华西耳鼻喉前沿学术速递——文献导读（第53期）
【Science】2024年08月09日-10月10日期刊论文导读
期刊介绍：
《Science》（科学杂志）是一份由美国科学促进会（American Association for the Advancement of Science，简称AAAS）出版的著名科学杂志。该杂志创建于1880年，是全球最具影响力和知名度的跨学科科学期刊之一。该杂志以发表高质量、原创性的科学研究论文和评论而闻名，涵盖了各个科学领域，包括生命科学、物理学、化学、地球与环境科学、工程技术等。它是科学领域内同行评议制度的支持者，通过评审和编辑确保所发表的研究具有学术水平和可靠性。除了研究论文外，该杂志还经常刊发关于科学前沿、科学政策、科学教育和科学社会问题的评论文章。《Science》还定期发布一些专题特刊，深入探讨特定主题或领域的最新进展。因其在科学界的声望和影响力，《Science》成为科学家们追求学术卓越和科学创新的重要平台之一。最新影响因子为44.7。
Volume 385| Issue 6709| 9 Aug 2024
在2024年8月9日，《Science》共发表文章35篇：
9篇NEWS
10篇INSIGHTS
其中4篇PERSPECTIVES，1篇POLICY FORUM，2本BOOK，3篇LETTERS
16篇RESEARCH
其中14篇RESEARCH ARTICLES，1篇REVIEWS
1.Regulation of the hematopoietic stem cell pool by C-Kit–associated trogocytosis【Blood stem cells】
C-Kit相关的胞啃作用对造血干细胞池的调节
第一单位：美国爱因斯坦医学院
Abstract
Hematopoietic stem cells (HSCs) are routinely mobilized from the bone marrow (BM) to the blood circulation for clinical transplantation. However, the precise mechanisms by which individual stem cells exit the marrow are not understood. This study identified cell-extrinsic and molecular determinants of a mobilizable pool of blood-forming stem cells. We found that a subset of HSCs displays macrophage-associated markers on their cell surface. Although fully functional, these HSCs are selectively niche-retained as opposed to stem cells lacking macrophage markers, which exit the BM upon forced mobilization. Macrophage markers on HSCs could be acquired through direct transfer by trogocytosis, regulated by receptor tyrosine-protein kinase C-Kit (CD117), from BM-resident macrophages in mouse and human settings. Our study provides proof of concept that adult stem cells utilize trogocytosis to rapidly establish and activate function-modulating molecular mechanisms.
造血干细胞（HSC）通常从骨髓（BM）动员到血液循环中用于临床移植。然而，单个干细胞离开骨髓的确切机制尚不清楚。这项研究确定了可动员的造血干细胞池的细胞外源性和分子决定因素。该研究发现，一部分的HSC的细胞表面有巨噬细胞相关的标记。尽管这些HSC功能完整，但这些HSC选择性地保留在细胞龛中，而缺乏巨噬细胞标记的干细胞在强制动员时会离开BM。HSC上的巨噬细胞标记物可以通过受体酪氨酸蛋白激酶C-Kit (CD117)调节的胞啃作用直接从小鼠和人类BM中的驻留巨噬细胞转移获取。该研究提供了概念验证，证明了成体干细胞利用胞啃作用快速建立和激活调节功能的分子机制。
C-Kit依赖的胞啃作用模型调节血液干细胞在细胞龛中的驻留
一部分造血干细胞（HSCs，标记为高C-Kit表达）可以通过胞啃作用（TROGPos HSC）从邻近的巨噬细胞获取膜材料，以增强其在骨髓细胞龛中的驻留。低C-Kit表达的干细胞则被动员到外周血中。
2.PI4P-mediated solid-like Merlin condensates orchestrate Hippo pathway regulation【Signal transduction】
由PI4P介导的类固体Merlin凝聚物调控Hippo信号通路
第一单位：美国德克萨斯大学西南医学中心
Abstract
Despite recent studies implicating liquid-like biomolecular condensates in diverse cellular processes, many biomolecular condensates exist in a solid-like state, and their function and regulation are less understood. We show that the tumor suppressor Merlin, an upstream regulator of the Hippo pathway, localizes to both cell junctions and medial apical cortex in Drosophila epithelia, with the latter forming solid-like condensates that activate Hippo signaling. Merlin condensation required phosphatidylinositol-4-phosphate (PI4P)–mediated plasma membrane targeting and was antagonistically controlled by Pez and cytoskeletal tension through plasma membrane PI4P regulation. The solid-like material properties of Merlin condensates are essential for physiological function and protect the condensates against external perturbations. Collectively, these findings uncover an essential role for solid-like condensates in normal physiology and reveal regulatory mechanisms for their formation and disassembly.
尽管最近的研究表明液态生物分子凝聚体参与了多种细胞过程，但许多生物分子凝聚体以类固态形式存在，其功能和调控机制尚不完全清楚。该研究发现，作为Hippo通路上游调控因子的肿瘤抑制因子Merlin在果蝇上皮细胞中定位于细胞连接处和顶端中部皮质，后者形成类固体凝聚体并激活Hippo信号通路。Merlin的凝聚依赖于磷脂酰肌醇-4-磷酸（PI4P）介导的质膜定位，并通过Pez和细胞骨架张力对质膜PI4P的调控进行拮抗控制。Merlin凝聚体的类固体材料特性对其生理功能至关重要，并能够保护其免受外部扰动。总体而言，这些发现揭示了类固体凝聚体在正常生理中的重要作用，并揭示了其形成和解体的调控机制。
类固体Merlin凝聚体在极化上皮中的功能和调控
Merlin在细胞连接处以动态、弱功能性的形式存在，而在顶端中部皮质处以非动态、强功能性的形式存在，形成类固体凝聚体，后者包含了Hippo通路的核心成分。类固体Merlin的凝聚受到Pez和细胞骨架张力通过PI4P调控的拮抗性控制。其类固体特性还能够保护Merlin凝聚体免受外界压力（如缺氧和低渗）的影响。
3.Brain region–specific action of ketamine as a rapid antidepressant【Neuroscience】
氯胺酮作为快速抗抑郁药在脑区的特异性作用
第一单位：浙江大学
Abstract
Ketamine has been found to have rapid and potent antidepressant activity. However, despite the ubiquitous brain expression of its molecular target, the N-methyl-d-aspartate receptor (NMDAR), it was not clear whether there is a selective, primary site for ketamine’s antidepressant action. We found that ketamine injection in depressive-like mice specifically blocks NMDARs in lateral habenular (LHb) neurons, but not in hippocampal pyramidal neurons. This regional specificity depended on the use-dependent nature of ketamine as a channel blocker, local neural activity, and the extrasynaptic reservoir pool size of NMDARs. Activating hippocampal or inactivating LHb neurons swapped their ketamine sensitivity. Conditional knockout of NMDARs in the LHb occluded ketamine’s antidepressant effects and blocked the systemic ketamine–induced elevation of serotonin and brain-derived neurotrophic factor in the hippocampus. This distinction of the primary versus secondary brain target(s) of ketamine should help with the design of more precise and efficient antidepressant treatments.
氯胺酮具有迅速且强效的抗抑郁活性。然而，其分子靶点N-甲基-D-天冬氨酸受体（NMDAR）在大脑中广泛表达，因此尚不清楚氯胺酮的抗抑郁作用是否有特异性的主要靶点。该研究发现，在抑郁样小鼠中，氯胺酮注射特异性地阻断了外侧缰核（LHb）神经元中的NMDAR，而对海马锥体神经元则无显著影响。这种区域特异性依赖于氯胺酮作为通道阻滞剂的使用依赖性特性、局部神经活动以及NMDAR的突触外储备池的大小。激活海马神经元或抑制LHb神经元会改变它们对氯胺酮的敏感性。对LHb中NMDAR的条件性敲除阻断了氯胺酮的抗抑郁效应，并抑制了氯胺酮系统性诱导的海马体内血清素和脑源性神经营养因子水平的升高。区分氯胺酮的主要与次要脑靶点有助于设计更加精确和高效的抗抑郁治疗方案。
氯胺酮在脑区的特异性作用
模型说明了为什么系统性的氯胺酮却能特异性地阻断抑郁样小鼠中外侧缰核（LHb）神经元中的NMDAR，而对海马CA1锥体神经元无显著影响。这种区域特异性依赖于氯胺酮作为通道阻滞剂的使用依赖性特征、局部神经活动以及NMDAR的突触外储存池的大小。
4.Engineered deletions of HIV replicate conditionally to reduce disease in nonhuman primates【HIV】
HIV的工程化删除变体在非人类灵长类动物中通过条件性复制减少疾病
第一单位：美国加州大学旧金山分校
Abstract
Antiviral therapies with reduced frequencies of administration and high barriers to resistance remain a major goal. For HIV, theories have proposed that viral-deletion variants, which conditionally replicate with a basic reproductive ratio [R0] > 1 (termed “therapeutic interfering particles” or “TIPs”), could parasitize wild-type virus to constitute single-administration, escape-resistant antiviral therapies. We report the engineering of a TIP that, in rhesus macaques, reduces viremia of a highly pathogenic model of HIV by >3log10 following a single intravenous injection. Animal lifespan was significantly extended, TIPs conditionally replicated and were continually detected for >6 months, and sequencing data showed no evidence of viral escape. A single TIP injection also suppressed virus replication in humanized mice and cells from persons living with HIV. These data provide proof of concept for a potential new class of single-administration antiviral therapies.
抗病毒治疗的主要目标是减少给药频率并提高抗药性壁垒。对于HIV，有理论提出，条件性复制且基本繁殖比 [R0] > 1的病毒删除变体（称为“治疗干扰粒子”或“TIPs”）可以寄生于野生型病毒，以构成单次给药、逃逸抗性强的抗病毒疗法。该研究报告了一种TIP的工程化，在猕猴中，通过单次静脉注射将高度致病性的HIV模型的病毒血症降低了超过3log10。动物寿命显著延长，TIPs在体内条件性复制并持续检测超过6个月，测序数据未发现病毒逃逸的证据。单次TIP注射也抑制了人源化小鼠以及HIV感染者细胞中的病毒复制。这些数据为一种潜在的新型单次给药抗病毒治疗提供了概念验证。
单次剂量HIV治疗：TIPs通过条件复制减少非人类灵长类动物中的疾病
(A) TIP概念概述。
(B) 在感染了高致病性SHIV的恒河猴中，TIP干预显著保护了疾病。
(C和D) TIP干预持久地减少了SHIV病毒载量约4log10 (C)，这归因于TIPs的持续条件复制 (D)。
5.Fetal exposure to the Ukraine famine of 1932–1933 and adult type 2 diabetes mellitus【Metabolism】
1932-1933年乌克兰饥荒的胎儿暴露与成年2型糖尿病
第一单位：哥伦比亚大学
Abstract
The short-term impact of famines on death and disease is well documented, but estimating their potential long-term impact is difficult. We used the setting of the man-made Ukrainian Holodomor famine of 1932–1933 to examine the relation between prenatal famine and adult type 2 diabetes mellitus (T2DM). This ecological study included 128,225 T2DM cases diagnosed from 2000 to 2008 among 10,186,016 male and female Ukrainians born from 1930 to 1938. Individuals who were born in the first half-year of 1934, and hence exposed in early gestation to the mid-1933 peak famine period, had a greater than twofold likelihood of T2DM compared with that of unexposed controls. There was a dose-response relationship between severity of famine exposure and increase in adult T2DM risk.
饥荒对死亡和疾病的短期影响已有充分记录，但估计其潜在的长期影响仍然困难。该研究利用1932-1933年人为制造的乌克兰大饥荒（Holodomor）作为背景，研究了产前饥荒与成年2型糖尿病（T2DM）之间的关系。这项生态学研究包括了2000年至2008年间诊断的128,225例T2DM病例，涉及10,186,016名1930年至1938年出生的乌克兰男性和女性。1934年上半年出生的个体，即在妊娠早期暴露于1933年中期饥荒高峰期的个体，其患T2DM的可能性是未暴露对照组的两倍以上。饥荒暴露的严重程度与成年T2DM风险增加之间存在剂量反应关系。
6.The β-d-manno-heptoses are immune agonists across kingdoms【Self and nonself】
β-d-甘露庚糖是跨界的免疫激动剂
第一单位：中国科学院微生物研究所
Abstract
Bacterial small molecule metabolites such as adenosine-diphosphate-D-glycero-β-D-manno-heptose (ADP-heptose) and their derivatives act as effective innate immune agonists in mammals. We show that functional nucleotide-diphosphate-heptose biosynthetic enzymes (HBEs) are distributed widely in bacteria, archaea, eukaryotes, and viruses. We identified a conserved STTR5 motif as a hallmark of heptose nucleotidyltransferases that can synthesize not only ADP-heptose but also cytidine-diphosphate (CDP)– and uridine-diphosphate (UDP)–heptose. Both CDP- and UDP-heptoses are agonists that trigger stronger alpha-protein kinase 1 (ALPK1)–dependent immune responses than ADP-heptose in human and mouse cells and mice. We also produced ADP-heptose in archaea and verified its innate immune agonist functions. Hence, the β-D-manno-heptoses are cross-kingdom, small-molecule, pathogen-associated molecular patterns that activate the ALPK1-dependent innate immune signaling cascade.
细菌的小分子代谢物，如腺苷二磷酸-d-甘油-β-D-甘露庚糖（ADP-heptose）及其衍生物，是哺乳动物中有效的先天免疫激动剂。该研究发现，功能性核苷酸二磷酸庚糖生物合成酶（HBEs）广泛分布于细菌、古菌、真核生物和病毒中。该研究确定了一个保守的STTR5基序，作为庚糖核苷酸转移酶的标志，该酶不仅能合成ADP-heptose，还能合成胞苷二磷酸（CDP）和尿苷二磷酸（UDP）-heptose。CDP和UDP-heptose都是激动剂，在人类和小鼠细胞以及小鼠中引发比ADP-heptose更强的α-蛋白激酶1（ALPK1）依赖的免疫反应。该研究还在古菌中生产了ADP-heptose，并验证了其先天免疫激动剂功能。因此，β-D-甘露庚糖是一种跨界的小分子病原体相关分子模式，可激活ALPK1依赖的先天免疫信号级联反应。
Volume 385| Issue 6710| 16 Aug 2024
在2024年8月16日，《Science》共发表文章33篇：
1篇EDITORIAL
6篇NEWS
11篇INSIGHTS
其中4篇PERSPECTIVES，1篇POLICY FORUM，2本BOOKS，4篇LETTERS
15篇RESEARCH ARTICLES
1.Hebbian instruction of axonal connectivity by endogenous correlated spontaneous activity【Neurodevelopment】
通过内源性相关自发活动对轴突连接进行赫布指导
第一单位：耶鲁大学医学院神经科学系
Abstract
Spontaneous activity refines neural connectivity prior to the onset of sensory experience, but it remains unclear how such activity instructs axonal connectivity with subcellular precision. We simultaneously measured spontaneous retinal waves and the activity of individual retinocollicular axons and tracked morphological changes in axonal arbors across hours in vivo in neonatal mice. We demonstrate that the correlation of an axon branch’s activity with neighboring axons or postsynaptic neurons predicts whether the branch will be added, stabilized, or eliminated. Desynchronizing individual axons from their local networks, changing the pattern of correlated activity, or blocking N-methyl-D-aspartate receptors all significantly altered single-axon morphology. These observations provide the first direct evidence in vivo that endogenous patterns of correlated neuronal activity instruct fine-scale refinement of axonal processes.
自发活动在感觉体验开始之前完善了神经连接，但目前尚不清楚这种活动如何以亚细胞精度指导轴突连接。该研究同时测量了新生小鼠体内的自发视网膜波和单个视网膜-下丘脑轴突的活动，并追踪了体内轴突树在数小时内的形态变化。研究证明了某一轴突分支的活动与其邻近轴突或突触后神经元的相关性可以预测该分支是否会增加、稳定还是被消除。将单个轴突与其局部网络去同步、改变相关活动模式或阻断N-甲基-D-天冬氨酸受体均显著改变了单个轴突的形态。这些观察首次直接证明了体内内源性神经活动模式可以指导轴突结构的精细化调整。
赫布轴突重塑
(A) 清醒小鼠中单个视网膜神经节细胞（RGC）轴突树的体内时间延迟成像示意图，以及轴突和邻近轴突自发活动的双色钙成像示意图。
(B) 单个轴突中GRS的体内成像。
(C) 由自发活动的时空模式介导的赫布轴突重塑示意图。ΔF/F，荧光变化分数；iGluSnFR3，荧光谷氨酸指示剂；tdTomato，红色荧光蛋白。
2.Structure and repair of replication-coupled DNA breaks【Molecular biology】
复制偶联DNA断裂的结构与修复
第一单位：美国国立卫生研究院
Abstract
Using CRISPR-Cas9 nicking enzymes, we examined the interaction between the replication machinery and single-strand breaks, one of the most common forms of endogenous DNA damage. We show that replication fork collapse at leading-strand nicks generates resected single-ended double-strand breaks (seDSBs) that are repaired by homologous recombination (HR). If these seDSBs are not promptly repaired, arrival of adjacent forks creates double-ended DSBs (deDSBs), which could drive genomic scarring in HR-deficient cancers. deDSBs can also be generated directly when the replication fork bypasses lagging-strand nicks. Unlike deDSBs produced independently of replication, end resection at nick-induced seDSBs and deDSBs is BRCA1-independent. Nevertheless, BRCA1 antagonizes 53BP1 suppression of RAD51 filament formation. These results highlight distinctive mechanisms that maintain replication fork stability.
该研究使用CRISPR-Cas9切口酶，探究了复制机制与单链断裂（最常见的内源性DNA损伤形式之一）之间的相互作用。该研究发现，前导链切口处的复制叉崩溃会产生被切除的单端双链断裂（seDSBs），这些断裂通过同源重组（HR）进行修复。如果这些seDSBs没有得到及时修复，相邻的复制叉的到达会产生双端双链断裂（deDSBs），这可能会导致HR缺陷癌症中的基因组瘢痕。当复制叉绕过后导链的切口时，也会直接产生deDSBs。与独立于复制产生的deDSBs不同，切口诱导的seDSBs和deDSBs的末端切除不依赖于BRCA1。然而，BRCA1会拮抗53BP1对RAD51丝状体形成的抑制。这些结果突显了维持复制叉稳定性的独特机制。
经典双链断裂和复制相关的双链断裂的处理方式不同
（右）在经典的DSBs中，末端切除机制和其他DNA损伤反应蛋白（如BRCA1和53BP1）会新招募到损伤部位。在这里，BRCA1和53BP1的促切除和抗切除活性直接竞争，调节切除过程和非同源末端连接和同源重组（HR）之间的选择。
（左）切除核酸酶与未受干扰的复制叉一起移动，并在复制叉崩溃时使细胞选择HR。BRCA1在切除之后起作用，限制53BP1对RAD51加载到切除末端的抑制作用。
3.Drivers of epidemic dynamics in real time from daily digital COVID-19 measurements【Coronavirus】
通过日常数字COVID-19测量实时了解流行病动态的驱动因素
第一单位：瑞士，巴塞尔大学生物中心
Abstract
Understanding the drivers of respiratory pathogen spread is challenging, particularly in a timely manner during an ongoing epidemic. In this work, we present insights that we obtained using daily data from the National Health Service COVID-19 app for England and Wales and that we shared with health authorities in almost real time. Our indicator of the reproduction number R(t) was available days earlier than other estimates, with an innovative capability to decompose R(t) into contact rates and probabilities of infection. When Omicron arrived, the main epidemic driver switched from contacts to transmissibility. We separated contacts and transmissions by day of exposure and setting and found pronounced variability over days of the week and during Christmas holidays and events. For example, during the Euro football tournament in 2021, days with England matches showed sharp spikes in exposures and transmissibility. Digital contact-tracing technologies can help control epidemics not only by directly preventing transmissions but also by enabling rapid analysis at scale and with unprecedented resolution.
了解呼吸道病原体传播的驱动因素颇具挑战，尤其是在疫情持续期间要做到及时了解更是难上加难。在该研究中，研究者从英格兰和威尔士国家医疗服务体系（National Health Service）的COVID-19应用程序获得每日数据，并几乎实时地与卫生部门进行了分享。研究提出的再生数R(t)指标比其他估计值提前数天得出，并具备将R(t)分解为接触率和感染概率的创新能力。当奥密克戎变异株出现时，疫情的主要驱动因素从接触性转变为传播性。研究根据暴露的天数和场所对接触性和传播性进行了区分，发现周末、圣诞节假期及活动期间存在显著的变化。例如，在2021年欧洲足球锦标赛期间，有英格兰队比赛的日子，暴露量和传播性均急剧上升。数字接触者追踪技术不仅可以直接预防传播，还能实现大规模且前所未有的高分辨率快速分析，从而助力疫情防控。
2020 年欧洲杯足球锦标赛期间英格兰基于应用程序的流行病监测示例
接触者（176,000）和COVID-19传播（7800）的最高峰值发生在决赛当天。这些线条显示应用程序检测到的每日接触者（蓝色，以100人为单位）和传播者（红色）的估计数量，以及涉及英格兰队、对手 [由国旗和国际足球联合会（FIFA）国家代码标识]和周末（灰色阴影）的比赛日期。
4.A hippocampal circuit mechanism to balance memory reactivation during sleep【Neuroscience】
一种海马回路机制，用于平衡睡眠期间的记忆重新激活
第一单位：康奈尔大学神经生物学和行为系
Abstract
Memory consolidation involves the synchronous reactivation of hippocampal cells active during recent experience in sleep sharp-wave ripples (SWRs). How this increase in firing rates and synchrony after learning is counterbalanced to preserve network stability is not understood. We discovered a network event generated by an intrahippocampal circuit formed by a subset of CA2 pyramidal cells to cholecystokinin-expressing (CCK+) basket cells, which fire a barrage of action potentials (“BARR”) during non–rapid eye movement sleep. CA1 neurons and assemblies that increased their activity during learning were reactivated during SWRs but inhibited during BARRs. The initial increase in reactivation during SWRs returned to baseline through sleep. This trend was abolished by silencing CCK+ basket cells during BARRs, resulting in higher synchrony of CA1 assemblies and impaired memory consolidation.
记忆巩固涉及到海马神经元在睡眠中的尖波涟漪（SWRs）同步激活，这些神经元在最近的经历中处于活跃状态。学习后，神经元的放电频率和同步性如何增加以达到平衡，从而维持网络稳定性，目前还不清楚。该研究发现，由CA2锥体细胞的一个子集和表达促胰液素（CCK）的篮状细胞组成的内侧海马回路，会在非快速眼动睡眠期间产生一种网络事件，即“BARR”（密集的行动电位）。在学习期间增加活动性的CA1神经元和神经元集群在SWR期间被重新激活，但在BARR期间被抑制。在SWR期间的初始激活增强趋势通过睡眠逐渐恢复到基线水平。这种趋势在BARR期间可以通过抑制CCK+篮状细胞而被消除，导致CA1神经元集群的同步性更高，记忆巩固受损。
5.Structure and inhibition of SARS-CoV-2 spike refolding in membranes【Coronavirus】
膜中 SARS-CoV-2 刺突再折叠的结构与抑制
第一单位：耶鲁大学
Abstract
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein binds the receptor angiotensin converting enzyme 2 (ACE2) and drives virus-host membrane fusion through refolding of its S2 domain. Whereas the S1 domain contains high sequence variability, the S2 domain is conserved and is a promising pan-betacoronavirus vaccine target. We applied cryo–electron tomography to capture intermediates of S2 refolding and understand inhibition by antibodies to the S2 stem-helix. Subtomogram averaging revealed ACE2 dimers cross-linking spikes before transitioning into S2 intermediates, which were captured at various stages of refolding. Pan-betacoronavirus neutralizing antibodies targeting the S2 stem-helix bound to and inhibited refolding of spike prehairpin intermediates. Combined with molecular dynamics simulations, these structures elucidate the process of SARS-CoV-2 entry and reveal how pan-betacoronavirus S2-targeting antibodies neutralize infectivity by arresting prehairpin intermediates.
严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的刺突蛋白与血管紧张素转换酶2（ACE2）受体结合，并通过其S2结构域的重新折叠驱动病毒-宿主膜融合。其中，S1结构域具有高序列变异性，而S2结构域则相对保守，是一个很有前景的泛β冠状病毒疫苗靶点。研究应用冷冻电子断层扫描技术捕捉S2结构域重新折叠的中间体，并了解针对S2茎螺旋的抗体的抑制作用。亚断层扫描平均分析显示，在转变为S2中间体之前，ACE2二聚体会交联刺突蛋白。研究在S2结构域重新折叠的不同阶段捕捉到了这些中间体。针对S2茎螺旋的泛β冠状病毒中和抗体与刺突蛋白前发夹中间体结合并抑制其重新折叠。结合分子动力学模拟，这些结构阐明了SARS-CoV-2的入侵过程，并揭示了泛β冠状病毒S2靶向抗体如何通过阻滞前发夹中间体来中和感染力。
6.Predictive grid coding in the medial entorhinal cortex【Neurogenesis】
内侧内嗅皮层的预测网格编码
第一单位：日本理化学研究所（RIKEN）脑科学研究中心
Abstract
The entorhinal cortex represents allocentric spatial geometry and egocentric speed and heading information required for spatial navigation. However, it remains unclear whether it contributes to the prediction of an animal’s future location. We discovered grid cells in the medial entorhinal cortex (MEC) that have grid fields representing future locations during goal-directed behavior. These predictive grid cells represented prospective spatial information by shifting their grid fields against the direction of travel. Predictive grid cells discharged at the trough phases of the hippocampal CA1 theta oscillation and, together with other types of grid cells, organized sequences of the trajectory from the current to future positions across each theta cycle. Our results suggest that the MEC provides a predictive map that supports forward planning in spatial navigation.
内嗅皮层负责表示空间导航所需的以环境为中心的空间几何信息和以自我为中心的速度及方向信息。然而，它是否有助于预测动物的未来位置尚不清楚。该研究发现内侧内嗅皮质(MEC)中的网格细胞具有代表目标导向行为期间未来位置的网格区域。这些预测性网格细胞通过使网格区域相对于行进方向发生偏移来表示未来的空间信息。预测性网格细胞在海马体CA1区的θ波振荡的波谷阶段放电，并与其他类型的网格细胞一起，在每个θ波周期内组织从当前位置到未来位置的轨迹序列。该研究的结果表明，内侧内嗅皮层提供了一个预测地图，支持空间导航的前瞻性规划。
Volume 385| Issue 6711| 23 AUGUST 2024
在2024年8月23日，《Science》共发表文章32篇：
6篇NEWS
10篇INSIGHTS
其中2本BOOK，4篇PERSPECTIVES，1篇POLICY FORUM， 3篇LETTERS
15篇RESEARCH ARTICLES
1篇EDITORIAL
1.Restoring hippocampal glucose metabolism rescues cognition across Alzheimer’s disease pathologies【Neurodegeneration】
恢复海马葡萄糖代谢可挽救阿尔茨海默病的认知功能
第一单位：斯坦福大学医学院，神经病学和神经科学系
Abstract
Impaired cerebral glucose metabolism is a pathologic feature of Alzheimer’s disease (AD), with recent proteomic studies highlighting disrupted glial metabolism in AD. We report that inhibition of indoleamine-2,3-dioxygenase 1 (IDO1), which metabolizes tryptophan to kynurenine (KYN), rescues hippocampal memory function in mouse preclinical models of AD by restoring astrocyte metabolism. Activation of astrocytic IDO1 by amyloid β and tau oligomers increases KYN and suppresses glycolysis in an aryl hydrocarbon receptor–dependent manner. In amyloid and tau models, IDO1 inhibition improves hippocampal glucose metabolism and rescues hippocampal long-term potentiation in a monocarboxylate transporter–dependent manner. In astrocytic and neuronal cocultures from AD subjects, IDO1 inhibition improved astrocytic production of lactate and uptake by neurons. Thus, IDO1 inhibitors presently developed for cancer might be repurposed for treatment of AD.
脑葡萄糖代谢障碍是阿尔茨海默病（AD）的病理特征之一，最近的蛋白质组学研究强调了AD中胶质细胞代谢的紊乱。吲哚胺-2,3-双加氧酶1 (IDO1)可将色氨酸代谢为犬尿氨酸(KYN)，抑制IDO1可恢复星形胶质细胞代谢，从而改善AD小鼠临床前模型的海马记忆功能。淀粉样β蛋白和tau寡聚体通过激活星形胶质细胞中的IDO1，以依赖芳烃受体（AhR）的方式增加KYN并抑制糖酵解。在淀粉样蛋白和tau模型中，抑制IDO1可改善海马葡萄糖代谢，并以依赖单羧酸转运体的方式挽救海马长时程增强。在来自AD患者的星形胶质细胞和神经元共培养体系中，抑制IDO1可改善星形胶质细胞的乳酸生成和神经元对乳酸的摄取。因此，目前为治疗癌症开发的IDO1抑制剂可能被用于AD的治疗。
2.The actin-spectrin submembrane scaffold restricts endocytosis along proximal axons【Cellular neuroscience】
肌动蛋白-血影蛋白亚膜支架限制了近端轴突的内吞作用
第一单位：索邦大学，法国国家健康与医学研究院
Abstract
Clathrin-mediated endocytosis has characteristic features in neuronal dendrites and presynapses, but how membrane proteins are internalized along the axon shaft remains unclear. We focused on clathrin-coated structures and endocytosis along the axon initial segment (AIS) and their relationship to the periodic actin-spectrin scaffold that lines the axonal plasma membrane. A combination of super-resolution microscopy and platinum-replica electron microscopy on cultured neurons revealed that AIS clathrin-coated pits form within “clearings”, circular areas devoid of actin-spectrin mesh. Actin-spectrin scaffold disorganization increased clathrin-coated pit formation. Cargo uptake and live-cell imaging showed that AIS clathrin-coated pits are particularly stable. Neuronal plasticity-inducing stimulation triggered internalization of the clathrin-coated pits through polymerization of branched actin around them. Thus, spectrin and actin regulate clathrin-coated pit formation and scission to control endocytosis at the AIS.
网格蛋白介导的内吞作用在神经元树突和突触前膜中具有典型的特征，但膜蛋白如何沿着轴突内吞仍不清楚。该研究重点关注轴突起始段（AIS）的网格蛋白包被结构和内吞作用，以及它们与轴突质膜内侧排列的周期性肌动蛋白-血影蛋白骨架的关系。对培养的神经元进行超分辨率显微镜和铂复型电子显微镜检查，发现AIS网格蛋白包被的凹坑在“间隙”内形成，这些空隙是缺乏肌动蛋白-血影蛋白网的圆形区域。肌动蛋白-血影蛋白骨架紊乱会增加网格蛋白包被凹坑的形成。货物摄取和活细胞成像显示，AIS的网格蛋白包被凹坑特别稳定。诱导神经元可塑性的刺激通过围绕网格蛋白包被凹坑的分支肌动蛋白聚合触发其内化。因此，血影蛋白和肌动蛋白通过调节网格蛋白包被凹坑的形成和分裂来控制AIS的内吞作用。
3.Mechanisms of minor pole–mediated spindle bipolarization in human oocytes【Reproduction】
人卵母细胞中次要极介导的纺锤体双极化的机制
第一单位：复旦大学附属儿科研究所
Abstract
Spindle bipolarization, the process of a microtubule mass transforming into a bipolar spindle, is a prerequisite for accurate chromosome segregation. In contrast to mitotic cells, the process and mechanism of spindle bipolarization in human oocytes remains unclear. Using high-resolution imaging in more than 1800 human oocytes, we revealed a typical state of multipolar intermediates that form during spindle bipolarization and elucidated the mechanism underlying this process. We found that the minor poles formed in multiple kinetochore clusters contribute to the generation of multipolar intermediates. We further determined the essential roles of HAUS6, KIF11, and KIF18A in spindle bipolarization and identified mutations in these genes in infertile patients characterized by oocyte or embryo defects. These results provide insights into the physiological and pathological mechanisms of spindle bipolarization in human oocytes.
纺锤体双极化，即微管团块转变为双极纺锤体的过程，是染色体精确分离的前提条件。与有丝分裂细胞相比，人类卵母细胞纺锤体双极化的过程和机制尚不清楚。通过对1800多个人类卵母细胞进行高分辨率成像，该研究揭示了纺锤体两极化过程中形成的多极中间体的典型状态，并阐明了这一过程的潜在机制。该研究发现，在多个着丝粒簇中形成的小极有助于多极中间体的产生。该研究进一步确定了HAUS6、KIF11和KIF18A在纺锤体双极化中的关键作用，并在以卵母细胞或胚胎缺陷为特征的不育患者中发现了这些基因的突变。这些结果为理解人类卵母细胞纺锤体双极化的生理和病理机制提供了新见解。
4.Structural insights into the human NuA4/TIP60 acetyltransferase and chromatin remodeling complex【Molecular biology】
人类NuA4/TIP60乙酰转移酶和染色质重塑复合物的结构研究
第一单位：加州大学伯克利分校，加利福尼亚定量生物科学研究所（QB3）
Abstract
The human nucleosome acetyltransferase of histone H4 (NuA4)/Tat-interactive protein, 60 kilodalton (TIP60) coactivator complex, a fusion of the yeast switch/sucrose nonfermentable related 1 (SWR1) and NuA4 complexes, both incorporates the histone variant H2A.Z into nucleosomes and acetylates histones H4, H2A, and H2A.Z to regulate gene expression and maintain genome stability. Our cryo–electron microscopy studies show that, within the NuA4/TIP60 complex, the E1A binding protein P400 (EP400) subunit serves as a scaffold holding the different functional modules in specific positions, creating a distinct arrangement of the actin-related protein (ARP) module. EP400 interacts with the transformation/transcription domain-associated protein (TRRAP) subunit by using a footprint that overlaps with that of the Spt-Ada-Gcn5 acetyltransferase (SAGA) complex, preventing the formation of a hybrid complex. Loss of the TRRAP subunit leads to mislocalization of NuA4/TIP60, resulting in the redistribution of H2A.Z and its acetylation across the genome, emphasizing the dual functionality of NuA4/TIP60 as a single macromolecular assembly.
人类组蛋白H4核小体乙酰转移酶（NuA4）/Tat互作蛋白60千道尔顿（TIP60）共激活复合物是酵母开关/蔗糖非发酵相关1（SWR1）复合物和NuA4复合物的融合体，该复合物可将组蛋白变体H2A.Z整合入核小体，并使组蛋白H4、H2A和H2A.Z乙酰化，以调控基因表达并维持基因组稳定性。该研究的冷冻电子显微镜结果显示，在NuA4/TIP60复合物中，E1A结合蛋白P400（EP400）亚基作为支架，将不同的功能模块固定在特定位置，从而形成肌动蛋白相关蛋白（ARP）模块的独特排列。EP400通过与变换/转录结构域相关蛋白（TRRAP）亚基相互作用，其结合位点与Spt-Ada-Gcn5乙酰转移酶（SAGA）复合物的结合位点重叠，从而阻止杂交复合物的形成。TRRAP亚基的缺失会导致NuA4/TIP60定位错误，进而导致H2A.Z及其乙酰化在整个基因组中重新分布，这强调了NuA4/TIP60作为单一大分子组装体的双重功能。
5.Massively parallel analysis of single-molecule dynamics on next-generation sequencing chips【Molecular biology】
在下一代测序芯片上进行单分子动力学的大规模并行分析
第一单位：乌普萨拉大学，生命科学实验室细胞与分子生物学系
Abstract
Single-molecule techniques are ideally poised to characterize complex dynamics but are typically limited to investigating a small number of different samples. However, a large sequence or chemical space often needs to be explored to derive a comprehensive understanding of complex biological processes. Here we describe multiplexed single-molecule characterization at the library scale (MUSCLE), a method that combines single-molecule fluorescence microscopy with next-generation sequencing to enable highly multiplexed observations of complex dynamics. We comprehensively profiled the sequence dependence of DNA hairpin properties and Cas9-induced target DNA unwinding-rewinding dynamics. The ability to explore a large sequence space for Cas9 allowed us to identify a number of target sequences with unexpected behaviors. We envision that MUSCLE will enable the mechanistic exploration of many fundamental biological processes.
单分子技术非常适用于表征复杂的动态学过程，但通常仅限于研究少量不同的样本。然而，为了全面理解复杂的生物过程，往往需要探索大量的序列或化学空间。该研究描述了一种名为MUSCLE的多重单分子分析方法，该方法将单分子荧光显微镜与下一代测序相结合，可实现对复杂动态过程的高通量观测。该研究全面分析了DNA发夹结构的序列依赖性以及Cas9诱导的目标DNA解旋-复位动态过程。由于Cas9能够在较大的序列空间中进行探索，该研究发现了多个具有意外行为的目标序列。因此该研究设想MUSCLE将能够用于探索许多基本生物过程的机制。
6.Single-molecule structural and kinetic studies across sequence space【Molecular biology】
跨序列空间的单分子结构和动力学研究
第一单位：荷兰，代尔夫特理工大学，卡夫利纳米科学研究所生物纳米科学系
Abstract
At the core of molecular biology lies the intricate interplay between sequence, structure, and function. Single-molecule techniques provide in-depth dynamic insights into structure and function, but laborious assays impede functional screening of large sequence libraries. We introduce high-throughput Single-molecule Parallel Analysis for Rapid eXploration of Sequence space (SPARXS), integrating single-molecule fluorescence with next-generation sequencing. We applied SPARXS to study the sequence-dependent kinetics of the Holliday junction, a critical intermediate in homologous recombination. By examining the dynamics of millions of Holliday junctions, covering thousands of distinct sequences, we demonstrated the ability of SPARXS to uncover sequence patterns, evaluate sequence motifs, and construct thermodynamic models. SPARXS emerges as a versatile tool for untangling the mechanisms that underlie sequence-specific processes at the molecular scale.
分子生物学的核心在于序列、结构与功能之间错综复杂的相互作用。单分子技术为结构和功能提供了深入的动态见解，但繁琐的检测阻碍了大规模序列文库的功能筛选。该研究引入了高通量单分子并行分析快速探索序列空间（SPARXS）技术，该技术将单分子荧光与下一代测序相结合。该研究应用SPARXS技术研究了Holliday连接体（同源重组中的一个关键中间体）的序列依赖性动力学。通过检测数百万个Holliday连接体的动力学，这些连接体涵盖了数千种不同的序列，该研究证明了SPARXS技术能够揭示序列模式、评估序列基序并构建热力学模型。SPARXS已成为一种多功能工具，用于解开分子尺度上序列特异性过程背后的机制。
Volume 385| Issue 6712| 30 AUGUST 2024
在2024年8月30日，《Science》共发表文章30篇：
6篇NEWS
9篇INSIGHTS
其中4篇PERSPECTIVES，1篇POLICY FORUM，2本BOOK，3篇LETTERS
14篇RESEARCH ARTICLES
1篇REVIEW
1.PERK-ATAD3A interaction provides a subcellular safe haven for protein synthesis during ER stress【Cell biology】
PERK-ATAD3A相互作用为内质网应激期间的蛋白质合成提供了一个亚细胞的安全庇护所
第一单位：英国剑桥科学研究所
Abstract
Endoplasmic reticulum (ER) stress induces the repression of protein synthesis throughout the cell. Attempts to understand how localized stress leads to widespread repression have been limited by difficulties in resolving translation rates at the subcellular level. Here, using live-cell imaging of reporter mRNA translation, we unexpectedly found that during ER stress, active translation at mitochondria was significantly protected. The mitochondrial protein ATPase family AAA domain-containing protein 3A (ATAD3A) interacted with protein kinase RNA–like endoplasmic reticulum kinase (PERK) and mediated this effect on localized translation by competing for binding with PERK's target, eukaryotic initiation factor 2 (eIF2). PERK-ATAD3A interactions increased during ER stress, forming mitochondria-ER contact sites. Furthermore, ATAD3A binding attenuated local PERK signaling and rescued the expression of some mitochondrial proteins. Thus, PERK-ATAD3A interactions can control translational repression at a subcellular level, mitigating the impact of ER stress on the cell.
内质网（ER）应激会诱导整个细胞内蛋白质合成的抑制。然而，由于难以解析亚细胞水平的翻译速率，理解局部应激如何导致广泛抑制的尝试一直受到限制。该研究利用报告基因mRNA翻译的活细胞成像技术，意外地发现，在内质网应激期间，线粒体上的活跃翻译得到了显著保护。线粒体蛋白ATP酶家族AAA结构域包含蛋白3A（ATAD3A）与RNA样内质网蛋白激酶（PERK）发生相互作用，并通过与PERK的靶基因真核细胞起始因子2 （eIF2）竞争结合来介导这种定位翻译效应。在内质网应激期间，PERK-ATAD3A相互作用增强，形成线粒体-内质网接触位点。此外，ATAD3A的结合减弱了局部的PERK信号，并挽救了部分线粒体蛋白的表达。因此，PERK-ATAD3A相互作用可以在亚细胞水平控制翻译抑制，从而减轻内质网应激对细胞的影响。
2.Micronuclear collapse from oxidative damage【Cell biology】
氧化损伤引起的微核崩溃
第一单位：美国纽约纪念斯隆-凯特琳癌症中心
Abstract
Chromosome-containing micronuclei are a hallmark of aggressive cancers. Micronuclei frequently undergo irreversible collapse, exposing their enclosed chromatin to the cytosol. Micronuclear rupture catalyzes chromosomal rearrangements, epigenetic abnormalities, and inflammation, yet mechanisms safeguarding micronuclear integrity are poorly understood. In this study, we found that mitochondria-derived reactive oxygen species (ROS) disrupt micronuclei by promoting a noncanonical function of charged multivesicular body protein 7 (CHMP7), a scaffolding protein for the membrane repair complex known as endosomal sorting complex required for transport III (ESCRT-III). ROS retained CHMP7 in micronuclei while disrupting its interaction with other ESCRT-III components. ROS-induced cysteine oxidation stimulated CHMP7 oligomerization and binding to the nuclear membrane protein LEMD2, disrupting micronuclear envelopes. Furthermore, this ROS-CHMP7 pathological axis engendered chromosome shattering known to result from micronuclear rupture. It also mediated micronuclear disintegrity under hypoxic conditions, linking tumor hypoxia with downstream processes driving cancer progression.
含染色体的微核是侵袭性癌症的一个标志。微核经常发生不可逆的崩解，使其封闭的染色质暴露于细胞质中。微核破裂会催化染色体重排、表观遗传异常和炎症反应，然而保护微核完整性的机制尚不清楚。该研究发现线粒体来源的活性氧（ROS）通过促进带电多泡体蛋白7（CHMP7）的非典型功能来破坏微核，CHMP7是膜修复复合体转运所需的内体分选复合体III（ESCRT-III）的支架蛋白。ROS使CHMP7滞留在微核内，同时破坏其与ESCRT-III其他组分的相互作用。ROS诱导的半胱氨酸氧化促进了CHMP7的低聚化及其与核膜蛋白LEMD2的结合，从而破坏了微核膜。此外，这一ROS-CHMP7病理轴导致了由微核破裂引起的染色体碎裂。它还介导了缺氧条件下的微核解体，将肿瘤缺氧与驱动癌症进展的下游过程联系起来。
3.A p62-dependent rheostat dictates micronuclei catastrophe and chromosome rearrangements【Cell biology】
p62依赖性调节器决定微核灾难和染色体重排
第一单位：意大利米兰，欧洲肿瘤研究所
Abstract
Chromosomal instability (CIN) generates micronuclei—aberrant extranuclear structures that catalyze the acquisition of complex chromosomal rearrangements present in cancer. Micronuclei are characterized by persistent DNA damage and catastrophic nuclear envelope collapse, which exposes DNA to the cytoplasm. We found that the autophagic receptor p62/SQSTM1 modulates micronuclear stability, influencing chromosome fragmentation and rearrangements. Mechanistically, proximity of micronuclei to mitochondria led to oxidation-driven homo-oligomerization of p62, limiting endosomal sorting complex required for transport (ESCRT)–dependent micronuclear envelope repair by triggering autophagic degradation. We also found that p62 levels correlate with increased chromothripsis across human cancer cell lines and with increased CIN in colorectal tumors. Thus, p62 acts as a regulator of micronuclei and may serve as a prognostic marker for tumors with high CIN.
染色体不稳定性（CIN）会产生位于细胞核外的异常结构——微核，而它能够催化癌症中出现的复杂染色体重排。微核的特征是持续的DNA损伤和灾难性的核膜崩解，从而使DNA暴露于细胞质中。该研究发现自噬受体p62/SQSTM1能够调节微核的稳定性，进而影响染色体的碎裂和重排。在机制上，微核与线粒体的靠近导致p62发生氧化驱动的同源低聚化，通过触发自噬降解来限制转运所需的内体分选复合体（ESCRT）依赖性的微核膜修复。该研究还发现，在人类癌细胞系中，p62的水平与增加的染色单体碎裂呈正相关，并且在结直肠癌中，p62的水平与增加的CIN也呈正相关。因此，p62作为微核的调节器，可能成为高CIN肿瘤的预后标志物。
4.Morphine-responsive neurons that regulate mechanical antinociception【Neuroscience】
吗啡反应神经元调节机械镇痛
第一单位：瑞典斯德哥尔摩，卡罗林斯卡医学院，医学生物化学和生物物理学系，分子神经生物学部
Abstract
Opioids are widely used, effective analgesics to manage severe acute and chronic pain, although they have recently come under scrutiny because of epidemic levels of abuse. While these compounds act on numerous central and peripheral pain pathways, the neuroanatomical substrate for opioid analgesia is not fully understood. By means of single-cell transcriptomics and manipulation of morphine-responsive neurons, we have identified an ensemble of neurons in the rostral ventromedial medulla (RVM) that regulates mechanical nociception in mice. Among these, forced activation or silencing of excitatory RVMBDNF projection neurons mimicked or completely reversed morphine-induced mechanical antinociception, respectively, via a brain-derived neurotrophic factor (BDNF)/tropomyosin receptor kinase B (TrkB)–dependent mechanism and activation of inhibitory spinal galanin-positive neurons. Our results reveal a specific RVM-spinal circuit that scales mechanical nociception whose function confers the antinociceptive properties of morphine.
阿片类药物是广泛使用的有效镇痛药，用于治疗严重的急性和慢性疼痛，但最近因其滥用情况而受到了审查。虽然这些化合物作用于众多中枢和外周疼痛传导通路，但阿片类药物镇痛的神经解剖学基础尚未完全阐明。该研究通过单细胞转录组学和吗啡反应神经元操控技术，确定了位于延髓头端腹外侧区（RVM）中的一组神经元，这组神经元可调节小鼠的机械性痛觉。其中，强制激活或沉默兴奋性RVMBDNF投射神经元分别通过脑源性神经营养因子 (BDNF) /原肌球蛋白受体激酶B（TrkB）依赖机制和激活抑制性脊髓甘丙肽阳性神经元来模拟或完全逆转吗啡诱导的机械性抗伤害感受。该研究的结果揭示了一个特定的RVM-脊髓环路，该环路可调节机械性痛觉，其功能赋予了吗啡的镇痛特性。
5.Aggregate-selective removal of pathological tau by clustering-activated degraders【Cellular neuroscience】
通过聚集激活的降解剂选择性去除病理性tau蛋白
第一单位：剑桥大学英国痴呆症研究所
Abstract
Selective degradation of pathological protein aggregates while sparing monomeric forms is of major therapeutic interest. The E3 ligase tripartite motif–containing protein 21 (TRIM21) degrades antibody-bound proteins in an assembly state–specific manner due to the requirement of TRIM21 RING domain clustering for activation, yet effective targeting of intracellular assemblies remains challenging. Here, we fused the RING domain of TRIM21 to a target-specific nanobody to create intracellularly expressed constructs capable of selectively degrading assembled proteins. We evaluated this approach against green fluorescent protein–tagged histone 2B (H2B-GFP) and tau, a protein that undergoes pathological aggregation in Alzheimer’s and other neurodegenerative diseases. RING-nanobody degraders prevented or reversed tau aggregation in culture and in vivo, with minimal impact on monomeric tau. This approach may have therapeutic potential for the many disorders driven by intracellular protein aggregation.
选择性降解病理性蛋白聚集体而保留单体形式具有重要的治疗意义。由于E3连接酶三结构域蛋白21（TRIM21）的激活需要TRIM21 RING结构域的聚类，因此TRIM21以组装状态特异性的方式降解抗体结合蛋白，但有效靶向胞内组装仍然具有挑战性。该研究将TRIM21的RING结构域与目标特异性的纳米抗体融合，创建了能够在细胞内表达且能选择性降解组装蛋白的构建体。该研究利用这种方法评估了绿色荧光蛋白标记的组蛋白2B（H2B-GFP）和tau蛋白（一种在阿尔茨海默病和其他神经退行性疾病中发生病理性聚集的蛋白）。RING-纳米抗体降解剂在培养条件下和体内均能够防止或逆转tau蛋白的聚集，而对单体tau蛋白的影响极小。这种方法对于许多由细胞内蛋白聚集驱动的疾病可能具有治疗潜力。
Volume 385| Issue 6713| 6 SEPTEMBER 2024
在2024年9月6日，《Science》共发表文章32篇：
6篇NEWS
10篇INSIGHTS
其中4篇PERSPECTIVES，1篇POLICY FORUM，2本BOOKS，3篇LETTERS
15篇RESEARCH ARTICLES
1篇REVIEW
1.The use of ectopic volar fibroblasts to modify skin identity 【Skin】
利用异位掌侧成纤维细胞改变皮肤特性
第一单位：约翰霍普金斯大学医学院皮肤科
Abstract
Skin identity is controlled by intrinsic features of the epidermis and dermis and their interactions. Modifying skin identity has clinical potential, such as the conversion of residual limb and stump (nonvolar) skin of amputees to pressure-responsive palmoplantar (volar) skin to enhance prosthesis use and minimize skin breakdown. Greater keratin 9 (KRT9) expression, higher epidermal thickness, keratinocyte cytoplasmic size, collagen length, and elastin are markers of volar skin and likely contribute to volar skin resiliency. Given fibroblasts’ capacity to modify keratinocyte differentiation, we hypothesized that volar fibroblasts influence these features. Bioprinted skin constructs confirmed the capacity of volar fibroblasts to induce volar keratinocyte features. A clinical trial of healthy volunteers demonstrated that injecting volar fibroblasts into nonvolar skin increased volar features that lasted up to 5 months, highlighting a potential cellular therapy.
皮肤特性是由表皮和真皮的内在特征及其相互作用所控制的。改变皮肤特性具有临床应用的潜力，例如，将截肢者的残肢和残端（非掌侧）皮肤转化为具有压力敏感的掌跖（掌侧）皮肤，以增强假肢的使用并最大限度地减少皮肤破损。掌侧皮肤的标志包括角蛋白9（KRT9）表达增加、表皮增厚、角质形成细胞胞质增大、胶原纤维变长以及弹性蛋白增多，这些特征可能有助于掌侧皮肤的韧性。鉴于成纤维细胞具有改变角质形成细胞分化的能力，该研究假设掌侧成纤维细胞能够影响这些特征。生物打印皮肤结构证实了掌侧成纤维细胞能够诱导掌侧角质形成细胞的特征。一项针对健康志愿者的临床试验表明，将掌侧成纤维细胞注射到非掌侧皮肤中，能够增加掌侧特征，且这些特征可持续长达5个月的时间，这凸显了一种潜在的细胞疗法。
2.Single-cell chromatin accessibility reveals malignant regulatory programs in primary human cancers【Cancer】
单细胞染色质可及性揭示原发性人类癌症中的恶性调控程序
第一单位：斯坦福大学医学院遗传学系
Abstract
To identify cancer-associated gene regulatory changes, we generated single-cell chromatin accessibility landscapes across eight tumor types as part of The Cancer Genome Atlas. Tumor chromatin accessibility is strongly influenced by copy number alterations that can be used to identify subclones, yet underlying cis-regulatory landscapes retain cancer type–specific features. Using organ-matched healthy tissues, we identified the “nearest healthy” cell types in diverse cancers, demonstrating that the chromatin signature of basal-like–subtype breast cancer is most similar to secretory-type luminal epithelial cells. Neural network models trained to learn regulatory programs in cancer revealed enrichment of model-prioritized somatic noncoding mutations near cancer-associated genes, suggesting that dispersed, nonrecurrent, noncoding mutations in cancer are functional. Overall, these data and interpretable gene regulatory models for cancer and healthy tissue provide a framework for understanding cancer-specific gene regulation.
为了识别与癌症相关的基因调控变化，该研究作为癌症基因组图谱（The Cancer Genome Atlas）项目的一部分，绘制了八种肿瘤类型的单细胞染色质可及性图谱。肿瘤染色质可及性受到拷贝数变异的强烈影响，这些变异可用于识别亚克隆，但潜在的顺式调控景观仍保留了癌症类型特异性的特征。通过使用与器官匹配的健康组织，该研究确定了多种癌症中“最接近健康”的细胞类型，证明基底细胞样亚型乳腺癌的染色质特征与分泌型管腔上皮细胞最为相似。该研究通过训练神经网络模型来学习癌症中的调控程序，发现模型优先排序的癌症相关基因附近存在体细胞非编码突变富集，这表明癌症中分散的、非重复的、非编码的突变是有功能的。总体而言，这些数据以及癌症和健康组织的可解释性基因调控模型为理解癌症特异性基因调控提供了一个框架。
全癌种单细胞染色质可及性基因调控图谱
该图谱基于基因规模的可及性模式，能够从其他细胞类型中识别出肿瘤细胞，并能够通过兆碱基规模的变化对肿瘤中的亚克隆异质性进行分析。神经网络模型揭示了癌症组织与健康组织之间特有的可及性染色质特征，并强调了非编码体细胞突变在癌症中的作用。scATAC-seq代表单细胞ATAC-seq；DL Pred.代表深度学习预测；Ref代表参考；Alt代表替代。
3.Role of protein kinase PLK1 in the epigenetic maintenance of centromeres【Cell cycle】
蛋白激酶 PLK1 在着丝粒表观遗传维持中的作用
第一单位：德国，马克斯普朗克分子生理学研究所，机械细胞生物学系
Abstract
The centromere, a chromosome locus defined by the histone H3–like protein centromeric protein A (CENP-A), promotes assembly of the kinetochore to bind microtubules during cell division. Centromere maintenance requires CENP-A to be actively replenished by dedicated protein machinery in the early G1 phase of the cell cycle to compensate for its dilution after DNA replication. Cyclin-dependent kinases (CDKs) limit CENP-A deposition to once per cell cycle and function as negative regulators outside of early G1. Antithetically, Polo-like kinase 1 (PLK1) promotes CENP-A deposition in early G1, but the molecular details of this process are still unknown. We reveal here a phosphorylation network that recruits PLK1 to the deposition machinery to control a conformational switch required for licensing the CENP-A deposition reaction. Our findings clarify how PLK1 contributes to the epigenetic maintenance of centromeres.
着丝粒是由组蛋白H3样蛋白着丝粒蛋白A（CENP-A）定义的一个染色体位点，它在细胞分裂期间促进着丝粒的组装以便与微管结合。着丝粒的维持需要在细胞周期的早期G1期由专门的蛋白质机器主动补充CENP-A，以补偿DNA复制后其浓度的稀释。细胞周期依赖性激酶（CDKs）将CENP-A的沉积限制在每个细胞周期内仅一次，并在早期G1期之外作为负调控因子发挥作用。相反，Polo样激酶1（PLK1）在早期G1期促进CENP-A的沉积，但这一过程的分子细节尚不清楚。该研究揭示了一个磷酸化网络，该网络将PLK1招募到沉积机器中，以控制CENP-A沉积反应所需的构象开关。该研究结果阐明了PLK1如何促进着丝粒的表观遗传维护。
4.PLK1-mediated phosphorylation cascade activates Mis18 complex to ensure centromere inheritance【Cell cycle】
PLK1 介导的磷酸化级联反应激活 Mis18 复合物以确保着丝粒遗传
第一单位：英国，爱丁堡大学，惠康细胞生物学中心
Abstract
Accurate chromosome segregation requires the attachment of microtubules to centromeres, epigenetically defined by the enrichment of CENP-A nucleosomes. During DNA replication, CENP-A nucleosomes undergo dilution. To preserve centromere identity, correct amounts of CENP-A must be restored in a cell cycle–controlled manner orchestrated by the Mis18 complex (Mis18α-Mis18β-Mis18BP1). We demonstrate here that PLK1 interacts with the Mis18 complex by recognizing self-primed phosphorylations of Mis18α (Ser54) and Mis18BP1 (Thr78 and Ser93) through its Polo-box domain. Disrupting these phosphorylations perturbed both centromere recruitment of the CENP-A chaperone HJURP and new CENP-A loading. Biochemical and functional analyses showed that phosphorylation of Mis18α and PLK1 binding were required to activate Mis18α-Mis18β and promote Mis18 complex-HJURP interaction. Thus, our study reveals key molecular events underpinning the licensing role of PLK1 in ensuring accurate centromere inheritance.
精确的染色体分离需要微管附着在由CENP-A核小体富集所定义的着丝粒上。在DNA复制期间，CENP-A核小体会减少。为了保持着丝粒的完整性，必须以细胞周期控制的方式在Mis18复合物（Mis18α-Mis18β-Mis18BP1）的协调下恢复适量的CENP-A。该研究发现，PLK1通过其Polo盒结构域识别Mis18α（Ser54）和Mis18BP1（Thr78和Ser93）的自发磷酸化，并与Mis18复合物相互作用。扰乱这些磷酸化会扰乱CENP-A伴侣HJURP的着丝粒招募和新CENP-A的装载。生化和功能分析显示，磷酸化Mis18α和PLK1结合是激活Mis18α-Mis18β并促进Mis18复合物-HJURP相互作用所必需的。因此，该研究揭示了PLK1在确保精确的着丝粒遗传中发挥许可作用的关键分子事件。
Volume 385| Issue 6713| 13 SEPTEMBER 2024
在2024年9月13日，《Science》共发表文章31篇：
6篇NEWS
10篇INSIGHTS
其中5篇PERSPECTIVES，1篇POLICY FORUM，1本BOOK，3篇LETTERS
14篇RESEARCH ARTICLES
1篇REVIEW
1.Autoregulated splicing of TRA2β programs T cell fate in response to antigen-receptor stimulation【Immunology】
抗原受体刺激下TRA2β的自发剪接调控T细胞命运
第一单位：美国，康涅狄格大学医学院，免疫学系
Abstract
T cell receptor (TCR) sensitivity to peptide–major histocompatibility complex (MHC) dictates T cell fate. Canonical models of TCR sensitivity cannot be fully explained by transcriptional regulation. In this work, we identify a posttranscriptional regulatory mechanism of TCR sensitivity that guides alternative splicing of TCR signaling transcripts through an evolutionarily ultraconserved poison exon (PE) in the RNA-binding protein (RBP) TRA2β in mouse and human. TRA2β-PE splicing, seen during cancer and infection, was required for TCR-induced effector T cell expansion and function. Tra2β-PE skipping enhanced T cell response to antigen by increasing TCR sensitivity. As antigen levels decreased, Tra2β-PE reinclusion allowed T cell survival. Finally, we found that TRA2β-PE was first included in the genome of jawed vertebrates that were capable of TCR gene rearrangements. We propose that TRA2β-PE splicing acts as a gatekeeper of TCR sensitivity to shape T cell fate.
T细胞受体（TCR）对肽-主要组织相容性复合体（MHC）的敏感性决定了T细胞的命运。传统的TCR敏感性模型无法完全用转录调控来解释。该研究鉴定了一种TCR敏感性的转录后调控机制，该机制通过小鼠和人类RNA结合蛋白（RBP）TRA2β中一个进化上高度保守的毒性外显子（PE）来指导TCR信号转录本的可变剪接。在癌症和感染期间观察到的TRA2β-PE剪接对于TCR诱导的效应T细胞扩增和功能至关重要。Tra2β-PE的跳过通过提高TCR敏感性而增强了T细胞对抗原的应答。随着抗原水平的降低，Tra2β-PE的再包涵使T细胞能够存活。最后，该研究发现TRA2β-PE首先出现在能够进行TCR基因重排的颌类脊椎动物基因组中。该研究提出，TRA2β-PE剪接作为TCR敏感性的守门人，塑造着T细胞的命运。
TRA2β的PE（加工元件）作为对抗原应答中T细胞受体（TCR）的守门人
在初始T细胞中，TRA2β的PE（TRA2β-PE）更频繁地包含在其转录本中，导致TRA2β蛋白的基础水平较低。抗原触发TCR会导致TRA2β-PE被跳过，从而增加TRA2β蛋白的表达并开启TCR信号的“闸门”。随后，TRA2β与TCR信号mRNA的可变剪接结合并进行调控，改变转录本异构体谱，以形成最佳效应T细胞。MHC I，即主要组织相容性复合体I类分子。
2.Transcripts of repetitive DNA elements signal to block phagocytosis of hematopoietic stem cells【Blood stem cells】
重复 DNA 元件的转录本信号阻断造血干细胞的吞噬作用
第一单位：美国，波士顿儿童医院，霍华德·休斯医学研究所
Abstract
Macrophages maintain hematopoietic stem cell (HSC) quality by assessing cell surface Calreticulin (Calr), an “eat-me” signal induced by reactive oxygen species (ROS). Using zebrafish genetics, we identified Beta-2-microglobulin (B2m) as a crucial “don’t eat-me” signal on blood stem cells. A chemical screen revealed inducers of surface Calr that promoted HSC proliferation without triggering ROS or macrophage clearance. Whole-genome CRISPR-Cas9 screening showed that Toll-like receptor 3 (Tlr3) signaling regulated b2m expression. Targeting b2m or tlr3 reduced the HSC clonality. Elevated B2m levels correlated with high expression of repetitive element (RE) transcripts. Overall, our data suggest that RE-associated double-stranded RNA could interact with TLR3 to stimulate surface expression of B2m on hematopoietic stem and progenitor cells. These findings suggest that the balance of Calr and B2m regulates macrophage-HSC interactions and defines hematopoietic clonality.
巨噬细胞通过评估细胞表面的钙网蛋白（Calr）来维持造血干细胞（HSC）的质量，这是一种由活性氧（ROS）诱导的“吃掉我”信号。利用斑马鱼遗传学方法，该研究确定了β2-微球蛋白（B2m）是血液干细胞上一个关键的“别吃我”信号。化学筛选方法发现了一种表面Calr的诱导剂，该物质能促进HSC增殖，而不会触发ROS或巨噬细胞清除。全基因组CRISPR-Cas9筛选显示，Toll样受体3（Tlr3）信号可调节b2m的表达。靶向b2m或tlr3降低了HSC的克隆性。B2m水平升高与重复元件（RE）转录本高表达相关。总体而言，该研究表明，与RE相关的双链RNA可能与TLR3相互作用，刺激造血干细胞和祖细胞表面B2m的表达。这些发现表明，Calr和B2m的平衡调节着巨噬细胞与HSC的相互作用，并决定了造血克隆性。
活性氧（ROS）和TLR3信号介导“吃掉我”和“别吃我”的信号，这些信号决定了巨噬细胞对干细胞的克隆选择。
图中所示为所提出模型的示意图，展示了由Calr介导的“吃掉我”信号和B2m介导的“别吃我”信号之间的平衡，这些信号影响着巨噬细胞的行为。
3.Inherent symmetry and flexibility in hepatitis B virus subviral particles【Structural biology】
乙型肝炎病毒亚病毒颗粒的固有对称性和灵活性
第一单位：中国，上海科技大学免疫化学研究所
Abstract
Chronic hepatitis B virus (HBV) infection poses a major global health challenge with massive morbidity and mortality. Despite a preventive vaccine, current treatments provide limited virus clearance, necessitating lifelong commitment. The HBV surface antigen (HBsAg) is crucial for diagnosis and prognosis, yet its high-resolution structure and assembly on the virus envelope remain elusive. Utilizing extensive datasets and advanced cryo–electron microscopy analysis, we present structural insights into HBsAg at a near-atomic resolution of 3.7 angstroms. HBsAg homodimers assemble into subviral particles with D2- and D4-like quasisymmetry, elucidating the dense-packing rules and structural adaptability of HBsAg. These findings provide insights into how HBsAg assembles into higher-order filaments and interacts with the capsid to form virions.
慢性乙型肝炎病毒（HBV）感染具有高发病率和高死亡率，是全球公共卫生领域面临的一项重大挑战。尽管已有预防性疫苗，但当前的治疗方法仅能实现有限的病毒清除，患者需要终身治疗。乙型肝炎病毒表面抗原（HBsAg）对于疾病的诊断和治疗预后至关重要，但其高分辨率结构以及在病毒包膜上的组装方式仍不清楚。该研究利用大量数据集和先进的冷冻电子显微镜分析技术，以3.7埃的近原子分辨率揭示了HBsAg的结构特征。HBsAg同源二聚体组装成具有D2和D4样准对称性的亚病毒颗粒，阐明了HBsAg的紧密排列规则和结构适应性。这些发现有助于了解HBsAg如何组装成高阶纤维，以及与衣壳相互作用形成病毒颗粒的机制。
Volume 385| Issue 6715| 20 SEPTEMBER 2024
在2024年9月20日，《Science》共发表文章34篇：
6篇NEWS
10篇INSIGHTS
其中5篇PERSPECTIVES，2本BOOKS，3篇LETTERS
13篇RESEARCH ARTICLES
1.Germline mutations in a G protein identify signaling cross-talk in T cells【Immunology】
G 蛋白中的种系突变可识别 T 细胞中的信号串扰
第一单位：美国国立卫生研究院(NIH)变态反应和传染病研究所(NIAID)
Abstract
Humans with monogenic inborn errors responsible for extreme disease phenotypes can reveal essential physiological pathways. We investigated germline mutations in GNAI2, which encodes Gαi2, a key component in heterotrimeric G protein signal transduction usually thought to regulate adenylyl cyclase–mediated cyclic adenosine monophosphate (cAMP) production. Patients with activating Gαi2 mutations had clinical presentations that included impaired immunity. Mutant Gαi2 impaired cell migration and augmented responses to T cell receptor (TCR) stimulation. We found that mutant Gαi2 influenced TCR signaling by sequestering the guanosine triphosphatase (GTPase)–activating protein RASA2, thereby promoting RAS activation and increasing downstream extracellular signal–regulated kinase (ERK)/mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI3K)–AKT S6 signaling to drive cellular growth and proliferation.
单基因先天缺陷可导致极端疾病表型，这些表型能够揭示关键的生理途径。该研究探究了编码Gαi2的GNAI2基因的生殖系突变。Gαi2是异源三聚体G蛋白信号转导的关键成分，通常认为它调节腺苷酸环化酶介导的环腺苷酸单磷酸（cAMP）的产生。携带激活性Gαi2突变的患者临床表现为免疫功能受损。突变的Gαi2影响细胞迁移并增强对T细胞受体（TCR）刺激的反应。该研究发现，突变的Gαi2通过扣留鸟苷三磷酸酶（GTPase）激活蛋白RASA2来影响TCR信号传导，从而促进RAS的激活，并增加下游细胞外信号调节激酶（ERK）/丝裂原活化蛋白激酶（MAPK）和磷脂酰肌醇3激酶（PI3K）-AKT S6信号传导，以驱动细胞生长和增殖。
2.Artificial kinetochore beads establish a biorientation-like state in the spindle【Synthetic biology】
人工着丝粒珠在纺锤体中建立双向样状态
第一单位：日本神户理化研究所，生物系统动力学研究中心，染色体分离实验室
Abstract
Faithful chromosome segregation requires biorientation, where the pair of kinetochores on the chromosome establish bipolar microtubule attachment. The integrity of the kinetochore, a macromolecular complex built on centromeric DNA, is required for biorientation, but components sufficient for biorientation remain unknown. Here, we show that tethering the outer kinetochore heterodimer NDC80-NUF2 to the surface of apolar microbeads establishes their biorientation-like state in mouse cells. NDC80-NUF2 microbeads align at the spindle equator and self-correct alignment errors. The alignment is associated with stable bipolar microtubule attachment and is independent of the outer kinetochore proteins SPC24-SPC25, KNL1, the Mis12 complex, inner kinetochore proteins, and Aurora. Larger microbeads align more rapidly, suggesting a size-dependent biorientation mechanism. This study demonstrates a biohybrid kinetochore design for synthetic biorientation of microscale particles in cells.
精确的染色体分离需要生物定向，即染色体上的一对着丝粒蛋白（kinetochores）建立起双极微管的附着。染色体着丝粒蛋白的完整性对于生物定向是必需的，但足以实现生物定向的成分尚未被发现。该研究展示了在微球表面固定外着丝粒异二聚体NDC80-NUF2可以使小鼠细胞中的微球进入类似于生物定向的状态。NDC80-NUF2微球在纺锤体赤道处排列并能自行纠正定位误差。这种排列与稳定的双极微管附着相关，并且与外着丝粒蛋白SPC24-SPC25、KNL1、Mis12复合物、内着丝粒蛋白以及Aurora无关。较大的微球排列速度更快，这表明存在一种与微球大小相关的生物定向机制。这项研究展示了一种生物混合型着丝粒设计的概念，可用于在细胞中对微尺度粒子进行合成生物定向。
3.Targeting cancer with small-molecule pan-KRAS degraders【Drug development】
使用小分子 pan-KRAS 降解剂靶向癌症
第一单位：英国，邓迪大学，生命科学学院，靶向蛋白质降解中心
Abstract
Mutations in the Kirsten rat sarcoma viral oncogene homolog (KRAS) protein are highly prevalent in cancer. However, small-molecule concepts that address oncogenic KRAS alleles remain elusive beyond replacing glycine at position 12 with cysteine (G12C), which is clinically drugged through covalent inhibitors. Guided by biophysical and structural studies of ternary complexes, we designed a heterobifunctional small molecule that potently degrades 13 out of 17 of the most prevalent oncogenic KRAS alleles. Compared with inhibition, KRAS degradation results in more profound and sustained pathway modulation across a broad range of KRAS mutant cell lines, killing cancer cells while sparing models without genetic KRAS aberrations. Pharmacological degradation of oncogenic KRAS was tolerated and led to tumor regression in vivo. Together, these findings unveil a new path toward addressing KRAS-driven cancers with small-molecule degraders.
KRAS（Kirsten鼠肉瘤病毒癌基因同源物）蛋白的突变在癌症中极为常见。然而，除了通过共价抑制剂将第12位的甘氨酸（G12C）替换为半胱氨酸而在临床上实现药物干预外，针对致癌性KRAS等位基因的小分子药物概念仍然难以捉摸。在三元复合物的生物物理和结构研究指导下，该研究设计了一种异双功能小分子，该小分子能够高效降解17种最常见的致癌性KRAS等位基因中的13种。与抑制KRAS活性相比，KRAS降解能够在广泛的KRAS突变细胞系中带来更深刻且持久的通路调节效果，既能杀死癌细胞，又不影响无KRAS基因突变的模型。致癌性KRAS的药理降解被证明是可耐受的，并在体内实验中导致了肿瘤消减。综上所述，这些发现为使用小分子降解剂治疗KRAS驱动的癌症开辟了一条新途径。
4.Large library docking identifies positive allosteric modulators of the calcium-sensing receptor【Drug discovery】
大文库对接可识别钙敏感受体的正变构调节剂
第一单位：美国，斯坦福大学，医学院，分子与细胞生理学教研室
Abstract
Positive allosteric modulator (PAM) drugs enhance the activation of the calcium-sensing receptor (CaSR) and suppress parathyroid hormone (PTH) secretion. Unfortunately, these hyperparathyroidism-treating drugs can induce hypocalcemia and arrhythmias. Seeking improved modulators, we docked libraries of 2.7 million and 1.2 billion molecules against the CaSR structure. The billion-molecule docking found PAMs with a 2.7-fold higher hit rate than the million-molecule library, with hits up to 37-fold more potent. Structure-based optimization led to nanomolar leads. In ex vivo organ assays, one of these PAMs was 100-fold more potent than the standard of care, cinacalcet, and reduced serum PTH levels in mice without the hypocalcemia typical of CaSR drugs. As determined from cryo–electron microscopy structures, the PAMs identified here promote CaSR conformations that more closely resemble the activated state than those induced by the established drugs.
正变构调节剂（PAM）药物可增强钙敏感受体（CaSR）的激活并抑制甲状旁腺激素（PTH）的分泌。然而，这些用于治疗甲状旁腺功能亢进的药物可能会引发低钙血症和心律失常。为了寻找更有效的调节剂，该研究将包含270万和12亿个分子的库与CaSR结构进行对接。十亿分子库对接发现的PAM的命中率比百万分子库高出2.7倍，且效力最高可达37倍。基于结构的优化产生了纳摩尔级别的先导化合物。在体外器官实验中，其中一种PAM的效力比标准治疗药物西那卡塞高出100倍，并能降低小鼠的血清PTH水平，且未出现CaSR药物典型的低钙血症。正如冷冻电子显微镜的结构测定所示，该研究发现的PAM促进的CaSR构象比现有药物诱导的构象更接近激活状态。
汇报人：毛敏姿
导师：赵宇
审核：胥飞宇、任建君
