Pub Date : 2024-08-08DOI: 10.1172/jci.insight.179680
J Natalie Howard, Thomas D Zaikos, Callie Levinger, Esteban Rivera, Elyse K McMahon, Carissa S Holmberg, Joshua Terao, Marta Sanz, Dennis C Copertino, Weisheng Wang, Natalia Soriano-Sarabia, R Brad Jones, Alberto Bosque
Nonreceptor tyrosine phosphatases (NTPs) play an important role regulating protein phosphorylation and have been proposed as attractive therapeutic targets for cancer and metabolic diseases. We have previously identified that 3-Hydroxy-1,2,3-benzotriazin-4(3H)-one (HODHBt) enhanced STAT activation upon cytokine stimulation leading to increased reactivation of latent HIV and effector functions of NK and CD8 T cells. Here, we demonstrated that HODHBt interacts with and inhibits the NTPs PTPN1 and PTPN2 through a mixed inhibition mechanism. We also confirmed that PTPN1 and PTPN2 specifically control the phosphorylation of different STATs. The small molecule ABBV-CLS-484 (AC-484) is an active site inhibitor of PTPN1 and PTPN2 currently in clinical trials for advanced solid tumors. We compared AC-484 and HODHBt and found similar effects on STAT5 and immune activation albeit with different mechanisms of action leading to varying effects on latency reversal. Our studies provide the first specific evidence that enhancing STAT phosphorylation via inhibition of PTPN1 and PTPN2 is an effective tool against HIV.
非受体酪氨酸磷酸酶(NTPs)在调节蛋白质磷酸化方面发挥着重要作用,并被认为是治疗癌症和代谢性疾病的有吸引力的靶点。我们之前发现,3-羟基-1,2,3-苯并三嗪-4(3H)-酮(HODHBt)在细胞因子刺激下可增强 STAT 的活化,从而增加潜伏 HIV 的再激活以及 NK 和 CD8 T 细胞的效应功能。在这里,我们证明了 HODHBt 通过混合抑制机制与 NTPs PTPN1 和 PTPN2 相互作用并对其产生抑制作用。我们还证实,PTPN1 和 PTPN2 能特异性地控制不同 STATs 的磷酸化。小分子 ABBV-CLS-484(AC-484)是 PTPN1 和 PTPN2 的活性位点抑制剂,目前正在进行晚期实体瘤的临床试验。我们比较了 AC-484 和 HODHBt,发现它们对 STAT5 和免疫激活的作用相似,但作用机制不同,导致潜伏期逆转的效果也不同。我们的研究首次提供了具体证据,证明通过抑制 PTPN1 和 PTPN2 来增强 STAT 磷酸化是抗击 HIV 的有效工具。
{"title":"The HIV latency reversing agent HODHBt inhibits the phosphatases PTPN1 and PTPN2.","authors":"J Natalie Howard, Thomas D Zaikos, Callie Levinger, Esteban Rivera, Elyse K McMahon, Carissa S Holmberg, Joshua Terao, Marta Sanz, Dennis C Copertino, Weisheng Wang, Natalia Soriano-Sarabia, R Brad Jones, Alberto Bosque","doi":"10.1172/jci.insight.179680","DOIUrl":"https://doi.org/10.1172/jci.insight.179680","url":null,"abstract":"<p><p>Nonreceptor tyrosine phosphatases (NTPs) play an important role regulating protein phosphorylation and have been proposed as attractive therapeutic targets for cancer and metabolic diseases. We have previously identified that 3-Hydroxy-1,2,3-benzotriazin-4(3H)-one (HODHBt) enhanced STAT activation upon cytokine stimulation leading to increased reactivation of latent HIV and effector functions of NK and CD8 T cells. Here, we demonstrated that HODHBt interacts with and inhibits the NTPs PTPN1 and PTPN2 through a mixed inhibition mechanism. We also confirmed that PTPN1 and PTPN2 specifically control the phosphorylation of different STATs. The small molecule ABBV-CLS-484 (AC-484) is an active site inhibitor of PTPN1 and PTPN2 currently in clinical trials for advanced solid tumors. We compared AC-484 and HODHBt and found similar effects on STAT5 and immune activation albeit with different mechanisms of action leading to varying effects on latency reversal. Our studies provide the first specific evidence that enhancing STAT phosphorylation via inhibition of PTPN1 and PTPN2 is an effective tool against HIV.</p>","PeriodicalId":14722,"journal":{"name":"JCI insight","volume":null,"pages":null},"PeriodicalIF":6.3,"publicationDate":"2024-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141906679","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The interleukin-17 (IL-17) family of cytokines has emerged as a critical player in autoimmune disease, including systemic lupus erythematosus (SLE). However, the role of IL-17B, a poorly understood cytokine, in the pathogenesis of SLE is still not clear. In this study, we investigated the role of IL-17B in the activation and differentiation of B cells, and the pathogenesis of SLE. Intriguingly, IL-17B deficiency aggravated disease in lupus-prone mice and promoted the activation of B cells and the differentiation of germinal center (GC) B cells and plasma cells, while recombinant mouse IL-17B (rmIL-17B) significantly alleviated disease in lupus-prone mice. Mechanistically, rmIL-17B inhibited the activation of the Toll-like receptor (TLR) and interferon (IFN) pathways in B cells by downregulating the FASN-mediated lipid metabolism. Loss of FASN significantly alleviated the disease in lupus-prone mice and inhibited the activation and differentiation of B cells. In addition, B cells had greater FASN expression and lower IL-17RB levels in patients with SLE than in healthy controls. Our study described the role of IL-17B in regulating B-cell activation and differentiation, and alleviating the onset of SLE. These findings will lay a theoretical foundation for further understanding of the pathogenesis of SLE.
白细胞介素-17(IL-17)家族细胞因子已成为自身免疫性疾病(包括系统性红斑狼疮)的关键因素。然而,IL-17B 这种鲜为人知的细胞因子在系统性红斑狼疮发病机制中的作用仍不明确。在这项研究中,我们探讨了 IL-17B 在 B 细胞活化和分化以及系统性红斑狼疮发病机制中的作用。有趣的是,IL-17B缺乏会加重狼疮易感小鼠的病情,并促进B细胞的活化和生殖中心(GC)B细胞及浆细胞的分化,而重组小鼠IL-17B(rmIL-17B)则能显著缓解狼疮易感小鼠的病情。从机理上讲,rmIL-17B 通过下调 FASN 介导的脂质代谢,抑制了 B 细胞中 Toll 样受体(TLR)和干扰素(IFN)通路的激活。FASN的缺失能明显缓解红斑狼疮易感小鼠的病情,并抑制B细胞的活化和分化。此外,与健康对照组相比,系统性红斑狼疮患者的 B 细胞中 FASN 表达更多,IL-17RB 水平更低。我们的研究描述了 IL-17B 在调节 B 细胞活化和分化以及缓解系统性红斑狼疮发病方面的作用。这些发现将为进一步了解系统性红斑狼疮的发病机制奠定理论基础。
{"title":"IL-17B alleviates the pathogenesis of systemic lupus erythematosus by inhibiting FASN-mediated differentiation of B cells.","authors":"Yucai Xiao, Yuxin Hu, Yangzhe Gao, Lin Wang, Lili Zhang, Qun Ma, Zhaochen Ning, Lu Yu, Haochen Li, Jiakun Liu, Junyu Wang, Yonghong Yang, Huabao Xiong, Guanjun Dong","doi":"10.1172/jci.insight.181906","DOIUrl":"https://doi.org/10.1172/jci.insight.181906","url":null,"abstract":"<p><p>The interleukin-17 (IL-17) family of cytokines has emerged as a critical player in autoimmune disease, including systemic lupus erythematosus (SLE). However, the role of IL-17B, a poorly understood cytokine, in the pathogenesis of SLE is still not clear. In this study, we investigated the role of IL-17B in the activation and differentiation of B cells, and the pathogenesis of SLE. Intriguingly, IL-17B deficiency aggravated disease in lupus-prone mice and promoted the activation of B cells and the differentiation of germinal center (GC) B cells and plasma cells, while recombinant mouse IL-17B (rmIL-17B) significantly alleviated disease in lupus-prone mice. Mechanistically, rmIL-17B inhibited the activation of the Toll-like receptor (TLR) and interferon (IFN) pathways in B cells by downregulating the FASN-mediated lipid metabolism. Loss of FASN significantly alleviated the disease in lupus-prone mice and inhibited the activation and differentiation of B cells. In addition, B cells had greater FASN expression and lower IL-17RB levels in patients with SLE than in healthy controls. Our study described the role of IL-17B in regulating B-cell activation and differentiation, and alleviating the onset of SLE. These findings will lay a theoretical foundation for further understanding of the pathogenesis of SLE.</p>","PeriodicalId":14722,"journal":{"name":"JCI insight","volume":null,"pages":null},"PeriodicalIF":6.3,"publicationDate":"2024-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141906677","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-08DOI: 10.1172/jci.insight.181968
Gabriela M Webb, Kristin A Sauter, Diana Takahashi, Melissa Kirigiti, Lindsay Bader, Sarah R Lindsley, Hannah M Blomenkamp, Cicely Zaro, Molly Shallman, Casey M McGuire, Heather Hofmeister, Uriel Avila, Cleiton Pessoa, Joseph M Hwang, Allyson J McCullen, Matthew Humkey, Jason Reed, Lina Gao, Lee Winchester, Courtney V Fletcher, Oleg Varlamov, Todd T Brown, Jonah B Sacha, Paul Kievit, Charles T Roberts
Current antiretroviral therapy (ART) regimens efficiently limit HIV replication, thereby improving life expectancy of people living with HIV, but also cause metabolic side effects. The ongoing obesity epidemic has resulted in more people with metabolic comorbidities at the time of HIV infection, yet the impact of pre-existing metabolic dysregulation on infection sequelae and response to ART is unclear. Here, to investigate the impact of preexisting obesity and insulin resistance on acute infection and subsequent long-term ART, we infected a cohort of lean and obese adult male macaques with SIV and administered ART. The responses of lean and obese macaques to SIV and ART were similar with respect to plasma and cell-associated viral loads, ART drug levels in plasma and tissues, SIV-specific immune responses, adipose tissue and islet morphology, and colon inflammation, with baseline differences between lean and obese groups largely maintained. Both groups exhibited a striking depletion of CD4+ T cells from adipose tissue that did not recover with ART. However, differential responses to SIV and ART were observed for body weight, omental adipocyte size, and the adiponectin/leptin ratio, a marker of cardiometabolic risk. Thus, obesity and insulin resistance had limited effects on multiple responses to acute SIV infection and ART, while several factors that underlie long-term metabolic comorbidities were influenced by prior obesity and insulin resistance. These studies provide the foundation for future investigations into the efficacy of adjunct therapies such as metformin and glucagon-like peptide-1 receptor agonists in the prevention of metabolic comorbidities in people living with HIV.
{"title":"Effect of metabolic status on response to SIV infection and antiretroviral therapy in nonhuman primates.","authors":"Gabriela M Webb, Kristin A Sauter, Diana Takahashi, Melissa Kirigiti, Lindsay Bader, Sarah R Lindsley, Hannah M Blomenkamp, Cicely Zaro, Molly Shallman, Casey M McGuire, Heather Hofmeister, Uriel Avila, Cleiton Pessoa, Joseph M Hwang, Allyson J McCullen, Matthew Humkey, Jason Reed, Lina Gao, Lee Winchester, Courtney V Fletcher, Oleg Varlamov, Todd T Brown, Jonah B Sacha, Paul Kievit, Charles T Roberts","doi":"10.1172/jci.insight.181968","DOIUrl":"https://doi.org/10.1172/jci.insight.181968","url":null,"abstract":"<p><p>Current antiretroviral therapy (ART) regimens efficiently limit HIV replication, thereby improving life expectancy of people living with HIV, but also cause metabolic side effects. The ongoing obesity epidemic has resulted in more people with metabolic comorbidities at the time of HIV infection, yet the impact of pre-existing metabolic dysregulation on infection sequelae and response to ART is unclear. Here, to investigate the impact of preexisting obesity and insulin resistance on acute infection and subsequent long-term ART, we infected a cohort of lean and obese adult male macaques with SIV and administered ART. The responses of lean and obese macaques to SIV and ART were similar with respect to plasma and cell-associated viral loads, ART drug levels in plasma and tissues, SIV-specific immune responses, adipose tissue and islet morphology, and colon inflammation, with baseline differences between lean and obese groups largely maintained. Both groups exhibited a striking depletion of CD4+ T cells from adipose tissue that did not recover with ART. However, differential responses to SIV and ART were observed for body weight, omental adipocyte size, and the adiponectin/leptin ratio, a marker of cardiometabolic risk. Thus, obesity and insulin resistance had limited effects on multiple responses to acute SIV infection and ART, while several factors that underlie long-term metabolic comorbidities were influenced by prior obesity and insulin resistance. These studies provide the foundation for future investigations into the efficacy of adjunct therapies such as metformin and glucagon-like peptide-1 receptor agonists in the prevention of metabolic comorbidities in people living with HIV.</p>","PeriodicalId":14722,"journal":{"name":"JCI insight","volume":null,"pages":null},"PeriodicalIF":6.3,"publicationDate":"2024-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141906675","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-08DOI: 10.1172/jci.insight.178050
Sophia M DiCesare, Antonio J Ortega, Gracen E Collier, Steffi Daniel, Krista N Thompson, Melissa K McCoy, Bruce A Posner, John D Hulleman
Malattia Leventinese/Doyne honeycomb retinal dystrophy (ML/DHRD) is an age-related macular degeneration-like (AMD-like) retinal dystrophy caused by an autosomal dominant R345W mutation in the secreted glycoprotein, fibulin-3 (F3). To identify new small molecules that reduce F3 production in retinal pigmented epithelium (RPE) cells, we knocked-in a luminescent peptide tag (HiBiT) into the endogenous F3 locus that enabled simple, sensitive, and high-throughput detection of the protein. The GSK3 inhibitor, CHIR99021 (CHIR), significantly reduced F3 burden (expression, secretion, and intracellular levels) in immortalized RPE and non-RPE cells. Low-level, long-term CHIR treatment promoted remodeling of the RPE extracellular matrix, reducing sub-RPE deposit-associated proteins (e.g., amelotin, complement component 3, collagen IV, and fibronectin), while increasing RPE differentiation factors (e.g., tyrosinase, and pigment epithelium-derived factor). In vivo, treatment of 8-month-old R345W+/+ knockin mice with CHIR (25 mg/kg i.p., 1 mo) was well tolerated and significantly reduced R345W F3-associated AMD-like basal laminar deposit number and size, thereby preventing the main pathological feature in these mice. This is an important demonstration of small molecule-based prevention of AMD-like pathology in ML/DHRD mice and may herald a rejuvenation of interest in GSK3 inhibition for the treatment of retinal degenerative diseases, including potentially AMD itself.
{"title":"GSK3 inhibition reduces ECM production and prevents age-related macular degeneration-like pathology.","authors":"Sophia M DiCesare, Antonio J Ortega, Gracen E Collier, Steffi Daniel, Krista N Thompson, Melissa K McCoy, Bruce A Posner, John D Hulleman","doi":"10.1172/jci.insight.178050","DOIUrl":"10.1172/jci.insight.178050","url":null,"abstract":"<p><p>Malattia Leventinese/Doyne honeycomb retinal dystrophy (ML/DHRD) is an age-related macular degeneration-like (AMD-like) retinal dystrophy caused by an autosomal dominant R345W mutation in the secreted glycoprotein, fibulin-3 (F3). To identify new small molecules that reduce F3 production in retinal pigmented epithelium (RPE) cells, we knocked-in a luminescent peptide tag (HiBiT) into the endogenous F3 locus that enabled simple, sensitive, and high-throughput detection of the protein. The GSK3 inhibitor, CHIR99021 (CHIR), significantly reduced F3 burden (expression, secretion, and intracellular levels) in immortalized RPE and non-RPE cells. Low-level, long-term CHIR treatment promoted remodeling of the RPE extracellular matrix, reducing sub-RPE deposit-associated proteins (e.g., amelotin, complement component 3, collagen IV, and fibronectin), while increasing RPE differentiation factors (e.g., tyrosinase, and pigment epithelium-derived factor). In vivo, treatment of 8-month-old R345W+/+ knockin mice with CHIR (25 mg/kg i.p., 1 mo) was well tolerated and significantly reduced R345W F3-associated AMD-like basal laminar deposit number and size, thereby preventing the main pathological feature in these mice. This is an important demonstration of small molecule-based prevention of AMD-like pathology in ML/DHRD mice and may herald a rejuvenation of interest in GSK3 inhibition for the treatment of retinal degenerative diseases, including potentially AMD itself.</p>","PeriodicalId":14722,"journal":{"name":"JCI insight","volume":null,"pages":null},"PeriodicalIF":6.3,"publicationDate":"2024-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141901816","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Organelle stress exacerbates podocyte injury, contributing to perturbed lipid metabolism. Simultaneous organelle stresses occur in kidney tissues; therefore, a thorough analysis of organelle communication is crucial for understanding the progression of kidney diseases. Although organelles closely interact with one another at membrane contact sites, limited studies have explored their involvement in kidney homeostasis. The endoplasmic reticulum (ER) protein, PDZ domain-containing 8 (PDZD8), is implicated in multiple organelle tethering processes and cellular lipid homeostasis. In this study, we aimed to elucidate the role of organelle communication in podocyte injury using podocyte-specific Pdzd8-knockout mice. Our findings demonstrated that Pdzd8 deletion exacerbated podocyte injury in an accelerated obesity-related kidney disease model. Proteomic analysis of isolated glomeruli revealed that Pdzd8 deletion exacerbated mitochondrial and endosomal dysfunction during podocyte lipotoxicity. Additionally, electron microscopy revealed the accumulation of "fatty abnormal endosomes" in Pdzd8-deficient podocytes during obesity-related kidney diseases. Lipidomic analysis indicated that glucosylceramide accumulated in Pdzd8-deficient podocytes, owing to accelerated production and decelerated degradation. Thus, the organelle-tethering factor, PDZD8, plays a crucial role in maintaining mitochondrial and endosomal homeostasis during podocyte lipotoxicity. Collectively, our findings highlight the importance of organelle communication at the three-way junction among the ER, mitochondria, and endosomes in preserving podocyte homeostasis.
{"title":"Organelle communication maintains mitochondrial and endosomal homeostasis during podocyte lipotoxicity.","authors":"Sho Hasegawa, Masaomi Nangaku, Yuto Takenaka, Chigusa Kitayama, Qi Li, Madina Saipidin, Yu Ah Hong, Jin Shang, Yusuke Hirabayashi, Naoto Kubota, Takashi Kadowaki, Reiko Inagi","doi":"10.1172/jci.insight.182534","DOIUrl":"https://doi.org/10.1172/jci.insight.182534","url":null,"abstract":"<p><p>Organelle stress exacerbates podocyte injury, contributing to perturbed lipid metabolism. Simultaneous organelle stresses occur in kidney tissues; therefore, a thorough analysis of organelle communication is crucial for understanding the progression of kidney diseases. Although organelles closely interact with one another at membrane contact sites, limited studies have explored their involvement in kidney homeostasis. The endoplasmic reticulum (ER) protein, PDZ domain-containing 8 (PDZD8), is implicated in multiple organelle tethering processes and cellular lipid homeostasis. In this study, we aimed to elucidate the role of organelle communication in podocyte injury using podocyte-specific Pdzd8-knockout mice. Our findings demonstrated that Pdzd8 deletion exacerbated podocyte injury in an accelerated obesity-related kidney disease model. Proteomic analysis of isolated glomeruli revealed that Pdzd8 deletion exacerbated mitochondrial and endosomal dysfunction during podocyte lipotoxicity. Additionally, electron microscopy revealed the accumulation of \"fatty abnormal endosomes\" in Pdzd8-deficient podocytes during obesity-related kidney diseases. Lipidomic analysis indicated that glucosylceramide accumulated in Pdzd8-deficient podocytes, owing to accelerated production and decelerated degradation. Thus, the organelle-tethering factor, PDZD8, plays a crucial role in maintaining mitochondrial and endosomal homeostasis during podocyte lipotoxicity. Collectively, our findings highlight the importance of organelle communication at the three-way junction among the ER, mitochondria, and endosomes in preserving podocyte homeostasis.</p>","PeriodicalId":14722,"journal":{"name":"JCI insight","volume":null,"pages":null},"PeriodicalIF":6.3,"publicationDate":"2024-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141906678","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-06DOI: 10.1172/jci.insight.181228
David G Tingay, Monique Fatmous, Kelly Kenna, Jack Chapman, Ellen Douglas, Arun Sett, Qi Hui Poh, Sophia I Dahm, Tuyen Kim Quach, Magdy Sourial, Haoyun Fang, David W Greening, Prue M Pereira-Fantini
Gas flow is fundamental for driving tidal ventilation and thus the speed of lung motion, but current bias flow settings to support the preterm lung after birth are without an evidence base. We aimed to determine the role of gas bias flow rates to generate positive pressure ventilation in initiating early lung injury pathways in the preterm lamb. Using slower speeds to inflate the lung during tidal ventilation (gas flow rates 4-6 L/min) did not impact lung mechanics, mechanical power or gas exchange compared to those currently used in clinical practice (8-10 L/min). Speed of pressure and volume change during inflation were faster with higher flow rates. Lower flow rates resulted in less bronchoalveolar fluid protein, better lung morphology and fewer detached epithelial cells. Overall, relative to unventilated fetal controls, there was greater protein change using 8-10 L/min, which was associated with enrichment of acute inflammatory and innate responses. Slowing the speed of lung motion by supporting the preterm lung from birth with lower flow rates than currently used clinically resulted in less lung injury without compromising tidal ventilation or gas exchange.
{"title":"Speed of lung inflation at birth influences the initiation of lung injury in preterm lambs.","authors":"David G Tingay, Monique Fatmous, Kelly Kenna, Jack Chapman, Ellen Douglas, Arun Sett, Qi Hui Poh, Sophia I Dahm, Tuyen Kim Quach, Magdy Sourial, Haoyun Fang, David W Greening, Prue M Pereira-Fantini","doi":"10.1172/jci.insight.181228","DOIUrl":"https://doi.org/10.1172/jci.insight.181228","url":null,"abstract":"<p><p>Gas flow is fundamental for driving tidal ventilation and thus the speed of lung motion, but current bias flow settings to support the preterm lung after birth are without an evidence base. We aimed to determine the role of gas bias flow rates to generate positive pressure ventilation in initiating early lung injury pathways in the preterm lamb. Using slower speeds to inflate the lung during tidal ventilation (gas flow rates 4-6 L/min) did not impact lung mechanics, mechanical power or gas exchange compared to those currently used in clinical practice (8-10 L/min). Speed of pressure and volume change during inflation were faster with higher flow rates. Lower flow rates resulted in less bronchoalveolar fluid protein, better lung morphology and fewer detached epithelial cells. Overall, relative to unventilated fetal controls, there was greater protein change using 8-10 L/min, which was associated with enrichment of acute inflammatory and innate responses. Slowing the speed of lung motion by supporting the preterm lung from birth with lower flow rates than currently used clinically resulted in less lung injury without compromising tidal ventilation or gas exchange.</p>","PeriodicalId":14722,"journal":{"name":"JCI insight","volume":null,"pages":null},"PeriodicalIF":6.3,"publicationDate":"2024-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141897453","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-06DOI: 10.1172/jci.insight.179292
Nicole E Gross, Zhehao Zhang, Jacob T Mitchell, Soren Charmsaz, Alexei G Hernandez, Erin M Coyne, Sarah M Shin, Diana Carolina Vargas Carvajal, Dimitrios N Sidiropoulos, Yeonju Cho, Guanglan Mo, Xuan Yuan, Courtney Cannon, Jayalaxmi Suresh Babu, Melissa R Lyman, Todd Armstrong, Luciane T Kagohara, Katherine M Bever, Dung T Le, Elizabeth M Jaffee, Elana J Fertig, Won Jin Ho
Pancreatic ductal adenocarcinoma (PDAC) is highly lethal and resistant to immunotherapy. Although immune recognition can be enhanced with immunomodulatory agents including checkpoint inhibitors and vaccines, few patients experience clinical efficacy because the tumor immune microenvironment (TiME) is dominated by immunosuppressive myeloid cells that impose T cell inhibition. Inhibition of phosphodiesterase-5 (PDE5) was reported to downregulate metabolic regulators arginase and iNOS in immunosuppressive myeloid cells and enhance immunity against immune-sensitive tumors including head and neck cancers. We show for the first time that combining a PDE5 inhibitor, tadalafil, with a mesothelin-specific vaccine, anti-PD1, and anti-CTLA4 yields antitumor efficacy even against immune-resistant PDAC. To determine immunologic advantages conferred by tadalafil, we profiled the TiME using mass cytometry and single-cell RNA analysis with Domino to infer intercellular signaling. Our analyses demonstrated that tadalafil reprograms myeloid cells to be less immunosuppressive. Moreover, tadalafil synergized with the vaccine, enhancing T cell activation including mesothelin-specific T cells. Tadalafil treatment was also associated with myeloid-T cell signaling axes important for antitumor responses (e.g., Cxcr3, Il12). Our study shows that PDE5 inhibition combined with vaccine-based immunotherapy promotes pro-inflammatory states of myeloid cells, activation of T cells, and enhanced myeloid-T cell crosstalk to yield antitumor efficacy against immune-resistant PDAC.
胰腺导管腺癌(PDAC)的致死率很高,而且对免疫疗法具有抗药性。虽然包括检查点抑制剂和疫苗在内的免疫调节药物可以增强免疫识别能力,但由于肿瘤免疫微环境(TiME)由免疫抑制性髓系细胞主导,对T细胞产生抑制作用,因此临床疗效不佳的患者寥寥无几。据报道,抑制磷酸二酯酶-5(PDE5)可下调免疫抑制性髓系细胞中的代谢调节因子精氨酸酶和iNOS,增强对包括头颈部癌症在内的免疫敏感性肿瘤的免疫力。我们首次发现,将PDE5抑制剂他达拉非与间皮素特异性疫苗、抗PD1和抗CTLA4结合使用,即使对免疫耐受的PDAC也能产生抗肿瘤疗效。为了确定他达拉非带来的免疫优势,我们使用质谱细胞计数法和 Domino 单细胞 RNA 分析法对 TiME 进行了分析,以推断细胞间信号转导。我们的分析表明,他达拉非能重编程髓系细胞,降低其免疫抑制性。此外,他达拉非与疫苗协同作用,增强了T细胞的活化,包括间皮素特异性T细胞。他达拉非治疗还与对抗肿瘤反应很重要的骨髓-T细胞信号轴(如Cxcr3、Il12)有关。我们的研究表明,PDE5抑制与基于疫苗的免疫疗法相结合可促进髓系细胞的促炎状态、T细胞的活化以及髓系-T细胞串联的增强,从而对免疫耐受的PDAC产生抗肿瘤疗效。
{"title":"Phosphodiesterase-5 inhibition collaborates with vaccine-based immunotherapy to reprogram myeloid cells in pancreatic ductal adenocarcinoma.","authors":"Nicole E Gross, Zhehao Zhang, Jacob T Mitchell, Soren Charmsaz, Alexei G Hernandez, Erin M Coyne, Sarah M Shin, Diana Carolina Vargas Carvajal, Dimitrios N Sidiropoulos, Yeonju Cho, Guanglan Mo, Xuan Yuan, Courtney Cannon, Jayalaxmi Suresh Babu, Melissa R Lyman, Todd Armstrong, Luciane T Kagohara, Katherine M Bever, Dung T Le, Elizabeth M Jaffee, Elana J Fertig, Won Jin Ho","doi":"10.1172/jci.insight.179292","DOIUrl":"https://doi.org/10.1172/jci.insight.179292","url":null,"abstract":"<p><p>Pancreatic ductal adenocarcinoma (PDAC) is highly lethal and resistant to immunotherapy. Although immune recognition can be enhanced with immunomodulatory agents including checkpoint inhibitors and vaccines, few patients experience clinical efficacy because the tumor immune microenvironment (TiME) is dominated by immunosuppressive myeloid cells that impose T cell inhibition. Inhibition of phosphodiesterase-5 (PDE5) was reported to downregulate metabolic regulators arginase and iNOS in immunosuppressive myeloid cells and enhance immunity against immune-sensitive tumors including head and neck cancers. We show for the first time that combining a PDE5 inhibitor, tadalafil, with a mesothelin-specific vaccine, anti-PD1, and anti-CTLA4 yields antitumor efficacy even against immune-resistant PDAC. To determine immunologic advantages conferred by tadalafil, we profiled the TiME using mass cytometry and single-cell RNA analysis with Domino to infer intercellular signaling. Our analyses demonstrated that tadalafil reprograms myeloid cells to be less immunosuppressive. Moreover, tadalafil synergized with the vaccine, enhancing T cell activation including mesothelin-specific T cells. Tadalafil treatment was also associated with myeloid-T cell signaling axes important for antitumor responses (e.g., Cxcr3, Il12). Our study shows that PDE5 inhibition combined with vaccine-based immunotherapy promotes pro-inflammatory states of myeloid cells, activation of T cells, and enhanced myeloid-T cell crosstalk to yield antitumor efficacy against immune-resistant PDAC.</p>","PeriodicalId":14722,"journal":{"name":"JCI insight","volume":null,"pages":null},"PeriodicalIF":6.3,"publicationDate":"2024-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141897452","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Antigen presentation by Major Histocompatibility Complex Class I (MHC-I) is crucial for T-cell-mediated killing, and aberrant surface MHC-I expression is tightly associated with immune evasion. To address MHC-I downregulation, we conducted a high-throughput flow cytometry screen, identifying bleomycin (BLM) as a potent inducer of cell surface MHC-I expression. BLM-induced MHC-I augmentation renders tumor cells more susceptible to T cells in co-culture assays and enhances anti-tumor responses in an adoptive cellular transfer mouse model. Mechanistically, BLM remodels the tumor immune microenvironment, inducing MHC-I expression in an ATM/ATR-NF-κB-dependent manner. Furthermore, BLM improves T-cell-dependent immunotherapeutic approaches, including bispecific antibodies therapy, immune checkpoint therapy (ICT), and autologous tumor-infiltrating lymphocytes (TILs) therapy. Importantly, low-dose BLM treatment in mouse models amplified the anti-tumor effect of immunotherapy without detectable pulmonary toxicity. In summary, our findings repurpose BLM as a potential inducer of MHC-I, enhancing its expression to improve the efficacy of T-cell-based immunotherapy.
主要组织相容性复合物 I 类(MHC-I)的抗原呈递对 T 细胞介导的杀伤至关重要,而 MHC-I 的异常表面表达与免疫逃避密切相关。为了解决 MHC-I 下调问题,我们进行了高通量流式细胞术筛选,确定博莱霉素(BLM)是细胞表面 MHC-I 表达的强效诱导剂。BLM诱导的MHC-I增强使肿瘤细胞在共培养试验中更易受T细胞的影响,并增强了小鼠采用性细胞转移模型中的抗肿瘤反应。从机理上讲,BLM 能重塑肿瘤免疫微环境,以 ATM/ATR-NF-κB 依赖性方式诱导 MHC-I 表达。此外,BLM 还能改善依赖 T 细胞的免疫治疗方法,包括双特异性抗体疗法、免疫检查点疗法(ICT)和自体肿瘤浸润淋巴细胞(TILs)疗法。重要的是,小鼠模型中的低剂量BLM治疗可增强免疫疗法的抗肿瘤效果,且不会产生可检测到的肺毒性。总之,我们的研究结果重新利用了BLM作为MHC-I的潜在诱导剂,增强了MHC-I的表达,从而提高了基于T细胞的免疫疗法的疗效。
{"title":"Pharmacological induction of MHC-I expression in tumor cells revitalizes T cell anti-tumor immunity.","authors":"Qian Yu, Yu Dong, Xiaobo Wang, Chenxuan Su, Runkai Zhang, Wei Xu, Shuai Jiang, Yongjun Dang, Wei Jiang","doi":"10.1172/jci.insight.177788","DOIUrl":"https://doi.org/10.1172/jci.insight.177788","url":null,"abstract":"<p><p>Antigen presentation by Major Histocompatibility Complex Class I (MHC-I) is crucial for T-cell-mediated killing, and aberrant surface MHC-I expression is tightly associated with immune evasion. To address MHC-I downregulation, we conducted a high-throughput flow cytometry screen, identifying bleomycin (BLM) as a potent inducer of cell surface MHC-I expression. BLM-induced MHC-I augmentation renders tumor cells more susceptible to T cells in co-culture assays and enhances anti-tumor responses in an adoptive cellular transfer mouse model. Mechanistically, BLM remodels the tumor immune microenvironment, inducing MHC-I expression in an ATM/ATR-NF-κB-dependent manner. Furthermore, BLM improves T-cell-dependent immunotherapeutic approaches, including bispecific antibodies therapy, immune checkpoint therapy (ICT), and autologous tumor-infiltrating lymphocytes (TILs) therapy. Importantly, low-dose BLM treatment in mouse models amplified the anti-tumor effect of immunotherapy without detectable pulmonary toxicity. In summary, our findings repurpose BLM as a potential inducer of MHC-I, enhancing its expression to improve the efficacy of T-cell-based immunotherapy.</p>","PeriodicalId":14722,"journal":{"name":"JCI insight","volume":null,"pages":null},"PeriodicalIF":6.3,"publicationDate":"2024-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141897367","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-01DOI: 10.1172/jci.insight.176212
Elsayed Metwally, Alfredo Sanchez Solano, Boris Lavanderos, Evan Yamasaki, Pratish Thakore, Conor McClenaghan, Natalia Rios, Rafael Radi, Yumei Feng Earley, Colin G Nichols, Scott Earley
Cantú syndrome is a multisystem disorder caused by gain-of-function (GOF) mutations in KCNJ8 and ABCC9, the genes encoding the pore-forming inward rectifier Kir6.1 and regulatory sulfonylurea receptor SUR2B subunits, respectively, of vascular ATP-sensitive K+ channels (KATP). In this study, we investigated changes in the vascular endothelium in mice in which Cantú syndrome -associated Kcnj8 or Abcc9 mutations were knocked-in to the endogenous loci. We found that endothelium-dependent dilation was impaired in small mesenteric arteries from Cantú mice. Loss of endothelium-dependent vasodilation led to increased vasoconstriction in response to intraluminal pressure or treatment with the adrenergic receptor agonist phenylephrine. We also found that either KATP GOF or acute activation of KATP channels with pinacidil increased the amplitude and frequency of wave-like Ca2+ events generated in the endothelium in response to the vasodilator agonist carbachol. Increased cytosolic Ca2+ signaling activity in arterial endothelial cells from Cantú mice was associated with elevated mitochondrial [Ca2+] and enhanced reactive oxygen species (ROS) and peroxynitrite levels. Scavenging intracellular or mitochondrial ROS restored endothelium-dependent vasodilation in the arteries of mice with KATP GOF mutations. We conclude that mitochondrial Ca2+ overload and ROS generation, which subsequently leads to nitric oxide consumption and peroxynitrite formation, cause endothelial dysfunction in mice with Cantú syndrome.
{"title":"Mitochondrial Ca2+-coupled generation of reactive oxygen species, peroxynitrite formation, and endothelial dysfunction in Cantú syndrome.","authors":"Elsayed Metwally, Alfredo Sanchez Solano, Boris Lavanderos, Evan Yamasaki, Pratish Thakore, Conor McClenaghan, Natalia Rios, Rafael Radi, Yumei Feng Earley, Colin G Nichols, Scott Earley","doi":"10.1172/jci.insight.176212","DOIUrl":"https://doi.org/10.1172/jci.insight.176212","url":null,"abstract":"<p><p>Cantú syndrome is a multisystem disorder caused by gain-of-function (GOF) mutations in KCNJ8 and ABCC9, the genes encoding the pore-forming inward rectifier Kir6.1 and regulatory sulfonylurea receptor SUR2B subunits, respectively, of vascular ATP-sensitive K+ channels (KATP). In this study, we investigated changes in the vascular endothelium in mice in which Cantú syndrome -associated Kcnj8 or Abcc9 mutations were knocked-in to the endogenous loci. We found that endothelium-dependent dilation was impaired in small mesenteric arteries from Cantú mice. Loss of endothelium-dependent vasodilation led to increased vasoconstriction in response to intraluminal pressure or treatment with the adrenergic receptor agonist phenylephrine. We also found that either KATP GOF or acute activation of KATP channels with pinacidil increased the amplitude and frequency of wave-like Ca2+ events generated in the endothelium in response to the vasodilator agonist carbachol. Increased cytosolic Ca2+ signaling activity in arterial endothelial cells from Cantú mice was associated with elevated mitochondrial [Ca2+] and enhanced reactive oxygen species (ROS) and peroxynitrite levels. Scavenging intracellular or mitochondrial ROS restored endothelium-dependent vasodilation in the arteries of mice with KATP GOF mutations. We conclude that mitochondrial Ca2+ overload and ROS generation, which subsequently leads to nitric oxide consumption and peroxynitrite formation, cause endothelial dysfunction in mice with Cantú syndrome.</p>","PeriodicalId":14722,"journal":{"name":"JCI insight","volume":null,"pages":null},"PeriodicalIF":6.3,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141874830","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-01DOI: 10.1172/jci.insight.182330
Mohd Parvez Khan, Elena Sabini, Katherine Beigel, Giulia Lanzolla, Brittany M Laslow, Dian Wang, Christophe Merceron, Amato Giaccia, Fanxin Long, Deanne M Taylor, Ernestina Schipani
Energy metabolism, through pathways such as oxidative phosphorylation (OxPhos) and glycolysis, plays a pivotal role in cellular differentiation and function. Our study investigates the impact of OxPhos disruption in cortical bone development by deleting Mitochondrial Transcription Factor A (TFAM). TFAM controls OxPhos by regulating the transcription of mitochondrial genes. The cortical bone, constituting the long bones' rigid shell, is sheathed by the periosteum, a connective tissue layer populated with skeletal progenitors that spawn osteoblasts, the bone-forming cells. TFAM-deficient mice presented with thinner cortical bone, spontaneous midshaft fractures, and compromised periosteal cell bioenergetics, characterized by reduced ATP levels. Additionally, they exhibited an enlarged periosteal progenitor cell pool with impaired osteoblast differentiation. Increasing Hypoxia-Inducible Factor 1a (HIF1) activity within periosteal cells significantly mitigated the detrimental effects induced by TFAM deletion. HIF1 is known to promote glycolysis in all cell types. Our findings underscore the indispensability of OxPhos for the proper accrual of cortical bone mass and indicate a compensatory mechanism between OxPhos and glycolysis in periosteal cells. The study opens new avenues for understanding the relationship between energy metabolism and skeletal health and suggests that modulating bioenergetic pathways may provide a therapeutic avenue for conditions characterized by bone fragility.
通过氧化磷酸化(OxPhos)和糖酵解等途径进行的能量代谢在细胞分化和功能中发挥着关键作用。我们的研究通过删除线粒体转录因子 A (TFAM),探讨了 OxPhos 中断对皮质骨发育的影响。TFAM 通过调节线粒体基因的转录来控制 OxPhos。骨皮质是长骨的坚硬外壳,由骨膜包裹,骨膜是一种结缔组织层,其中充满了骨骼祖细胞,这些祖细胞可催生成骨细胞(骨形成细胞)。缺乏 TFAM 的小鼠皮质骨较薄,会出现自发性中轴骨折,骨膜细胞生物能受损,表现为 ATP 水平降低。此外,它们还表现出骨膜祖细胞池扩大,成骨细胞分化受损。提高骨膜细胞中缺氧诱导因子1a(HIF1)的活性可显著减轻TFAM缺失诱导的有害影响。众所周知,HIF1能促进所有类型细胞的糖酵解。我们的研究结果强调了 OxPhos 对皮质骨量的正常累积的不可或缺性,并表明了骨膜细胞中 OxPhos 和糖酵解之间的补偿机制。这项研究为了解能量代谢与骨骼健康之间的关系开辟了新的途径,并表明调节生物能途径可为骨质脆弱的病症提供治疗途径。
{"title":"HIF1 activation safeguards cortical bone formation against impaired oxidative phosphorylation.","authors":"Mohd Parvez Khan, Elena Sabini, Katherine Beigel, Giulia Lanzolla, Brittany M Laslow, Dian Wang, Christophe Merceron, Amato Giaccia, Fanxin Long, Deanne M Taylor, Ernestina Schipani","doi":"10.1172/jci.insight.182330","DOIUrl":"https://doi.org/10.1172/jci.insight.182330","url":null,"abstract":"<p><p>Energy metabolism, through pathways such as oxidative phosphorylation (OxPhos) and glycolysis, plays a pivotal role in cellular differentiation and function. Our study investigates the impact of OxPhos disruption in cortical bone development by deleting Mitochondrial Transcription Factor A (TFAM). TFAM controls OxPhos by regulating the transcription of mitochondrial genes. The cortical bone, constituting the long bones' rigid shell, is sheathed by the periosteum, a connective tissue layer populated with skeletal progenitors that spawn osteoblasts, the bone-forming cells. TFAM-deficient mice presented with thinner cortical bone, spontaneous midshaft fractures, and compromised periosteal cell bioenergetics, characterized by reduced ATP levels. Additionally, they exhibited an enlarged periosteal progenitor cell pool with impaired osteoblast differentiation. Increasing Hypoxia-Inducible Factor 1a (HIF1) activity within periosteal cells significantly mitigated the detrimental effects induced by TFAM deletion. HIF1 is known to promote glycolysis in all cell types. Our findings underscore the indispensability of OxPhos for the proper accrual of cortical bone mass and indicate a compensatory mechanism between OxPhos and glycolysis in periosteal cells. The study opens new avenues for understanding the relationship between energy metabolism and skeletal health and suggests that modulating bioenergetic pathways may provide a therapeutic avenue for conditions characterized by bone fragility.</p>","PeriodicalId":14722,"journal":{"name":"JCI insight","volume":null,"pages":null},"PeriodicalIF":6.3,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141874827","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}