华西耳鼻喉前沿学术速递——文献导读（第50期）
【Cell】2024年7月-2024年8月论文导读
（187卷，15-17期）
期刊介绍：
Cell是Cell Press细胞出版社旗下的旗舰刊，创办于1974年，由Elsevier公司出版发行。这是一本多学科期刊，包括但不限于细胞生物学、分子生物学、神经科学、免疫学、病毒学和微生物学、癌症、人类遗传学、系统生物学、信号传导和疾病机制和疾病治疗。该期刊为双周刊，最新影响因子为45.5。
本期文献导读将呈现2024年7月-2024年8月的主要刊物内容。
Aug 22, 2024
Volume 187，Issue 17，p4431-4812
在2024年8月22日，共发表25篇文章，其中包括8篇leading edge，1篇voices，2篇Preview，13篇Articles，1篇Resources。
1.Intestinal Blastocystis is linked to healthier diets and more favorable cardiometabolic outcomes in 56,989 individuals from 32 countries
在来自 32 个国家/地区的 56,989 名受试者中，肠道芽囊原虫与更健康的饮食和更佳的心脏代谢结果有关
Diet impacts human health, influencing body adiposity and the risk of developing cardiometabolic diseases. The gut microbiome is a key player in the diet-health axis, but while its bacterial fraction is widely studied, the role of micro-eukaryotes, including Blastocystis, is underexplored. We performed a global-scale analysis on 56,989 metagenomes and showed that human Blastocystis exhibits distinct prevalence patterns linked to geography, lifestyle, and dietary habits. Blastocystis presence defined a specific bacterial signature and was positively associated with more favorable cardiometabolic profiles and negatively with obesity (p＜1e-16) and disorders linked to altered gut ecology (p＜1e-8). In a diet intervention study involving 1,124 individuals, improvements in dietary quality were linked to weight loss and increases in Blastocystis prevalence (p=0.003) and abundance (p＜1e-7). Our findings suggest a potentially beneficial role for Blastocystis, which may help explain personalized host responses to diet and downstream disease etiopathogenesis.
饮食影响人类健康，影响身体肥胖和患心脏代谢疾病的风险。肠道微生物组是饮食-健康轴中的关键参与者，但尽管其细菌部分已被广泛研究，但包括芽囊原虫在内的微型真核生物的作用尚未得到充分探索。本研究对 56,989 个宏基因组进行了全球范围的分析，结果显示人类芽囊原虫表现出与地理、生活方式和饮食习惯相关的不同流行模式。芽囊原虫的存在定义了一种特定的细菌特征，其与更有利的心脏代谢特征呈正相关，与肥胖（p＜1e-16）和与肠道生态改变相关的疾病（p < 1e-8）呈负相关。在一项涉及1,124人的饮食干预研究中，饮食质量的改善与体重减轻以及芽囊原虫流行率（p=0.003）和丰度（p＜1e-7）的增加有关。本研究的结果表明芽囊原虫具有潜在的有益作用，这可能有助于解释宿主对饮食的个性化反应以及下游疾病的病因。
2.Host and environmental factors shape upper airway microbiota and respiratory health across the human lifespan
宿主和环境因素塑造整个人类生命周期中的上呼吸道微生物群和呼吸系统健康
Our understanding of the normal variation in the upper respiratory tract (URT) microbiota across the human lifespan and how these relate to host, environment, and health is limited. We studied the microbiota of 3,104 saliva (<10 year-olds)/oropharynx (≥10 year-olds) and 2,485 nasopharynx samples of 3,160 Dutch individuals 0-87 years of age, participating in a cross-sectional population-wide study (PIENTER-3) using 16S-rRNA sequencing. The microbiota composition was strongly related to age, especially in the nasopharynx, with maturation occurring throughout childhood and adolescence. Clear niche- and age-specific associations were found between the microbiota composition and host/environmental factors and health outcomes. Among others, social interaction, sex, and season were associated with the nasopharyngeal microbial community. By contrast, the oral microbiota was more related to antibiotics, tobacco, and alcohol use. We present an atlas of the URT microbiota across the lifespan in association with environment and health, establishing a baseline for future research.
现有研究对人类生命周期中上呼吸道 (URT) 微生物群的正常变化以及这些变化与宿主、环境和健康的关系的理解有限。本研究利用16S-rRNA测序技术，探索了来自3,160名参与横断面全民研究项目（PIENTER-3）、年龄为0-87岁的荷兰人的3,104个唾液（<10 岁）/口咽（≥10 岁）和2,485个鼻咽样本的微生物群。研究发现，微生物群组成与年龄密切相关，尤其是在鼻咽部，其成熟过程发生在整个儿童期和青少年期。在微生物群组成与宿主/环境因素和健康结果之间发现了明显的生态位特异性关联和年龄特异性关联。其中，社交互动、性别和季节与鼻咽微生物群落有关。相比之下，口腔微生物群与抗生素、烟草和酒精的使用更相关。本研究展示了与环境和健康相关的整个生命周期中的URT微生物群图谱，为未来的研究奠定了基础。
3.High expression of oleoyl-ACP hydrolase underpins life-threatening respiratory viral diseases
油酰-ACP水解酶的高表达是危及生命的呼吸道病毒性疾病的基础
Respiratory infections cause significant morbidity and mortality, yet it is unclear why some individuals succumb to severe disease. In patients hospitalized with avian A(H7N9) influenza, we investigated early drivers underpinning fatal disease. Transcriptomics strongly linked oleoyl-acyl-carrier-protein (ACP) hydrolase (OLAH), an enzyme mediating fatty acid production, with fatal A(H7N9) early after hospital admission, persisting until death. Recovered patients had low OLAH expression throughout hospitalization. High OLAH levels were also detected in patients hospitalized with life-threatening seasonal influenza, COVID-19, respiratory syncytial virus (RSV), and multisystem inflammatory syndrome in children (MIS-C) but not during mild disease. In olah−/− mice, lethal influenza infection led to survival and mild disease as well as reduced lung viral loads, tissue damage, infection-driven pulmonary cell infiltration, and inflammation. This was underpinned by differential lipid droplet dynamics as well as reduced viral replication and virus-induced inflammation in macrophages. Supplementation of oleic acid, the main product of OLAH, increased influenza replication in macrophages and their inflammatory potential. Our findings define how the expression of OLAH drives life-threatening viral disease.
呼吸道感染具有较高的发病率和死亡率，但尚不清楚为什么有些人会死于严重疾病。本研究在因禽流感A(H7N9)住院的患者中探索了导致致命疾病的早期驱动因素。转录组学将油酰酰基载体蛋白(ACP)水解酶(OLAH)（一种介导脂肪酸产生的酶）与入院后早期致命的A(H7N9)紧密关联，并持续到死亡。康复患者在住院期间的OLAH表达较低。在因危及生命的重度季节性流感、COVID-19、呼吸道合胞病毒 (RSV) 和儿童多系统炎症综合征(MIS-C)住院的患者中也检测到了高OLAH水平，但在轻症中没有检测到。在olah−/−小鼠中，致命的流感感染后的生存率提高，疾病症状减轻，同时肺病毒载量、组织损伤、感染驱动的肺细胞浸润和炎症减少。这是由不同的脂滴动力学以及巨噬细胞中病毒复制和病毒诱导炎症的减少所致。补充油酸（OLAH 的主要产物）可增加巨噬细胞中的流感复制及其炎症潜力。本研究的结果确定了OLAH的表达如何驱动危及生命的病毒性疾病。
4.The germline coordinates mitokine signaling
生殖系协调线粒体因子信号传导
The ability of mitochondria to coordinate stress responses across tissues is critical for health. In C. elegans, neurons experiencing mitochondrial stress elicit an inter-tissue signaling pathway through the release of mitokine signals, such as serotonin or the Wnt ligand EGL-20, which activate the mitochondrial unfolded protein response (UPRMT) in the periphery to promote organismal health and lifespan. We find that germline mitochondria play a surprising role in neuron-to-periphery UPRMT signaling. Specifically, we find that germline mitochondria signal downstream of neuronal mitokines, Wnt and serotonin, and upstream of lipid metabolic pathways in the periphery to regulate UPRMT activation. We also find that the germline tissue itself is essential for UPRMT signaling. We propose that the germline has a central signaling role in coordinating mitochondrial stress responses across tissues, and germline mitochondria play a defining role in this coordination because of their inherent roles in germline integrity and inter-tissue signaling.
线粒体协调组织间应激反应的能力对健康至关重要。在秀丽隐杆线虫中，经历线粒体应激的神经元通过释放线粒体因子信号（例如血清素或Wnt配体 EGL-20）引发组织间信号通路，这些信号激活外周的线粒体未折叠蛋白反应 (UPRMT)，以促进生物体健康和寿命。本研究发现生殖系线粒体在神经元到外周的URMT信号传导中发挥着令人惊讶的作用。具体而言，本研究发现生殖系线粒体在神经元线粒体因子、Wnt和血清素的下游以及外周脂质代谢途径的上游发出信号，以调节URMT激活。本研究还发现生殖系组织本身对于URMT信号传导至关重要。本研究认为，生殖系在协调跨组织线粒体应激反应方面具有核心信号传导作用，而生殖系线粒体由于其在生殖系完整性和组织间信号传导中的固有作用，在这种协调中起着决定性的作用。
5.Extensive structural rearrangement of intraflagellar transport trains underpins bidirectional cargo transport
鞭毛内运输列车的广泛结构重排是双向货物运输的基础
Bidirectional transport in cilia is carried out by polymers of the IFTA and IFTB protein complexes, called anterograde and retrograde intraflagellar transport (IFT) trains. Anterograde trains deliver cargoes from the cell to the cilium tip, then convert into retrograde trains for cargo export. We set out to understand how the IFT complexes can perform these two directly opposing roles before and after conversion. We use cryoelectron tomography and in situ cross-linking mass spectrometry to determine the structure of retrograde IFT trains and compare it with the known structure of anterograde trains. The retrograde train is a 2-fold symmetric polymer organized around a central thread of IFTA complexes. We conclude that anterograde-to-retrograde remodeling involves global rearrangements of the IFTA/B complexes and requires complete disassembly of the anterograde train. Finally, we describe how conformational changes to cargo-binding sites facilitate unidirectional cargo transport in a bidirectional system.
纤毛中的双向运输由IFTA和IFTB蛋白复合物的聚合物进行，称为顺行和逆行鞭毛内运输 (IFT) 列车。顺行列车将货物从细胞运送到纤毛尖端，然后转换为逆行列车以排出货物。本研究着手了解IFT复合物在转换前后如何发挥这两个方向相反的作用。本研究使用冷冻电子断层扫描和原位交联质谱法来确定逆行IFT列车的结构，并将其与已知的顺行列车结构进行比较。逆行列车是一种围绕IFTA复合物的中心线组织的2倍对称聚合物。本研究得出结论，顺行到逆行的重塑涉及IFTA/B复合物的整体重排，并需要完全拆解顺行列车。最后，本研究描述了货物结合位点的构象变化如何促进双向系统中的单向货物运输。
6.Innate immune memory after brain injury drives inflammatory cardiac dysfunction
脑损伤后的先天免疫记忆导致炎症性心脏功能障碍
The medical burden of stroke extends beyond the brain injury itself and is largely determined by chronic comorbidities that develop secondarily. We hypothesized that these comorbidities might share a common immunological cause, yet chronic effects post-stroke on systemic immunity are underexplored. Here, we identify myeloid innate immune memory as a cause of remote organ dysfunction after stroke. Single-cell sequencing revealed persistent pro-inflammatory changes in monocytes/macrophages in multiple organs up to 3 months after brain injury, notably in the heart, leading to cardiac fibrosis and dysfunction in both mice and stroke patients. IL-1β was identified as a key driver of epigenetic changes in innate immune memory. These changes could be transplanted to naive mice, inducing cardiac dysfunction. By neutralizing post-stroke IL-1β or blocking pro-inflammatory monocyte trafficking with a CCR2/5 inhibitor, we prevented post-stroke cardiac dysfunction. Such immune-targeted therapies could potentially prevent various IL-1β-mediated comorbidities, offering a framework for secondary prevention immunotherapy.
中风的医疗负担不仅限于脑损伤本身，很大程度上取决于继发的慢性合并症。本研究假设这些合并症可能具有共同的免疫学原因，但中风对全身免疫的慢性影响尚未得到充分探索。本研究将髓系先天免疫记忆确定为中风后远期器官功能障碍的原因。单细胞测序显示，在脑损伤后长达3个月内，多个器官中的单核细胞/巨噬细胞（尤其是心脏）持续存在促炎性变化，导致小鼠和中风患者的心脏纤维化和功能障碍。IL-1β被确定为先天免疫记忆表观遗传变化的关键驱动因素。这些变化可以转移到未受影响的小鼠身上，诱发心脏功能障碍。通过中和中风后IL-1β或用CCR2/5抑制剂阻断促炎性单核细胞运输，可预防中风后的心脏功能障碍。这种免疫靶向疗法可能可以预防各种IL-1β介导的合并症，为二级预防免疫疗法提供框架。
7.Stress-dependent condensate formation regulated by the ubiquitin-related modifier Urm1
受泛素相关修饰因子Urm1调控的应激依赖性凝聚物形成
The ability of proteins and RNA to coalesce into phase-separated assemblies, such as the nucleolus and stress granules, is a basic principle in organizing membraneless cellular compartments. While the constituents of biomolecular condensates are generally well documented, the mechanisms underlying their formation under stress are only partially understood. Here, we show in yeast that covalent modification with the ubiquitin-like modifier Urm1 promotes the phase separation of a wide range of proteins. We find that the drop in cellular pH induced by stress triggers Urm1 self-association and its interaction with both target proteins and the Urm1-conjugating enzyme Uba4. Urmylation of stress-sensitive proteins promotes their deposition into stress granules and nuclear condensates. Yeast cells lacking Urm1 exhibit condensate defects that manifest in reduced stress resilience. We propose that Urm1 acts as a reversible molecular adhesive to drive protective phase separation of functionally critical proteins under cellular stress.
蛋白质和RNA能够聚集成相分离组件，例如核仁和应激颗粒，这是组织无膜细胞区室的基本原理。虽然生物分子凝聚体的成分通常都有很好的记录，但它们在应激下形成的机制却只被部分理解。本研究在酵母中表明，泛素样修饰剂Urm1的共价修饰可促进多种蛋白质的相分离。研究者发现，应激引起的细胞pH值下降会触发Urm1自我结合及其与靶蛋白和Urm1结合酶Uba4的相互作用。应激敏感蛋白的Urmylation（模素化）会促进它们沉积到应激颗粒和核凝聚体中。缺乏Urm1的酵母细胞表现出凝聚体缺陷，表现为应激弹性降低。本研究提出，Urm1充当可逆分子“粘合剂”，以驱动细胞应激下功能关键蛋白质的保护性相分离。
8.All-RNA-mediated targeted gene integration in mammalian cells with rationally engineered R2 retrotransposons
利用合理设计的R2逆转录转座子在哺乳动物细胞中进行全RNA介导的靶向基因整合
All-RNA-mediated targeted gene integration methods, rendering reduced immunogenicity, effective deliverability with non-viral vehicles, and a low risk of random mutagenesis, are urgently needed for next-generation gene addition technologies. Naturally occurring R2 retrotransposons hold promise in this context due to their site-specific integration profile. Here, we systematically analyzed the biodiversity of R2 elements and screened several R2 orthologs capable of full-length gene insertion in mammalian cells. Robust R2 system gene integration efficiency was attained using combined donor RNA and protein engineering. Importantly, the all-RNA-delivered engineered R2 system showed effective integration activity, with efficiency over 60% in mouse embryos. Unbiased high-throughput sequencing demonstrated that the engineered R2 system exhibited high on-target integration specificity (99%). In conclusion, our study provides engineered R2 tools for applications based on hit-and-run targeted DNA integration and insights for further optimization of retrotransposon systems.
下一代基因添加技术迫切需要全RNA介导的靶向基因整合方法，这种方法可以降低免疫原性，通过非病毒载体实现有效传递，并降低随机突变风险。天然存在的R2逆转录转座子由于其位点特异性整合特性而具有良好的前景。本研究系统地分析了R2元件的生物多样性，并筛选了几种能够在哺乳动物细胞中实现全长基因插入的R2直系同源物。使用供体RNA和蛋白质联合工程实现了高效率的R2系统基因整合。重要的是，全RNA传递的工程化R2系统表现出有效的整合活性，在小鼠胚胎中的效率超过60%。无偏倚高通量测序表明，工程化R2系统表现出较高的靶向整合特异性(99%)。总之，本研究为基于命中即跑靶向DNA整合的应用提供了工程化R2工具，并为进一步优化逆转录转座子系统提供了见解。
9.Schwann cell-secreted PGE2 promotes sensory neuron excitability during development
施万细胞分泌的 PGE2 促进发育过程中感觉神经元的兴奋性
Electrical excitability-the ability to fire and propagate action potentials-is a signature feature of neurons. How neurons become excitable during development and whether excitability is an intrinsic property of neurons remain unclear. Here, we demonstrate that Schwann cells, the most abundant glia in the peripheral nervous system, promote somatosensory neuron excitability during development. We find that Schwann cells secrete prostaglandin E2, which is necessary and sufficient to induce developing somatosensory neurons to express normal levels of genes required for neuronal function, including voltage-gated sodium channels, and to fire action potential trains. Inactivating this signaling pathway in Schwann cells impairs somatosensory neuron maturation, causing multimodal sensory defects that persist into adulthood. Collectively, our studies uncover a neurodevelopmental role for prostaglandin E2 distinct from its established role in inflammation, revealing a cell non-autonomous mechanism by which glia regulate neuronal excitability to enable the development of normal sensory functions.
电兴奋性（激发和传播动作电位的能力）是神经元的标志性特征。神经元在发育过程中如何变得可兴奋以及兴奋性是否是神经元的固有属性仍不清楚。本研究证明施万细胞（周围神经系统中最丰富的神经胶质细胞）在发育过程中促进体感神经元的兴奋性。本研究发现施万细胞分泌前列腺素E2，这是诱导发育中的体感神经元表达正常水平的神经元功能所需基因（包括电压门控钠通道）和激发动作电位序列的必要和充分条件。在施万细胞中失活该信号通路会损害体感神经元的成熟，导致持续到成年的多模态感觉缺陷。总之，本研究揭示了前列腺素E2在神经发育中的作用，不同于其在炎症中已确定的作用，揭示了一种细胞非自主机制，神经胶质细胞通过该机制调节神经元兴奋性，从而促进正常感觉功能的发育。
10.Progestogen-driven B7-H4 contributes to onco-fetal immune tolerance
孕激素驱动的B7-H4有助于癌-胚免疫耐受
Immune tolerance mechanisms are shared in cancer and pregnancy. Through cross-analyzing single-cell RNA-sequencing data from multiple human cancer types and the maternal-fetal interface, we found B7-H4 (VTCN1) is an onco-fetal immune tolerance checkpoint. We showed that genetic deficiency of B7-H4 resulted in immune activation and fetal resorption in allogeneic pregnancy models. Analogously, B7-H4 contributed to MPA/DMBA-induced breast cancer progression, accompanied by CD8+ T cell exhaustion. Female hormone screening revealed that progesterone stimulated B7-H4 expression in placental and breast cancer cells. Mechanistically, progesterone receptor (PR) bound to a newly identified -58 kb enhancer, thereby mediating B7-H4 transcription via the PR-P300-BRD4 axis. PR antagonist or BRD4 degrader potentiated immunotherapy in a murine B7-H4+ breast cancer model. Thus, our work unravels a mechanistic and biological connection of a female sex hormone (progesterone) to onco-fetal immune tolerance via B7-H4 and suggests that the PR-P300-BRD4 axis is targetable for treating B7-H4+ cancer.
癌症和妊娠具有共同的免疫耐受机制。通过交叉分析多种人类癌症类型和母胎界面的单细胞RNA测序数据，本研究发现B7-H4 (VTCN1)是“癌-胚”免疫耐受检查点。本研究发现B7-H4的遗传缺陷导致同种异体妊娠模型中的免疫激活和胎儿吸收。类似地，B7-H4导致MPA/DMBA诱导的乳腺癌进展，并伴有CD8+ T细胞耗竭。女性激素筛查显示孕酮刺激胎盘和乳腺癌细胞中的B7-H4表达。从机制上讲，孕酮受体(PR)与新发现的-58 kb增强子结合，从而通过PR-P300-BRD4轴介导B7-H4转录。PR拮抗剂或BRD4降解剂增强了小鼠B7-H4+乳腺癌模型中的免疫治疗潜力。因此，本研究揭示了雌性激素（孕酮）通过B7-H4而与“癌-胚”免疫耐受联系起来的机制和生物学联系，并表明PR-P300-BRD4轴是治疗B7-H4+癌症的潜在靶点。
11.Early rhombic lip Protogenin+ve stem cells in a human-specific neurovascular niche initiate and maintain group 3 medulloblastoma
人类特异性神经血管生态位的早期菱形唇Protogenin+ve干细胞引发并维持3型髓母细胞瘤
We identify a population of Protogenin-positive (PRTG+ve) MYChigh NESTINlow stem cells in the four-week-old human embryonic hindbrain that subsequently localizes to the ventricular zone of the rhombic lip (RLVZ). Oncogenic transformation of early Prtg+ve rhombic lip stem cells initiates group 3 medulloblastoma (Gr3-MB)-like tumors. PRTG+ve stem cells grow adjacent to a human-specific interposed vascular plexus in the RLVZ, a phenotype that is recapitulated in Gr3-MB but not in other types of medulloblastoma. Co-culture of Gr3-MB with endothelial cells promotes tumor stem cell growth, with the endothelial cells adopting an immature phenotype. Targeting the PRTGhigh compartment of Gr3-MB in vivo using either the diphtheria toxin system or chimeric antigen receptor T cells constitutes effective therapy. Human Gr3-MBs likely arise from early embryonic RLVZ PRTG+ve stem cells inhabiting a specific perivascular niche. Targeting the PRTGhigh compartment and/or the perivascular niche represents an approach to treat children with Gr3-MB.
本研究在四周大的人类胚胎后脑中发现了一群Protogenin阳性(PRTG+ve) MYChigh NESTINlow干细胞，这些干细胞随后定位到菱形唇的脑室区(RLVZ)。早期 Prtg+ve 菱形唇干细胞的致癌转化引发了3型髓母细胞瘤(Gr3-MB)样肿瘤。PRTG+ve 干细胞在RLVZ中人类特异性介入血管丛附近生长，这种表型在Gr3-MB中重现，但在其他类型的髓母细胞瘤中没有重现。Gr3-MB与内皮细胞共培养可促进肿瘤干细胞生长，内皮细胞呈现未成熟表型。使用白喉毒素系统或嵌合抗原受体T细胞在体内靶向Gr3-MB的PRTGhigh区室是一种有效的治疗方法。人类Gr3-MB可能源自早期胚胎RLVZ PRTG+ve干细胞，这些干细胞栖息于特定的血管周围微环境中。针对PRTGhigh区室和/或血管周围微环境代表了一种治疗患有Gr3-MB的儿童的方法。
12.Molecular mechanism of distinct chemokine engagement and functional divergence of the human Duffy antigen receptor
人类Duffy抗原受体不同趋化因子结合和功能分化的分子机制
The Duffy antigen receptor is a seven-transmembrane (7TM) protein expressed primarily at the surface of red blood cells and displays strikingly promiscuous binding to multiple inflammatory and homeostatic chemokines. It serves as the basis of the Duffy blood group system in humans and also acts as the primary attachment site for malarial parasite Plasmodium vivax and pore-forming toxins secreted by Staphylococcus aureus. Here, we comprehensively profile transducer coupling of this receptor, discover potential non-canonical signaling pathways, and determine the cryoelectron microscopy (cryo-EM) structure in complex with the chemokine CCL7. The structure reveals a distinct binding mode of chemokines, as reflected by relatively superficial binding and a partially formed orthosteric binding pocket. We also observe a dramatic shortening of TM5 and 6 on the intracellular side, which precludes the formation of the docking site for canonical signal transducers, thereby providing a possible explanation for the distinct pharmacological and functional phenotype of this receptor.
Duffy抗原受体是一种七跨膜(7TM)蛋白，主要表达于红细胞表面，与多种炎症和稳态趋化因子表现出惊人的混杂结合。它是人类Duffy血型系统的基础，也是间日疟原虫（疟疾寄生虫）和金黄色葡萄球菌分泌的成孔毒素的主要附着位点。本研究全面分析了这种受体的传感器耦合，发现了潜在的非典型信号通路，并确定了与趋化因子CCL7复合的低温电子显微镜(cryo-EM)结构。该结构揭示了趋化因子的独特结合模式，反映在相对表面的结合和部分形成的正构结合口袋中。本研究还观察到细胞内侧的TM5和TM6急剧缩短，这阻止了典型信号转导的对接位点的形成，从而为该受体独特的药理学和功能表型提供了可能的解释。
13.A non-canonical role for a small nucleolar RNA in ribosome biogenesis and senescence
小核仁RNA在核糖体生物发生和衰老中的非典型作用
Cellular senescence is an irreversible state of cell-cycle arrest induced by various stresses, including aberrant oncogene activation, telomere shortening, and DNA damage. Through a genome-wide screen, we discovered a conserved small nucleolar RNA (snoRNA), SNORA13, that is required for multiple forms of senescence in human cells and mice. Although SNORA13 guides the pseudouridylation of a conserved nucleotide in the ribosomal decoding center, loss of this snoRNA minimally impacts translation. Instead, we found that SNORA13 negatively regulates ribosome biogenesis. Senescence-inducing stress perturbs ribosome biogenesis, resulting in the accumulation of free ribosomal proteins (RPs) that trigger p53 activation. SNORA13 interacts directly with RPL23, decreasing its incorporation into maturing 60S subunits and, consequently, increasing the pool of free RPs, thereby promoting p53-mediated senescence. Thus, SNORA13 regulates ribosome biogenesis and the p53 pathway through a non-canonical mechanism distinct from its role in guiding RNA modification. These findings expand our understanding of snoRNA functions and their roles in cellular signaling.
细胞衰老是一种不可逆的细胞周期停滞状态，由各种应激因素引起，包括异常的致癌基因激活、端粒缩短和DNA损伤。通过全基因组筛选，本研究发现了一种保守的小核仁RNA(snoRNA)，即SNORA13，它是人类细胞和小鼠多种衰老形式所必需的。尽管SNORA13指导核糖体解码中心保守核苷酸的假尿苷化，但这种snoRNA的丢失对翻译的影响很小。相反，本研究发现SNORA13负向调节核糖体的生物合成。诱导衰老的应激会扰乱核糖体的生物合成，导致游离核糖体蛋白(RP)的积累，从而触发p53激活。SNORA13直接与RPL23相互作用，降低其在成熟60S亚基中的整合，从而增加游离RP池，从而促进p53介导的衰老。因此，SNORA13通过一种非典型机制调节核糖体生物合成和p53通路，这种机制不同于其在引导RNA修饰中的作用。这些发现扩展了对snoRNA功能及其在细胞信号传导中的作用的理解。
Aug 08, 2024
Volume 187Issue 16p4129-4430
在2024年8月8日，共发表21篇文章，其中包括1篇leading edge，1篇primer，2篇Preview，13篇Articles，2篇Resources，2篇Correction。
1.Loss of transient receptor potential channel 5 causes obesity and postpartum depression
瞬时受体电位通道5的缺失导致肥胖和产后抑郁
Hypothalamic neural circuits regulate instinctive behaviors such as food seeking, the fight/flight response, socialization, and maternal care. Here, we identified microdeletions on chromosome Xq23 disrupting the brain-expressed transient receptor potential (TRP) channel 5 (TRPC5). This family of channels detects sensory stimuli and converts them into electrical signals interpretable by the brain. Male TRPC5 deletion carriers exhibited food seeking, obesity, anxiety, and autism, which were recapitulated in knockin male mice harboring a human loss-of-function TRPC5 mutation. Women carrying TRPC5 deletions had severe postpartum depression. As mothers, female knockin mice exhibited anhedonia and depression-like behavior with impaired care of offspring. Deletion of Trpc5 from oxytocin neurons in the hypothalamic paraventricular nucleus caused obesity in both sexes and postpartum depressive behavior in females, while Trpc5 overexpression in oxytocin neurons in knock-in mice reversed these phenotypes. We demonstrate that TRPC5 plays a pivotal role in mediating innate human behaviors fundamental to survival, including food seeking and maternal care.
下丘脑神经回路调节本能行为，如觅食、战斗/逃跑反应、社交和母性照料。本研究发现Xq23染色体上的微缺失会破坏大脑表达的瞬时受体电位(TRP)通道5(TRPC5)。该通道家族能够检测感官刺激并将其转换为可被大脑解读的电信号。男性TRPC5缺失携带者表现出觅食、肥胖、焦虑和自闭症，这些症状在携带人类功能丧失TRPC5突变的敲入雄性小鼠中重现。携带TRPC5缺失的女性患有严重的产后抑郁症。作为母亲，雌性敲入小鼠表现出快感缺乏和抑郁样行为，降低对后代的照料。下丘脑室旁核催产素神经元中Trpc5的缺失导致男女肥胖和女性产后抑郁行为，而敲入小鼠催产素神经元中Trpc5的过表达则逆转了这些表型。本研究证明TRPC5在介导与生存相关的人类先天行为（包括寻食和母性照料）方面发挥着关键作用。
2.Microglial-derived C1q integrates into neuronal ribonucleoprotein complexes and impacts protein homeostasis in the aging brain
小胶质细胞衍生的C1q整合到神经元核糖核蛋白复合物中并影响衰老大脑中的蛋白质稳态
Neuroimmune interactions mediate intercellular communication and underlie critical brain functions. Microglia, CNS-resident macrophages, modulate the brain through direct physical interactions and the secretion of molecules. One such secreted factor, the complement protein C1q, contributes to complement-mediated synapse elimination in both developmental and disease models, yet brain C1q protein levels increase significantly throughout aging. Here, we report that C1q interacts with neuronal ribonucleoprotein (RNP) complexes in an age-dependent manner. Purified C1q protein undergoes RNA-dependent liquid-liquid phase separation (LLPS) in vitro, and the interaction of C1q with neuronal RNP complexes in vivo is dependent on RNA and endocytosis. Mice lacking C1q have age-specific alterations in neuronal protein synthesis in vivo and impaired fear memory extinction. Together, our findings reveal a biophysical property of C1q that underlies RNA- and age-dependent neuronal interactions and demonstrate a role of C1q in critical intracellular neuronal processes.
神经免疫相互作用介导细胞间通讯，是大脑关键功能的基础。小胶质细胞是中枢神经系统驻留的巨噬细胞，通过直接的物理相互作用和分子分泌来调节大脑。补体蛋白C1q是这样一种分泌因子，它在发育和疾病模型中都有助于补体介导的突触消除，但大脑C1q蛋白水平在整个衰老过程中显著增加。本研究报告C1q以年龄依赖的方式与神经元核糖核蛋白(RNP)复合物相互作用。纯化的C1q蛋白在体外经历RNA依赖的液-液相分离 (LLPS)，并且C1q与体内神经元RNP复合物的相互作用依赖于RNA和内吞作用。缺乏C1q的小鼠体内神经元蛋白质合成会发生年龄特异性改变，恐惧记忆消退受损。总之，本研究揭示了C1q的生物物理特性，它是RNA和年龄依赖性的神经元相互作用的基础，并证明了C1q在关键的细胞内神经元过程中的作用。
3.Integrated cryoEM structure of a spumaretrovirus reveals cross-kingdom evolutionary relationships and the molecular basis for assembly and virus entry
泡沫逆转录病毒的综合低温电子显微镜结构揭示了跨界进化关系以及组装和病毒入侵的分子基础
Foamy viruses (FVs) are an ancient lineage of retroviruses, with an evolutionary history spanning over 450 million years. Vector systems based on Prototype Foamy Virus (PFV) are promising candidates for gene and oncolytic therapies. Structural studies of PFV contribute to the understanding of the mechanisms of FV replication, cell entry and infection, and retroviral evolution. Here we combine cryoEM and cryoET to determine high-resolution in situ structures of the PFV icosahedral capsid (CA) and envelope glycoprotein (Env), including its type III transmembrane anchor and membrane-proximal external region (MPER), and show how they are organized in an integrated structure of assembled PFV particles. The atomic models reveal an ancient retroviral capsid architecture and an unexpected relationship between Env and other class 1 fusion proteins of the Mononegavirales. Our results represent the de novo structure determination of an assembled retrovirus particle.
泡沫病毒 (FV) 是一种古老的逆转录病毒谱系，其进化史跨越了4.5亿年。基于原型泡沫病毒 (PFV) 的载体系统是基因和溶瘤疗法的有希望的候选者。PFV的结构研究有助于了解FV复制、细胞进入和感染以及逆转录病毒进化的机制。本研究结合cryoEM和cryoET来确定PFV二十面体衣壳(CA)和包膜糖蛋白(Env)的高分辨率原位结构，包括其III型跨膜锚和膜近端外部区域(MPER)，并展示PFV颗粒整合结构中这些结构是如何组织的。原子模型揭示了一种古老的逆转录病毒衣壳结构以及Env与其他单股反式病毒1类融合蛋白之间意想不到的关系。本研究的结果代表了组装逆转录病毒颗粒的全新结构测定。
4.Human coronavirus HKU1 recognition of the TMPRSS2 host receptor
人类冠状病毒HKU1识别TMPRSS2宿主受体
The human coronavirus HKU1 spike (S) glycoprotein engages host cell surface sialoglycans and transmembrane protease serine 2 (TMPRSS2) to initiate infection. The molecular basis of HKU1 binding to TMPRSS2 and determinants of host receptor tropism remain elusive. We designed an active human TMPRSS2 construct enabling high-yield recombinant production in human cells of this key therapeutic target. We determined a cryo-electron microscopy structure of the HKU1 RBD bound to human TMPRSS2, providing a blueprint of the interactions supporting viral entry and explaining the specificity for TMPRSS2 among orthologous proteases. We identified TMPRSS2 orthologs from five mammalian orders promoting HKU1 S-mediated entry into cells along with key residues governing host receptor usage. Our data show that the TMPRSS2 binding motif is a site of vulnerability to neutralizing antibodies and suggest that HKU1 uses S conformational masking and glycan shielding to balance immune evasion and receptor engagement.
人类冠状病毒HKU1刺突(S)糖蛋白与宿主细胞表面唾液酸聚糖和跨膜蛋白酶丝氨酸2(TMPRSS2)结合，从而引发感染。HKU1与TMPRSS2结合的分子基础和宿主受体趋向性的决定因素仍然难以捉摸。本研究设计了一种激活的人类TMPRSS2构建体，能够在人类细胞中高产地重组生产这种关键治疗靶点。本研究确定了与人类TMPRSS2结合的HKU1 RBD的低温电子显微镜结构，为支持病毒进入的相互作用提供了蓝图，并解释了直系同源蛋白酶中TMPRSS2的特异性。本研究从五种哺乳动物目中鉴定了TMPRSS2直系同源物，它们促进HKU1 S介导的细胞进入，以及控制宿主受体使用的关键残基。本研究的数据表明，TMPRSS2结合基序是中和抗体的脆弱位点，并表明HKU1使用S构象掩蔽和聚糖屏蔽来平衡免疫逃避和受体结合。
5.Structural basis of TMPRSS2 zymogen activation and recognition by the HKU1 seasonal coronavirus
HKU1季节性冠状病毒激活并识别TMPRSS2酶原的结构基础
The human seasonal coronavirus HKU1-CoV, which causes common colds worldwide, relies on the sequential binding to surface glycans and transmembrane serine protease 2 (TMPRSS2) for entry into target cells. TMPRSS2 is synthesized as a zymogen that undergoes autolytic activation to process its substrates. Several respiratory viruses, in particular coronaviruses, use TMPRSS2 for proteolytic priming of their surface spike protein to drive membrane fusion upon receptor binding. We describe the crystal structure of the HKU1-CoV receptor binding domain in complex with TMPRSS2, showing that it recognizes residues lining the catalytic groove. Combined mutagenesis of interface residues and comparison across species highlight positions 417 and 469 as determinants of HKU1-CoV host tropism. The structure of a receptor-blocking nanobody in complex with zymogen or activated TMPRSS2 further provides the structural basis of TMPRSS2 activating conformational change, which alters loops recognized by HKU1-CoV and dramatically increases binding affinity.
人类季节性冠状病毒HKU1-CoV可引起全球流行的普通感冒，它依靠与表面聚糖和跨膜丝氨酸蛋白酶2(TMPRSS2)的顺序结合以进入靶细胞。TMPRSS2是以酶原形式合成的，经过自溶激活来处理其底物。多种呼吸道病毒，特别是冠状病毒，使用TMPRSS2来催化其表面刺突蛋白的蛋白水解，以在受体结合后驱动膜融合。本研究描述了HKU1-CoV受体结合域与TMPRSS2复合物的晶体结构，表明它可以识别催化槽内的残基。界面残基的联合诱变和跨物种比较突出了417和469位点是HKU1-CoV宿主趋向性的决定因素。受体阻断纳米抗体与酶原或活化的TMPRSS2复合物的结构进一步为TMPRSS2激活构象变化提供了结构基础，从而改变了HKU1-CoV识别的环并显著增加了结合亲和力。
6.TMPRSS2 and glycan receptors synergistically facilitate coronavirus entry
TMPRSS2 和聚糖受体协同促进冠状病毒进入
The entry of coronaviruses is initiated by spike recognition of host cellular receptors, involving proteinaceous and/or glycan receptors. Recently, TMPRSS2 was identified as the proteinaceous receptor for HCoV-HKU1 alongside sialoglycan as a glycan receptor. However, the underlying mechanisms for viral entry remain unknown. Here, we investigated the HCoV-HKU1C spike in the inactive, glycan-activated, and functionally anchored states, revealing that sialoglycan binding induces a conformational change of the NTD and promotes the neighboring RBD of the spike to open for TMPRSS2 recognition, exhibiting a synergistic mechanism for the entry of HCoV-HKU1. The RBD of HCoV-HKU1 features an insertion subdomain that recognizes TMPRSS2 through three previously undiscovered interfaces. Furthermore, structural investigation of HCoV-HKU1A in combination with mutagenesis and binding assays confirms a conserved receptor recognition pattern adopted by HCoV-HKU1. These studies advance our understanding of the complex viral-host interactions during entry, laying the groundwork for developing new therapeutics against coronavirus-associated diseases.
冠状病毒的入侵始于刺突对宿主细胞受体的识别，包括蛋白质和/或聚糖受体。最近，TMPRSS2被鉴定为HCoV-HKU1的蛋白质受体，唾液酸聚糖则为聚糖受体。然而，病毒入侵的潜在机制仍然未知。本研究研究了HCoV-HKU1C刺突在非激活、聚糖激活和功能锚定状态下的变化，发现唾液酸聚糖结合会诱导NTD的构象变化，并促使刺突的邻近RBD打开以供TMPRSS2识别，从而表现出HCoV-HKU1进入的协同机制。HCoV-HKU1的RBD具有插入子域，可通过三个以前未发现的界面识别TMPRSS2。此外，对HCoV-HKU1A的结构研究结合诱变和结合试验证实了HCoV-HKU1采用的保守受体识别模式。这些研究加深了对病毒入侵过程中复杂的病毒-宿主相互作用的理解，为开发针对冠状病毒相关疾病的新疗法奠定了基础。
7.The WDR11 complex is a receptor for acidic-cluster-containing cargo proteins
WDR11复合物是含有酸性簇货物蛋白的受体
Vesicle trafficking is a fundamental process that allows for the sorting and transport of specific proteins (i.e., “cargoes”) to different compartments of eukaryotic cells. Cargo recognition primarily occurs through coats and the associated proteins at the donor membrane. However, it remains unclear whether cargoes can also be selected at other stages of vesicle trafficking to further enhance the fidelity of the process. The WDR11-FAM91A1 complex functions downstream of the clathrin-associated AP-1 complex to facilitate protein transport from endosomes to the TGN. Here, we report the cryo-EM structure of human WDR11-FAM91A1 complex. WDR11 directly and specifically recognizes a subset of acidic clusters, which we term super acidic clusters (SACs). WDR11 complex assembly and its binding to SAC-containing proteins are indispensable for the trafficking of SAC-containing proteins and proper neuronal development in zebrafish. Our studies thus uncover that cargo proteins could be recognized in a sequence-specific manner downstream of a protein coat.
囊泡运输是一种基本过程，它允许对特定蛋白质（即“货物”）进行分类和运输到真核细胞的不同区域。货物识别主要通过供体膜上的外壳和相关蛋白质进行。然而，目前尚不清楚是否也可以在囊泡运输的其他阶段选择货物以进一步提高该过程的准确性。WDR11-FAM91A1复合物在网格蛋白相关AP-1复合物的下游发挥作用，以促进蛋白质从内体到TGN的运输。本研究报告了人类WDR11-FAM91A1复合物的低温电子显微镜结构。WDR11直接且特异性地识别一组酸性簇，本研究将其称为超酸性簇(SACs)。WDR11复合物组装及其与含SAC蛋白质的结合对于含SAC蛋白质的运输和斑马鱼的正常神经元发育是必不可少的。因此，本研究发现，货物蛋白可以在蛋白质外壳下游以序列特异性的方式被识别。
8.The extracellular matrix integrates mitochondrial homeostasis
细胞外基质整合线粒体稳态
Cellular homeostasis is intricately influenced by stimuli from the microenvironment, including signaling molecules, metabolites, and pathogens. Functioning as a signaling hub within the cell, mitochondria integrate information from various intracellular compartments to regulate cellular signaling and metabolism. Multiple studies have shown that mitochondria may respond to various extracellular signaling events. However, it is less clear how changes in the extracellular matrix (ECM) can impact mitochondrial homeostasis to regulate animal physiology. We find that ECM remodeling alters mitochondrial homeostasis in an evolutionarily conserved manner. Mechanistically, ECM remodeling triggers a TGF-? response to induce mitochondrial fission and the unfolded protein response of the mitochondria (UPRMT). At the organismal level, ECM remodeling promotes defense of animals against pathogens through enhanced mitochondrial stress responses. We postulate that this ECM-mitochondria crosstalk represents an ancient immune pathway, which detects infection- or mechanical-stress-induced ECM damage, thereby initiating adaptive mitochondria-based immune and metabolic responses.
细胞稳态受到微环境刺激的密切影响，包括信号分子、代谢物和病原体。线粒体作为细胞内的信号枢纽，整合来自不同细胞内区室的信息，以调节细胞信号和代谢。多项研究表明，线粒体可能对各种细胞外信号事件作出反应。然而，尚不清楚细胞外基质(ECM)的变化如何影响线粒体稳态以调节动物生理。本研究发现ECM重塑以进化保守的方式改变线粒体稳态。从机制上讲，ECM重塑会触发TGF-β反应，从而诱导线粒体裂变和线粒体的未折叠蛋白反应 (UPRMT)。从生物体层面上讲，ECM重塑通过增强线粒体应激反应促进动物对病原体的防御。本研究推测这种ECM-线粒体串扰代表一种古老的免疫途径，它可以检测感染或机械应力引起的ECM损伤，从而启动基于线粒体的适应性免疫和代谢反应。
9.Preclinical proof of principle for orally delivered Th17 antagonist miniproteins
口服Th17拮抗剂微蛋白的临床前原理验证
Interleukin (IL)-23 and IL-17 are well-validated therapeutic targets in autoinflammatory diseases. Antibodies targeting IL-23 and IL-17 have shown clinical efficacy but are limited by high costs, safety risks, lack of sustained efficacy, and poor patient convenience as they require parenteral administration. Here, we present designed miniproteins inhibiting IL-23R and IL-17 with antibody-like, low picomolar affinities at a fraction of the molecular size. The minibinders potently block cell signaling in vitro and are extremely stable, enabling oral administration and low-cost manufacturing. The orally administered IL-23R minibinder shows efficacy better than a clinical anti-IL-23 antibody in mouse colitis and has a favorable pharmacokinetics (PK) and biodistribution profile in rats. This work demonstrates that orally administered de novo-designed minibinders can reach a therapeutic target past the gut epithelial barrier. With high potency, gut stability, and straightforward manufacturability, de novo-designed minibinders are a promising modality for oral biologics.
白细胞介素(IL)-23和IL-17是自身炎症疾病中经过充分验证的治疗靶点。针对IL-23和IL-17的抗体已显示出临床疗效，但因其需要肠外给药而受到成本高、存在安全风险、缺乏持续疗效以及患者不方便等因素的限制。本研究展示了设计出的抑制IL-23R和IL-17的微型蛋白，它们具有类似抗体的低至皮摩尔级别的亲和力，而分子大小仅为其一小部分。微型结合剂可在体外有效阻断细胞信号传导，并且非常稳定，因此可以口服给药并实现低成本制造。口服IL-23R微型结合剂在小鼠结肠炎中显示出比临床抗IL-23抗体更好的疗效，并且在大鼠中具有良好的药代动力学(PK)和生物分布特征。这项研究表明，口服的全新设计的微型结合剂可以穿过肠道上皮屏障到达治疗靶点。由于具有高效力、肠道稳定性和简单的可制造性，全新设计的微型结合剂是口服生物制剂的一种很有前途的方式。
10.The life-saving benefit of dexamethasone in severe COVID-19 is linked to a reversal of monocyte dysregulation
地塞米松在重症新冠肺炎患者中的救命作用与单核细胞失调的逆转有关
Dexamethasone is a life-saving treatment for severe COVID-19, yet its mechanism of action is unknown, and many patients deteriorate or die despite timely treatment initiation. Here, we identify dexamethasone treatment-induced cellular and molecular changes associated with improved survival in COVID-19 patients. We observed a reversal of transcriptional hallmark signatures in monocytes associated with severe COVID-19 and the induction of a monocyte substate characterized by the expression of glucocorticoid-response genes. These molecular responses to dexamethasone were detected in circulating and pulmonary monocytes, and they were directly linked to survival. Monocyte single-cell RNA sequencing (scRNA-seq)-derived signatures were enriched in whole blood transcriptomes of patients with fatal outcome in two independent cohorts, highlighting the potential for identifying non-responders refractory to dexamethasone. Our findings link the effects of dexamethasone to specific immunomodulation and reversal of monocyte dysregulation, and they highlight the potential of single-cell omics for monitoring in vivo target engagement of immunomodulatory drugs and for patient stratification for precision medicine approaches.
地塞米松是一种可以挽救严重COVID-19患者生命的药物，但其作用机制尚不清楚，而且尽管及时开始治疗，许多患者病情仍会恶化或死亡。本研究确定了地塞米松治疗引起的细胞和分子变化与COVID-19患者生存率提高有关。本研究观察到与严重COVID-19相关的单核细胞转录标志特征的逆转，以及以糖皮质激素反应基因表达为特征的单核细胞亚状态的诱导。在外周循环和肺的单核细胞中检测到了对地塞米松的这些分子反应，它们与生存直接相关。在两个独立队列中，单核细胞单细胞RNA测序(scRNA-seq)衍生的特征在死亡患者的全血转录组中富集，突出了识别对地塞米松有耐药性的无反应者的潜力。本研究将地塞米松的作用与特定的免疫调节和单核细胞失调的逆转联系起来，并强调了单细胞组学在监测免疫调节药物的体内靶标参与以及精准医疗方法的患者分层方面的潜力。
11.LAG-3 sustains TOX expression and regulates the CD94/NKG2-Qa-1b axis to govern exhausted CD8 T cell NK receptor expression and cytotoxicity
LAG-3 维持 TOX 表达并调节 CD94/NKG2-Qa-1b 轴以控制耗竭的 CD8 T 细胞 NK 受体表达和细胞毒性
Exhausted CD8 T (Tex) cells in chronic viral infection and cancer have sustained co-expression of inhibitory receptors (IRs). Tex cells can be reinvigorated by blocking IRs, such as PD-1, but synergistic reinvigoration and enhanced disease control can be achieved by co-targeting multiple IRs including PD-1 and LAG-3. To dissect the molecular changes intrinsic when these IR pathways are disrupted, we investigated the impact of loss of PD-1 and/or LAG-3 on Tex cells during chronic infection. These analyses revealed distinct roles of PD-1 and LAG-3 in regulating Tex cell proliferation and effector functions, respectively. Moreover, these studies identified an essential role for LAG-3 in sustaining TOX and Tex cell durability as well as a LAG-3-dependent circuit that generated a CD94/NKG2+ subset of Tex cells with enhanced cytotoxicity mediated by recognition of the stress ligand Qa-1b, with similar observations in humans. These analyses disentangle the non-redundant mechanisms of PD-1 and LAG-3 and their synergy in regulating Tex cells.
慢性病毒感染和癌症中耗竭的 CD8 T (Tex) 细胞具有持续的抑制性受体 (IR) 共表达。Tex 细胞可以通过阻断IR（例如 PD-1）来恢复活力，但通过同时靶向多个IR（包括 PD-1 和 LAG-3）可以实现协同恢复活力和增强疾病控制。为了剖析这些 IR 通路被破坏时内在的分子变化，本研究探索了慢性感染期间 PD-1 和/或 LAG-3 的缺失对 Tex 细胞的影响。这些分析揭示了 PD-1 和 LAG-3 在调节 Tex 细胞增殖和效应功能方面的不同作用。此外，这些研究确定了 LAG-3 在维持 TOX 和 Tex 细胞耐久性方面的重要作用，以及一个 LAG-3 依赖性回路，该回路产生了一个 CD94/NKG2+ 亚群的 Tex 细胞，该亚群具有通过识别应激配体 Qa-1b 介导的增强细胞毒性，在人类中也有类似的观察结果。这些分析揭示了 PD-1 和 LAG-3 的非冗余机制及其在调节 Tex 细胞中的协同作用。
12.LAG-3 and PD-1 synergize on CD8+ T cells to drive T cell exhaustion and hinder autocrine IFN-γ-dependent anti-tumor immunity
LAG-3 和 PD-1 协同作用于 CD8+ T细胞，以驱动T细胞耗竭并阻碍自分泌 IFN-γ依赖的抗肿瘤免疫
Overcoming immune-mediated resistance to PD-1 blockade remains a major clinical challenge. Enhanced efficacy has been demonstrated in melanoma patients with combined nivolumab (anti-PD-1) and relatlimab (anti-LAG-3) treatment, the first in its class to be FDA approved. However, how these two inhibitory receptors synergize to hinder anti-tumor immunity remains unknown. Here, we show that CD8+ T cells deficient in both PD-1 and LAG-3, in contrast to CD8+ T cells lacking either receptor, mediate enhanced tumor clearance and long-term survival in mouse models of melanoma. PD-1- and LAG-3-deficient CD8+ T cells were transcriptionally distinct, with broad TCR clonality and enrichment of effector-like and interferon-responsive genes, resulting in enhanced IFN-γ release indicative of functionality. LAG-3 and PD-1 combined to drive T cell exhaustion, playing a dominant role in modulating TOX expression. Mechanistically, autocrine, cell-intrinsic IFN-γ signaling was required for PD-1- and LAG-3-deficient CD8+ T cells to enhance anti-tumor immunity, providing insight into how combinatorial targeting of LAG-3 and PD-1 enhances efficacy.
克服免疫介导的对 PD-1 阻断的抵抗力仍然是一项重大的临床挑战。已证明，联合使用 nivolumab（抗 PD-1）和 relatlimab（抗 LAG-3）治疗黑色素瘤患者的疗效有所提高，这是此类药物中第一种被FDA批准的。然而，这两种抑制性受体如何协同作用阻碍抗肿瘤免疫仍不清楚。本研究表明，与缺乏任一受体的 CD8+ T 细胞相比，缺乏 PD-1 和 LAG-3 的 CD8+ T 细胞在小鼠黑色素瘤模型中能促进更强的肿瘤清除和长期生存。缺乏 PD-1 和 LAG-3 的 CD8+ T 细胞在转录上是不同的，其具有广泛的 TCR 克隆性以及效应基因和干扰素反应基因的富集，导致IFN-γ释放增强，提示其功能性。LAG-3 和 PD-1 共同驱动 T 细胞耗竭，在调节 TOX 表达中起主导作用。从机制上看，PD-1 和 LAG-3 缺陷的 CD8+ T 细胞需要自分泌、细胞内在的IFN-γ信号传导来增强抗肿瘤免疫力，这为 LAG-3 和 PD-1 的组合靶向如何增强疗效提供了见解。
13.Blockade of LAG-3 and PD-1 leads to co-expression of cytotoxic and exhaustion gene modules in CD8+ T cells to promote antitumor immunity
阻断 LAG-3 和 PD-1 可导致 CD8+T 细胞中细胞毒性和耗竭基因模块共同表达，从而促进抗肿瘤免疫
Relatlimab (rela; anti-LAG-3) plus nivolumab (nivo; anti-PD-1) is safe and effective for treatment of advanced melanoma. We designed a trial (NCT03743766) where advanced melanoma patients received rela, nivo, or rela+nivo to interrogate the immunologic mechanisms of rela+nivo. Analysis of biospecimens from this ongoing trial demonstrated that rela+nivo led to enhanced capacity for CD8+ T cell receptor signaling and altered CD8+ T cell differentiation, leading to heightened cytotoxicity despite the retention of an exhaustion profile. Co-expression of cytotoxic and exhaustion signatures was driven by PRDM1, BATF, ETV7, and TOX. Effector function was upregulated in clonally expanded CD8+ T cells that emerged after rela+nivo. A rela+nivo intratumoral CD8+ T cell signature was associated with a favorable prognosis. This intratumoral rela+nivo signature was validated in peripheral blood as an elevated frequency of CD38+TIM3+CD8+ T cells. Overall, we demonstrated that cytotoxicity can be enhanced despite the retention of exhaustion signatures, which will inform future therapeutic strategies.
Relatlimab (rela；抗 LAG-3) 加 nivolumab (nivo；抗 PD-1) 可安全有效地治疗晚期黑色素瘤。本研究设计了一项试验 (NCT03743766)，让晚期黑色素瘤患者接受 rela、nivo 或 rela+nivo 治疗，以探究 rela+nivo 的免疫机制。对这项正在进行的试验的生物样本的分析表明，rela+nivo 可增强 CD8+ T 细胞受体信号传导能力并改变 CD8+ T 细胞分化，从而导致细胞毒性增强，尽管其仍保留了耗竭特征。细胞毒性和耗竭特征的共表达由 PRDM1、BATF、ETV7 和 TOX 驱动。在 rela+nivo 治疗后出现的克隆扩增的 CD8+ T 细胞中效应子功能上调。rela+nivo 肿瘤内 CD8+ T 细胞特征与良好的预后相关。这种肿瘤内相关特征在外周血中被证实为 CD38+TIM3+CD8+ T 细胞频率升高。总体而言，本研究证明了尽管耗竭特征仍然存在，但细胞毒性仍可增强，这将为未来的治疗策略提供参考。
Jul 25, 2024
Volume 187，Issue 15，p3787-4128
在2024年7月25日，共发表24篇文章，其中包括9篇leading edge，1篇Preview，13篇Articles，1篇Resources。
1.Correction of age-associated defects in dendritic cells enables CD4+T cells to eradicate tumors
纠正树突状细胞中与年龄相关的缺陷可使 CD4+ T 细胞消灭肿瘤
Defective host defenses later in life are associated with changes in immune cell activities, suggesting that age-specific considerations are needed in immunotherapy approaches. In this study, we found that PD-1 and CTLA4-based cancer immunotherapies are unable to eradicate tumors in elderly mice. This defect in anti-tumor activity correlated with two known age-associated immune defects: diminished abundance of systemic naive CD8+ T cells and weak migratory activities of dendritic cells (DCs). We identified a vaccine adjuvant, referred to as a DC hyperactivator, which corrects DC migratory defects in the elderly. Vaccines containing tumor antigens and DC hyperactivators induced T helper type 1 (TH1) CD4+ T cells with cytolytic activity that drive anti-tumor immunity in elderly mice. When administered early in life, DC hyperactivators were the only adjuvant identified that elicited anti-tumor CD4+ T cells that persisted into old age. These results raise the possibility of correcting age-associated immune defects through DC manipulation.
生命后期宿主防御缺陷与免疫细胞活性变化有关，这表明免疫治疗方法需要考虑年龄相关的因素。本研究发现基于 PD-1 和 CTLA4 的癌症免疫疗法无法根除老年小鼠的肿瘤。这种抗肿瘤活性缺陷与两种已知的与年龄相关的免疫缺陷有关：全身未成熟 CD8+ T 细胞丰度减少和树突状细胞 (DC) 迁移活性减弱。本研究发现了一种疫苗佐剂，称为DC超活化剂，可纠正老年小鼠的DC迁移缺陷。含有肿瘤抗原和 DC 超活化剂的疫苗诱导老年小鼠产生具有细胞溶解活性的辅助性T细胞1型 (TH1) CD4+ T细胞，从而驱动抗肿瘤免疫。当在生命早期使用时，DC超活化剂是唯一确定的可诱导抗肿瘤CD4+ T细胞持续到老年的佐剂。这些结果提高了通过DC操作来纠正与年龄相关的免疫缺陷的可能性。
2.The rate and nature of mitochondrial DNA mutations in human pedigrees
人类谱系中线粒体 DNA 突变的发生率和性质
We examined the rate and nature of mitochondrial DNA (mtDNA) mutations in humans using sequence data from 64,806 contemporary Icelanders from 2,548 matrilines. Based on 116,663 mother-child transmissions, 8,199 mutations were detected, providing robust rate estimates by nucleotide type, functional impact, position, and different alleles at the same position. We thoroughly document the true extent of hypermutability in mtDNA, mainly affecting the control region but also some coding-region variants. The results reveal the impact of negative selection on viable deleterious mutations, including rapidly mutating disease-associated 3243A>G and 1555A>G and pre-natal selection that most likely occurs during the development of oocytes. Finally, we show that the fate of new mutations is determined by a drastic germline bottleneck, amounting to an average of 3 mtDNA units effectively transmitted from mother to child.
本研究使用来自 2,548 个母系的 64,806 名当代冰岛人的序列数据检查了人类线粒体 DNA (mtDNA) 突变的发生率和性质。基于 116,663 次母子传递，检测到8,199个突变，通过核苷酸类型、功能影响、位置和同一位置的不同等位基因提供了可靠的速率估计。本研究全面记录了mtDNA中高突变性的真实程度，主要影响控制区，但也影响一些编码区变体。结果揭示了负选择对可行的有害突变的影响，包括快速突变的与疾病相关的3243A>G和1555A>G以及最有可能在卵母细胞发育过程中发生的产前选择。最后，本研究表明新突变的命运由极端的生殖系瓶颈决定，平均有3个mtDNA单元有效地从母亲传递给孩子。
3.Systematic mapping of organism-scale gene-regulatory networks in aging using population asynchrony
利用群体异步性系统地绘制衰老过程中生物体规模的基因调控网络
In aging, physiologic networks decline in function at rates that differ between individuals, producing a wide distribution of lifespan. Though 70% of human lifespan variance remains unexplained by heritable factors, little is known about the intrinsic sources of physiologic heterogeneity in aging. To understand how complex physiologic networks generate lifespan variation, new methods are needed.Here, we present Asynch-seq, an approach that uses gene-expression heterogeneity within isogenic populations to study the processes generating lifespan variation. By collecting thousands of single-individual transcriptomes, we capture the Caenorhabditis elegans “pan-transcriptome”-a highly resolved atlas of non-genetic variation. We use our atlas to guide a large-scale perturbation screen that identifies the decoupling of total mRNA content between germline and soma as the largest source of physiologic heterogeneity in aging, driven by pleiotropic genes whose knockdown dramatically reduces lifespan variance. Our work demonstrates how systematic mapping of physiologic heterogeneity can be applied to reduce inter-individual disparities in aging.
在衰老过程中，生理网络功能衰退的速度因人而异，导致寿命分布广泛。尽管 70% 的人类寿命差异仍无法用遗传因素来解释，但人们对衰老中生理异质性的内在来源知之甚少。要了解复杂的生理网络如何产生寿命差异，需要新的方法。本研究提出了异步测序，这是一种利用同源种群内基因表达异质性来研究产生寿命差异的过程的方法。通过收集数千个单个个体的转录组数据，本研究捕获了秀丽隐杆线虫的“全转录组”——一个高度解析的非遗传变异图谱。使用该图谱来指导大规模扰动筛选实验，该筛选确定了生殖细胞和体细胞之间总mRNA含量的分离是衰老中生理异质性的最大来源，这是由多效性基因驱动的，这些基因的敲除会显著降低寿命差异。本研究展示了如何系统地绘制生理异质性以减少个体间衰老差异。
4.Amplification editing enables efficient and precise duplication of DNA from short sequence to megabase and chromosomal scale
扩增编辑能够高效、精确地实现DNA从短序列到兆碱基和染色体规模的复制
Duplication is a foundation of molecular evolution and a driver of genomic and complex diseases. Here, we develop a genome editing tool named Amplification Editing (AE) that enables programmable DNA duplication with precision at chromosomal scale. AE can duplicate human genomes ranging from 20 bp to 100 Mb, a size comparable to human chromosomes. AE exhibits activity across various cell types, encompassing diploid, haploid, and primary cells. AE exhibited up to 73.0% efficiency for 1 Mb and 3.4% for 100 Mb duplications, respectively. Whole-genome sequencing and deep sequencing of the junctions of edited sequences confirm the precision of duplication. AE can create chromosomal microduplications within disease-relevant regions in embryonic stem cells, indicating its potential for generating cellular and animal models. AE is a precise and efficient tool for chromosomal engineering and DNA duplication, broadening the landscape of precision genome editing from an individual genetic locus to the chromosomal scale.
复制是分子进化的基础，也是基因组和复杂疾病的驱动因素。本研究开发了一种名为扩增编辑 (AE) 的基因组编辑工具，该工具可以在染色体规模上精确地实现可编程的 DNA 复制。AE 可以复制从 20 bp 到 100 Mb 的人类基因组，其大小与人类染色体相当。AE 在各种细胞类型中表现出活性，包括二倍体、单倍体和原始细胞。AE 对1 Mb的复制效率高达73.0%，对100 Mb的复制效率为3.4%。全基因组测序和编辑序列连接处的深度测序证实了复制的精确度。AE 可以在胚胎干细胞中与疾病相关的区域内产生染色体微复制，表明其具有生成细胞和动物模型的潜力。AE是一种精确、有效的染色体工程和DNA复制工具，将精准基因组编辑的范围从单个基因位点拓展到染色体规模。
5.Open-ST: High-resolution spatial transcriptomics in 3D
Open-ST：3D 高分辨率空间转录组学
Spatial transcriptomics (ST) methods unlock molecular mechanisms underlying tissue development, homeostasis, or disease. However, there is a need for easy-to-use, high-resolution, cost-efficient, and 3D-scalable methods. Here, we report Open-ST, a sequencing-based, open-source experimental and computational resource to address these challenges and to study the molecular organization of tissues in 2D and 3D. In mouse brain, Open-ST captured transcripts at subcellular resolution and reconstructed cell types. In primary head-and-neck tumors and patient-matched healthy/metastatic lymph nodes, Open-ST captured the diversity of immune, stromal, and tumor populations in space, validated by imaging-based ST. Distinct cell states were organized around cell-cell communication hotspots in the tumor but not the metastasis. Strikingly, the 3D reconstruction and multimodal analysis of the metastatic lymph node revealed spatially contiguous structures not visible in 2D and potential biomarkers precisely at the 3D tumor/lymph node boundary. All protocols and software are available at https://rajewsky-lab.github.io/openst.
空间转录组学(ST)方法揭示了组织发育、体内平衡或疾病背后的分子机制。然而，仍需要一种易于使用、高分辨率、经济高效且可扩展至3D的方法。本研究报告了Open-ST，这是一种基于测序的开源实验和计算资源，用于解决这些挑战并研究组织在2D和3D中的分子结构。在小鼠大脑中，Open-ST以亚细胞分辨率捕获转录本并重建细胞类型。在原发性头颈部肿瘤和患者匹配的健康/转移性淋巴结中，Open-ST捕获了空间中免疫、基质和肿瘤群体的多样性，并通过基于成像的ST进行了验证。不同的细胞状态围绕肿瘤中的细胞间通讯热点组织，但不是转移。令人惊讶的是，转移性淋巴结的3D重建和多模态分析揭示了2D中不可见的空间连续结构，以及精确位于3D肿瘤/淋巴结边界的潜在生物标志物。所有协议和软件均可在https://rajewsky-lab.github.io/openst上找到。
6.Ring-shaped odor coding in the antennal lobe of migratory locusts
飞蝗触角叶中的环状气味编码
The representation of odors in the locust antennal lobe with its >2,000 glomeruli has long remained a perplexing puzzle. We employed the CRISPR-Cas9 system to generate transgenic locusts expressing the genetically encoded calcium indicator GCaMP in olfactory sensory neurons. Using two-photon functional imaging, we mapped the spatial activation patterns representing a wide range of ecologically relevant odors across all six developmental stages. Our findings reveal a functionally ring-shaped organization of the antennal lobe composed of specific glomerular clusters. This configuration establishes an odor-specific chemotopic representation by encoding different chemical classes and ecologically distinct odors in the form of glomerular rings. The ring-shaped glomerular arrangement, which we confirm by selective targeting of OR70a-expressing sensory neurons, occurs throughout development, and the odor-coding pattern within the glomerular population is consistent across developmental stages. Mechanistically, this unconventional spatial olfactory code reflects the locust-specific and multiplexed glomerular innervation pattern of the antennal lobe.
蝗虫触角叶中含有超过 2,000 个球状体，而其气味表征长期以来一直是个令人困惑的难题。本研究利用 CRISPR-Cas9 系统生成了转基因蝗虫，这些蝗虫在嗅觉传感神经元中表达遗传编码的钙指示剂 GCaMP。利用双光子功能成像，绘制了代表所有六个发育阶段的各种生态相关气味的空间激活模式。本研究揭示了触角叶的功能性环状组织，由特定的球状体簇组成。这种结构通过以球状体环的形式编码不同的化学类别和生态上不同的气味，建立了气味特异性的化学定位表征。本研究通过选择性靶向表达 OR70a 的传感神经元证实了这种环状球状排列，这种排列在整个发育过程中都会发生，并且球状体群中的气味编码模式在各个发育阶段都是一致的。从机制上看，这种非常规的空间嗅觉代码反映了蝗虫特有的、多路复用的触角叶球状体神经支配模式。
7.Single-molecule imaging reveals the mechanism of bidirectional replication initiation in metazoa
单分子成像揭示后生动物双向复制起始机制
Metazoan genomes are copied bidirectionally from thousands of replication origins. Replication initiation entails the assembly and activation of two CMG helicases (Cdc45⋅Mcm2-7⋅GINS) at each origin. This requires several replication firing factors (including TopBP1, RecQL4, and DONSON) whose exact roles are still under debate. How two helicases are correctly assembled and activated at each origin is a long-standing question. By visualizing the recruitment of GINS, Cdc45, TopBP1, RecQL4, and DONSON in real time, we uncovered that replication initiation is surprisingly dynamic. First, TopBP1 transiently binds to the origin and dissociates before the start of DNA synthesis. Second, two Cdc45 are recruited together, even though Cdc45 alone cannot dimerize. Next, two copies of DONSON and two GINS simultaneously arrive at the origin, completing the assembly of two CMG helicases. Finally, RecQL4 is recruited to the CMG⋅DONSON⋅DONSON⋅CMG complex and promotes DONSON dissociation and CMG activation via its ATPase activity.
后生动物基因组从数千个复制起点双向复制。复制起始需要在每个复制起点组装和激活两个CMG解旋酶 (Cdc45-Mcm2-7-GINS)。这需要几个复制启动因子（包括 TopBP1、RecQL4 和 DONSON），它们的确切作用仍有争议。两个解旋酶如何在每个复制起点正确组装和激活是一个长期存在的问题。通过实时可视化 GINS、Cdc45、TopBP1、RecQL4 和 DONSON 的募集，本研究发现复制起始具有令人惊讶的动态性。首先，TopBP1暂时与复制起点结合并在DNA合成开始前解离。其次，两个Cdc45一起被募集，尽管Cdc45本身不能二聚化。接下来，两个DONSON和两个GINS同时到达复制起点，完成两个CMG解旋酶的组装。最后，RecQL4被招募到CMG⋅DONSON⋅DONSON⋅CMG复合体，并通过其ATPase活性促进DONSON解离和CMG激活。
8.The primitive endoderm supports lineage plasticity to enable regulative development
原始内胚层支持谱系可塑性，从而实现调节性发育
Mammalian blastocyst formation involves the specification of the trophectoderm followed by the differentiation of the inner cell mass into embryonic epiblast and extra-embryonic primitive endoderm (PrE). During this time, the embryo maintains a window of plasticity and can redirect its cellular fate when challenged experimentally. In this context, we found that the PrE alone was sufficient to regenerate a complete blastocyst and continue post-implantation development. We identify an in vitro population similar to the early PrE in vivo that exhibits the same embryonic and extra-embryonic potency and can form complete stem cell-based embryo models, termed blastoids. Commitment in the PrE is suppressed by JAK/STAT signaling, collaborating with OCT4 and the sustained expression of a subset of pluripotency-related transcription factors that safeguard an enhancer landscape permissive for multi-lineage differentiation. Our observations support the notion that transcription factor persistence underlies plasticity in regulative development and highlight the importance of the PrE in perturbed development.
哺乳动物囊胚的形成涉及滋养外胚层的形成，随后内细胞团分化为胚胎外胚层和胚胎外原始内胚层 (PrE)。在此期间，胚胎保持可塑性，在受到实验挑战时可以改变其细胞命运。在这种情况下，本研究发现仅 PrE 就足以再生完整的囊胚并继续植入后发育。本研究在体外发现了一种类似于体内早期 PrE 的群体，该群体具有与胚胎和外胚层相同的全能性，可以形成完整的干细胞胚胎模型，称为胚状体。PrE 中的分化受到 JAK/STAT 信号抑制，与 OCT4 和一组多能性相关转录因子的持续表达协同作用，这些转录因子可保护允许多谱系分化的增强子环境。本研究的观察支持了以下观点：转录因子持久性是调节发育中可塑性的基础，并强调了PrE在受干扰发育中的重要性。
9.TERT activation targets DNA methylation and multiple aging hallmarks
TERT激活靶向DNA甲基化和多种衰老特征
Insufficient telomerase activity, stemming from low telomerase reverse transcriptase (TERT) gene transcription, contributes to telomere dysfunction and aging pathologies. Besides its traditional function in telomere synthesis, TERT acts as a transcriptional co-regulator of genes pivotal in aging and age-associated diseases. Here, we report the identification of a TERT activator compound (TAC) that upregulates TERT transcription via the MEK/ERK/AP-1 cascade. In primary human cells and naturally aged mice, TAC-induced elevation of TERT levels promotes telomere synthesis, blunts tissue aging hallmarks with reduced cellular senescence and inflammatory cytokines, and silences p16INK4a expression via upregulation of DNMT3B-mediated promoter hypermethylation. In the brain, TAC alleviates neuroinflammation, increases neurotrophic factors, stimulates adult neurogenesis, and preserves cognitive function without evident toxicity, including cancer risk. Together, these findings underscore TERT?s critical role in aging processes and provide preclinical proof of concept for physiological TERT activation as a strategy to mitigate multiple aging hallmarks and associated pathologies.
端粒酶活性不足是由低端粒酶逆转录酶 (TERT) 基因转录水平引起的，会导致端粒功能障碍和衰老病理。除了传统的端粒合成功能外，TERT 还充当衰老和与年龄相关的疾病中关键基因的转录共调节剂。本研究报告了一种 TERT 激活剂化合物 (TAC)，它通过 MEK/ERK/AP-1级联上调TERT转录。在原代人类细胞和自然衰老的小鼠中，TAC诱导的TERT水平升高可促进端粒合成，通过减少细胞衰老和炎性细胞因子来减弱组织衰老特征，并通过上调DNMT3B介导的启动子高甲基化来抑制p16INK4a表达。在大脑中，TAC可缓解神经炎症、增加神经营养因子、刺激成人神经发生并保持认知功能，且无明显毒性（包括癌症风险）。总之，这些发现强调了TERT在衰老过程中的关键作用，并为生理性TERT激活作为缓解多种衰老特征和相关病理的策略提供了临床前概念证明。
10.STING orchestrates the neuronal inflammatory stress response in multiple sclerosis
STING 调控多发性硬化症中的神经元炎症应激反应
Inflammation-induced neurodegeneration is a defining feature of multiple sclerosis (MS), yet the underlying mechanisms remain unclear. By dissecting the neuronal inflammatory stress response, we discovered that neurons in MS and its mouse model induce the stimulator of interferon genes (STING). However, activation of neuronal STING requires its detachment from the stromal interaction molecule 1 (STIM1), a process triggered by glutamate excitotoxicity. This detachment initiates non-canonical STING signaling, which leads to autophagic degradation of glutathione peroxidase 4 (GPX4), essential for neuronal redox homeostasis and thereby inducing ferroptosis. Both genetic and pharmacological interventions that target STING in neurons protect against inflammation-induced neurodegeneration. Our findings position STING as a central regulator of the detrimental neuronal inflammatory stress response, integrating inflammation with glutamate signaling to cause neuronal cell death, and present it as a tractable target for treating neurodegeneration in MS.
炎症引起的神经退行性病变是多发性硬化症 (MS) 的典型特征，但其潜在机制仍不清楚。通过分析神经元炎症应激反应，本研究发现 MS 及其小鼠模型中的神经元会诱导干扰素基因刺激因子 (STING)。然而，神经元 STING 的激活需要其与基质相互作用分子1(STIM1)分离，这一过程由谷氨酸兴奋毒性触发。这种分离会启动非典型STING信号传导，导致谷胱甘肽过氧化物酶4 (GPX4) 的自噬降解（这对神经元氧化还原稳态至关重要），从而诱导铁死亡。针对神经元中STING的基因和药物干预均能预防炎症引起的神经退行性病变。本研究将STING定位为有害的神经元炎症应激反应的中心调节器，将炎症与谷氨酸信号相结合导致神经元细胞死亡，并将其作为治疗MS中神经退行性变的可处理靶点。
11.NLRC5 senses NAD+ depletion, forming a PANoptosome and driving PANoptosis and inflammation
NLRC5感知NAD+耗竭，形成PANoptosome（广泛凋亡体）并驱动PANoptosis（广泛凋亡）和炎症
NLRs constitute a large, highly conserved family of cytosolic pattern recognition receptors that are central to health and disease, making them key therapeutic targets. NLRC5 is an enigmatic NLR with mutations associated with inflammatory and infectious diseases, but little is known about its function as an innate immune sensor and cell death regulator. Therefore, we screened for NLRC5’s role in response to infections, PAMPs, DAMPs, and cytokines. We identified that NLRC5 acts as an innate immune sensor to drive inflammatory cell death, PANoptosis, in response to specific ligands, including PAMP/heme and heme/cytokine combinations. NLRC5 interacted with NLRP12 and PANoptosome components to form a cell death complex, suggesting an NLR network forms similar to those in plants. Mechanistically, TLR signaling and NAD+ levels regulated NLRC5 expression and ROS production to control cell death. Furthermore, NLRC5-deficient mice were protected in hemolytic and inflammatory models, suggesting that NLRC5 could be a potential therapeutic target.
NLR构成一个庞大且高度保守的细胞质模式识别受体家族，对健康和疾病至关重要，因此成为关键的治疗靶点。NLRC5是一种神秘的NLR，其突变与炎症和传染病有关，但人们对其作为先天免疫传感器和细胞死亡调节剂的功能知之甚少。因此，本研究筛选了NLRC5在应对感染、PAMP、DAMP和细胞因子方面的作用。本研究发现NLRC5充当先天免疫传感器，以响应特定配体（包括PAMP/血红素和血红素/细胞因子组合）来驱动炎症细胞凋亡（PANoptosis）。NLRC5与NLRP12和PANoptosome（广泛凋亡体）成分相互作用形成细胞死亡复合物，表明形成的 NLR 网络与植物中的网络相似。从机制上讲，TLR信号和NAD+水平调节NLRC5表达和ROS产生以控制细胞死亡。此外，NLRC5缺陷小鼠在溶血和炎症模型中受到保护，这表明NLRC5可能是一个潜在的治疗靶点。
12.Therapeutic potential of co-signaling receptor modulation in hepatitis B
共信号受体调节在乙肝治疗中的潜力
Reversing CD8+ T cell dysfunction is crucial in treating chronic hepatitis B virus (HBV) infection, yet specific molecular targets remain unclear. Our study analyzed co-signaling receptors during hepatocellular priming and traced the trajectory and fate of dysfunctional HBV-specific CD8+ T cells. Early on, these cells upregulate PD-1, CTLA-4, LAG-3, OX40, 4-1BB, and ICOS. While blocking co-inhibitory receptors had minimal effect, activating 4-1BB and OX40 converted them into antiviral effectors. Prolonged stimulation led to a self-renewing, long-lived, heterogeneous population with a unique transcriptional profile. This includes dysfunctional progenitor/stem-like (TSL) cells and two distinct dysfunctional tissue-resident memory (TRM) populations. While 4-1BB expression is ubiquitously maintained, OX40 expression is limited to TSL. In chronic settings, only 4-1BB stimulation conferred antiviral activity. In HBeAg+ chronic patients, 4-1BB activation showed the highest potential to rejuvenate dysfunctional CD8+ T cells. Targeting all dysfunctional T cells, rather than only stem-like precursors, holds promise for treating chronic HBV infection.
逆转CD8+ T细胞功能障碍对于治疗慢性乙型肝炎病毒(HBV)感染至关重要，但具体的分子靶点仍不清楚。本研究分析了肝细胞启动过程中的共信号受体，并追踪了功能失调的 HBV 特异性 CD8+ T 细胞的轨迹和命运。早期，这些细胞上调PD-1、CTLA-4、LAG-3、OX40、4-1BB 和 ICOS。虽然阻断共抑制受体的效果很小，但激活4-1BB和OX40会将它们转化为抗病毒效应物。长时间的刺激会产生具有独特转录谱的自我更新、长寿、异质性群体。这包括功能失调的祖细胞/干细胞样(TSL)细胞和两个不同的功能失调的组织驻留记忆(TRM)群体。虽然4-1BB表达普遍存在，但OX40表达仅限于TSL细胞。在慢性病中，只有4-1BB刺激才能产生抗病毒活性。在HBeAg+慢性患者中，4-1BB激活显示出恢复功能失调的CD8+ T细胞的最大潜力。针对所有功能失调的T细胞（而不仅仅是干细胞样前体细胞）有望治疗慢性HBV感染。
13.A metabolomics pipeline highlights microbial metabolism in bloodstream infections
代谢组学流程揭示血液感染中的微生物代谢
The growth of antimicrobial resistance (AMR) highlights an urgent need to identify bacterial pathogenic functions that may be targets for clinical intervention. Although severe infections profoundly alter host metabolism, prior studies have largely ignored microbial metabolism in this context. Here, we describe an iterative, comparative metabolomics pipeline to uncover microbial metabolic features in the complex setting of a host and apply it to investigate gram-negative bloodstream infection (BSI) in patients. We find elevated levels of bacterially derived acetylated polyamines during BSI and discover the enzyme responsible for their production (SpeG). Blocking SpeG activity reduces bacterial proliferation and slows pathogenesis. Reduction of SpeG activity also enhances bacterial membrane permeability and increases intracellular antibiotic accumulation, allowing us to overcome AMR in culture and in vivo. This study highlights how tools to study pathogen metabolism in the natural context of infection can reveal and prioritize therapeutic strategies for addressing challenging infections.
抗菌素耐药性(AMR)的增长凸显了迫切需要确定可能成为临床干预目标的细菌致病功能。尽管严重感染会极大地改变宿主代谢，但先前的研究在很大程度上忽略了这种背景下的微生物代谢。本研究描述了一种迭代的比较代谢组学流程，以揭示宿主复杂环境中的微生物代谢特征，并将其应用于研究患者的革兰氏阴性血流感染(BSI)。发现本研究BSI期间细菌衍生的乙酰化多胺水平升高，并发现了负责其产生的酶(SpeG)。阻断SpeG活性可减缓细菌增殖并减缓发病。降低SpeG活性还会增强细菌膜通透性并增加细胞内抗生素的积累，使得在培养物中和在体内克服AMR成为可能。这项研究强调了在感染的自然环境中研究病原体代谢的工具如何揭示和优先考虑治疗策略，以应对具有挑战性的感染。
汇报人：李俊虹
导师：赵宇
审核：张宇阳、庞文都、任建君
