Pub Date : 2024-11-08DOI: 10.1172/jci.insight.182664
Amy Y Sato, Meloney Cregor, Kevin McAndrews, Charles A Schurman, Eric Schaible, Jennifer Shutter, Punit Vyas, Bhawana Adhikari, Monte S Willis, Marjan Boerma, Tamara Alliston, Teresita Bellido
Despite their beneficial actions as immunosuppressants, glucocorticoids (GC) have devastating effects on the musculoskeletal and cardiac systems, as long-term treated patients exhibit high incidence of falls, bone fractures, and cardiovascular events. Herein, we show that GC upregulate simultaneously in bone, skeletal muscle, and the heart the expression of E3 ubiquitin ligases (atrogenes), known to stimulate the proteasomal degradation of proteins. Activation of vitamin D receptor (VDR) signaling with the VDR ligands calcitriol or eldecalcitol prevented GC-induced atrogene upregulation in vivo and ex vivo in bone/muscle organ cultures and preserved tissue structure/mass and function of the 3 tissues in vivo. Direct pharmacologic inhibition of the proteasome with carfilzomib also conferred musculoskeletal protection. Genetic loss of the atrogene MuRF1-mediated protein ubiquitination in ΔRING mice afforded temporary or sustained protection from GC excess in bone or skeletal and heart muscle. We concluded that the atrogene pathway downstream of MuRF1 underlies GC action in bone, muscle, and the heart, and it can be pharmacologically or genetically targeted to confer protection against the damaging actions of GC simultaneously in the 3 tissues.
{"title":"Pharmacologic or genetic interference with atrogene signaling protects against glucocorticoid-induced musculoskeletal and cardiac disease.","authors":"Amy Y Sato, Meloney Cregor, Kevin McAndrews, Charles A Schurman, Eric Schaible, Jennifer Shutter, Punit Vyas, Bhawana Adhikari, Monte S Willis, Marjan Boerma, Tamara Alliston, Teresita Bellido","doi":"10.1172/jci.insight.182664","DOIUrl":"10.1172/jci.insight.182664","url":null,"abstract":"<p><p>Despite their beneficial actions as immunosuppressants, glucocorticoids (GC) have devastating effects on the musculoskeletal and cardiac systems, as long-term treated patients exhibit high incidence of falls, bone fractures, and cardiovascular events. Herein, we show that GC upregulate simultaneously in bone, skeletal muscle, and the heart the expression of E3 ubiquitin ligases (atrogenes), known to stimulate the proteasomal degradation of proteins. Activation of vitamin D receptor (VDR) signaling with the VDR ligands calcitriol or eldecalcitol prevented GC-induced atrogene upregulation in vivo and ex vivo in bone/muscle organ cultures and preserved tissue structure/mass and function of the 3 tissues in vivo. Direct pharmacologic inhibition of the proteasome with carfilzomib also conferred musculoskeletal protection. Genetic loss of the atrogene MuRF1-mediated protein ubiquitination in ΔRING mice afforded temporary or sustained protection from GC excess in bone or skeletal and heart muscle. We concluded that the atrogene pathway downstream of MuRF1 underlies GC action in bone, muscle, and the heart, and it can be pharmacologically or genetically targeted to confer protection against the damaging actions of GC simultaneously in the 3 tissues.</p>","PeriodicalId":14722,"journal":{"name":"JCI insight","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2024-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142465874","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 : 2024-11-08DOI: 10.1172/jci.insight.180081
Jennifer H Lawrence, Asha Patel, Melvin W King, Collin J Nadarajah, Richard Daneman, Erik S Musiek
The blood-brain barrier (BBB) is critical for maintaining brain homeostasis but is susceptible to inflammatory dysfunction. While transporter-dependent efflux of some lipophilic substrates across the BBB shows circadian variation due to rhythmic transporter expression, basal transporter-independent permeability and leakage is nonrhythmic. Whether daily timing influences BBB permeability in response to inflammation is unknown. Here, we induced systemic inflammation through repeated LPS injections either in the morning (ZT1) or evening (ZT13) under standard lighting conditions; we then examined BBB permeability to a polar molecule that is not a transporter substrate, sodium fluorescein. We observed clear diurnal variation in inflammatory BBB permeability, with a striking increase in paracellular leak across the BBB specifically following evening LPS injection. Evening LPS led to persisting glia activation as well as inflammation in the brain that was not observed in the periphery. The exaggerated evening neuroinflammation and BBB disruption were suppressed by microglial depletion or through keeping mice in constant darkness. Our data show that diurnal rhythms in microglial inflammatory responses to LPS drive daily variability in BBB breakdown and reveal time of day as a key regulator of inflammatory BBB disruption.
{"title":"Microglia drive diurnal variation in susceptibility to inflammatory blood-brain barrier breakdown.","authors":"Jennifer H Lawrence, Asha Patel, Melvin W King, Collin J Nadarajah, Richard Daneman, Erik S Musiek","doi":"10.1172/jci.insight.180081","DOIUrl":"10.1172/jci.insight.180081","url":null,"abstract":"<p><p>The blood-brain barrier (BBB) is critical for maintaining brain homeostasis but is susceptible to inflammatory dysfunction. While transporter-dependent efflux of some lipophilic substrates across the BBB shows circadian variation due to rhythmic transporter expression, basal transporter-independent permeability and leakage is nonrhythmic. Whether daily timing influences BBB permeability in response to inflammation is unknown. Here, we induced systemic inflammation through repeated LPS injections either in the morning (ZT1) or evening (ZT13) under standard lighting conditions; we then examined BBB permeability to a polar molecule that is not a transporter substrate, sodium fluorescein. We observed clear diurnal variation in inflammatory BBB permeability, with a striking increase in paracellular leak across the BBB specifically following evening LPS injection. Evening LPS led to persisting glia activation as well as inflammation in the brain that was not observed in the periphery. The exaggerated evening neuroinflammation and BBB disruption were suppressed by microglial depletion or through keeping mice in constant darkness. Our data show that diurnal rhythms in microglial inflammatory responses to LPS drive daily variability in BBB breakdown and reveal time of day as a key regulator of inflammatory BBB disruption.</p>","PeriodicalId":14722,"journal":{"name":"JCI insight","volume":"9 21","pages":""},"PeriodicalIF":6.3,"publicationDate":"2024-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142604485","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 : 2024-11-08DOI: 10.1172/jci.insight.182876
Heike J Wobst, Andreu Viader, Giovanni Muncipinto, Ryan Hollibaugh, Daniel van Kalken, Christopher T Burkhart, Susan M Cantin, Rachel M Bates, Yannik Regimbald-Dumas, Liam Gross, Mitchell T Antalek, Joshua E Zweig, Frank Wu, T Justin Rettenmaier, Matthew T Labenski, Nicholas Pullen, Heather S Blanchette, Jaclyn L Henderson, Haoling H Weng, Toby A Vaughn, Dean G Brown, John P Throup, Joel C Barrish
BACKGROUNDThe toxic accumulation of phenylalanine (Phe) in the brain underlies the neurological presentation of phenylketonuria (PKU). Solute carrier family 6 member 19 (SLC6A19) is the major transporter responsible for the (re)absorption of Phe in the kidney and intestine. Here, we describe the characterization of the first small molecule SLC6A19 inhibitor to enter clinical development for the treatment of PKU.METHODSC57Bl/6J WT and Pahenu2 mice were dosed with an inhibitor of SLC6A19 to investigate the effects on urinary amino acids and plasma Phe. In a phase 1 study, healthy human volunteers were dosed with JNT-517, an investigational oral inhibitor of SLC6A19. The primary objective of the study was safety. Secondary objectives included pharmacokinetic and pharmacodynamic studies.RESULTSInhibition of SLC6A19 increased the urinary excretion of Phe in a mouse model of PKU, thereby reducing plasma Phe levels. JNT-517, an investigational oral SLC6A19 inhibitor, was found to be safe and well tolerated and increased the urinary excretion of Phe in a phase 1 healthy volunteer study.CONCLUSIONSThese data indicate that pharmacological inhibition of SLC6A19 presents a promising approach to lower toxic elevated levels of amino acids found in PKU and related amino acid metabolism disorders by facilitating their renal elimination.TRIAL REGISTRATIONAustralian New Zealand Clinical Trials Registry (ANZCTR), ACTRN12622001222730.FUNDINGThe studies in this paper were funded by Jnana Therapeutics.
{"title":"SLC6A19 inhibition facilitates urinary neutral amino acid excretion and lowers plasma phenylalanine.","authors":"Heike J Wobst, Andreu Viader, Giovanni Muncipinto, Ryan Hollibaugh, Daniel van Kalken, Christopher T Burkhart, Susan M Cantin, Rachel M Bates, Yannik Regimbald-Dumas, Liam Gross, Mitchell T Antalek, Joshua E Zweig, Frank Wu, T Justin Rettenmaier, Matthew T Labenski, Nicholas Pullen, Heather S Blanchette, Jaclyn L Henderson, Haoling H Weng, Toby A Vaughn, Dean G Brown, John P Throup, Joel C Barrish","doi":"10.1172/jci.insight.182876","DOIUrl":"https://doi.org/10.1172/jci.insight.182876","url":null,"abstract":"<p><p>BACKGROUNDThe toxic accumulation of phenylalanine (Phe) in the brain underlies the neurological presentation of phenylketonuria (PKU). Solute carrier family 6 member 19 (SLC6A19) is the major transporter responsible for the (re)absorption of Phe in the kidney and intestine. Here, we describe the characterization of the first small molecule SLC6A19 inhibitor to enter clinical development for the treatment of PKU.METHODSC57Bl/6J WT and Pahenu2 mice were dosed with an inhibitor of SLC6A19 to investigate the effects on urinary amino acids and plasma Phe. In a phase 1 study, healthy human volunteers were dosed with JNT-517, an investigational oral inhibitor of SLC6A19. The primary objective of the study was safety. Secondary objectives included pharmacokinetic and pharmacodynamic studies.RESULTSInhibition of SLC6A19 increased the urinary excretion of Phe in a mouse model of PKU, thereby reducing plasma Phe levels. JNT-517, an investigational oral SLC6A19 inhibitor, was found to be safe and well tolerated and increased the urinary excretion of Phe in a phase 1 healthy volunteer study.CONCLUSIONSThese data indicate that pharmacological inhibition of SLC6A19 presents a promising approach to lower toxic elevated levels of amino acids found in PKU and related amino acid metabolism disorders by facilitating their renal elimination.TRIAL REGISTRATIONAustralian New Zealand Clinical Trials Registry (ANZCTR), ACTRN12622001222730.FUNDINGThe studies in this paper were funded by Jnana Therapeutics.</p>","PeriodicalId":14722,"journal":{"name":"JCI insight","volume":"9 21","pages":""},"PeriodicalIF":6.3,"publicationDate":"2024-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142604503","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 : 2024-11-08DOI: 10.1172/jci.insight.178644
Nune Markosyan, Il-Kyu Kim, Charu Arora, Liz Quinones-Ware, Nikhil Joshi, Noah Cheng, Emma Y Schechter, John W Tobias, Joseph E Hochberg, Emily Corse, Kang Liu, Varenka Rodriguez DiBlasi, Li-Chuan Eric Chan, Emer M Smyth, Garret A FitzGerald, Ben Z Stanger, Robert H Vonderheide
Tumor cell-derived prostaglandin E2 (PGE2) is a tumor cell-intrinsic factor that supports immunosuppression in the tumor microenvironment (TME) by acting on the immune cells, but the impact of PGE2 signaling in tumor cells on the immunosuppressive TME is unclear. We demonstrate that deleting the PGE2 synthesis enzyme or disrupting autocrine PGE2 signaling through EP4 receptors on tumor cells reverses the T cell-low, myeloid cell-rich TME, activates T cells, and suppresses tumor growth. Knockout (KO) of Ptges (the gene encoding the PGE2 synthesis enzyme mPGES-1) or the EP4 receptor gene (Ptger4) in KPCY (KrasG12D P53R172H Yfp CrePdx) pancreatic tumor cells abolished growth of implanted tumors in a T cell-dependent manner. Blockade of the EP4 receptor in combination with immunotherapy, but not immunotherapy alone, induced complete tumor regressions and immunological memory. Mechanistically, Ptges- and Ptger4-KO tumor cells exhibited altered T and myeloid cell attractant chemokines, became more susceptible to TNF-α-induced killing, and exhibited reduced adenosine synthesis. In hosts treated with an adenosine deaminase inhibitor, Ptger4-KO tumor cells accumulated adenosine and gave rise to tumors. These studies reveal an unexpected finding - a nonredundant role for the autocrine mPGES-1/PGE2/EP4 signaling axis in pancreatic cancer cells, further nominating mPGES-1 inhibition and EP4 blockade as immune-sensitizing therapy in cancer.
肿瘤细胞衍生的前列腺素 E2(PGE2)是一种肿瘤细胞内在因子,它通过作用于免疫细胞支持肿瘤微环境(TME)中的免疫抑制,但肿瘤细胞中的 PGE2 信号传导对免疫抑制性 TME 的影响尚不清楚。我们证明,删除肿瘤细胞上的 PGE2 合成酶或破坏通过 EP4 受体的自分泌 PGE2 信号,可逆转 T 细胞低下、髓样细胞丰富的 TME,激活 T 细胞并抑制肿瘤生长。KPCY(KrasG12D/P53R172H/Yfp/CrePdx)胰腺肿瘤细胞中Ptges(编码PGE2合成酶mPGES-1的基因)或EP4受体基因(Ptger4)的敲除(KO)以T细胞依赖的方式抑制了植入肿瘤的生长。EP4受体的阻断与免疫疗法相结合,而不是单独使用免疫疗法,可诱导肿瘤完全消退和免疫记忆。从机理上讲,Ptges 和 Ptger4 KO 肿瘤细胞表现出改变的 T 细胞和骨髓细胞吸引趋化因子,更容易被 TNF-α 杀死,并表现出腺苷合成减少。在用腺苷脱氨酶抑制剂处理的宿主中,Ptger4 KO 肿瘤细胞会积累腺苷并产生肿瘤。这些研究揭示了一个意想不到的发现--mPGES1-PGE2-EP4 信号轴在胰腺癌细胞中的非冗余作用--进一步确定了 mPGES-1 抑制剂和 EP4 阻断剂作为癌症免疫增敏疗法的地位。
{"title":"Pivotal roles for cancer cell-intrinsic mPGES-1 and autocrine EP4 signaling in suppressing antitumor immunity.","authors":"Nune Markosyan, Il-Kyu Kim, Charu Arora, Liz Quinones-Ware, Nikhil Joshi, Noah Cheng, Emma Y Schechter, John W Tobias, Joseph E Hochberg, Emily Corse, Kang Liu, Varenka Rodriguez DiBlasi, Li-Chuan Eric Chan, Emer M Smyth, Garret A FitzGerald, Ben Z Stanger, Robert H Vonderheide","doi":"10.1172/jci.insight.178644","DOIUrl":"10.1172/jci.insight.178644","url":null,"abstract":"<p><p>Tumor cell-derived prostaglandin E2 (PGE2) is a tumor cell-intrinsic factor that supports immunosuppression in the tumor microenvironment (TME) by acting on the immune cells, but the impact of PGE2 signaling in tumor cells on the immunosuppressive TME is unclear. We demonstrate that deleting the PGE2 synthesis enzyme or disrupting autocrine PGE2 signaling through EP4 receptors on tumor cells reverses the T cell-low, myeloid cell-rich TME, activates T cells, and suppresses tumor growth. Knockout (KO) of Ptges (the gene encoding the PGE2 synthesis enzyme mPGES-1) or the EP4 receptor gene (Ptger4) in KPCY (KrasG12D P53R172H Yfp CrePdx) pancreatic tumor cells abolished growth of implanted tumors in a T cell-dependent manner. Blockade of the EP4 receptor in combination with immunotherapy, but not immunotherapy alone, induced complete tumor regressions and immunological memory. Mechanistically, Ptges- and Ptger4-KO tumor cells exhibited altered T and myeloid cell attractant chemokines, became more susceptible to TNF-α-induced killing, and exhibited reduced adenosine synthesis. In hosts treated with an adenosine deaminase inhibitor, Ptger4-KO tumor cells accumulated adenosine and gave rise to tumors. These studies reveal an unexpected finding - a nonredundant role for the autocrine mPGES-1/PGE2/EP4 signaling axis in pancreatic cancer cells, further nominating mPGES-1 inhibition and EP4 blockade as immune-sensitizing therapy in cancer.</p>","PeriodicalId":14722,"journal":{"name":"JCI insight","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2024-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142287564","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 : 2024-11-08DOI: 10.1172/jci.insight.179126
Hannah Bd Duffy, Colleen Byrnes, Hongling Zhu, Galina Tuymetova, Y Terry Lee, Frances M Platt, Richard L Proia
Gaucher disease, the most prevalent lysosomal storage disease, is caused by homozygous mutations at the GBA gene, which is responsible for encoding the enzyme glucocerebrosidase. Neuronopathic Gaucher disease is associated with microgliosis, astrogliosis, and neurodegeneration. However, the role that microglia, astrocytes, and neurons play in the disease remains to be determined. In the current study, we developed inducible, cell-type-specific Gba-KO mice to better understand the individual impacts of Gba deficiencies on microglia and neurons. Gba was conditionally knocked out either exclusively in microglia or neurons or throughout the body. These mouse models were developed using a tamoxifen-inducible Cre system, with tamoxifen administration commencing at weaning. Microglia-specific Gba-KO mice showed no signs of disease. However, the neuron-specific Gba KO resulted in a shortened lifespan, severe weight loss, and ataxia. These mice also had significant neurodegeneration, microgliosis, and astrogliosis accompanied by the accumulation of glucosylceramide and glucosylsphingosine, recapitulating Gaucher disease-like symptoms. These surprising findings reveal that, unlike the neuron-specific Gba deficiency, microglia-specific Gba deficiency alone does not induce disease. The neuronal Gaucher disease mouse model, with a median survival of 16 weeks, may be useful for future studies of pathogenesis and the evaluation of therapies.
戈谢病是最常见的溶酶体贮积病,由负责编码葡萄糖脑苷脂酶的 GBA 基因的同源突变引起。神经病变性戈谢病与小胶质细胞增生、星形胶质细胞增生和神经变性有关。然而,小胶质细胞、星形胶质细胞和神经元在该病中所起的作用仍有待确定。在目前的研究中,我们开发了新型、可诱导、细胞类型特异的 GBA KO 小鼠,以了解 GBA 缺陷对小胶质细胞和神经元的个体影响。GBA 在小胶质细胞或神经元中被有条件地敲除,或在全身被敲除。这些新型小鼠模型是利用他莫昔芬诱导的Cre系统开发的,断奶时开始服用他莫昔芬。小胶质细胞特异性 GBA KO 小鼠没有表现出疾病迹象。然而,神经元特异性 GBA KO 会导致寿命缩短、体重严重下降和共济失调。这些小鼠还出现了明显的神经变性、小胶质细胞病变和星形胶质细胞病变,并伴有葡萄糖甘油酰胺和葡萄糖鞘氨醇的积累,再现了类似戈谢病的症状。这些令人惊讶的发现表明,与神经元特异性 GBA 缺乏症不同,小胶质细胞特异性 GBA 缺乏症不会诱发疾病。这种新型神经元戈谢病小鼠模型的中位存活期为16周,可能有助于未来的发病机制研究和疗法评估。
{"title":"Deletion of Gba in neurons, but not microglia, causes neurodegeneration in a Gaucher mouse model.","authors":"Hannah Bd Duffy, Colleen Byrnes, Hongling Zhu, Galina Tuymetova, Y Terry Lee, Frances M Platt, Richard L Proia","doi":"10.1172/jci.insight.179126","DOIUrl":"10.1172/jci.insight.179126","url":null,"abstract":"<p><p>Gaucher disease, the most prevalent lysosomal storage disease, is caused by homozygous mutations at the GBA gene, which is responsible for encoding the enzyme glucocerebrosidase. Neuronopathic Gaucher disease is associated with microgliosis, astrogliosis, and neurodegeneration. However, the role that microglia, astrocytes, and neurons play in the disease remains to be determined. In the current study, we developed inducible, cell-type-specific Gba-KO mice to better understand the individual impacts of Gba deficiencies on microglia and neurons. Gba was conditionally knocked out either exclusively in microglia or neurons or throughout the body. These mouse models were developed using a tamoxifen-inducible Cre system, with tamoxifen administration commencing at weaning. Microglia-specific Gba-KO mice showed no signs of disease. However, the neuron-specific Gba KO resulted in a shortened lifespan, severe weight loss, and ataxia. These mice also had significant neurodegeneration, microgliosis, and astrogliosis accompanied by the accumulation of glucosylceramide and glucosylsphingosine, recapitulating Gaucher disease-like symptoms. These surprising findings reveal that, unlike the neuron-specific Gba deficiency, microglia-specific Gba deficiency alone does not induce disease. The neuronal Gaucher disease mouse model, with a median survival of 16 weeks, may be useful for future studies of pathogenesis and the evaluation of therapies.</p>","PeriodicalId":14722,"journal":{"name":"JCI insight","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2024-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142307753","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 : 2024-11-08DOI: 10.1172/jci.insight.182589
Benjamin I Burke, Ahmed Ismaeel, Douglas E Long, Lauren A Depa, Peyton T Coburn, Jensen Goh, Tolulope P Saliu, Bonnie J Walton, Ivan J Vechetti, Bailey D Peck, Taylor R Valentino, C Brooks Mobley, Hasiyet Memetimin, Dandan Wang, Brian S Finlin, Philip A Kern, Charlotte A Peterson, John J McCarthy, Yuan Wen
Extracellular vesicles (EVs) have emerged as important mediators of intertissue signaling and exercise adaptations. In this human study, we provide evidence that muscle-specific microRNA-1 (miR-1) was transferred to adipose tissue via EVs following an acute bout of resistance exercise. Using a multimodel machine learning automation tool, we discovered muscle primary miR-1 transcript and CD63+ EV count in circulation as top explanatory features for changes in adipose miR-1 levels in response to resistance exercise. RNA-Seq and in-silico prediction of miR-1 target genes identified caveolin 2 (CAV2) and tripartite motif containing 6 (TRIM6) as miR-1 target genes downregulated in the adipose tissue of a subset of participants with the highest increases in miR-1 levels following resistance exercise. Overexpression of miR-1 in differentiated human adipocyte-derived stem cells downregulated these miR-1 targets and enhanced catecholamine-induced lipolysis. These data identify a potential EV-mediated mechanism by which skeletal muscle communicates with adipose tissue and modulates lipolysis via miR-1.
{"title":"Extracellular vesicle transfer of miR-1 to adipose tissue modifies lipolytic pathways following resistance exercise.","authors":"Benjamin I Burke, Ahmed Ismaeel, Douglas E Long, Lauren A Depa, Peyton T Coburn, Jensen Goh, Tolulope P Saliu, Bonnie J Walton, Ivan J Vechetti, Bailey D Peck, Taylor R Valentino, C Brooks Mobley, Hasiyet Memetimin, Dandan Wang, Brian S Finlin, Philip A Kern, Charlotte A Peterson, John J McCarthy, Yuan Wen","doi":"10.1172/jci.insight.182589","DOIUrl":"10.1172/jci.insight.182589","url":null,"abstract":"<p><p>Extracellular vesicles (EVs) have emerged as important mediators of intertissue signaling and exercise adaptations. In this human study, we provide evidence that muscle-specific microRNA-1 (miR-1) was transferred to adipose tissue via EVs following an acute bout of resistance exercise. Using a multimodel machine learning automation tool, we discovered muscle primary miR-1 transcript and CD63+ EV count in circulation as top explanatory features for changes in adipose miR-1 levels in response to resistance exercise. RNA-Seq and in-silico prediction of miR-1 target genes identified caveolin 2 (CAV2) and tripartite motif containing 6 (TRIM6) as miR-1 target genes downregulated in the adipose tissue of a subset of participants with the highest increases in miR-1 levels following resistance exercise. Overexpression of miR-1 in differentiated human adipocyte-derived stem cells downregulated these miR-1 targets and enhanced catecholamine-induced lipolysis. These data identify a potential EV-mediated mechanism by which skeletal muscle communicates with adipose tissue and modulates lipolysis via miR-1.</p>","PeriodicalId":14722,"journal":{"name":"JCI insight","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2024-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142346981","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 : 2024-11-08DOI: 10.1172/jci.insight.164762
Chandra M Menendez, Jonathan Zuccolo, Susan E Swedo, Sean Reim, Brian Richmand, Hilla Ben-Pazi, Abraham Kovoor, Madeleine W Cunningham
Despite growing recognition, neuropsychiatric diseases associated with infections are a major unsolved problem worldwide. Group A streptococcal (GAS) infections can cause autoimmune sequelae characterized by movement disorders, such as Sydenham chorea, and neuropsychiatric disorders. The molecular mechanisms underlying these diseases are not fully understood. Our previous work demonstrates that autoantibodies (AAbs) can target dopaminergic neurons and increase dopamine D2 receptor (D2R) signaling. However, AAb influence on dopamine D1 receptor (D1R) activity is underexplored. We found evidence that suggests GAS-induced cross-reactive AAbs promote autoimmune encephalitis of the basal ganglia, a region of high dopamine receptor density. Here, we report a mechanism whereby neuropsychiatric syndromes are distinguished from movement disorders by differences in D1R and D2R AAb titers, signaling, receiver operating characteristic curves, and immunoreactivity with D1R and D2R autoreactive epitopes. D1R AAb signaling was observed through patient serum AAbs and novel patient-derived monoclonal antibodies (mAbs), which induced both D1R G protein- and β-arrestin-transduced signals. Furthermore, patient AAbs and mAbs enhanced D1R signaling mechanisms mediated by the neurotransmitter dopamine. Our findings suggest that AAb-mediated D1R signaling may contribute to the pathogenesis of neuropsychiatric sequelae and inform new options for diagnosis and treatment of GAS sequelae and related disorders.
{"title":"Dopamine receptor autoantibody signaling in infectious sequelae differentiates movement versus neuropsychiatric disorders.","authors":"Chandra M Menendez, Jonathan Zuccolo, Susan E Swedo, Sean Reim, Brian Richmand, Hilla Ben-Pazi, Abraham Kovoor, Madeleine W Cunningham","doi":"10.1172/jci.insight.164762","DOIUrl":"10.1172/jci.insight.164762","url":null,"abstract":"<p><p>Despite growing recognition, neuropsychiatric diseases associated with infections are a major unsolved problem worldwide. Group A streptococcal (GAS) infections can cause autoimmune sequelae characterized by movement disorders, such as Sydenham chorea, and neuropsychiatric disorders. The molecular mechanisms underlying these diseases are not fully understood. Our previous work demonstrates that autoantibodies (AAbs) can target dopaminergic neurons and increase dopamine D2 receptor (D2R) signaling. However, AAb influence on dopamine D1 receptor (D1R) activity is underexplored. We found evidence that suggests GAS-induced cross-reactive AAbs promote autoimmune encephalitis of the basal ganglia, a region of high dopamine receptor density. Here, we report a mechanism whereby neuropsychiatric syndromes are distinguished from movement disorders by differences in D1R and D2R AAb titers, signaling, receiver operating characteristic curves, and immunoreactivity with D1R and D2R autoreactive epitopes. D1R AAb signaling was observed through patient serum AAbs and novel patient-derived monoclonal antibodies (mAbs), which induced both D1R G protein- and β-arrestin-transduced signals. Furthermore, patient AAbs and mAbs enhanced D1R signaling mechanisms mediated by the neurotransmitter dopamine. Our findings suggest that AAb-mediated D1R signaling may contribute to the pathogenesis of neuropsychiatric sequelae and inform new options for diagnosis and treatment of GAS sequelae and related disorders.</p>","PeriodicalId":14722,"journal":{"name":"JCI insight","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2024-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142346970","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 : 2024-11-08DOI: 10.1172/jci.insight.179982
Joris L Vos, Joleen Jh Traets, Xiaohang Qiao, Iris M Seignette, Dennis Peters, Michel Wjm Wouters, Erik Hooijberg, Annegien Broeks, Jacqueline E van der Wal, M Baris Karakullukcu, W Martin C Klop, Arash Navran, Marc van Beurden, Oscar R Brouwer, Luc Gt Morris, Mariette Ie van Poelgeest, Ellen Kapiteijn, John Bag Haanen, Christian U Blank, Charlotte L Zuur
Mucosal melanoma (MucM) is a rare cancer with a poor prognosis and low response rate to immune checkpoint inhibition (ICI) compared with cutaneous melanoma (CM). To explore the immune microenvironment and potential drivers of MucM's relative resistance to ICI drugs, we characterized 101 MucM tumors (43 head and neck [H&N], 31 female urogenital, 13 male urogenital, 11 anorectal, and 3 other gastrointestinal) using bulk RNA-Seq and immunofluorescence. RNA-Seq data show that MucM has a significantly lower IFN-γ signature levels than CM. MucM tumors of the H&N region show a significantly greater abundance of CD8+ T cells, cytotoxic cells, and higher IFN-γ signature levels than MucM from lower body sites. In the subcohort of 35 patients with MucM treated with ICI, hierarchical clustering reveals clusters with a high and low degree of immune infiltration, with a differential ICI response rate. Immune-associated gene sets were enriched in responders. Signatures associated with cancer-associated fibroblasts, macrophages, and TGF-β signaling may be higher in immune-infiltrated, but ICI-unresponsive tumors, suggesting a role for these resistance mechanisms in MucM. Our data show organ region-specific differences in immune infiltration and IFN-γ signature levels in MucM, with H&N MucM displaying the most favorable immune profile. Our study might offer a starting point for developing more personalized treatment strategies for this disease.
{"title":"Diversity of the immune microenvironment and response to checkpoint inhibitor immunotherapy in mucosal melanoma.","authors":"Joris L Vos, Joleen Jh Traets, Xiaohang Qiao, Iris M Seignette, Dennis Peters, Michel Wjm Wouters, Erik Hooijberg, Annegien Broeks, Jacqueline E van der Wal, M Baris Karakullukcu, W Martin C Klop, Arash Navran, Marc van Beurden, Oscar R Brouwer, Luc Gt Morris, Mariette Ie van Poelgeest, Ellen Kapiteijn, John Bag Haanen, Christian U Blank, Charlotte L Zuur","doi":"10.1172/jci.insight.179982","DOIUrl":"https://doi.org/10.1172/jci.insight.179982","url":null,"abstract":"<p><p>Mucosal melanoma (MucM) is a rare cancer with a poor prognosis and low response rate to immune checkpoint inhibition (ICI) compared with cutaneous melanoma (CM). To explore the immune microenvironment and potential drivers of MucM's relative resistance to ICI drugs, we characterized 101 MucM tumors (43 head and neck [H&N], 31 female urogenital, 13 male urogenital, 11 anorectal, and 3 other gastrointestinal) using bulk RNA-Seq and immunofluorescence. RNA-Seq data show that MucM has a significantly lower IFN-γ signature levels than CM. MucM tumors of the H&N region show a significantly greater abundance of CD8+ T cells, cytotoxic cells, and higher IFN-γ signature levels than MucM from lower body sites. In the subcohort of 35 patients with MucM treated with ICI, hierarchical clustering reveals clusters with a high and low degree of immune infiltration, with a differential ICI response rate. Immune-associated gene sets were enriched in responders. Signatures associated with cancer-associated fibroblasts, macrophages, and TGF-β signaling may be higher in immune-infiltrated, but ICI-unresponsive tumors, suggesting a role for these resistance mechanisms in MucM. Our data show organ region-specific differences in immune infiltration and IFN-γ signature levels in MucM, with H&N MucM displaying the most favorable immune profile. Our study might offer a starting point for developing more personalized treatment strategies for this disease.</p>","PeriodicalId":14722,"journal":{"name":"JCI insight","volume":"9 21","pages":""},"PeriodicalIF":6.3,"publicationDate":"2024-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142604478","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 : 2024-11-08DOI: 10.1172/jci.insight.181067
Mohadeseh Hasanpourghadi, Mikhail Novikov, Robert Ambrose, Arezki Chekaoui, Dakota Newman, ZhiQuan Xiang, Andrew D Luber, Sue L Currie, XiangYang Zhou, Hildegund Cj Ertl
In patients who progress from acute hepatitis B virus (HBV) infection to a chronic HBV (CHB) infection, CD8+ T cells fail to eliminate the virus and become impaired. A functional cure of CHB likely requires CD8+ T cell responses different from those induced by the infection. Here we report preclinical immunogenicity and efficacy of an HBV therapeutic vaccine that includes herpes simplex virus (HSV) glycoprotein D (gD), a checkpoint modifier of early T cell activation, that augments CD8+ T cell responses. The vaccine is based on a chimpanzee adenovirus serotype 6 (AdC6) vector, called AdC6-gDHBV2, which targets conserved and highly immunogenic regions of the viral polymerase and core antigens fused to HSV gD. The vaccine was tested with and without gD in mice for immunogenicity, and in an AAV8-1.3HBV vector model of antiviral efficacy. The vaccine encoding the HBV antigens within gD stimulates potent and broad CD8+ T cell responses. In a surrogate model of HBV infection, a single intramuscular injection achieved pronounced and sustained declines of circulating HBV DNA copies and HBV surface antigen; both inversely correlated with HBV-specific CD8+ T cell frequencies in spleen and liver.
在从急性乙型肝炎病毒(HBV)感染发展为慢性 HBV(CHB)感染的患者中,CD8+ T 细胞无法清除病毒并受损。慢性乙型肝炎的功能性治愈可能需要与感染诱导的CD8+ T细胞反应不同的新的高功能CD8+ T细胞反应。在此,我们报告了一种 HBV 治疗性疫苗的临床前免疫原性和疗效,该疫苗包括单纯疱疹病毒 (HSV) 糖蛋白 D (gD),这是一种早期 T 细胞活化的检查点调节剂,可增强、扩大和延长 CD8+ T 细胞应答。我们开发了一种基于黑猩猩腺病毒血清型 6 (AdC6) 载体的治疗性 HBV 疫苗,称为 AdC6-gDHBV2,其靶标是病毒聚合酶 (pol) 的保守和高免疫原性区域以及融合到 HSV gD 中的核心抗原。该疫苗在小鼠体内进行了含 gD 和不含 gD 的免疫原性测试,并在腺相关病毒 (AAV)8-1.3HBV 载体模型中进行了抗病毒效力测试。在 gD 中编码 HBV 抗原的疫苗可激发强效、广泛的 CD8+ T 细胞反应。在代理 HBV 感染模型中,单次肌肉注射 AdC6-gDHBV2 可使循环 HBV DNA 拷贝(cps)和 HBV 表面抗原(HBsAg)显著持续下降;两者均与脾脏和肝脏中的 HBV 特异性 CD8+ T 细胞频率成反比。AdC6-gDHBV2 是首个单独使用就能显著降低 HBV 基因组拷贝和 HBsAg 水平的治疗性疫苗,即使在感染后数月才接种疫苗也是如此。
{"title":"A therapeutic HBV vaccine containing a checkpoint modifier enhances CD8+ T cell and antiviral responses.","authors":"Mohadeseh Hasanpourghadi, Mikhail Novikov, Robert Ambrose, Arezki Chekaoui, Dakota Newman, ZhiQuan Xiang, Andrew D Luber, Sue L Currie, XiangYang Zhou, Hildegund Cj Ertl","doi":"10.1172/jci.insight.181067","DOIUrl":"10.1172/jci.insight.181067","url":null,"abstract":"<p><p>In patients who progress from acute hepatitis B virus (HBV) infection to a chronic HBV (CHB) infection, CD8+ T cells fail to eliminate the virus and become impaired. A functional cure of CHB likely requires CD8+ T cell responses different from those induced by the infection. Here we report preclinical immunogenicity and efficacy of an HBV therapeutic vaccine that includes herpes simplex virus (HSV) glycoprotein D (gD), a checkpoint modifier of early T cell activation, that augments CD8+ T cell responses. The vaccine is based on a chimpanzee adenovirus serotype 6 (AdC6) vector, called AdC6-gDHBV2, which targets conserved and highly immunogenic regions of the viral polymerase and core antigens fused to HSV gD. The vaccine was tested with and without gD in mice for immunogenicity, and in an AAV8-1.3HBV vector model of antiviral efficacy. The vaccine encoding the HBV antigens within gD stimulates potent and broad CD8+ T cell responses. In a surrogate model of HBV infection, a single intramuscular injection achieved pronounced and sustained declines of circulating HBV DNA copies and HBV surface antigen; both inversely correlated with HBV-specific CD8+ T cell frequencies in spleen and liver.</p>","PeriodicalId":14722,"journal":{"name":"JCI insight","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2024-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142125761","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 : 2024-11-08DOI: 10.1172/jci.insight.183400
Ye Chen, Maogen Chen, Yu Liu, Qiang Li, Youqiu Xue, Liu Liu, Rongzhen Liang, Yiding Xiong, Jun Zhao, Jingrong Chen, Weidong Lin, Julie Wang, Yun Feng Pan, William Stohl, Song Guo Zheng
T follicular helper (Tfh) cells represent an important subset of CD4+ T cells that is crucial to the maturation and differentiation of B cells and the production of high-affinity antibodies. Because B cell activating-factor (BAFF), a vital B cell survival factor, is also crucial to B cell maturation and differentiation, we assessed the effects of BAFF on Tfh cell development and function. We demonstrated that deficiency of BAFF, but not of APRIL, markedly inhibited Tfh cell development, germinal center (GC) formation, and antigen-specific antibody production. The promoting effect of BAFF on Tfh cell development was dependent on expression of BR3 on T cells, and its promoting effect on GC formation was dependent on expression of BR3 on both T cells and B cells. BAFF directly promoted expression of the Tfh cell-characteristic genes via NF-κB signaling. This effect did need BR3 expression. Thus, BAFF not only has direct effects on B cells, but it also has direct effects on Tfh cell differentiation via engagement of BR3, which collectively promoted GC formation and production of high-affinity antibodies. This dual effect of BAFF on B cells and Tfh cells may help explain the clinical utility of BAFF antagonists in the management of certain autoimmune diseases.
T 滤泡辅助细胞(Tfh)是 CD4+ T 细胞的一个重要亚群,对 B 细胞的成熟和分化以及高亲和力抗体的产生至关重要。BAFF是一种重要的B细胞存活因子,对B细胞的成熟和分化也至关重要,因此我们评估了BAFF对Tfh细胞发育和功能的影响。我们证明,缺乏 BAFF(而非 APRIL)会明显抑制 Tfh 细胞的发育、生殖中心(GC)的形成和抗原特异性抗体的产生。BAFF对Tfh细胞发育的促进作用依赖于T细胞上BR3的表达,而其对GC形成的促进作用依赖于T细胞和B细胞上BR3的表达。BAFF 通过 NF-κB 信号直接促进 Tfh 细胞特征基因的表达。这种作用需要 BR3 的表达。因此,BAFF 不仅对 B 细胞有直接作用,而且还通过 BR3 的参与对 Tfh 细胞的分化有直接作用,从而共同促进 GC 的形成和高亲和性抗体的产生。BAFF 对 B 细胞和 Tfh 细胞的这种双重作用可能有助于解释 BAFF 拮抗剂在治疗某些自身免疫性疾病方面的临床用途。
{"title":"BAFF promotes follicular helper T cell development and germinal center formation through BR3 signal.","authors":"Ye Chen, Maogen Chen, Yu Liu, Qiang Li, Youqiu Xue, Liu Liu, Rongzhen Liang, Yiding Xiong, Jun Zhao, Jingrong Chen, Weidong Lin, Julie Wang, Yun Feng Pan, William Stohl, Song Guo Zheng","doi":"10.1172/jci.insight.183400","DOIUrl":"10.1172/jci.insight.183400","url":null,"abstract":"<p><p>T follicular helper (Tfh) cells represent an important subset of CD4+ T cells that is crucial to the maturation and differentiation of B cells and the production of high-affinity antibodies. Because B cell activating-factor (BAFF), a vital B cell survival factor, is also crucial to B cell maturation and differentiation, we assessed the effects of BAFF on Tfh cell development and function. We demonstrated that deficiency of BAFF, but not of APRIL, markedly inhibited Tfh cell development, germinal center (GC) formation, and antigen-specific antibody production. The promoting effect of BAFF on Tfh cell development was dependent on expression of BR3 on T cells, and its promoting effect on GC formation was dependent on expression of BR3 on both T cells and B cells. BAFF directly promoted expression of the Tfh cell-characteristic genes via NF-κB signaling. This effect did need BR3 expression. Thus, BAFF not only has direct effects on B cells, but it also has direct effects on Tfh cell differentiation via engagement of BR3, which collectively promoted GC formation and production of high-affinity antibodies. This dual effect of BAFF on B cells and Tfh cells may help explain the clinical utility of BAFF antagonists in the management of certain autoimmune diseases.</p>","PeriodicalId":14722,"journal":{"name":"JCI insight","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2024-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142346969","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}