensitive to ICB. These findings highlight the potential of combination cytokine inhibition and ICB as an immunotherapeutic strategy in TNBC.
{"title":"Inhibition of Notch enhances efficacy of immune checkpoint blockade in triple-negative breast cancer","authors":"Qiang Shen, Kiichi Murakami, Valentin Sotov, Marcus Butler, Pamela S. Ohashi, Michael Reedijk","doi":"10.1126/sciadv.ado8275","DOIUrl":"10.1126/sciadv.ado8275","url":null,"abstract":"<div >Aberrant Notch, which is a defining feature of triple-negative breast cancer (TNBC) cells, regulates intercellular communication in the tumor immune microenvironment (TIME). This includes tumor-associated macrophage (TAM) recruitment through Notch-dependent cytokine secretion, contributing to an immunosuppressive TIME. Despite the low response rate of TNBC to immune checkpoint blockade (ICB), here, we report that inhibition of Notch-driven cytokine-mediated programs reduces TAMs and induces responsiveness to sequentially delivered ICB. This is characterized by the emergence of GrB<sup>+</sup> cytotoxic T lymphocytes (CTLs) in the primary tumor. A more impressive effect of sequential treatment is observed in the lung where TAM depletion and increased CTLs are accompanied by near-complete abolition of metastases. This is due to (i) therapeutic reduction in Notch-dependent, prometastatic circulating factors released by the primary tumor, and (ii) elevated PD ligand 1 (PD-L1) in lung metastases, rendering them profoundly sensitive to ICB. These findings highlight the potential of combination cytokine inhibition and ICB as an immunotherapeutic strategy in TNBC.</div>","PeriodicalId":21609,"journal":{"name":"Science Advances","volume":null,"pages":null},"PeriodicalIF":11.7,"publicationDate":"2024-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.science.org/doi/reader/10.1126/sciadv.ado8275","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142547198","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Asmaysinh A. Gharia, Clinton J. Bradfield, Elise P. W. Jenkins, Iain D. C. Fraser, George G. Malliaras
Precise and efficient delivery of macromolecules into cells enhances basic biology research and therapeutic applications in cell therapies, drug delivery, and personalized medicine. While pulsed electric field electroporation effectively permeabilizes cell membranes to deliver payloads without the need for toxic chemical or viral transduction agents, conventional bulk electroporation devices face major challenges with cell viability and heterogeneity due to variations in fields generated across cells and electrochemistry at the electrode-electrolyte interface. Here, we introduce the use of microfabricated electrodes based on the conducting polymer poly(3,4-ethylenedioxythiophene) doped with polystyrene sulfonate (PEDOT:PSS), which substantially increases cell viability and transfection efficiency. As a proof of concept, we demonstrate the enhanced delivery of Cas9 protein, guide RNA, and plasmid DNA into cell lines and primary cells. This use of PEDOT:PSS enables rapid modification of difficult-to-transfect cell types to accelerate their study and use as therapeutic platforms.
将大分子精确、高效地输送到细胞中，可促进基础生物学研究以及细胞疗法、药物输送和个性化医疗方面的治疗应用。脉冲电场电穿孔能有效渗透细胞膜，无需使用有毒的化学或病毒转导剂就能递送有效载荷，但传统的散装电穿孔设备在细胞存活率和异质性方面面临着重大挑战，这是因为跨细胞产生的电场和电极-电解质界面的电化学存在差异。在这里，我们介绍了基于掺杂聚苯乙烯磺酸盐的导电聚合物聚（3,4-亚乙二氧基噻吩）（PEDOT:PSS）的微细加工电极的使用，这种电极可大幅提高细胞存活率和转染效率。作为概念验证，我们展示了向细胞系和原代细胞输送 Cas9 蛋白、引导 RNA 和质粒 DNA 的增强效果。使用 PEDOT:PSS 可以快速改造难以转染的细胞类型，从而加快研究并将其用作治疗平台。
{"title":"Efficient electroporation in primary cells with PEDOT:PSS electrodes","authors":"Asmaysinh A. Gharia, Clinton J. Bradfield, Elise P. W. Jenkins, Iain D. C. Fraser, George G. Malliaras","doi":"10.1126/sciadv.ado5042","DOIUrl":"10.1126/sciadv.ado5042","url":null,"abstract":"<div >Precise and efficient delivery of macromolecules into cells enhances basic biology research and therapeutic applications in cell therapies, drug delivery, and personalized medicine. While pulsed electric field electroporation effectively permeabilizes cell membranes to deliver payloads without the need for toxic chemical or viral transduction agents, conventional bulk electroporation devices face major challenges with cell viability and heterogeneity due to variations in fields generated across cells and electrochemistry at the electrode-electrolyte interface. Here, we introduce the use of microfabricated electrodes based on the conducting polymer poly(3,4-ethylenedioxythiophene) doped with polystyrene sulfonate (PEDOT:PSS), which substantially increases cell viability and transfection efficiency. As a proof of concept, we demonstrate the enhanced delivery of Cas9 protein, guide RNA, and plasmid DNA into cell lines and primary cells. This use of PEDOT:PSS enables rapid modification of difficult-to-transfect cell types to accelerate their study and use as therapeutic platforms.</div>","PeriodicalId":21609,"journal":{"name":"Science Advances","volume":null,"pages":null},"PeriodicalIF":11.7,"publicationDate":"2024-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.science.org/doi/reader/10.1126/sciadv.ado5042","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142489375","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Diego Bernal-Bernal, David Pantoja-Uceda, Jorge Pedro López-Alonso, Alfonso López-Rojo, José Antonio López-Ruiz, Marisa Galbis-Martínez, Borja Ochoa-Lizarralde, Igor Tascón, Montserrat Elías-Arnanz, Iban Ubarretxena-Belandia, S. Padmanabhan
How CRISPR-Cas and cyclic oligonucleotide–based antiphage signaling systems (CBASS) are coordinately deployed against invaders remains unclear. We show that a locus containing two CBASS and one type III-B CRISPR-Cas system, regulated by the transmembrane anti-σ DdvA and its cognate extracytoplasmic function (ECF) σ DdvS, can defend Myxococcus xanthus against a phage. Cryo–electron microscopy reveals DdvA-DdvS pairs assemble as arrow-shaped transmembrane dimers. Each DdvA periplasmic domain adopts a separase/craspase-type tetratricopeptide repeat (TPR)–caspase HetF–associated with TPR (TPR-CHAT) architecture with an incomplete His-Cys active site, lacking three α-helices conserved among CHAT domains. Each active site faces the dimer interface, raising the possibility that signal-induced caspase-like DdvA autoproteolysis in trans precedes RseP-mediated intramembrane proteolysis and DdvS release. Nuclear magnetic resonance reveals a DdvA cytoplasmic CHCC-type zinc-bound three-helix bundle that binds to DdvS σ2 and σ4 domains, undergoing σ4-induced helix extension to trap DdvS. Altogether, we provide structural-mechanistic insights into membrane anti–σ-ECF σ regulation of an antiviral CBASS-CRISPR-Cas defense island.
{"title":"Structural basis for regulation of a CBASS-CRISPR-Cas defense island by a transmembrane anti-σ factor and its ECF σ partner","authors":"Diego Bernal-Bernal, David Pantoja-Uceda, Jorge Pedro López-Alonso, Alfonso López-Rojo, José Antonio López-Ruiz, Marisa Galbis-Martínez, Borja Ochoa-Lizarralde, Igor Tascón, Montserrat Elías-Arnanz, Iban Ubarretxena-Belandia, S. Padmanabhan","doi":"10.1126/sciadv.adp1053","DOIUrl":"10.1126/sciadv.adp1053","url":null,"abstract":"<div >How CRISPR-Cas and cyclic oligonucleotide–based antiphage signaling systems (CBASS) are coordinately deployed against invaders remains unclear. We show that a locus containing two CBASS and one type III-B CRISPR-Cas system, regulated by the transmembrane anti-σ DdvA and its cognate extracytoplasmic function (ECF) σ DdvS, can defend <i>Myxococcus xanthus</i> against a phage. Cryo–electron microscopy reveals DdvA-DdvS pairs assemble as arrow-shaped transmembrane dimers. Each DdvA periplasmic domain adopts a separase/craspase-type tetratricopeptide repeat (TPR)–caspase HetF–associated with TPR (TPR-CHAT) architecture with an incomplete His-Cys active site, lacking three α-helices conserved among CHAT domains. Each active site faces the dimer interface, raising the possibility that signal-induced caspase-like DdvA autoproteolysis in trans precedes RseP-mediated intramembrane proteolysis and DdvS release. Nuclear magnetic resonance reveals a DdvA cytoplasmic CHCC-type zinc-bound three-helix bundle that binds to DdvS σ<sub>2</sub> and σ<sub>4</sub> domains, undergoing σ<sub>4</sub>-induced helix extension to trap DdvS. Altogether, we provide structural-mechanistic insights into membrane anti–σ-ECF σ regulation of an antiviral CBASS-CRISPR-Cas defense island.</div>","PeriodicalId":21609,"journal":{"name":"Science Advances","volume":null,"pages":null},"PeriodicalIF":11.7,"publicationDate":"2024-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.science.org/doi/reader/10.1126/sciadv.adp1053","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142489383","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
扫码关注我们
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号