EMBO Reports最新文献

英文中文

3R centres contributions to change animal experimentation : A field report from Charité 3R, the 3R centre of a medical faculty. 3R 中心对改变动物实验的贡献：来自 Charité 3R（一个医学院的 3R 中心）的实地报告。

IF 7.7 1区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

EMBO Reports

Pub Date : 2024-09-13 DOI: 10.1038/s44319-024-00262-y

Ida Retter,Laura Behm,Lisa Grohmann,Karin Schmelz,Jennifer Rosowski,Stefan Hippenstiel

引用次数: 0

Glutaminolysis provides nucleotides and amino acids to regulate osteoclast differentiation in mice. 谷氨酰胺溶解为调节小鼠破骨细胞分化提供了核苷酸和氨基酸。

IF 7.7 1区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

EMBO Reports

Pub Date : 2024-09-13 DOI: 10.1038/s44319-024-00255-x

Guoli Hu,Yilin Yu,Yinshi Ren,Robert J Tower,Guo-Fang Zhang,Courtney M Karner

Osteoclasts are bone resorbing cells that are essential to maintain skeletal integrity and function. While many of the growth factors and molecular signals that govern osteoclastogenesis are well studied, how the metabolome changes during osteoclastogenesis is unknown. Using a multifaceted approach, we identified a metabolomic signature of osteoclast differentiation consisting of increased amino acid and nucleotide metabolism. Maintenance of the osteoclast metabolic signature is governed by elevated glutaminolysis. Mechanistically, glutaminolysis provides amino acids and nucleotides which are essential for osteoclast differentiation and bone resorption in vitro. Genetic experiments in mice found that glutaminolysis is essential for osteoclastogenesis and bone resorption in vivo. Highlighting the therapeutic implications of these findings, inhibiting glutaminolysis using CB-839 prevented ovariectomy induced bone loss in mice. Collectively, our data provide strong genetic and pharmacological evidence that glutaminolysis is essential to regulate osteoclast metabolism, promote osteoclastogenesis and modulate bone resorption in mice.

破骨细胞是骨吸收细胞，对维持骨骼的完整性和功能至关重要。尽管对支配破骨细胞生成的许多生长因子和分子信号进行了深入研究，但破骨细胞生成过程中代谢组如何变化尚不清楚。我们采用多方面的方法，确定了破骨细胞分化的代谢组特征，包括氨基酸和核苷酸代谢的增加。破骨细胞代谢特征的维持受制于谷氨酰胺溶解的增加。从机理上讲，谷氨酰胺分解提供了破骨细胞分化和体外骨吸收所必需的氨基酸和核苷酸。小鼠遗传实验发现，谷氨酰胺分解对体内破骨细胞生成和骨吸收至关重要。利用 CB-839 抑制谷氨酰胺溶解可预防卵巢切除术诱发的小鼠骨质流失，这凸显了这些发现的治疗意义。总之，我们的数据提供了强有力的遗传学和药理学证据，证明谷氨酰胺溶解对调节小鼠破骨细胞代谢、促进破骨细胞生成和调节骨吸收至关重要。

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引用次数: 0

FBXO38 is dispensable for PD-1 regulation. FBXO38 对于 PD-1 的调控是必不可少的。

IF 7.7 1区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

EMBO Reports

Pub Date : 2024-09-12 DOI: 10.1038/s44319-024-00220-8

Nikol Dibus,Eva Salyova,Karolina Kolarova,Alikhan Abdirov,Michele Pagano,Ondrej Stepanek,Lukas Cermak

SKP1-CUL1-F-box protein (SCF) ubiquitin ligases are versatile protein complexes that mediate the ubiquitination of protein substrates. The direct substrate recognition relies on a large family of F-box-domain-containing subunits. One of these substrate receptors is FBXO38, which is encoded by a gene found mutated in families with early-onset distal motor neuronopathy. SCFFBXO38 ubiquitin ligase controls the stability of ZXDB, a nuclear factor associated with the centromeric chromatin protein CENP-B. Loss of FBXO38 in mice results in growth retardation and defects in spermatogenesis characterized by deregulation of the Sertoli cell transcription program and compromised centromere integrity. Moreover, it was reported that SCFFBXO38 mediates the degradation of PD-1, a key immune-checkpoint inhibitor in T cells. Here, we have re-addressed the link between SCFFBXO38 and PD-1 proteolysis. Our data do not support the notion that SCFFBXO38 directly or indirectly controls the abundance and stability of PD-1 in T cells.

SKP1-CUL1-F-box 蛋白（SCF）泛素连接酶是介导蛋白质底物泛素化的多功能蛋白质复合体。底物的直接识别依赖于一个庞大的含 F-box 域的亚基家族。FBXO38 是这些底物受体之一，在早发性远端运动神经元病家族中，该基因发生了突变。SCFFBXO38 泛素连接酶控制着与中心染色质蛋白 CENP-B 相关的核因子 ZXDB 的稳定性。小鼠缺失 FBXO38 会导致生长迟缓和精子发生缺陷，其特征是 Sertoli 细胞转录程序失调和中心粒完整性受损。此外，有报道称 SCFFBXO38 能介导 T 细胞中关键免疫检查点抑制剂 PD-1 的降解。在这里，我们重新探讨了 SCFFBXO38 与 PD-1 蛋白解质之间的联系。我们的数据不支持 SCFFBXO38 直接或间接控制 T 细胞中 PD-1 的丰度和稳定性的观点。

引用次数: 0

CaMKII suppresses proteotoxicity by phosphorylating BAG3 in response to proteasomal dysfunction. CaMKII 在蛋白酶体功能失调时通过磷酸化 BAG3 来抑制蛋白毒性。

IF 7.7 1区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

EMBO Reports

Pub Date : 2024-09-11 DOI: 10.1038/s44319-024-00248-w

Chenliang Zhang,Huanji Xu,Qiulin Tang,Yichun Duan,Hongwei Xia,Huixi Huang,Di Ye,Feng Bi

Protein quality control serves as the primary defense mechanism for cells against proteotoxicity induced by proteasome dysfunction. While cells can limit the build-up of ubiquitinated misfolded proteins during proteasome inhibition, the precise mechanism is unclear. Here, we find that protein kinase Ca2+/Calmodulin (CaM)-dependent protein kinase II (CaMKII) maintains proteostasis during proteasome inhibition. We show that proteasome inhibition activates CaMKII, which phosphorylates B-cell lymphoma 2 (Bcl-2)-associated athanogene 3 (BAG3) at residues S173, S377, and S386. Phosphorylated BAG3 activates the heme-regulated inhibitor (HRI)- eukaryotic initiation factor-2α (eIF2α) signaling pathway, suppressing protein synthesis and the production of aggregated ubiquitinated misfolded proteins, ultimately mitigating the proteotoxic crisis. Inhibition of CaMKII exacerbates the accumulation of aggregated misfolded proteins and paraptosis induced by proteasome inhibitors. Based on these findings, we validate that combined targeting of proteasome and CaMKII accelerates tumor cell death and enhances the efficacy of proteasome inhibitors in tumor treatment. Our data unveil a new proteasomal inhibition-induced misfolded protein quality control mechanism and propose a novel therapeutic intervention for proteasome inhibitor-mediated tumor treatment.

蛋白质质量控制是细胞抵御蛋白酶体功能障碍引起的蛋白质毒性的主要防御机制。虽然细胞在蛋白酶体抑制过程中可以限制泛素化错误折叠蛋白的积累，但其确切机制尚不清楚。在这里，我们发现蛋白激酶 Ca2+/钙调蛋白（CaM）依赖性蛋白激酶 II（CaMKII）能在蛋白酶体抑制过程中维持蛋白稳态。我们发现蛋白酶体抑制激活了 CaMKII，CaMKII 使 B 细胞淋巴瘤 2（Bcl-2）相关的 athanogene 3（BAG3）在残基 S173、S377 和 S386 处磷酸化。磷酸化的 BAG3 会激活血红素调节抑制因子（HRI）-真核启动因子-2α（eIF2α）信号通路，抑制蛋白质合成和聚集泛素化错误折叠蛋白的产生，最终缓解蛋白毒性危机。抑制 CaMKII 会加剧蛋白酶体抑制剂诱导的错误折叠蛋白聚集和凋亡。基于这些发现，我们验证了联合靶向蛋白酶体和 CaMKII 能加速肿瘤细胞死亡并提高蛋白酶体抑制剂治疗肿瘤的疗效。我们的数据揭示了一种新的蛋白酶体抑制诱导的错误折叠蛋白质量控制机制，并为蛋白酶体抑制剂介导的肿瘤治疗提出了一种新的治疗干预方法。

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引用次数: 0

Terminal α1,2-fucosylation of glycosphingolipids by FUT1 is a key regulator in early cell-fate decisions. FUT1对糖磷脂的末端α1,2-岩藻糖基化是早期细胞命运决定的关键调节因子。

IF 7.7 1区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

EMBO Reports

Pub Date : 2024-09-10 DOI: 10.1038/s44319-024-00243-1

Saray Chen,Dana Hayoun-Neeman,Michal Nagar,Sapir Pinyan,Limor Hadad,Liat Yaacobov,Lilach Alon,Liraz Efrat Shachar,Tair Swissa,Olga Kryukov,Orly Gershoni-Yahalom,Benyamin Rosental,Smadar Cohen,Rachel G Lichtenstein

The embryonic cell surface is rich in glycosphingolipids (GSLs), which change during differentiation. The reasons for GSL subgroup variation during early embryogenesis remain elusive. By combining genomic approaches, flow cytometry, confocal imaging, and transcriptomic data analysis, we discovered that α1,2-fucosylated GSLs control the differentiation of human pluripotent cells (hPCs) into germ layer tissues. Overexpression of α1,2-fucosylated GSLs disrupts hPC differentiation into mesodermal lineage and reduces differentiation into cardiomyocytes. Conversely, reducing α1,2-fucosylated groups promotes hPC differentiation and mesoderm commitment in response to external signals. We find that bone morphogenetic protein 4 (BMP4), a mesodermal gene inducer, suppresses α1,2-fucosylated GSL expression. Overexpression of α1,2-fucosylated GSLs impairs SMAD activation despite BMP4 presence, suggesting α-fucosyl end groups as BMP pathway regulators. Additionally, the absence of α1,2-fucosylated GSLs in early/late mesoderm and primitive streak stages in mouse embryos aligns with the hPC results. Thus, α1,2-fucosylated GSLs may regulate early cell-fate decisions and embryo development by modulating cell signaling.

胚胎细胞表面富含糖磷脂（GSLs），这些物质在分化过程中会发生变化。早期胚胎发生过程中 GSL 亚群变化的原因仍然难以捉摸。通过结合基因组学方法、流式细胞术、共聚焦成像和转录组数据分析，我们发现α1,2-岩藻糖基化的GSLs控制着人类多能细胞（hPCs）向生殖层组织的分化。过量表达α1,2-岩藻糖基 GSL 会破坏 hPC 向中胚层的分化，并减少向心肌细胞的分化。相反，减少α1,2-岩藻糖基化基团可促进 hPC 分化和中胚层对外界信号的承诺。我们发现，中胚层基因诱导剂骨形态发生蛋白 4（BMP4）会抑制α1,2-岩藻糖基化 GSL 的表达。尽管存在 BMP4，α1,2-岩藻糖基化 GSL 的过表达仍会损害 SMAD 的激活，这表明α-岩藻糖基末端基团是 BMP 通路的调节因子。此外，小鼠胚胎早期/晚期中胚层和原始条纹阶段缺乏α1,2-岩藻糖基化的GSL，这与hPC的结果一致。因此，α1,2-岩藻糖基化的GSL可能通过调节细胞信号传导来调控早期细胞命运决定和胚胎发育。

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引用次数: 0

O-GlcNAcylation mediates Wnt-stimulated bone formation by rewiring aerobic glycolysis. O-GlcNAcylation通过重新连接有氧糖酵解介导Wnt刺激的骨形成。

IF 7.7 1区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

EMBO Reports

Pub Date : 2024-09-10 DOI: 10.1038/s44319-024-00237-z

Chengjia You,Fangyuan Shen,Puying Yang,Jingyao Cui,Qiaoyue Ren,Moyu Liu,Yujie Hu,Boer Li,Ling Ye,Yu Shi

Wnt signaling is an important target for anabolic therapies in osteoporosis. A sclerostin-neutralizing antibody (Scl-Ab), that blocks the Wnt signaling inhibitor (sclerostin), has been shown to promote bone mass in animal models and clinical studies. However, the cellular mechanisms by which Wnt signaling promotes osteogenesis remain to be further investigated. O-GlcNAcylation, a dynamic post-translational modification of proteins, controls multiple critical biological processes including transcription, translation, and cell fate determination. Here, we report that Wnt3a either induces O-GlcNAcylation rapidly via the Ca2+-PKA-Gfat1 axis, or increases it in a Wnt-β-catenin-dependent manner following prolonged stimulation. Importantly, we find O-GlcNAcylation indispensable for osteoblastogenesis both in vivo and in vitro. Genetic ablation of O-GlcNAcylation in the osteoblast-lineage diminishes bone formation and delays bone fracture healing in response to Wnt stimulation in vivo. Mechanistically, Wnt3a induces O-GlcNAcylation at Serine 174 of PDK1 to stabilize the protein, resulting in increased glycolysis and osteogenesis. These findings highlight O-GlcNAcylation as an important mechanism regulating Wnt-induced glucose metabolism and bone anabolism.

Wnt 信号是骨质疏松症同化疗法的一个重要靶点。在动物模型和临床研究中，一种能阻断 Wnt 信号抑制剂（硬骨素）的硬骨素中和抗体（Scl-Ab）已被证明能促进骨量。然而，Wnt 信号促进成骨的细胞机制仍有待进一步研究。O-GlcNA酰化是蛋白质的一种动态翻译后修饰，它控制着多种关键的生物过程，包括转录、翻译和细胞命运决定。在此，我们报告了 Wnt3a 通过 Ca2+-PKA-Gfat1 轴快速诱导 O-GlcNAcylation 或在长时间刺激后以 Wnt-β-catenin 依赖性方式增加 O-GlcNAcylation 的情况。重要的是，我们发现 O-GlcNAcylation 对体内和体外成骨细胞的形成都是不可或缺的。遗传性消减成骨细胞系中的 O-GlcNAcylation 会减少骨形成并延迟体内 Wnt 刺激下的骨折愈合。从机理上讲，Wnt3a诱导PDK1丝氨酸174处的O-GlcNAcylation以稳定该蛋白，从而导致糖酵解和骨生成增加。这些发现强调了O-GlcNAcylation是调节Wnt诱导的糖代谢和骨合成代谢的重要机制。

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引用次数: 0

O-GlcNAcylation in the osteoblast lineage-boosting the complexity of Wnt-stimulated bone formation. 成骨细胞谱系中的 O-GlcNAcylation 促进了 Wnt 刺激骨形成的复杂性。

IF 7.7 1区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

EMBO Reports

Pub Date : 2024-09-10 DOI: 10.1038/s44319-024-00242-2

Sandra Pohl,Thorsten Schinke

引用次数: 0

Gene therapy for epilepsy targeting neuropeptide Y and its Y2 receptor to dentate gyrus granule cells. 针对齿状回颗粒细胞神经肽 Y 及其 Y2 受体的癫痫基因疗法。

IF 7.7 1区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

EMBO Reports

Pub Date : 2024-09-09 DOI: 10.1038/s44319-024-00244-0

Stefano Cattaneo,Barbara Bettegazzi,Lucia Crippa,Laila Asth,Maria Regoni,Marie Soukupova,Silvia Zucchini,Alessio Cantore,Franca Codazzi,Flavia Valtorta,Michele Simonato

Gene therapy is emerging as an alternative option for individuals with drug-resistant focal epilepsy. Here, we explore the potential of a novel gene therapy based on Neuropeptide Y (NPY), a well-known endogenous anticonvulsant. We develop a lentiviral vector co-expressing NPY with its inhibitory receptor Y2 in which, for the first time, both transgenes are placed under the control of the minimal CamKIIa(0.4) promoter, biasing expression toward excitatory neurons and allowing autoregulation of neuronal excitability by Y2 receptor-mediated inhibition. Vector-induced NPY and Y2 expression and safety are first assessed in cultures of hippocampal neurons. In vivo experiments demonstrate efficient and nearly selective overexpression of both genes in granule cell mossy fiber terminals following vector administration in the dentate gyrus. Telemetry video-EEG monitoring reveals a reduction in the frequency and duration of seizures in the synapsin triple KO model. This study shows that targeting a small subset of neurons (hippocampal granule cells) with a combined overexpression of NPY and Y2 receptor is sufficient to reduce the occurrence of spontaneous seizures.

基因疗法正成为耐药性局灶性癫痫患者的另一种选择。在这里，我们探索了一种基于神经肽 Y（NPY）的新型基因疗法的潜力，NPY是一种众所周知的内源性抗惊厥剂。我们开发了一种共同表达 NPY 及其抑制性受体 Y2 的慢病毒载体，首次将这两种转基因置于最小 CamKIIa(0.4) 启动子的控制之下，使其偏向兴奋性神经元表达，并通过 Y2 受体介导的抑制作用自动调节神经元的兴奋性。首先在海马神经元培养物中评估了载体诱导的 NPY 和 Y2 表达及其安全性。体内实验证明，在齿状回施用载体后，这两种基因在颗粒细胞苔藓纤维末梢中都得到了高效和近乎选择性的过表达。遥测视频脑电图监测显示，在突触素三重 KO 模型中，癫痫发作的频率和持续时间都有所减少。这项研究表明，针对一小部分神经元（海马颗粒细胞）联合过表达 NPY 和 Y2 受体足以减少自发性癫痫发作的发生。

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引用次数: 0

SPIN1 facilitates chemoresistance and HR repair by promoting Tip60 binding to H3K9me3. SPIN1 通过促进 Tip60 与 H3K9me3 的结合，促进化疗抗性和 HR 修复。

IF 6.5 1区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

EMBO Reports

Pub Date : 2024-09-01 Epub Date: 2024-08-01 DOI: 10.1038/s44319-024-00219-1

Yukun Wang, Mengyao Li, Yuhan Chen, Yuhan Jiang, Ziyu Zhang, Zhenzhen Yan, Xiuhua Liu, Chen Wu

The tandem Tudor-like domain-containing protein Spindlin1 (SPIN1) is a transcriptional coactivator with critical functions in embryonic development and emerging roles in cancer. However, the involvement of SPIN1 in DNA damage repair has remained unclear. Our study shows that SPIN1 is recruited to DNA lesions through its N-terminal disordered region that binds to Poly-ADP-ribose (PAR), and facilitates homologous recombination (HR)-mediated DNA damage repair. SPIN1 promotes H3K9me3 accumulation at DNA damage sites and enhances the interaction between H3K9me3 and Tip60, thereby promoting the activation of ATM and HR repair. We also show that SPIN1 increases chemoresistance. These findings reveal a novel role for SPIN1 in the activation of H3K9me3-dependent DNA repair pathways, and suggest that SPIN1 may contribute to cancer chemoresistance by modulating the efficiency of double-strand break (DSB) repair.

含串联都铎样结构域的蛋白Spindlin1（SPIN1）是一种转录辅激活因子，在胚胎发育中具有关键功能，并在癌症中发挥着新的作用。然而，SPIN1参与DNA损伤修复的情况仍不清楚。我们的研究表明，SPIN1通过其N端无序区与聚ADP核糖（Poly-ADP-ribose，PAR）结合，被招募到DNA损伤处，促进同源重组（HR）介导的DNA损伤修复。SPIN1 可促进 H3K9me3 在 DNA 损伤位点的积累，并增强 H3K9me3 与 Tip60 之间的相互作用，从而促进 ATM 的激活和 HR 修复。我们还发现，SPIN1 增加了化疗抗性。这些发现揭示了 SPIN1 在激活 H3K9me3 依赖性 DNA 修复途径中的新作用，并表明 SPIN1 可能通过调节双链断裂（DSB）修复的效率来促进癌症的化疗耐受性。

引用次数: 0

Identification of resistance mechanisms to small-molecule inhibition of TEAD-regulated transcription. 鉴定小分子抑制 TEAD 调控转录的抗性机制。

IF 6.5 1区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

EMBO Reports

Pub Date : 2024-09-01 Epub Date: 2024-08-05 DOI: 10.1038/s44319-024-00217-3

Aishwarya Kulkarni, Varshini Mohan, Tracy T Tang, Leonard Post, Yih-Chih Chan, Murray Manning, Niko Thio, Benjamin L Parker, Mark A Dawson, Joseph Rosenbluh, Joseph Ha Vissers, Kieran F Harvey

The Hippo tumor suppressor pathway controls transcription by regulating nuclear abundance of YAP and TAZ, which activate transcription with the TEAD1-TEAD4 DNA-binding proteins. Recently, several small-molecule inhibitors of YAP and TEADs have been reported, with some entering clinical trials for different cancers with Hippo pathway deregulation, most notably, mesothelioma. Using genome-wide CRISPR/Cas9 screens we reveal that mutations in genes from the Hippo, MAPK, and JAK-STAT signaling pathways all modulate the response of mesothelioma cell lines to TEAD palmitoylation inhibitors. By exploring gene expression programs of mutant cells, we find that MAPK pathway hyperactivation confers resistance to TEAD inhibition by reinstating expression of a subset of YAP/TAZ target genes. Consistent with this, combined inhibition of TEAD and the MAPK kinase MEK, synergistically blocks proliferation of multiple mesothelioma and lung cancer cell lines and more potently reduces the growth of patient-derived lung cancer xenografts in vivo. Collectively, we reveal mechanisms by which cells can overcome small-molecule inhibition of TEAD palmitoylation and potential strategies to enhance the anti-tumor activity of emerging Hippo pathway targeted therapies.

Hippo肿瘤抑制通路通过调节YAP和TAZ的核丰度来控制转录，YAP和TAZ与TEAD1-TEAD4 DNA结合蛋白一起激活转录。最近，有报道称出现了几种YAP和TEADs的小分子抑制剂，其中一些已进入临床试验阶段，可用于治疗Hippo通路失调的不同癌症，尤其是间皮瘤。我们利用全基因组 CRISPR/Cas9 筛选发现，Hippo、MAPK 和 JAK-STAT 信号通路基因的突变都会调节间皮瘤细胞系对 TEAD 棕榈酰化抑制剂的反应。通过探索突变细胞的基因表达程序，我们发现 MAPK 通路的过度激活通过恢复 YAP/TAZ 靶基因子集的表达，赋予了细胞对 TEAD 抑制的抗性。与此相一致的是，联合抑制 TEAD 和 MAPK 激酶 MEK 能协同阻断多种间皮瘤和肺癌细胞系的增殖，并能更有效地减少源自患者的肺癌异种移植物在体内的生长。总之，我们揭示了细胞克服小分子抑制 TEAD 棕榈酰化的机制，以及增强新兴 Hippo 通路靶向疗法抗肿瘤活性的潜在策略。

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引用次数: 0

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