Biochimica et biophysica acta. Molecular cell research最新文献_第9页

Histone deacetylase 11 regulates stress granule formation to promote endothelial-to-mesenchymal transition in atherosclerosis 组蛋白去乙酰化酶11调节应激颗粒形成，促进动脉粥样硬化中内皮向间质转化。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-07-19 DOI: 10.1016/j.bbamcr.2025.120026

Lingxuan Ren , Yizhen Liu , Danli Chen , Xiaoyu Rong , Jiazheng Wen , Zihan Zheng , Lifang Chen , Jianyu He , Weirong Wang , Rong Lin

Histone deacetylase 11 (HDAC11) is the only member of the class IV HDAC family and is involved in cardiovascular diseases (CVDs). Stress granule (SG) is non-membranous cytoplasmic foci induced by various stress conditions, and also has emerged as a key player for CVDs. However, the regulatory role of HDAC11 in SG formation and underlying mechanism during atherosclerosis remain elusive. Therefore, we aimed to investigate the effect of HDAC11 on SG in ApoE^−/− mice fed with a HFD and HUVECs induced by H₂O₂. Firstly, we found that the expression levels of SG core proteins G3BP1/2 and HDAC11 were increased in the aorta of ApoE^−/− mice fed with a HFD for 12w via analyses of Western blotting, Real-time PCR and immunofluorescence staining. In addition, endothelial-to-mesenchymal transition (EndMT) was occurred in the aorta of ApoE^−/− mice. Then, in vitro experiments demonstrated that treatment of HUVECs with H₂O₂ resulted in SG formation, HDAC11 upregulation, and EndMT occurrence. Furthermore, knockdown of HDAC11 by siRNA significantly attenuated SG formation and EndMT activation in HUVECs induced by H₂O₂. Silencing of HDAC11 suppressed H₂O₂-induced EndMT activation in HUVECs, which may be attributed to increased acetylation of G3BP1/2 and the consequent impairment of SG formation. Further studies found that suppression of SG formation not only facilitated the expression of endothelial markers, but also decreased the levels of mesenchymal cell markers. Taken together, these findings identified that HDAC11 may regulate SG formation to promote EndMT in atherosclerosis, targeting SG could represent a novel therapeutic strategy for addressing the underlying mechanisms of atherosclerosis.

组蛋白去乙酰化酶11 （HDAC11）是IV类HDAC家族中唯一的成员，与心血管疾病（cvd）有关。应激颗粒（Stress particle， SG）是由各种应激条件诱导的非膜性细胞质病灶，也是心血管疾病的重要发病机制。然而，在动脉粥样硬化过程中，HDAC11在SG形成中的调节作用及其机制尚不清楚。因此，我们旨在研究HDAC11对H2O2诱导的HFD和HUVECs对ApoE-/-小鼠SG的影响。首先，我们通过Western blotting、Real-time PCR和免疫荧光染色分析发现，经HFD喂养12w的ApoE-/-小鼠主动脉中SG核心蛋白G3BP1/2和HDAC11的表达水平升高。此外，在ApoE-/-小鼠的主动脉中发生了内皮到间质转化（EndMT）。然后，体外实验表明，H2O2处理HUVECs可导致SG形成、HDAC11上调和EndMT的发生。此外，siRNA敲低HDAC11可显著减弱H2O2诱导的HUVECs中SG的形成和EndMT的激活。HDAC11的沉默抑制了h2o2诱导的HUVECs中EndMT的激活，这可能是由于G3BP1/2乙酰化降低，从而损害了SG的形成。进一步的研究发现，抑制SG的形成促进内皮细胞标志物的表达降低，但降低间充质细胞标志物的水平。综上所述，这些发现表明HDAC11可能调节SG的形成以促进动脉粥样硬化中的EndMT，靶向SG可能是解决动脉粥样硬化潜在机制的一种新的治疗策略。

{"title":"Histone deacetylase 11 regulates stress granule formation to promote endothelial-to-mesenchymal transition in atherosclerosis","authors":"Lingxuan Ren , Yizhen Liu , Danli Chen , Xiaoyu Rong , Jiazheng Wen , Zihan Zheng , Lifang Chen , Jianyu He , Weirong Wang , Rong Lin","doi":"10.1016/j.bbamcr.2025.120026","DOIUrl":"10.1016/j.bbamcr.2025.120026","url":null,"abstract":"<div><div>Histone deacetylase 11 (HDAC11) is the only member of the class IV HDAC family and is involved in cardiovascular diseases (CVDs). Stress granule (SG) is non-membranous cytoplasmic foci induced by various stress conditions, and also has emerged as a key player for CVDs. However, the regulatory role of HDAC11 in SG formation and underlying mechanism during atherosclerosis remain elusive. Therefore, we aimed to investigate the effect of HDAC11 on SG in ApoE<sup>−/−</sup> mice fed with a HFD and HUVECs induced by H<sub>2</sub>O<sub>2</sub>. Firstly, we found that the expression levels of SG core proteins G3BP1/2 and HDAC11 were increased in the aorta of ApoE<sup>−/−</sup> mice fed with a HFD for 12w via analyses of Western blotting, Real-time PCR and immunofluorescence staining. In addition, endothelial-to-mesenchymal transition (EndMT) was occurred in the aorta of ApoE<sup>−/−</sup> mice. Then, in vitro experiments demonstrated that treatment of HUVECs with H<sub>2</sub>O<sub>2</sub> resulted in SG formation, HDAC11 upregulation, and EndMT occurrence. Furthermore, knockdown of HDAC11 by siRNA significantly attenuated SG formation and EndMT activation in HUVECs induced by H<sub>2</sub>O<sub>2</sub>. Silencing of HDAC11 suppressed H<sub>2</sub>O<sub>2</sub>-induced EndMT activation in HUVECs, which may be attributed to increased acetylation of G3BP1/2 and the consequent impairment of SG formation. Further studies found that suppression of SG formation not only facilitated the expression of endothelial markers, but also decreased the levels of mesenchymal cell markers. Taken together, these findings identified that HDAC11 may regulate SG formation to promote EndMT in atherosclerosis, targeting SG could represent a novel therapeutic strategy for addressing the underlying mechanisms of atherosclerosis.</div></div>","PeriodicalId":8754,"journal":{"name":"Biochimica et biophysica acta. Molecular cell research","volume":"1872 7","pages":"Article 120026"},"PeriodicalIF":4.6,"publicationDate":"2025-07-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144681929","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The thylakoidal Tat receptor complex appears as a homo-trimeric TatC core with three associated TatB subunits 类囊体Tat受体复合物表现为具有三个相关TatB亚基的同源三聚体TatC核心。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-07-18 DOI: 10.1016/j.bbamcr.2025.120024

Matthias Reimers , Mario Jakob , Ralf Bernd Klösgen

The Twin-arginine translocation (Tat) machinery, which is found in most cellular membranes containing a respiratory or photosynthetic electron transport chain, is characterized by its unique ability to catalyze membrane transport of folded proteins without impairing the membrane potential. In plant thylakoids, Tat machinery consists of three subunits, TatA, TatB, and TatC, with the latter two, TatB and TatC, forming membrane-integral multimeric TatBC receptor complexes. Here we have analyzed the stability and the subunit composition of these complexes after solubilization of thylakoids with the mild detergent digitonin as well as after additional affinity-purification. Employing different detergent combinations and/or heat treatment (40 °C) followed by BN-PAGE and Western analysis we could identify four distinct Tat complexes with apparent molecular masses ranging from approximately 230 kDa to 620 kDa. Treatment of the largest Tat complex with either heat or detergents like DDM or Triton X-114 led to its stepwise breakdown into the three smaller complexes resulting from the successive release of TatB subunits from a relatively stable TatC core complex. From these data we postulate that the fully assembled, physiologically active TatBC receptor complex consists of a stable, trimeric TatC core to which three TatB subunits are bound independently from each other.

双精氨酸易位（Tat）机制存在于大多数含有呼吸或光合电子传递链的细胞膜中，其特点是其独特的催化折叠蛋白的膜运输能力而不损害膜电位。在植物类囊体中，Tat机制由三个亚基TatA、TatB和TatC组成，后两个亚基TatB和TatC形成膜积分多聚体TatBC受体复合物。在这里，我们分析了这些配合物的稳定性和亚基组成与温和的洗涤剂洋地黄苷溶解后的类囊体，并经过额外的亲和纯化。采用不同的洗涤剂组合和/或热处理（40 °C），然后进行BN-PAGE和Western分析，我们可以鉴定出四种不同的Tat配合物，其表观分子质量约为230 kDa至620 kDa。用热或DDM或Triton X-114等洗涤剂处理最大的Tat配合物，导致其逐步分解为三个较小的配合物，这是由于相对稳定的TatC核心配合物连续释放TatB亚基。根据这些数据，我们假设完全组装的生理活性TatBC受体复合物由一个稳定的三聚体TatC核心组成，其中三个TatB亚基彼此独立结合。

{"title":"The thylakoidal Tat receptor complex appears as a homo-trimeric TatC core with three associated TatB subunits","authors":"Matthias Reimers , Mario Jakob , Ralf Bernd Klösgen","doi":"10.1016/j.bbamcr.2025.120024","DOIUrl":"10.1016/j.bbamcr.2025.120024","url":null,"abstract":"<div><div>The Twin-arginine translocation (Tat) machinery, which is found in most cellular membranes containing a respiratory or photosynthetic electron transport chain, is characterized by its unique ability to catalyze membrane transport of folded proteins without impairing the membrane potential. In plant thylakoids, Tat machinery consists of three subunits, TatA, TatB, and TatC, with the latter two, TatB and TatC, forming membrane-integral multimeric TatBC receptor complexes. Here we have analyzed the stability and the subunit composition of these complexes after solubilization of thylakoids with the mild detergent digitonin as well as after additional affinity-purification. Employing different detergent combinations and/or heat treatment (40 °C) followed by BN-PAGE and Western analysis we could identify four distinct Tat complexes with apparent molecular masses ranging from approximately 230 kDa to 620 kDa. Treatment of the largest Tat complex with either heat or detergents like DDM or Triton X-114 led to its stepwise breakdown into the three smaller complexes resulting from the successive release of TatB subunits from a relatively stable TatC core complex. From these data we postulate that the fully assembled, physiologically active TatBC receptor complex consists of a stable, trimeric TatC core to which three TatB subunits are bound independently from each other.</div></div>","PeriodicalId":8754,"journal":{"name":"Biochimica et biophysica acta. Molecular cell research","volume":"1872 7","pages":"Article 120024"},"PeriodicalIF":4.6,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144673877","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

M6A-mediated uc003pes.1 stability regulates proliferation and migration of human coronary artery smooth muscle cells via scaffolding STAT1 and USP10 M6A-mediated uc003pes。1稳定性通过支架STAT1和USP10调控人冠状动脉平滑肌细胞的增殖和迁移。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-07-15 DOI: 10.1016/j.bbamcr.2025.120023

Rongli Jiang , Qiaowei Jia , Chengcheng Li , Shu He , Hanxiao Zhou , Mengmeng Ren , Lanyu Liang , Enzhi Jia

Background

The uc003pes.1 has been identified as a potential diagnostic biomarker for coronary artery disease (CAD) and is associated with the risk of CAD. However, the precise function and intricate mechanism of uc003pes.1 in CAD remain elusive.

Methods

To elucidate the underlying mechanism we employed various techniques including RNA pull-down assays coupled with mass spectrometry analysis, RIP, RNA MeRIP, as well as luciferase reporter gene analysis.

Results

Overexpression of uc003pes.1 suppressed the proliferation and migration of HCASMCs. The upregulation of uc003pes.1 can be attributed to m6A modification mediated by the methylase RBM15. In terms of downstream mechanisms, uc003pes.1 acts as a molecular scaffold facilitating the interaction between STAT1 and the USP10, thereby impeding ubiquitination degradation of STAT1 and promoting phosphorylation levels at Tyr701 and Ser727, ultimately influencing HCASMCs' proliferation and migration.

Conclusion

Our study has discovered, for the first time, that uc003pes.1, regulated by m6A modification, functions as a molecular scaffold for STAT1 and USP10. This interaction plays a crucial role in regulating the proliferation and migration of HCASMCs, thereby offering novel insights into the diagnosis and treatment of CAD.

背景：uc003pes。1已被确定为冠状动脉疾病（CAD）的潜在诊断生物标志物，并与CAD的风险相关。然而，uc003pes的精确功能和复杂机制。1在CAD仍然难以捉摸。方法：为了阐明潜在的机制，我们采用了多种技术，包括RNA下拉试验结合质谱分析、RIP、RNA MeRIP和荧光素酶报告基因分析。结果：uc003pes过表达。1抑制HCASMCs的增殖和迁移。uc003pes的上调。1可归因于甲基化酶RBM15介导的m6A修饰。在下游机制方面，uc003pes。1作为分子支架，促进STAT1与USP10相互作用，从而阻碍STAT1的泛素化降解，促进Tyr701和Ser727的磷酸化水平，最终影响HCASMCs的增殖和迁移。结论：我们的研究首次发现了uc003pe。1，受m6A修饰调控，作为STAT1和USP10的分子支架。这种相互作用在调节HCASMCs的增殖和迁移中起着至关重要的作用，从而为CAD的诊断和治疗提供了新的见解。

{"title":"M6A-mediated uc003pes.1 stability regulates proliferation and migration of human coronary artery smooth muscle cells via scaffolding STAT1 and USP10","authors":"Rongli Jiang , Qiaowei Jia , Chengcheng Li , Shu He , Hanxiao Zhou , Mengmeng Ren , Lanyu Liang , Enzhi Jia","doi":"10.1016/j.bbamcr.2025.120023","DOIUrl":"10.1016/j.bbamcr.2025.120023","url":null,"abstract":"<div><h3>Background</h3><div>The <em>uc003pes.1</em> has been identified as a potential diagnostic biomarker for coronary artery disease (CAD) and is associated with the risk of CAD. However, the precise function and intricate mechanism of <em>uc003pes.1</em> in CAD remain elusive.</div></div><div><h3>Methods</h3><div>To elucidate the underlying mechanism we employed various techniques including RNA pull-down assays coupled with mass spectrometry analysis, RIP, RNA MeRIP, as well as luciferase reporter gene analysis.</div></div><div><h3>Results</h3><div>Overexpression of uc003pes.1 suppressed the proliferation and migration of HCASMCs. The upregulation of uc003pes.1 can be attributed to m6A modification mediated by the methylase RBM15. In terms of downstream mechanisms, uc003pes.1 acts as a molecular scaffold facilitating the interaction between STAT1 and the USP10, thereby impeding ubiquitination degradation of STAT1 and promoting phosphorylation levels at Tyr701 and Ser727, ultimately influencing HCASMCs' proliferation and migration.</div></div><div><h3>Conclusion</h3><div>Our study has discovered, for the first time, that uc003pes.1, regulated by m6A modification, functions as a molecular scaffold for STAT1 and USP10. This interaction plays a crucial role in regulating the proliferation and migration of HCASMCs, thereby offering novel insights into the diagnosis and treatment of CAD.</div></div>","PeriodicalId":8754,"journal":{"name":"Biochimica et biophysica acta. Molecular cell research","volume":"1872 7","pages":"Article 120023"},"PeriodicalIF":4.6,"publicationDate":"2025-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144648360","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

GRK5 regulates mitotic progression and promotes resistance against anti-mitotic agents in a CDK1 and AKT1 dependent manner GRK5以CDK1和AKT1依赖的方式调节有丝分裂进程并促进对抗有丝分裂药物的抗性。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-07-13 DOI: 10.1016/j.bbamcr.2025.120020

Nusrat Nabi , Syed Qaaifah Gillani , Marjan Fatima , Misbah Un Nisa , Zarka Sarwar , Sameer Ahmed Bhat , Irfana Reshi , Shareen Bashir , Fazl Q Parray , Shaida Andrabi

G protein receptor kinase 5 (GRK5) is a serine/threonine protein kinase that belongs to the family of G protein receptor kinases (GRKs), which are important regulators of G protein-coupled receptor (GPCR) functions. GRK5 regulates signaling by binding to various receptors on the plasma membrane or by regulating transcription within the nucleus. It also has been found to critically regulate several physiological processes including vascular remodelling, invasion, metastasis and migration of the cells. Although its role in cancer progression and metastasis is known, its role in cell division, and particularly in mitosis has not been investigated much. Here, we report that GRK5 is an important mitotic protein and is regulated by well-known cellular proteins that have a critical role in cell cycle regulation, especially mitosis. In particular, we show that GRK5 is regulated by two key players of mitosis, AKT1 and CDK1, which regulate GRK5 by interacting with it. We also provide evidence that GRK5 protein levels fluctuate throughout the cell cycle and reach their maximum during mitosis. Further, we report that overexpression of GRK5 promotes resistance against cell death that is induced by polyomavirus small T (PolST) antigen and different chemotherapeutic drugs including paclitaxel. Additionally, we found that GRK5 levels are upregulated in colorectal cancers supporting a potential role in tumor progression. Our findings thus add GRK5 to the growing list of mitotic kinases, which play a role in the regulation of cell cycle and promotion of drug resistance in cancer.

G蛋白受体激酶5 （GRK5）是一种丝氨酸/苏氨酸蛋白激酶，属于G蛋白受体激酶（GRKs）家族，是G蛋白偶联受体（GPCR）功能的重要调节器。GRK5通过与质膜上的各种受体结合或通过调节细胞核内的转录来调节信号传导。它还被发现对血管重构、细胞的侵袭、转移和迁移等生理过程具有重要的调控作用。虽然它在癌症进展和转移中的作用是已知的，但它在细胞分裂中的作用，特别是在有丝分裂中的作用尚未得到太多的研究。在这里，我们报道了GRK5是一种重要的有丝分裂蛋白，受众所周知的细胞蛋白的调控，这些蛋白在细胞周期调控，特别是有丝分裂中起着关键作用。特别是，我们发现GRK5受到有丝分裂的两个关键参与者AKT1和CDK1的调节，它们通过与GRK5相互作用来调节GRK5。我们还提供证据表明GRK5蛋白水平在整个细胞周期中波动，并在有丝分裂期间达到最大值。此外，我们报道了GRK5的过表达促进了对多瘤病毒小T （PolST）抗原和不同化疗药物（包括紫杉醇）诱导的细胞死亡的抵抗。此外，我们发现GRK5水平在结直肠癌中上调，支持在肿瘤进展中的潜在作用。因此，我们的研究结果将GRK5添加到不断增长的有丝分裂激酶列表中，这些激酶在调节细胞周期和促进癌症耐药中发挥作用。

{"title":"GRK5 regulates mitotic progression and promotes resistance against anti-mitotic agents in a CDK1 and AKT1 dependent manner","authors":"Nusrat Nabi , Syed Qaaifah Gillani , Marjan Fatima , Misbah Un Nisa , Zarka Sarwar , Sameer Ahmed Bhat , Irfana Reshi , Shareen Bashir , Fazl Q Parray , Shaida Andrabi","doi":"10.1016/j.bbamcr.2025.120020","DOIUrl":"10.1016/j.bbamcr.2025.120020","url":null,"abstract":"<div><div>G protein receptor kinase 5 (GRK5) is a serine/threonine protein kinase that belongs to the family of G protein receptor kinases (GRKs), which are important regulators of G protein-coupled receptor (GPCR) functions. GRK5 regulates signaling by binding to various receptors on the plasma membrane or by regulating transcription within the nucleus. It also has been found to critically regulate several physiological processes including vascular remodelling, invasion, metastasis and migration of the cells. Although its role in cancer progression and metastasis is known, its role in cell division, and particularly in mitosis has not been investigated much. Here, we report that GRK5 is an important mitotic protein and is regulated by well-known cellular proteins that have a critical role in cell cycle regulation, especially mitosis. In particular, we show that GRK5 is regulated by two key players of mitosis, AKT1 and CDK1, which regulate GRK5 by interacting with it. We also provide evidence that GRK5 protein levels fluctuate throughout the cell cycle and reach their maximum during mitosis. Further, we report that overexpression of GRK5 promotes resistance against cell death that is induced by polyomavirus small T (PolST) antigen and different chemotherapeutic drugs including paclitaxel. Additionally, we found that GRK5 levels are upregulated in colorectal cancers supporting a potential role in tumor progression. Our findings thus add GRK5 to the growing list of mitotic kinases, which play a role in the regulation of cell cycle and promotion of drug resistance in cancer.</div></div>","PeriodicalId":8754,"journal":{"name":"Biochimica et biophysica acta. Molecular cell research","volume":"1872 7","pages":"Article 120020"},"PeriodicalIF":4.6,"publicationDate":"2025-07-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144641644","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Synthesis, characterization and size-dependent cytotoxicity of magnesium ammonium phosphate hexahydrate crystals of different sizes on renal epithelial cells 不同大小磷酸铵镁六水晶体的合成、表征及其对肾上皮细胞的细胞毒性。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-07-12 DOI: 10.1016/j.bbamcr.2025.120022

Run-Min Tan , Xin-Yi Tong , Wen-Xuan Dai, Jia-Yi Zhang, Jian-Ming Ouyang

Magnesium ammonium phosphate hexahydrate (MAP) crystals with sizes of 98.5 ± 20.6 nm, 310 ± 67 nm, 1.12 ± 0.34 μm, and 3.23 ± 0.90 μm were synthesized and characterized. These crystals can cause damage to renal tubular epithelial cells (HK−2), which is manifested by crystal-induced cell morphological changes, cell viability, a decrease in superoxide dismutase and mitochondrial membrane potential. In addition, there were crystal-induced increases in reactive oxygen species, lactate dehydrogenase, and malondialdehyde levels, as well as phosphatidylserine ectropion. That is, MAP crystals can lead to cell necrosis and apoptosis, and promote the release of inflammatory cytokines IL-18 and IL-6. The cytotoxicity of MAP crystals has a size effect, that is, the cytotoxicity is: MAP-100 nm > MAP-300 nm > MAP-1 μm > MAP-3 μm. The factors enhancing the cytotoxicity of MAP include small size, large specific surface area, and a more negative crystal surface zeta potential. Nano-crystal MAP-100 nm mainly causes cell death by inducing extensive cell necrosis. When the larger-sized MAP-300 nm, MAP-1 μm and MAP-3 μm crystals acted on HK-2 cells, cell necrosis, apoptosis and autophagy occurred simultaneously. Investigating the relationship between MAP crystal size and cytotoxicity may provide insights into elucidating the mechanism of MAP stone (struvite) formation and preventing its occurrence.

磷酸镁铵六水合物(MAP)晶体大小为98.5 ±20.6  nm, 310 ± 67 海里, 1.12±0.34  μm和3.23 ±0.90  μm是合成和特征。这些晶体可引起肾小管上皮细胞（HK-2）的损伤，表现为晶体诱导的细胞形态改变、细胞活力降低、超氧化物歧化酶和线粒体膜电位降低。此外，晶体诱导活性氧、乳酸脱氢酶和丙二醛水平的增加，以及磷脂酰丝氨酸外翻。即MAP晶体可导致细胞坏死和凋亡，促进炎性细胞因子IL-18和IL-6的释放。地图的细胞毒性晶体尺寸效应,也就是说,细胞毒性是:地图- 100 nm > 地图- 300 nm > MAP-1 μm > MAP-3 μm。增强MAP细胞毒性的因素包括体积小、比表面积大、晶体表面zeta电位更负。纳米晶体MAP-100 nm主要通过诱导细胞广泛坏死导致细胞死亡。当较大尺寸的MAP-300 nm、MAP-1 μm和MAP-3 μm晶体作用于HK-2细胞时，细胞同时发生坏死、凋亡和自噬。研究MAP晶体大小与细胞毒性的关系，有助于阐明MAP结石（鸟粪石）的形成机制和预防其发生。

{"title":"Synthesis, characterization and size-dependent cytotoxicity of magnesium ammonium phosphate hexahydrate crystals of different sizes on renal epithelial cells","authors":"Run-Min Tan , Xin-Yi Tong , Wen-Xuan Dai, Jia-Yi Zhang, Jian-Ming Ouyang","doi":"10.1016/j.bbamcr.2025.120022","DOIUrl":"10.1016/j.bbamcr.2025.120022","url":null,"abstract":"<div><div>Magnesium ammonium phosphate hexahydrate (MAP) crystals with sizes of 98.5 ± 20.6 nm, 310 ± 67 nm, 1.12 ± 0.34 μm, and 3.23 ± 0.90 μm were synthesized and characterized. These crystals can cause damage to renal tubular epithelial cells (HK−2), which is manifested by crystal-induced cell morphological changes, cell viability, a decrease in superoxide dismutase and mitochondrial membrane potential. In addition, there were crystal-induced increases in reactive oxygen species, lactate dehydrogenase, and malondialdehyde levels, as well as phosphatidylserine ectropion. That is, MAP crystals can lead to cell necrosis and apoptosis, and promote the release of inflammatory cytokines IL-18 and IL-6. The cytotoxicity of MAP crystals has a size effect, that is, the cytotoxicity is: MAP-100 nm > MAP-300 nm > MAP-1 μm > MAP-3 μm. The factors enhancing the cytotoxicity of MAP include small size, large specific surface area, and a more negative crystal surface zeta potential. Nano-crystal MAP-100 nm mainly causes cell death by inducing extensive cell necrosis. When the larger-sized MAP-300 nm, MAP-1 μm and MAP-3 μm crystals acted on HK-2 cells, cell necrosis, apoptosis and autophagy occurred simultaneously. Investigating the relationship between MAP crystal size and cytotoxicity may provide insights into elucidating the mechanism of MAP stone (struvite) formation and preventing its occurrence.</div></div>","PeriodicalId":8754,"journal":{"name":"Biochimica et biophysica acta. Molecular cell research","volume":"1872 7","pages":"Article 120022"},"PeriodicalIF":4.6,"publicationDate":"2025-07-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144636064","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Deubiquitinases in metabolic diseases, fibrosis and cancer of the liver 去泛素酶在代谢性疾病、纤维化和肝癌中的作用

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-07-11 DOI: 10.1016/j.bbamcr.2025.120021

Ananya Thakur, Kateryna Kubaichuk, Thomas Kietzmann

Obesity is a key driver of the progression from metabolic dysfunction-associated steatotic liver disease (MASLD), to metabolic dysfunction-associated steatohepatitis (MASH), liver fibrosis, and hepatocellular carcinoma (HCC). Excess adiposity and elevated circulating fatty acids disrupt metabolic, inflammatory, and signaling pathways, creating conditions permissive for hepatic injury, inflammation, fibrogenesis, and carcinogenesis. One of the mechanisms that regulates these pathways is ubiquitylation, a post-translational modification that controls protein degradation and cellular signaling. Deubiquitinases (DUBs) counterbalance this process by removing ubiquitin chains, thereby maintaining cellular homeostasis.

This review examines the role of DUBs in obesity-induced MASLD progression, focusing on how dysregulated DUB expression affects insulin signaling, lipogenesis, inflammation, and oxidative stress. While DUBs have been extensively studied in cancer and metabolic syndrome, their therapeutic potential in obesity-related MASH, liver fibrosis, and HCC has not been fully explored.

By synthesizing clinical and experimental evidence, we highlight DUBs as promising precision medicine tools for both patient stratification and therapeutic intervention. This review emphasizes the critical need for further translational research to develop DUB-targeted strategies for early intervention in MASLD progression.

肥胖是代谢功能障碍相关脂肪性肝病（MASLD）发展为代谢功能障碍相关脂肪性肝炎（MASH）、肝纤维化和肝细胞癌（HCC）的关键驱动因素。过量的脂肪和升高的循环脂肪酸会破坏代谢、炎症和信号通路，为肝损伤、炎症、纤维化和癌变创造有利条件。调控这些途径的机制之一是泛素化，这是一种控制蛋白质降解和细胞信号传导的翻译后修饰。去泛素酶（DUBs）通过去除泛素链来平衡这一过程，从而维持细胞稳态。这篇综述探讨了DUB在肥胖诱导的MASLD进展中的作用，重点关注DUB表达失调如何影响胰岛素信号、脂肪生成、炎症和氧化应激。虽然DUBs在癌症和代谢综合征方面已被广泛研究，但其在肥胖相关的MASH、肝纤维化和HCC方面的治疗潜力尚未得到充分探索。通过综合临床和实验证据，我们强调dub是有前途的精准医学工具，用于患者分层和治疗干预。这篇综述强调，迫切需要进一步的转化研究，以制定针对dub的MASLD进展早期干预策略。

{"title":"Deubiquitinases in metabolic diseases, fibrosis and cancer of the liver","authors":"Ananya Thakur, Kateryna Kubaichuk, Thomas Kietzmann","doi":"10.1016/j.bbamcr.2025.120021","DOIUrl":"10.1016/j.bbamcr.2025.120021","url":null,"abstract":"<div><div>Obesity is a key driver of the progression from metabolic dysfunction-associated steatotic liver disease (MASLD), to metabolic dysfunction-associated steatohepatitis (MASH), liver fibrosis, and hepatocellular carcinoma (HCC). Excess adiposity and elevated circulating fatty acids disrupt metabolic, inflammatory, and signaling pathways, creating conditions permissive for hepatic injury, inflammation, fibrogenesis, and carcinogenesis. One of the mechanisms that regulates these pathways is ubiquitylation, a post-translational modification that controls protein degradation and cellular signaling. Deubiquitinases (DUBs) counterbalance this process by removing ubiquitin chains, thereby maintaining cellular homeostasis.</div><div>This review examines the role of DUBs in obesity-induced MASLD progression, focusing on how dysregulated DUB expression affects insulin signaling, lipogenesis, inflammation, and oxidative stress. While DUBs have been extensively studied in cancer and metabolic syndrome, their therapeutic potential in obesity-related MASH, liver fibrosis, and HCC has not been fully explored.</div><div>By synthesizing clinical and experimental evidence, we highlight DUBs as promising precision medicine tools for both patient stratification and therapeutic intervention. This review emphasizes the critical need for further translational research to develop DUB-targeted strategies for early intervention in MASLD progression.</div></div>","PeriodicalId":8754,"journal":{"name":"Biochimica et biophysica acta. Molecular cell research","volume":"1872 7","pages":"Article 120021"},"PeriodicalIF":4.6,"publicationDate":"2025-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144623698","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Modulation of mitochondrial quality control through autophagic pathway in familial Alzheimer's disease 家族性阿尔茨海默病自噬途径对线粒体质量控制的调节。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-07-07 DOI: 10.1016/j.bbamcr.2025.120019

Adriana Limone , Clelia Di Napoli , Giusy De Rosa , Silvia Bagnoli , Antonella Izzo , Claudio Procaccini , Giuseppe Matarese , Benedetta Nacmias , Antonio Lavecchia , Daniela Sarnataro

Autophagy is a highly conserved cellular catabolic process recognized as an essential pathway for the maintenance of cellular homeostasis. Growing evidence implicates autophagic dysfunction in the pathogenesis of several neurodegenerative disorders, including Alzheimer's disease (AD), thus its modulation might represent an interesting therapeutic tool. Searching for a compound that stimulates autophagic pathway, led us to identify the inhibitor of RPSA receptor, NSC47924. In this study, we show that, NSC47924 down-modulated Akt-mTOR-axis pathway, the master regulator of autophagy, which was abnormally hyperactivated in fibroblasts from genetic AD-affected patients. Consistently, by monitoring the conversion of LC3, we found that inhibition of RPSA enhanced and restored the compromised autophagic flux. Moreover, by qRT-PCR analysis we found that inhibitor treatment upregulated the expression of autophagy-linked genes.

Importantly, AD-affected fibroblasts exhibited massive mitochondrial network fragmentation and mitophagy defects, which were restored through the stimulation of autophagy induced by RPSA inhibition. Consistent with an efficient elimination of dysfunctional mitochondria, we found that the turnover of both the mitophagy regulators PINK1 and Parkin and the autophagic receptors p62, NDP52, OPTN, was modulated, thus restoring a highly interconnected organelle's network. In addition, the improvement of mitochondrial morphology correlated with a functional recovery, as assessed by Seahorse analysis and mitochondrial ROS production evaluation. Collectively, our findings suggest that RPSA inhibition stimulates an autophagic pathway promoting the efficient removal of damaged mitochondria, favouring the recovery of cellular homeostasis, and counteracting crucial AD pathogenic mechanisms.

自噬是一种高度保守的细胞分解代谢过程，被认为是维持细胞稳态的重要途径。越来越多的证据表明，自噬功能障碍在包括阿尔茨海默病（AD）在内的几种神经退行性疾病的发病机制中起作用，因此其调节可能是一种有趣的治疗工具。为了寻找刺激自噬通路的化合物，我们确定了RPSA受体的抑制剂NSC47924。在这项研究中，我们发现NSC47924下调了自噬的主要调节因子akt - mtor轴通路，该通路在遗传性ad患者的成纤维细胞中异常亢进。同样，通过监测LC3的转化，我们发现抑制RPSA可以增强和恢复受损的自噬通量。此外，通过qRT-PCR分析，我们发现抑制剂处理上调自噬相关基因的表达。重要的是，受ad影响的成纤维细胞表现出大量线粒体网络断裂和线粒体自噬缺陷，这些缺陷通过抑制RPSA诱导的自噬刺激得以恢复。与功能失调线粒体的有效消除一致，我们发现线粒体自噬调节因子PINK1和Parkin以及自噬受体p62， NDP52， OPTN的更替都被调节，从而恢复了高度互联的细胞器网络。此外，海马分析和线粒体ROS生成评估表明，线粒体形态的改善与功能恢复相关。总的来说，我们的研究结果表明，RPSA抑制刺激自噬途径，促进受损线粒体的有效清除，有利于细胞稳态的恢复，并抵消AD的关键致病机制。

{"title":"Modulation of mitochondrial quality control through autophagic pathway in familial Alzheimer's disease","authors":"Adriana Limone , Clelia Di Napoli , Giusy De Rosa , Silvia Bagnoli , Antonella Izzo , Claudio Procaccini , Giuseppe Matarese , Benedetta Nacmias , Antonio Lavecchia , Daniela Sarnataro","doi":"10.1016/j.bbamcr.2025.120019","DOIUrl":"10.1016/j.bbamcr.2025.120019","url":null,"abstract":"<div><div>Autophagy is a highly conserved cellular catabolic process recognized as an essential pathway for the maintenance of cellular homeostasis. Growing evidence implicates autophagic dysfunction in the pathogenesis of several neurodegenerative disorders, including Alzheimer's disease (AD), thus its modulation might represent an interesting therapeutic tool. Searching for a compound that stimulates autophagic pathway, led us to identify the inhibitor of RPSA receptor, NSC47924. In this study, we show that, NSC47924 down-modulated Akt-mTOR-axis pathway, the master regulator of autophagy, which was abnormally hyperactivated in fibroblasts from genetic AD-affected patients. Consistently, by monitoring the conversion of LC3, we found that inhibition of RPSA enhanced and restored the compromised autophagic flux. Moreover, by qRT-PCR analysis we found that inhibitor treatment upregulated the expression of autophagy-linked genes.</div><div>Importantly, AD-affected fibroblasts exhibited massive mitochondrial network fragmentation and mitophagy defects, which were restored through the stimulation of autophagy induced by RPSA inhibition. Consistent with an efficient elimination of dysfunctional mitochondria, we found that the turnover of both the mitophagy regulators PINK1 and Parkin and the autophagic receptors p62, NDP52, OPTN, was modulated, thus restoring a highly interconnected organelle's network. In addition, the improvement of mitochondrial morphology correlated with a functional recovery, as assessed by Seahorse analysis and mitochondrial ROS production evaluation. Collectively, our findings suggest that RPSA inhibition stimulates an autophagic pathway promoting the efficient removal of damaged mitochondria, favouring the recovery of cellular homeostasis, and counteracting crucial AD pathogenic mechanisms.</div></div>","PeriodicalId":8754,"journal":{"name":"Biochimica et biophysica acta. Molecular cell research","volume":"1872 7","pages":"Article 120019"},"PeriodicalIF":4.6,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144599237","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Epigenetic targeting of DNA damage response (DDR)-related mechanisms to overcome acquired cisplatin resistance of tumor cells 表观遗传靶向DNA损伤反应（DDR）相关机制克服肿瘤细胞获得性顺铂耐药。

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-07-02 DOI: 10.1016/j.bbamcr.2025.120018

Marlena Sekeres , Daniel Lengle , Lena Abbey , Matthias Ulrich Kassack , Fabian Fischer , Thomas Kurz , Gerhard Fritz

Mechanisms of the DNA damage response (DDR) are considered as useful targets to overcome tumor cell resistance. We investigated the impact of various classes of histone deacetylase inhibitors (HDACi) (i.e. broad-spectrum HDACi (vorinostat), class I HDACi (entinostat), preferential class IIb HDAC6i (ricolinostat) and dual HDAC class I/IIb inhibitors (HDAC1/6i)) on mechanisms of the DDR using parental (J82^WT) and cisplatin (CisPt)-resistant bladder carcinoma cells (J82^CisR). Mono-treatment with entinostat revealed relatively low genotoxic and DDR-activating potency, while showing similar antiproliferative, cytotoxic and pro-apoptotic activities as the other HDACi. Despite its low DNA double-strand break (DSB) forming potency with mono-treatment, entinostat conferred the highest synergistic cytotoxicity (CI ≤ 0.8) in J82^CisR following co-treatment with cisplatin. Notably, this effect is independent of Pt-(GpG)-DNA-intrastrand-crosslink formation. Entinostat increased the level of CisPt-induced DNA strand-breaks, promotes replication and transcription blockage, pRPA32 foci formation, PARP- and caspase-7 cleavage, influenced the activation of DDR-related factors (e.g. γH2AX, pp53, pRPA32, pKap1), reverted CisPt-induced p53 acetylation and blocked drug-stimulated mRNA expression of multiple DSB (HR, NHEJ)- and BER repair-related factors. Summarizing, the class I-selective HDACi entinostat reveals the lowest DDR-activating potency in mono-treatment and the highest anticancer efficacy in combination with CisPt, likely involving inhibition of HDAC3. This is due to amplification of replicative and transcriptional stress caused by CisPt treatment as well as interference with mechanisms of DDR and DNA repair, eventually promoting apoptosis. Thus, epigenetic targeting of DDR-related death pathways by class I HDACi is useful to overcome acquired CisPt resistance of tumor cells.

DNA损伤反应（DDR）的机制被认为是克服肿瘤细胞耐药的有效靶点。我们研究了不同类别的组蛋白去乙酰化酶抑制剂（HDACi）（即广谱HDACi（伏立诺他），I类HDACi（恩替诺他），优先IIb类HDAC6i（立可立他）和双重HDAC类I/IIb抑制剂（HDAC1/6i））对亲代（J82WT）和顺铂（CisPt）耐药膀胱癌细胞（J82CisR）的DDR机制的影响。恩替诺他单药治疗显示出相对较低的基因毒性和ddr激活效力，同时显示出与其他HDACi相似的抗增殖、细胞毒性和促凋亡活性。尽管单独治疗时，恩替司他的DNA双链断裂（DSB）形成能力较低，但与顺铂联合治疗后，在J82CisR中，恩替司他具有最高的协同细胞毒性（CI ≤ 0.8）。值得注意的是，这种效应与Pt-(GpG)- dna -链内交联的形成无关。Entinostat增加cispt诱导的DNA链断裂水平，促进复制和转录阻断，pRPA32灶形成，PARP-和caspase-7切割，影响ddr相关因子（如γH2AX， pp53, pRPA32, pKap1）的激活，逆转cispt诱导的p53乙酰化，阻断药物刺激的多种DSB (HR， NHEJ)-和BER修复相关因子的mRNA表达。综上所述，i类选择性HDACi entinostat单用时ddr激活效力最低，与CisPt合用时抗癌效果最高，可能与抑制HDAC3有关。这是由于CisPt治疗引起的复制和转录应激扩增，以及对DDR机制和DNA修复的干扰，最终促进细胞凋亡。因此，通过I类HDACi的表观遗传靶向ddr相关的死亡途径有助于克服肿瘤细胞获得性CisPt耐药。

{"title":"Epigenetic targeting of DNA damage response (DDR)-related mechanisms to overcome acquired cisplatin resistance of tumor cells","authors":"Marlena Sekeres , Daniel Lengle , Lena Abbey , Matthias Ulrich Kassack , Fabian Fischer , Thomas Kurz , Gerhard Fritz","doi":"10.1016/j.bbamcr.2025.120018","DOIUrl":"10.1016/j.bbamcr.2025.120018","url":null,"abstract":"<div><div>Mechanisms of the DNA damage response (DDR) are considered as useful targets to overcome tumor cell resistance. We investigated the impact of various classes of histone deacetylase inhibitors (HDACi) (i.e. broad-spectrum HDACi (vorinostat), class I HDACi (entinostat), preferential class IIb HDAC6i (ricolinostat) and dual HDAC class I/IIb inhibitors (HDAC1/6i)) on mechanisms of the DDR using parental (J82<sup>WT</sup>) and cisplatin (CisPt)-resistant bladder carcinoma cells (J82<sup>CisR</sup>). Mono-treatment with entinostat revealed relatively low genotoxic and DDR-activating potency, while showing similar antiproliferative, cytotoxic and pro-apoptotic activities as the other HDACi. Despite its low DNA double-strand break (DSB) forming potency with mono-treatment, entinostat conferred the highest synergistic cytotoxicity (CI ≤ 0.8) in J82<sup>CisR</sup> following co-treatment with cisplatin. Notably, this effect is independent of Pt-(GpG)-DNA-intrastrand-crosslink formation. Entinostat increased the level of CisPt-induced DNA strand-breaks, promotes replication and transcription blockage, pRPA32 foci formation, PARP- and caspase-7 cleavage, influenced the activation of DDR-related factors (e.g. γH2AX, pp53, pRPA32, pKap1), reverted CisPt-induced p53 acetylation and blocked drug-stimulated mRNA expression of multiple DSB (HR, NHEJ)- and BER repair-related factors. Summarizing, the class I-selective HDACi entinostat reveals the lowest DDR-activating potency in mono-treatment and the highest anticancer efficacy in combination with CisPt, likely involving inhibition of HDAC3. This is due to amplification of replicative and transcriptional stress caused by CisPt treatment as well as interference with mechanisms of DDR and DNA repair, eventually promoting apoptosis. Thus, epigenetic targeting of DDR-related death pathways by class I HDACi is useful to overcome acquired CisPt resistance of tumor cells.</div></div>","PeriodicalId":8754,"journal":{"name":"Biochimica et biophysica acta. Molecular cell research","volume":"1872 7","pages":"Article 120018"},"PeriodicalIF":4.6,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144564322","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

USP15 promotes brain cell disulfidptosis in mouse subjected to ischemic stroke through a mechanism involving deubiquitination of SETD1B USP15通过与SETD1B去泛素化有关的机制促进缺血性中风小鼠的脑细胞二亢

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-07-01 DOI: 10.1016/j.bbamcr.2025.120016

Hong-Rui Liu , Lin Wu , Kai-Jia Wang , Yan-Xi Che , Wen-Jun Zhu , Xi-Sheng Li , Yu Han , Guang-Rong Wei , Yi-Yue Zhang , Xiu-Ju Luo

Disulfidptosis due to excessive accumulation of disulfides is a novel form of regulated cell death. Whether disulfidptosis occurs in ischemic stroke and the underlying mechanisms remain elusive. RNA transcriptomics sequencing (RNA-seq) reveales that knockdown of SET domain containing 1B (SETD1B), a histone lysine methyltransferase, decreases the expression of disulfidptosis-related genes. Using Ubibrowser database, Ubiquitin-specific protease 15 (USP15) is predicted to be a deubiquitinase (DUB) for SETD1B. This study investigates whether SETD1B promote disulfidptosis in ischemic stroke via upregulating disulfidptosis-related genes and whether USP15 deubiquitinates SETD1B. A C57BL/6 J mouse model of ischemic stroke was established, which results in brain injury and upregulation of USP15 and SETD1B, concomitant with the increased disulfidptosis, as indicated by the increased level of NADP⁺/NADPH ratio, elevated protein levels of NCK associated protein 1 like (NCKAP1L) and WASP family protein member 2 (WAVE-2), disulfide bond accumulation, and cytoskeleton detachment from the cytoplasmic membrane. Similar results were observed in cultured HT22 cells subjected to oxygen-glucose deprivation plus reoxygenation (OGD/R), and USP15 or SETD1B siRNAs reversed these phenomena. Mechanistically, knockdown of SETD1B prevented H3K4me3 enrichment at the Nckap1l and Wasf2 promoters and reduced Nckap1l and Wasf2 expression. Moreover, knockdown of USP15 increased the ubiquitination level of SETD1B thus decreasing its protein level. Based on these findings, we concluded that SETD1B can promote disulfidptosis in stroke brain cells via a mechanism involving facilitating H3K4me3 enrichment at the Nckap1l and Wasf2 promoters, while USP15 is able to deubiquitinate SETD1B and increase SETD1B level in ischemic stroke mice.

由于过量积累的二硫化物导致的二硫下垂是一种新的细胞死亡形式。缺血性脑卒中是否发生双睑下垂及其潜在机制尚不清楚。RNA转录组学测序（RNA-seq）显示，组蛋白赖氨酸甲基转移酶（SET）结构域1B （SETD1B）的敲低可降低二硫分解相关基因的表达。利用Ubibrowser数据库，我们预测泛素特异性蛋白酶15 （USP15）是SETD1B的去泛素酶（DUB）。本研究探讨SETD1B是否通过上调二硫塌陷相关基因促进缺血性卒中的二硫塌陷，以及USP15是否去泛素化SETD1B。建立C57BL/ 6j小鼠缺血性卒中模型，脑损伤，USP15和SETD1B水平上调，伴有二硫质下沉增加，表现为NADP+/NADPH比水平升高，NCK相关蛋白1样蛋白（NCKAP1L）和WASP家族蛋白成员2（波浪-2）蛋白水平升高，二硫键积累，细胞骨架脱离细胞质膜。在氧-葡萄糖剥夺加再氧化（OGD/R）的HT22细胞中也观察到类似的结果，USP15或SETD1B sirna逆转了这些现象。从机制上讲，敲低SETD1B可阻止H3K4me3在Nckap1l和Wasf2启动子上的富集，降低Nckap1l和Wasf2的表达。此外，USP15的敲除增加了SETD1B的泛素化水平，从而降低了其蛋白水平。基于这些发现，我们得出结论，SETD1B可以通过促进Nckap1l和Wasf2启动子上H3K4me3富集的机制促进脑卒中细胞的二硫醇凋亡，而USP15可以使缺血性脑卒中小鼠的SETD1B去泛素化并增加SETD1B水平。

{"title":"USP15 promotes brain cell disulfidptosis in mouse subjected to ischemic stroke through a mechanism involving deubiquitination of SETD1B","authors":"Hong-Rui Liu , Lin Wu , Kai-Jia Wang , Yan-Xi Che , Wen-Jun Zhu , Xi-Sheng Li , Yu Han , Guang-Rong Wei , Yi-Yue Zhang , Xiu-Ju Luo","doi":"10.1016/j.bbamcr.2025.120016","DOIUrl":"10.1016/j.bbamcr.2025.120016","url":null,"abstract":"<div><div>Disulfidptosis due to excessive accumulation of disulfides is a novel form of regulated cell death. Whether disulfidptosis occurs in ischemic stroke and the underlying mechanisms remain elusive. RNA transcriptomics sequencing (RNA-seq) reveales that knockdown of SET domain containing 1B (SETD1B), a histone lysine methyltransferase, decreases the expression of disulfidptosis-related genes. Using Ubibrowser database, Ubiquitin-specific protease 15 (USP15) is predicted to be a deubiquitinase (DUB) for SETD1B. This study investigates whether SETD1B promote disulfidptosis in ischemic stroke via upregulating disulfidptosis-related genes and whether USP15 deubiquitinates SETD1B. A C57BL/6 J mouse model of ischemic stroke was established, which results in brain injury and upregulation of USP15 and SETD1B, concomitant with the increased disulfidptosis, as indicated by the increased level of NADP<sup>+</sup>/NADPH ratio, elevated protein levels of NCK associated protein 1 like (NCKAP1L) and WASP family protein member 2 (WAVE-2), disulfide bond accumulation, and cytoskeleton detachment from the cytoplasmic membrane. Similar results were observed in cultured HT22 cells subjected to oxygen-glucose deprivation plus reoxygenation (OGD/R), and USP15 or SETD1B siRNAs reversed these phenomena. Mechanistically, knockdown of SETD1B prevented H3K4me3 enrichment at the <em>Nckap1l</em> and <em>Wasf2</em> promoters and reduced <em>Nckap1l</em> and <em>Wasf2</em> expression. Moreover, knockdown of USP15 increased the ubiquitination level of SETD1B thus decreasing its protein level. Based on these findings, we concluded that SETD1B can promote disulfidptosis in stroke brain cells via a mechanism involving facilitating H3K4me3 enrichment at the <em>Nckap1l</em> and <em>Wasf2</em> promoters, while USP15 is able to deubiquitinate SETD1B and increase SETD1B level in ischemic stroke mice.</div></div>","PeriodicalId":8754,"journal":{"name":"Biochimica et biophysica acta. Molecular cell research","volume":"1872 7","pages":"Article 120016"},"PeriodicalIF":4.6,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144534744","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

GlycoRNA-L and glycoRNA-S mediate human monocyte adhesion via binding to Siglec-5 GlycoRNA-L和glycoRNA-S通过与siglece -5结合介导人单核细胞粘附

IF 4.6 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Molecular cell research

Pub Date : 2025-07-01 DOI: 10.1016/j.bbamcr.2025.120017

Yong Li , Yisong Qian , Evan Huang , Zain Schwarz , Hannah Tai , Katherine Tillock , Tianhua Lei , Xiaofeng Yang , Mingui Fu

It was recently reported that RNAs can be glycosylated, and such glycosylated RNAs (referred to as glycoRNAs) are located on the outer cell surface. We here reported that there are two forms of glycoRNAs, named as glycoRNA-L and glycoRNA-S, robustly expressed in human monocytes. We verified that the glycoRNA-S specifically detected in human monocytes is synthesized by enzyme-catalyzed conjugation, but not artificial products of labelling probe. RNase-treatment removed both glycoRNA-L and glycoRNA-S, suggesting that they are localized on cell surface. Removing glycoRNAs significantly suppressed the interaction of human monocytes with endothelial cells, suggesting that glycoRNAs mediate human monocyte adhesion. Using flow cytometry, immunoprecipitation and northern blotting we identified Siglec-5 as the binding receptor of glycoRNAs. Siglec-5 is expressed in human endothelial cells but presented on endothelial cell surface when endothelial cells are activated. We observed that glycoRNA-L was heavily labeled with sialic acid, whereas glycoRNA-S was heavily labeled with N-acetylgalactosamine and N-acetylglucosamine. Together, these results demonstrate that two forms of glycoRNAs exist in human monocytes, which may play significant role in controlling the interaction of human monocytes and endothelial cells and contribute to the pathogenesis of inflammatory diseases.

最近有报道称rna可以被糖基化，这种糖基化的rna（称为glycoRNAs）位于细胞外表面。我们在这里报道了两种形式的glycorna，分别命名为glycoRNA-L和glycoRNA-S，在人类单核细胞中强烈表达。我们证实了glycoRNA-S在人单核细胞中特异性检测是通过酶催化偶联合成的，而不是标记探针的人工产物。rnase处理去除了glycoRNA-L和glycoRNA-S，表明它们定位于细胞表面。去除糖cornas可显著抑制人单核细胞与内皮细胞的相互作用，提示糖cornas介导人单核细胞粘附。通过流式细胞术、免疫沉淀和northern blotting，我们鉴定出siglece -5是glycoRNAs的结合受体。siglece -5在人内皮细胞中表达，但在内皮细胞被激活时出现在内皮细胞表面。我们观察到glycoRNA-L被唾液酸大量标记，而glycoRNA-S被n -乙酰半乳糖胺和n -乙酰氨基葡萄糖胺大量标记。综上所述，这些结果表明，人单核细胞中存在两种形式的glycoRNAs，它们可能在控制人单核细胞与内皮细胞的相互作用中发挥重要作用，并参与炎症性疾病的发病机制。

{"title":"GlycoRNA-L and glycoRNA-S mediate human monocyte adhesion via binding to Siglec-5","authors":"Yong Li , Yisong Qian , Evan Huang , Zain Schwarz , Hannah Tai , Katherine Tillock , Tianhua Lei , Xiaofeng Yang , Mingui Fu","doi":"10.1016/j.bbamcr.2025.120017","DOIUrl":"10.1016/j.bbamcr.2025.120017","url":null,"abstract":"<div><div>It was recently reported that RNAs can be glycosylated, <em>and</em> such glycosylated RNAs (referred to as glycoRNAs) are located on the outer cell surface. We here reported that there are two forms of glycoRNAs, named as glycoRNA-L and glycoRNA-S, robustly expressed in human monocytes. We verified that the glycoRNA-S specifically detected in human monocytes is synthesized by enzyme-catalyzed conjugation, but not artificial products of labelling probe. RNase-treatment removed both glycoRNA-L and glycoRNA-S, suggesting that they are localized on cell surface. Removing glycoRNAs significantly suppressed the interaction of human monocytes with endothelial cells, suggesting that glycoRNAs mediate human monocyte adhesion. Using flow cytometry, immunoprecipitation and northern blotting we identified Siglec-5 as the binding receptor of glycoRNAs. Siglec-5 is expressed in human endothelial cells but presented on endothelial cell surface when endothelial cells are activated. We observed that glycoRNA-L was heavily labeled with sialic acid, whereas glycoRNA-S was heavily labeled with <em>N</em>-acetylgalactosamine and <em>N</em>-acetylglucosamine. Together, these results demonstrate that two forms of glycoRNAs exist in human monocytes, which may play significant role in controlling the interaction of human monocytes and endothelial cells and contribute to the pathogenesis of inflammatory diseases.</div></div>","PeriodicalId":8754,"journal":{"name":"Biochimica et biophysica acta. Molecular cell research","volume":"1872 7","pages":"Article 120017"},"PeriodicalIF":4.6,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144534745","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0