更新于：2024-09-05

EF-1A

更新于：2024-09-05

基本信息

别名

DKFZp434g247、EF-1a、eRF3c

+ [7]

简介

GTPase component of the Pelota-HBS1L complex, a complex that recognizes stalled ribosomes and triggers the No-Go Decay (NGD) pathway (PubMed:21448132, PubMed:23667253, PubMed:27863242). The Pelota-HBS1L complex recognizes ribosomes stalled at the 3' end of an mRNA and engages stalled ribosomes by destabilizing mRNA in the mRNA channel (PubMed:27863242). Following mRNA extraction from stalled ribosomes by the SKI complex, the Pelota-HBS1L complex promotes recruitment of ABCE1, which drives the disassembly of stalled ribosomes, followed by degradation of damaged mRNAs as part of the NGD pathway (PubMed:21448132, PubMed:32006463).

关联

项与 EF-1A 相关的药物

FFT-135

靶点

EF-1A x GSPT1 x GSPT2

作用机制

EF-1A降解剂 [+2]

在研机构

F5 Therapeutics, Inc.

原研机构

F5 Therapeutics, Inc.

在研适应症

结直肠癌 [+2]

非在研适应症-

最高研发阶段临床前

首次获批国家/地区-

首次获批日期1800-01-20

100 项与 EF-1A 相关的临床结果

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100 项与 EF-1A 相关的转化医学

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0 项与 EF-1A 相关的专利（医药）

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764

项与 EF-1A 相关的文献（医药）

2024-02-16·International journal of molecular sciences

Novel Enhanced Mammalian Cell Transient Expression Vector via Promoter Combination.

Article

作者: SunKyung Yoon ; SeJin Park ; JuneWoo Lee ; Byoungguk Kim ; WonSeok Gwak

During the emergence of infectious diseases, evaluating the efficacy of newly developed vaccines requires antigen proteins. Available methods enhance antigen protein productivity; however, structural modifications may occur. Therefore, we aimed to construct a novel transient overexpression vector capable of rapidly producing large quantities of antigenic proteins in mammalian cell lines. This involved expanding beyond the exclusive use of the human cytomegalovirus (CMV) promoter, and was achieved by incorporating a transcriptional enhancer (CMV enhancer), a translational enhancer (woodchuck hepatitis virus post-transcriptional regulatory element), and a promoter based on the CMV promoter. Twenty novel transient expression vectors were constructed, with the vector containing the human elongation factor 1-alpha (EF-1a) promoter showing the highest efficiency in expressing foreign proteins. This vector exhibited an approximately 27-fold higher expression of enhanced green fluorescent protein than the control vector containing only the CMV promoter. It also expressed the highest level of severe acute respiratory syndrome coronavirus 2 receptor-binding domain protein. These observations possibly result from the simultaneous enhancement of the transcriptional activity of the CMV promoter and the human EF-1a promoter by the CMV enhancer. Additionally, the synergistic effect between the CMV and human EF-1a promoters likely contributed to the further enhancement of protein expression.

2024-02-10·Human molecular genetics

The genetic dissection of fetal haemoglobin persistence in sickle cell disease in Nigeria.

Article

作者: Oyesola O Ojewunmi ; Titilope A Adeyemo ; Ajoke I Oyetunji ; Bassey Inyang ; Afolashade Akinrindoye ; Baraka S Mkumbe ; Kate Gardner ; Helen Rooks ; John Brewin ; Hamel Patel ; Sang Hyuck Lee ; Raymond Chung ; Sara Rashkin ; Guolian Kang ; Reuben Chianumba ; Raphael Sangeda ; Liberata Mwita ; Hezekiah Isa ; Uche-Nnebe Agumadu ; Rosemary Ekong ; Jamilu A Faruk ; Bello Y Jamoh ; Niyi M Adebiyi ; Ismail A Umar ; Abdulaziz Hassan ; Christopher Grace ; Anuj Goel ; Baba P D Inusa ; Mario Falchi ; Siana Nkya ; Julie Makani ; Hafsat R Ahmad ; Obiageli Nnodu ; John Strouboulis ; Stephan Menzel

The clinical severity of sickle cell disease (SCD) is strongly influenced by the level of fetal haemoglobin (HbF) persistent in each patient. Three major HbF loci (BCL11A, HBS1L-MYB, and Xmn1-HBG2) have been reported, but a considerable hidden heritability remains. We conducted a genome-wide association study for HbF levels in 1006 Nigerian patients with SCD (HbSS/HbSβ0), followed by a replication and meta-analysis exercise in four independent SCD cohorts (3,582 patients). To dissect association signals at the major loci, we performed stepwise conditional and haplotype association analyses and included public functional annotation datasets. Association signals were detected for BCL11A (lead SNP rs6706648, β = -0.39, P = 4.96 × 10-34) and HBS1L-MYB (lead SNP rs61028892, β = 0.73, P = 1.18 × 10-9), whereas the variant allele for Xmn1-HBG2 was found to be very rare. In addition, we detected three putative new trait-associated regions. Genetically, dissecting the two major loci BCL11A and HBS1L-MYB, we defined trait-increasing haplotypes (P < 0.0001) containing so far unidentified causal variants. At BCL11A, in addition to a haplotype harbouring the putative functional variant rs1427407-'T', we identified a second haplotype, tagged by the rs7565301-'A' allele, where a yet-to-be-discovered causal DNA variant may reside. Similarly, at HBS1L-MYB, one HbF-increasing haplotype contains the likely functional small indel rs66650371, and a second tagged by rs61028892-'C' is likely to harbour a presently unknown functional allele. Together, variants at BCL11A and HBS1L-MYB SNPs explained 24.1% of the trait variance. Our findings provide a path for further investigation of the causes of variable fetal haemoglobin persistence in sickle cell disease.

2024-02-06·Plant disease

Rhizopus oryzae causes the leaf rot disease of Epimedium sagittatum in Guizhou, China.

Article

作者: Shihua Zhou ; Yanping Gao ; Jiuchun An ; Lanping Guo ; Weike Jiang ; Xiaohong Ou ; Lang Qin ; Tao Zhou ; Qing-Song Yuan

Epimedium sagittatum is a collective term for herbaceous plants belonging to the family Berberidaceae. Their dried leaves and stems have significant therapeutic effects on tumor inhibition, hypertension control, and coronary heart disease (Ke et al. 2023; Zhao et al. 2019). In 2021 and 2022, plants with similar leaf rot symptoms ranging from 30% to 55% was observed on E. sagittatum in Congjiang County, Guizhou province. The initial symptoms of the disease manifest locally on the leaf, with yellowing on the surface edge of the affected tissue, browning in the middle part, and brown-white discoloration in the innermost part (Supplementary Figure S1B). As the disease progresses, the entire infected leaf gradually softens, while the veins remain intact (Supplementary Figure S1C). Ultimately, the leaf withers and dehisces. The nine samples with typical symptoms were collected from Congjiang County, Guizhou province (26.598°N, 106.707°E). Twenty-seven fungi were isolated, including ten isolates of Rhizopus and seventeen isolates of seven other genera. On isolate YYH-CJ-17 many sporangia were formed and turned to a brown-gray to black color on potato dextrose agar medium (PDA) after culturing 5 days under dark at 25 ℃ (Supplementary Figure S2A and S2B). The branches of mycelium were finger-shaped or root-shaped. The sporangium was spherical or nearly spherical, 60-250 µm in diameter, and sporangiospores were elliptical or spherical and 4-8 µm in diameter. The obtained 547 bp ITS fragment (accession OR225970) and 1231 bp EF-1α region (accession OR242258) from isolate YYH-CJ-17 were compared with NR database using the BLAST tool provided by NCBI, which revealed more than 99.5% identity (query cover more than 98%) with the sequences of ITS (accessions MF522822.1) and EF-1α (accession AB281541.1) of Rhizopus oryzae Went & H.C. Prinsen Geerlings (Gao et al. 2022; Zhang et al. 2022). The phylogenetic tree constructed with the ITS and EF-1α gene sequences demonstrates that strain YYH-CJ-17 clusters with R. oryzae in the same branch and the bootstrap value was greater than 99% (Supplementary Figure S3). Based on the morphological characteristics and ITS and EF-1a sequences, the isolate YYH-CJ-17 is identified as R. oryzae. Pathogenicity tests were performed on detached healthy leaves and living plants of E. sagittatum. Healthy leaves of E. sagittatum were subjected to inoculation with isolate YYH-CJ-17 with 5 × 105 CFU mL-1 concentration in sterile culture dishes. The progression of the disease was marked by the gradual softening of the infected leaves and the expansion of the lesions, which ultimately produced black-brown sporangium (Supplementary Figure S4A). Furthermore, the E. sagittatum living plants were sprayed with 5 × 105 CFU mL-1 conidial suspension of isolate YYH-CJ-17, with ddH2O as a negative control, and then were cultivated at 25℃ and 90% humidity for 21 days in the greenhouse. This assay found that the E. sagittatum leaves treated with isolate YYH-CJ-17 exhibited the same symptoms observed on plants in fields (Supplementary Figure S4B). The fungus re-isolated from the inoculated leaves were identified as R. oryzae by ITS sequencing and were blasted with NR database, which highest matched with the sequence of ITS (accessions MF522822.1) mentioned above, thus fulfilling Koch's postulates. R. oryzae has been identified as a causative agent of a diverse array of host diseases, including leaf mildew of tobacco, fruit rot of yellow oleander and pears, and soft rot of bananas (Farooq et al. 2017; Khokhar et al. 2019; Kwon et al. 2012; Pan et al. 2021). To the best of our knowledge, this is the first report of leaf rot on E. sagittatum caused by R. oryzae in China, which will provide clear prevention and management target for the leaf rot disease of E. sagittatum.

项与 EF-1A 相关的新闻（医药）

2022-10-30

·BiG生物创新社

重大发现 | 高密度CAR或将显著限制CAR-T疗效！附临床分析

引言嵌合抗原受体（CAR）T细胞疗法已成为一种有前途的癌症治疗工具，彻底改变了癌症免疫疗法。尽管缓解率很高，但并非每个患者在CAR-T细胞治疗后都能达到完全缓解（CR）。此外，大量患者会经历复发，特别是MM患者[1]。是什么影响了CAR-T细胞的临床疗效呢？随着基因组学和基因调控网络的技术进步，如单细胞测序等，帮助我们在理解CAR-T细胞的基因组景观方面取得显著进展 [2]。近日，来自纳瓦拉大学的团队的研究表明CAR密度在CAR-T活性中起着重要作用，对临床反应有着显著影响。 [4]。作者 | 小鱼☆ 本文重点：1. 揭示了高、低表达CAR分子的CAR-T细胞之间的不同特征；2. 定义了与CAR密度相关的分子特征，用以预测临床反应；3. 高表达CAR的CAR-T细胞增加CAR信号，上调耗竭标志物，增加体外细胞毒性，减少体内BM浸润，限制CAR-T疗法的功效；4. 在不同的血液系统恶性肿瘤中，CARHighT水平升高的患者表现出明显较差的临床反应。☆ 在此之前，我们已知影响CAR-T细胞功效的因素包括：1. 外在因素，如抗原密度或肿瘤负荷信号；2. CAR结构相关的因素，如共刺激域或铰链长度；3. 内在细胞因子，如T细胞的分化状态，CD4 / CD8比率或T细胞多功能性；4. 具有耗竭表型的T细胞的增加可能导致CAR-T细胞的持久性降低，临床反应下降。☆ 本文在此基础上做了以下几项工作：1. 讨论CAR密度对B细胞成熟抗原（BCMA）CAR-T细胞功能的影响；2. 正式探讨CAR-T细胞膜中CAR分子的密度对临床疗效的影响。以下附上正文详解：01CAR-T细胞表面的CAR密度影响CAR介导的信号传导目前的CAR-T细胞产物是使用逆转录病毒/慢病毒（LV）载体产生的，这些载体在细胞内产生不同水平的转导和转基因表达，从而观察到转导细胞表面上的各种不同的CAR分子密度（图1）。图1：CAR-T细胞在膜表面呈现出不同的CAR密度为了确定CAR密度是否会影响CAR介导的信号传导，从而影响CAR-T细胞功效。研究人员首先在Jurkat细胞中使用三重参数报告器（TPR）系统，通过流式细胞术测量CAR介导的主要信号通路的激活。Jurkat-TPR细胞靶向BCMA， 4-1BB作为共刺激结构域，用报告基因（EGFRt）进一步修饰，便于测量CAR水平（图2）。图2：CAR构建然后，根据EGFRt的荧光强度(FI)区分不同水平CAR的Jurkat -TPR细胞亚群(称为CARHigh和CARLow细胞)(图3)。图3：CARHigh和CARLow细胞分群并与表达不同BCMA的MM细胞系共培养后进行分析，我们观察到CAR内三种途径的激活水平显著增加（图4）。图4：CAR密度影响CAR介导的信号02CAR密度影响靶向BCMA的CAR-T细胞的抗肿瘤反应为了进一步分析CAR密度对抗肿瘤功效的影响，表征了来自10个健康供体的CAR-T细胞，这些细胞采用BCMA靶向CAR构建体（来自ARI-0002h）共表达蓝色荧光蛋白（BFP）（图5）。图5：CAR构建基于BFP表达，将CAR-T细胞分选为CARHigh和CARLow细胞亚群（图6）。图6：分类示意图CARHigh T细胞：细胞毒性活性增加，干扰素-γ（IFN-γ）、白细胞介素-2（IL-2）、肿瘤坏死因子-α（TNFα）和颗粒酶B（GZMB）的产生水平更高（图7）。图7：CARHighT和CARLow T细胞毒性活性和上清液中IFN-γ水平接下来，研究人员分析了CAR的表型。观察CARHigh T和CARLow T细胞在用肿瘤细胞刺激之前和之后的表型变化，发现CARHighT细胞的基础激活增加，人白细胞抗原（HLA）-DR和CD137细胞的水平显著升高（图8E、F），以及表达两种或更多种耗竭标志物（LAG3，TIM3和/或PD1）组合的CD8+T细胞的百分比显著升高（图8G）。图8：CARHigh T细胞和CARLowT细胞耗竭表型比较用肿瘤细胞连续重复体外刺激21天，发现CARHigh T细胞和CARLow T细胞分化和耗竭表型差异显著，并且CARHighT细胞损失严重（图9）。图9：CARHighT细胞和CARLow T细胞数量比较总体而言，这些结果表明 CARHigh T细胞增加细胞毒性和CAR信号传导。此外，在CARHigh T细胞中观察到的分化和耗竭的表型更多，导致在刺激条件下体内抗肿瘤作用降低，这可能对临床反应产生影响。03CARHighT细胞显示出不同的转录组和染色质景观深入研究CAR-T细胞的两个亚群（CARHighT细胞和CARLow T细胞）的转录组和表观遗传景观，CARHigh T细胞显示出与淋巴细胞活化相关的基因（如HLA-DRA，CIITA和CD74）以及共刺激分子（如CTLA-4，TNFRSF4（OX40）和TNFRSF9（4-1BB）的表达增加（图10）。图10：CARHighT细胞和CARLow T细胞基因表达定量分析表现出差异和染色质可及性的基因主要与T细胞活化和共刺激有关（图11）。图11：UCSC基因组浏览器分析总之，这些结果表明，与CAR水平升高相关的基因表达和染色质可及性的微小差异可以显著改变CAR-T细胞的整体表型和功能特征。04单细胞测序揭示了CARHigh T细胞的CAR特异性分布为了更好地了解CAR-T细胞的异质性以及CAR水平对其转录组学的影响，研究人员对来自三个独立生产的43981个CAR-T细胞进行了单细胞转录组分析。在质量控制和过滤后，研究人员进行综合分析，并确定了23个CAR-T细胞亚群集群。根据表型结果，研究人员鉴定出一个预先耗竭的CD8 CAR-T细胞簇，其特征在于细胞毒性基因的表达以及抑制性受体，如LAG3和TIGIT（图12）。图12：在单细胞水平上表征CAR-T细胞05CAR密度与监管网络的差异激活有关为了阐明CAR的分子调控机制，研究人员应用了SimiC，这是一种用于scRNA-seq数据的新型GRN推理算法，在共同推断每个特定细胞状态的GRN时施加相似性约束。SimiC的GRN分析提供了表型和功能差异背后的监管网络（图13）。图13：CAR密度与监管网络的差异激活有关06CARHighT基因特征与临床反应相关研究人员推断CAR密度可能对CAR-T细胞疗法在临床反应中产生影响。由图观察到，CD4+和CD8+CARHighT细胞在来自临床反应不佳的患者（PR/NR）中均显示出更高的分数（图14）。图14：CD4+和CD8+CARHigh T细胞特征评分研究人员还评估了来自24名DLBCL患者的抗CD19 CAR-T输注产物的T细胞特征。发现，来自无反应患者（PR/NR）的输注产物显著富集CD8+CARHighT细胞，支持CAR密度与临床反应之间的相关性（图15）。图15：CD8+CARHighT细胞比例与临床反应与严格意义完全缓解（sCR）的患者相比，部分缓解（PR）的患者CARHigh T细胞数量增加（图16）。图16：CARHighT细胞比例与临床反应总体而言，我们的数据表明，在膜上具有高CAR密度的CAR-T治疗产品会对临床反应产生负面影响。总结与展望CAR-T细胞的功能依赖于肿瘤和工程T细胞之间的相互作用。然而，作为活的药物，CAR T细胞是异质产物，其中内在和外在因素可以影响其功能，从而对其临床疗效产生显著影响。本文确定了影响CAR-T细胞功能的新决定因素——CAR表达水平。高水平的CAR表达与增强的CAR信号和细胞耗竭表型有关，其特征在于多种抑制性受体的表达，如PD1，CTLA4，LAG3，TIM3和TIGIT等，本文研究也表明在不同的血液系统恶性肿瘤中，CARHigh T水平升高的患者临床反应下降。如何防止CAR增加？使用生理启动子：大多数CAR-T构建体是使用具有强启动子的逆转录/ LV载体生成的。这些启动子诱发高水平的CAR表达，可导致CAR信号传导和过早耗竭。通过较弱的启动子以及通过更多的生理启动子减少基因表达，从而减少CAR信号传导。MND启动子的使用可以降低CAR表面密度，而EF1a启动子增加了其密度，导致细胞毒性活性增强，细胞因子产生增强和耗竭标志物表达。因此，使用生理启动子将防止CARHighT细胞增加，从而减少了CAR-T细胞的衰竭，有利于维持长期持久的抗肿瘤功效。如何正确识别CAR？对CD4+和CD8+ CAR-T细胞的转录组学分析，研究人员定义了与CAR密度增加相关的分子特征，该特征与细胞表面CAR水平的相关性高于RNA水平。单细胞数据进行GRN分析，识别与CAR-T细胞耗尽相关的关键区域。总之， CAR密度在CAR-T活性中起着重要作用，对临床反应有影响。此外，在单细胞水平上理解CAR密度驱动的调节机制，可以用于改进CAR-T疗法。最后，CAR密度增加相关的基因特征有望预测患者对CAR-T细胞治疗的反应。参考文献1. Berdeja, Jesus G et al. “Ciltacabtagene autoleucel, a B-cell maturation antigen-directed chimeric antigen receptor T-cell therapy in patients with relapsed or refractory multiple myeloma (CARTITUDE-1): a phase 1b/2 open-label study.” Lancet (London, England) vol. 398,10297 (2021): 314-324. doi:10.1016/S0140-6736(21)00933-82. Chen, Gregory M et al. “Integrative Bulk and Single-Cell Profiling of Premanufacture T-cell Populations Reveals Factors Mediating Long-Term Persistence of CAR T-cell Therapy.” Cancer discovery vol. 11,9 (2021): 2186-2199. doi:10.1158/2159-8290.CD-20-16773. Deng, Qing et al. “Characteristics of anti-CD19 CAR T cell infusion products associated with efficacy and toxicity in patients with large B cell lymphomas.” Nature medicine vol. 26,12 (2020): 1878-1887. doi:10.1038/s41591-020-1061-74. Rodriguez-Marquez, Paula et al. “CAR density influences antitumoral efficacy of BCMA CAR T cells and correlates with clinical outcome.” Science advances vol. 8,39 (2022): eabo0514. doi:10.1126/sciadv.abo0514扫码加入BiG生物创新社读者交流群，分享、交流纯粹的行业知识，非诚勿扰！BiGScientific Driven, Making Impact!创新生态丨医药论坛丨行业分析媒体公关丨BiG Webinar联系我们商务：Max 18662346610媒体：Kathy 17621909690

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