第3期 | Countstar与科研之间的故事

发布日期：2021-06-14 09:12:00浏览次数：2660

本周CountStar在最新的应用发表文献中选取了6篇分享给大家。第1篇是关于COVID-19单细胞测序相关的研究，文章通过对感染者和健康者的免疫细胞的转录谱测定，揭示了发病是机体免疫反应的动态性质。第3篇文章介绍了LncRNA-DILA1对于乳腺癌的调控机理。第4篇文章在免疫治疗领域新型化学材料的应用前景探究。

1.Single-celllandscape of immunological responses in COVID-19 patient

COVID-19患者免疫应答的单细胞研究

[Nat Immunol]      IF=20.479     PMID:32788748

Abstract ：In COVID-19 caused bySARS-CoV-2 infection, the relationship between disease severity and the hostimmune response is not fully understood. Here we performed single-cell RNAsequencing in peripheral blood samples of five healthy donors and 13 COVID-19patients including moderate, severe and convalescent cases. Through determiningthe transcriptional profiles of immune cells, coupled with assembled T cellreceptor and B cell receptor sequences, we analyzed the functional propertiesof immune cells. Most cell types in COVID-19 patients showed a stronginterferon-alpha response, and an overall acute inflammatory response.Moreover, intensive expansion of highly cytotoxic effector T cell subsets, suchas CD4+ Effector-GNLY (Granulysin), CD8+ Effector-GNLY and NKT CD160, wasassociated with convalescence in moderate patients. In severe patients, theimmune landscape featured a deranged interferon response, profound immuneexhaustion with skewed T cell receptor repertoire and broad T cell expansion.These findings illustrate the dynamic nature of immune responses during thedisease progression.

摘要：在SARS-CoV-2感染引起的COVID-19中，疾病严重程度与宿主免疫应答之间的关系尚不完全清楚。在这里，我们对5名健康者和13名COVID-19患者（包括中、重度和康复期患者）的外周血样本进行了单细胞RNA测序。通过测定免疫细胞的转录谱，结合组装的T细胞受体和B细胞受体的序列，分析了免疫细胞的功能特性。COVID-19患者的大多数细胞类型显示出强烈的干扰素-α反应和整体急性炎症反应。此外，高度细胞毒性效应T细胞亚群，如CD4+效应子GNLY（Granulysin）、CD8+效应子GNLY和NKT-CD160的密集扩增与中度患者的恢复有关。在重症患者中，免疫系统的特点是干扰素反应紊乱，严重的免疫衰竭，T细胞受体库倾斜，T细胞广泛扩张。这些发现说明了疾病发展过程中免疫反应的动态性质。

2.Wnt/β‐catenin‐mediated heat exposure inhibitsintestinal epithelial cell proliferation and stem cell expansion throughendoplasmic reticulum stress

Wnt/β-catenin介导的热暴露通过内质网应激抑制肠上皮细胞增殖和干细胞扩张

[J Cell Physiol]        IF=5.546        PMID:31960439

Abstract：Heat stress induced by continuous high ambient temperaturesor strenuous exercise in humans and animals leads to intestinal epithelialdamage through the induction of intracellular stress response. However, theprecise mechanisms involved in the regulation of intestinal epithelial cellinjury, especially intestinal stem cells (ISCs), remain unclear. Thereby, invitro a confluent monolayer of IPEC‐J2 cells was exposed to the high temperatures (39, 40,and 41°C), the IPEC‐J2 cell proliferation, apoptosis, differentiation, andbarrier were determined, as well as the expression of GRP78, which is a markerprotein of endoplasmic reticulum stress (ERS). The Wnt/β‐catenin pathway‐ mediated regenerative response was validated using R‐spondin 1(Rspo1). And ex‐vivo, three‐dimensional cultured enteroids were developed from pigletjejunal crypt and employed to assess the ISC activity under heat exposure. Theresults showed that exposure to 41°C for 72 hr, rather than 39°C and 40°C,decreased IPEC‐J2 cell viability, inhibited cell proliferation anddifferentiation, induced ERS and cell apoptosis, damaged barrier function andrestricted the Wnt/β‐cateninpathway. Nevertheless, Wnt/β‐cateninreactivation via Rspo1 protects the intestinal epithelium from heat exposure‐inducedinjury. Furthermore, exposure to 41°C for 24 hr reduced ISC activity,stimulated crypt‐cell apoptosis, upregulated the expression of GRP78 andcaspase‐3, and downregulated the expression of β‐catenin, Lgr5, Bmi1, Ki67, KRT20, ZO‐1,occludin, and claudin‐1. Taken together, we conclude that heat exposure inducesERS and downregulates the Wnt/β‐cateninsignaling pathway to disrupt epithelial integrity by inhibiting the intestinalepithelial cell proliferation and stem cell expansion.

摘要：持续高温或剧烈运动引起的热应激通过诱导细胞内应激反应导致肠上皮损伤。然而，肠上皮细胞损伤，特别是肠干细胞（ISCs）的机制尚不清楚。因此，体外将融合的IPEC-J2细胞单层暴露于高温（39、40和41°C）检测IPEC-J2细胞增殖、凋亡和分化，以及内质网应激（ERS）标志蛋白GRP78的表达。结果表明，暴露于41°C 72小时，而不是39°C和40°C、降低IPEC-J2细胞活力，抑制细胞增殖和分化，诱导内质网和细胞凋亡，破坏屏障功能，限制Wnt/β‐连环蛋白途径。尽管如此，Wnt/β‐通过Rspo1激活连环蛋白保护肠上皮免受热暴露诱导的损伤。此外，暴露于41°C持续24小时可降低ISC活性，刺激细胞凋亡，上调GRP78和caspase-3的表达，下调GRP78和caspase-3的表达β‐catenin、Lgr5、Bmi1、Ki67、KRT20、ZO-1、occludin和claudin-1。综上所述，我们得出结论，热暴露诱导和下调Wnt/β‐连环蛋白信号通路通过抑制肠上皮细胞增殖和干细胞扩张破坏上皮完整性。

3.LncRNA DILA1 inhibits Cyclin D1 degradation andcontributes to tamoxifen resistance in breast cancer

LncRNA-DILA1抑制细胞周期蛋白D1降解并促进乳腺癌对三苯氧胺的耐药性

[Nat Commun]        IF=12.121        PMID: 33139730

Abstract：Cyclin D1 is one of the most important oncoproteins thatdrives cancer cell proliferation and associates with tamoxifen resistance inbreast cancer. Here, we identify a lncRNA, DILA1, which interacts with CyclinD1 and is overexpressed in tamoxifen-resistant breast cancer cells.Mechanistically, DILA1 inhibits the phosphorylation of Cyclin D1 at Thr286 bydirectly interacting with Thr286 and blocking its subsequent degradation,leading to overexpressed Cyclin D1 protein in breast cancer. Knocking downDILA1 decreases Cyclin D1 protein expression, inhibits cancer cell growth andrestores tamoxifen sensitivity both in vitro and in vivo. High expression ofDILA1 is associated with overexpressed Cyclin D1 protein and poor prognosis inbreast cancer patients who received tamoxifen treatment. This study shows the previouslyunappreciated importance of post-translational dysregulation of Cyclin D1 con tributingto tamoxifen resistance in breast cancer. Moreover, it reveals the novelmechanism of DILA1 in regulating Cyclin D1 protein stability and suggests DILA1is a specifific therapeutic target to downregulate Cyclin D1 protein andreverse tamoxifen resistance in treating breast cancer.

摘要：周期蛋白D1是乳腺癌中最重要的癌蛋白之一，它能促进癌细胞增殖，并与三苯氧胺的耐药有关。在此，我们鉴定了一种与cyclind1相互作用的lncRNA，DILA1，它在三苯氧胺耐药的乳腺癌细胞中过度表达。DILA1通过直接与Thr286相互作用并阻断其随后的降解，抑制Thr286处cyclind1的磷酸化，导致乳腺癌中cyclind1蛋白的过度表达。敲除DILA1可降低细胞周期蛋白D1的表达，在体内外抑制癌细胞生长并恢复三苯氧胺的敏感性。在接受三苯氧胺治疗的乳腺癌患者中，DILA1的高表达与细胞周期蛋白D1的过度表达和不良预后相关。这项研究显示了先前未被重视的细胞周期蛋白D1翻译后失调对乳腺癌三苯氧胺耐药的重要性。此外，还揭示了DILA1调节cyclind1蛋白稳定性的新机制，提示DILA1是下调cyclind1蛋白和逆转三苯氧胺耐药治疗乳腺癌的特异性治疗靶点。

4.Nanoparticle-enhanced chemo-immunotherapy to triggerrobust antitumor immunity

纳米颗粒增强化学免疫治疗激发抗肿瘤免疫

[Sci Adv]         IF=13.113         PMID: 32923651

Abstract：Mounting evidence suggests that immunotherapies are apromising new class of anticancer therapies. However, the immunosuppressivetumor microenvironment (TME), poor immunogenicity, and off-target toxicityhinder the broader implementation of immunotherapies. Here, we describe a novelstrategy combining chemotherapy and immunotherapy to modulate the TME bysystemically and concurrently delivering the chemotherapeutic agent SN38(7-ethyl-10-hydroxycamptothecin) and the STING agonist DMXAA(5,6-dimethylxanthenone-4-acetic acid) into tumors using triblock copolymernanoparticles, named PS3D1@DMXAA, which enhances antigen cross-presentation andinduces the conversion of the immunosuppressive TME to immunogenic TME throughthe newly found synergistic function between SN38 and STING activation.PS3D1@DMXAA thus shows potent therapeutic efficacy in three mice tumor modelsand elicits remarkable therapeutic benefit when combined with anti–PD-1therapy. Our engineered nanosystem offers a rational design of an effectiveimmunotherapy combination regimen to convert uninflamed “cold” tumors into“hot” tumors, addressing the major challenges immunotherapies faced.

摘要：越来越多的证据表明免疫疗法是一种很有前途的新型抗癌疗法。然而，免疫抑制性肿瘤微环境（TME）、免疫原性差、靶向毒性等因素阻碍了免疫治疗的广泛应用。在这里，我们阐述了一种结合化疗和免疫治疗的新策略，通过使用三嵌段共聚物纳米颗粒将SN38（7-乙基-10-羟基喜树碱）和DMXAA（5,6-二甲基黄嘌呤酮-4-乙酸）系统地和同时地递送到肿瘤中来调节TME，命名PS3D1@DMXAA。通过新发现的SN38与STING激活之间的协同作用，诱导免疫抑制性TME转化为免疫原性TME。在三种小鼠肿瘤模型中显示出有效的治疗效果，并且与抗PD-1治疗相结合时产生显著的治疗效果。我们的工程纳米系统提供了一个有效的免疫治疗组合方案的合理设计，将未燃烧的“冷”肿瘤转化为“热”肿瘤，解决了免疫治疗面临的主要挑战。

5.Intrinsic ColorSensing System Allows for Real-Time Observable Functional Changes on Human InducedPluripotent Stem Cell-Derived Cardiomyocytes

固有颜色传感系统允许实时观察到人类诱导的多能干细胞衍生心肌细胞的功能变化

[ACS Nano]         IF=14.588        PMID: 32609489

Abstract：Stem-cell based in vitro difffferentiation for diseasemodeling offffers great value to explore the molecular and functionalunderpinnings driving many types of cardiomyopathy and congenital heartdiseases. Nevertheless, one major caveat in the application of in vitrodifffferentiation of human induced pluripotent stem cell (hiPSC)-derivedcardiomyocytes (hiPSC-CMs) involves the immature phenotype of the CMs. Most ofthe existing methods need complex apparatus and require laborious procedures inorder to monitor the cardiac difffferentiation/maturation process and oftenresult in cell death. Here we developed an intrinsic color sensing systemutilizing a microgroove structural color methacrylated gelatin fifilm, whichallows us to monitor the cardiac difffferentiation  process of hiPSC-derived cardiac progenitorcells in real time. Subsequently this system can be employed as an assay systemto live monitor induced functional changes on hiPSC-CMs stemming from drugtreatment, the effffects of which are simply revealed through color diversity.Our research shows that early intervention of cardiac difffferentiation throughsimple physical cues can enhance cardiac difffferentiation and maturation tosome extent. Our system also simplififies the previous complex experimentalprocesses for evaluating the physiological effffects of successfuldifffferentiation and drug treatment and lays a solid foundation for futuretransformational applications.

摘要：干细胞为基础的体外分化的疾病模型对于探索驱动多种类型心肌病和先天性心脏病的分子和功能基础具有重要价值。然而，在人诱导多能干细胞（hiPSC）衍生的心肌细胞（hiPSC-CMs）体外分化应用中的一个主要注意事项涉及CMs的不成熟表型。大多数现有的方法需要复杂的仪器和费力的程序，以监测心脏分化/成熟过程，往往导致细胞死亡。在这里，我们开发了一个固有的颜色传感系统，利用微槽结构颜色甲基丙烯酸明胶薄膜，使我们能够实时监测hiPSC衍生的心脏祖细胞的心脏分化过程。随后，该系统可以作为一个检测系统，实时监测药物治疗引起的hiPSC-CMs功能变化，其效果可以通过颜色多样性简单地显示出来。我们的研究表明，通过简单的物理线索对心脏分化的早期干预可以在一定程度上促进心脏的分化和成熟。我们的系统还简化了以前复杂的实验过程，以评估成功的分化和药物治疗的生理效应，为将来的转化应用奠定了坚实的基础。

6.Mitochondrial UQCC3 Modulates Hypoxia Adaptation

byOrchestrating OXPHOS and Glycolysis in Hepatocellular Carcinoma

线粒体UQCC3通过协调OXPHOS和糖酵解调节肝细胞癌的缺氧适应

[Cell Rep]       IF=8.109  PMID: 33147459

Abstract：Bioenergetic reprogramming during hypoxia adaption iscritical to promote hepatocellular carcinoma (HCC) growth and progression.However, the mechanism underlying the orchestration of mitochondrial OXPHOS(oxidative phosphorylation) and glycolysis in hypoxia is not fully understood.Here, we report that mitochondrial UQCC3 (C11orf83) expression increases inhypoxia and correlates with the poor prognosis of HCC patients. Loss of UQCC3impairs HCC cell proliferation in hypoxia in vitro and in vivo.Mechanistically, UQCC3 forms a positive feedback loop with mitochondrialreactive oxygen species (ROS) to sustain UQCC3 expression and ROS generation inhypoxic HCC cells and subsequently maintains mitochondrial structure andfunction and stabilizes HIF-1a expression to enhance glycolysis under hypoxia.Thus, UQCC3 plays an indispensable role for bioenergetic reprogramming of HCCcells during hypoxia adaption by simultaneously regulating OXPHOS andglycolysis. The positive feedback between UQCC3 and ROS indicates aself-modulating model within mitochondria that initiates the adaptation of HCCto hypoxic stress.

摘要：低氧适应过程中的重编程序是促进肝细胞癌（HCC）生长和发展的关键。然而，线粒体OXPHOS（oxyphosphylation）和糖酵解在缺氧条件下的协调机制尚不完全清楚。在这里，我们阐述了线粒体UQCC3（C11orf83）在缺氧条件下的表达增加，并与肝癌患者的不良预后相关。缺氧条件下UQCC3缺失对肝癌细胞增殖的影响。在机制上，UQCC3与线粒体活性氧（ROS）形成正反馈环，维持UQCC3在缺氧肝癌细胞中的表达和ROS的产生，进而维持线粒体的结构和功能，稳定HIF-1a的表达，增强缺氧条件下的糖酵解。因此，UQCC3通过同时调节OXPHOS和糖酵解，在肝癌细胞缺氧适应过程中的重编程中起着不可或缺的作用。UQCC3和ROS之间的正反馈表明线粒体内存在一个自我调节模型，启动HCC对缺氧应激的自适应。

Countstar Fluorescence Cell Analyzer