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Olink Proteomics-Based Exploration of Immuno-Oncology-Related Biomarkers Leading to Lung Adenocarcinoma Progression.
IF 3.8 2区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-19 DOI: 10.1021/acs.jproteome.4c00377
Shiwen Yu, Liangwei Yang, Jianfeng Shu, Tian Zhao, Liyuan Han, Ting Cai, Guofang Zhao

Introduction: It is crucial to investigate the distinct proteins that contribute to the advancement of lung cancer.

Material and methods: We analyzed the expression levels of 92 immuno-oncology-related proteins in 96 pairs of lung adenocarcinoma tissue samples using Olink proteomics. The differentially expressed proteins (DEPs) were successively screened in tumor and paraneoplastic groups, early and intermediate-late groups by a nonparametric rank sum test, and the distribution and expression levels of DEPs were determined by volcano and heat maps, etc., and the area under the curve was calculated.

Results: A total of 24 DEPs were identified in comparisons between tumor and paracancerous tissues. Among them, interleukin-8 (IL8) and chemokine (C-C motif) ligand 20 (CCL20) as potential markers for distinguishing tumor tissues. Through further screening, it was found that interleukin-6 (IL6) and vascular endothelial growth factor A (VEGFA) may be able to lead to tumor progression through the JaK-STAT signaling pathway, Toll-like receptor signaling pathway and PI3K/AKT signaling pathway. Interestingly, our study revealed a down-regulation of IL6 and VEGFA in tumor tissues compared to paracancerous tissues.

Conclusions: IL8 + CCL20 (AUC: 0.7056) have the potential to differentiate tumor tissue from paracancerous tissue; IL6 + VEGFA (AUC: 0.7531) are important protein markers potentially responsible for tumor progression.

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引用次数: 0
Spatial Proteomic Profiling of Colorectal Cancer Revealed Its Tumor Microenvironment Heterogeneity. 结直肠癌的空间蛋白质组分析揭示了肿瘤微环境的异质性
IF 3.8 2区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-18 DOI: 10.1021/acs.jproteome.3c00719
Lifen Xie, Qian Kong, Meiling Ai, An He, Bin Yao, Luobin Zhang, Keren Zhang, Chaowei Zhu, Yangqiu Li, Ligang Xia, Ruijun Tian, Ruilian Xu

Colorectal cancer is a predominant malignancy with a second mortality worldwide. Despite its prevalence, therapeutic options remain constrained and surgical operation is still the most useful therapy. In this regard, a comprehensive spatially resolved quantitative proteome atlas was constructed to explore the functional proteomic landscape of colorectal cancer. This strategy integrates histopathological analysis, laser capture microdissection, and proteomics. Spatial proteome profiling of 200 tissue section samples facilitated by the fully integrated sample preparation technology SISPROT enabled the identification of more than 4000 proteins on the Orbitrap Exploris 240 from 2 mm2 × 10 μm tissue sections. Compared with normal adjacent tissues, we identified a spectrum of cancer-associated proteins and dysregulated pathways across various regions of colorectal cancer including ascending colon, transverse colon, descending colon, sigmoid colon, and rectum. Additionally, we conducted proteomic analysis on tumoral epithelial cells and paracancerous epithelium from early to advanced stages in hallmark rectum cancer and sigmoid colon cancer. Bioinformatics analysis revealed functional proteins and cell-type signatures associated with different regions of colorectal tumors, suggesting potential clinical implications. Overall, this study provides a comprehensive spatially resolved functional proteome landscape of colorectal cancer, serving as a valuable resource for exploring potential biomarkers and therapeutic targets.

结肠直肠癌是一种主要的恶性肿瘤,死亡率居全球第二位。尽管其发病率很高,但治疗方法仍然有限,外科手术仍然是最有效的治疗方法。为此,我们构建了一个全面的空间分辨定量蛋白质组图谱,以探索结直肠癌的功能蛋白质组图谱。这一策略整合了组织病理学分析、激光捕获显微切割和蛋白质组学。利用完全集成的样品制备技术 SISPROT 对 200 份组织切片样品进行了空间蛋白质组图谱分析,在 Orbitrap Exploris 240 上从 2 mm2 × 10 μm 组织切片中鉴定出了 4000 多种蛋白质。与邻近的正常组织相比,我们在结直肠癌的各个区域(包括升结肠、横结肠、降结肠、乙状结肠和直肠)鉴定出了一系列癌症相关蛋白和失调通路。此外,我们还对标志性直肠癌和乙状结肠癌从早期到晚期的肿瘤上皮细胞和癌旁上皮细胞进行了蛋白质组学分析。生物信息学分析揭示了与结直肠癌不同区域相关的功能蛋白和细胞类型特征,并提出了潜在的临床意义。总之,这项研究提供了全面的结直肠癌空间分辨功能蛋白质组图谱,是探索潜在生物标记物和治疗靶点的宝贵资源。
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引用次数: 0
Quantitative Proteomics Reveal the Mechanism of MiR-138-5p Suppressing Cervical Cancer via Targeting ZNF385A. 定量蛋白质组学揭示 MiR-138-5p 通过靶向 ZNF385A 抑制宫颈癌的机制
IF 3.8 2区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-18 DOI: 10.1021/acs.jproteome.4c00349
Qihang Peng, Yiting Deng, Guopan Li, Jingda Li, Peng Zheng, Qian Xiong, Jin Li, Ying Chen, Feng Ge

MicroRNAs are short, noncoding RNA molecules that exert pivotal roles in cancer development and progression by modulating various target genes. There is growing evidence that miR-138-5p is significantly involved in cervical cancer (CC). However, its precise molecular mechanism has yet to be fully understood. In the current investigation, a quantitative proteomics approach was utilized to detect possible miR-138-5p targets in HeLa cells systematically. In total, 364 proteins were downregulated, and 150 were upregulated after miR-138-5p overexpression. Bioinformatic analysis of these differentially expressed proteins (DEPs) revealed significant enrichment in several cancer-related pathways. Zinc finger protein 385A (ZNF385A) was determined as a novel direct target of miR-138-5p and discovered to facilitate the proliferation, migration, and cell cycle progression of HeLa cells. SFN and Fas cell surface death receptor(FAS) were then identified as functional downstream effectors of ZNF385A and miR-138-5p. Moreover, a tumor xenograft experiment was conducted to validate the association of miR-138-5p-ZNF385A-SFN/FAS axis with the development of CC in vivo. Our findings have collectively established a catalog of proteins mediated by miR-138-5p and have provided an in-depth comprehension of the molecular mechanisms responsible for the inhibitory effect of miR-138-5p on CC. The miR-138-5p-ZNF385A-SFN/FAS axis could also be beneficial to the identification of new therapeutic targets.

微RNA是一种短小的非编码RNA分子,通过调节各种靶基因在癌症的发生和发展过程中发挥关键作用。越来越多的证据表明,miR-138-5p 与宫颈癌(CC)密切相关。然而,其确切的分子机制尚未完全明了。本研究利用定量蛋白质组学方法系统检测了 HeLa 细胞中 miR-138-5p 的可能靶标。在 miR-138-5p 过表达后,共有 364 个蛋白质下调,150 个蛋白质上调。对这些差异表达蛋白(DEPs)进行的生物信息学分析表明,它们在几种癌症相关通路中都有显著的富集。锌指蛋白 385A(ZNF385A)被确定为 miR-138-5p 的一个新的直接靶标,并被发现能促进 HeLa 细胞的增殖、迁移和细胞周期进展。随后,SFN 和 Fas 细胞表面死亡受体(FAS)被确定为 ZNF385A 和 miR-138-5p 的功能性下游效应物。此外,我们还进行了肿瘤异种移植实验,以验证 miR-138-5p-ZNF385A-SFN/FAS 轴与 CC 在体内发展的关系。我们的研究结果共同建立了一个由miR-138-5p介导的蛋白质目录,并深入理解了miR-138-5p对CC产生抑制作用的分子机制。miR-138-5p-ZNF385A-SFN/FAS轴还有助于确定新的治疗靶点。
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引用次数: 0
Chronic Exposure to Petroleum-Derived Hydrocarbons Alters Human Skin Microbiome and Metabolome Profiles: A Pilot Study. 慢性接触石油衍生碳氢化合物会改变人类皮肤微生物组和代谢组图谱:一项试点研究。
IF 3.8 2区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-18 DOI: 10.1021/acs.jproteome.4c00256
Alan G Hernández-Melgar, Abraham Guerrero, Aldo Moreno-Ulloa

Petroleum-derived substances, like industrial oils and grease, are ubiquitous in our daily lives. Comprised of petroleum hydrocarbons (PH), these substances can come into contact with our skin, potentially causing molecular disruptions and contributing to the development of chronic disease. In this pilot study, we employed mass spectrometry-based untargeted metabolomics and 16S rRNA gene sequencing analyses to explore these effects. Superficial skin samples were collected from subjects with and without chronic dermal exposure to PH at two anatomical sites: the fingers (referred to as the hand) and arms (serving as an intersubject variability control). Exposed hands exhibited higher bacterial diversity (Shannon and Simpson indices) and an enrichment of oil-degrading bacteria (ODB), including Dietzia, Paracoccus, and Kocuria. Functional prediction suggested enriched pathways associated with PH degradation in exposed hands vs non-exposed hands, while no differences were observed when comparing the arms. Furthermore, carboxylic acids, glycerophospholipids, organooxygen compounds, phenol ethers, among others, were found to be more abundant in exposed hands. We observed positive correlations among multiple ODB and xenobiotics, suggesting a chemical remodeling of the skin favorable for ODB thriving. Overall, our study offers insights into the complex dysregulation of bacterial communities and the chemical milieu induced by chronic dermal exposure to PH.

石油衍生物质,如工业油和油脂,在我们的日常生活中无处不在。这些物质由石油碳氢化合物(PH)组成,会与我们的皮肤接触,可能会造成分子干扰,导致慢性疾病的发生。在这项试验性研究中,我们采用了基于质谱的非靶向代谢组学和 16S rRNA 基因测序分析来探讨这些影响。我们在两个解剖部位采集了长期皮肤暴露于 PH 的受试者和未暴露于 PH 的受试者的表皮样本:手指(称为手部)和手臂(作为受试者间变异对照)。暴露的手部表现出更高的细菌多样性(香农指数和辛普森指数)和丰富的油脂降解细菌(ODB),包括 Dietzia、Paracoccus 和 Kocuria。功能预测表明,暴露的手与未暴露的手相比,与 PH 降解相关的途径更丰富,而比较两只手则未观察到差异。此外,我们还发现羧酸、甘油磷脂、有机氧化合物、苯酚醚等物质在暴露的双手中含量更高。我们观察到多种 ODB 与异种生物之间存在正相关,这表明皮肤的化学重塑有利于 ODB 的茁壮成长。总之,我们的研究为了解长期皮肤暴露于 PH 引起的细菌群落和化学环境的复杂失调提供了见解。
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引用次数: 0
Taurine Alleviates Ferroptosis-Induced Metabolic Impairments in C2C12 Myoblasts by Stabilizing the Labile Iron Pool and Improving Redox Homeostasis. 牛磺酸通过稳定易失铁池和改善氧化还原稳态缓解铁中毒诱导的 C2C12 肌细胞代谢损伤
IF 3.8 2区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-18 DOI: 10.1021/acs.jproteome.4c00123
Xi Liu, Yu Zhou, Zhen Qi, Caihua Huang, Donghai Lin

Ferroptosis adversely affects the viability, differentiation, and metabolic integrity of C2C12 myoblasts, contributing to the decline in skeletal muscle health. The intricate mechanisms behind this process are not fully understood. In this study, we induced ferroptosis in myoblasts using targeted inducers and found a marked decrease in specific redox metabolites, particularly taurine. Taurine supplementation effectively reversed the deleterious effects of ferroptosis, significantly increased cellular glutathione levels, reduced MDA and ROS levels, and rejuvenated impaired myogenic differentiation. Furthermore, taurine downregulated HO-1 expression and decreased intracellular Fe2+ levels, thereby stabilizing the labile iron pool. Using NMR metabolomic analysis, we observed that taurine profoundly promoted glycerophospholipid metabolism, which is critical for cell membrane repair, and enhanced mitochondrial bioenergetics, thereby increasing the energy reserves essential for muscle satellite cell regeneration. These results suggest that taurine is a potent ferroptosis inhibitor that attenuates key drivers of this process, strengthens oxidative defenses, and improves redox homeostasis. This combined effect protects cells from ferroptosis-induced damage. This study highlights the potential of taurine as a valuable ferroptosis inhibitor that protects skeletal muscle from ferroptosis-induced damage and provides a basis for therapeutic strategies to rejuvenate and facilitate the regeneration of aging skeletal muscle.

铁中毒对 C2C12 肌母细胞的活力、分化和新陈代谢的完整性产生不利影响,导致骨骼肌健康状况下降。这一过程背后的复杂机制尚不完全清楚。在本研究中,我们使用靶向诱导剂诱导了肌母细胞的铁凋亡,并发现特定氧化还原代谢物(尤其是牛磺酸)明显减少。补充牛磺酸能有效逆转铁变态反应的有害影响,显著提高细胞谷胱甘肽水平,降低 MDA 和 ROS 水平,使受损的成肌细胞分化恢复活力。此外,牛磺酸还能下调HO-1的表达,降低细胞内Fe2+的水平,从而稳定易溶铁池。通过核磁共振代谢组学分析,我们观察到牛磺酸能显著促进对细胞膜修复至关重要的甘油磷脂代谢,并增强线粒体生物能,从而增加肌肉卫星细胞再生所必需的能量储备。这些结果表明,牛磺酸是一种强效的铁跃迁抑制剂,可减轻铁跃迁过程的关键驱动因素,增强氧化防御能力,改善氧化还原平衡。这种综合效应可保护细胞免受铁氧化诱导的损伤。这项研究强调了牛磺酸作为一种有价值的铁突变抑制剂的潜力,它能保护骨骼肌免受铁突变诱导的损伤,并为恢复衰老骨骼肌的活力和促进其再生的治疗策略提供了基础。
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引用次数: 0
Full Native timsTOF PASEF-Enabled Quantitative Proteomics with the i2MassChroQ Software Package. 采用 i2MassChroQ 软件包的全原生 timsTOF PASEF 定量蛋白质组学。
IF 3.8 2区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-17 DOI: 10.1021/acs.jproteome.3c00732
Olivier Langella, Thomas Renne, Thierry Balliau, Marlène Davanture, Sven Brehmer, Michel Zivy, Mélisande Blein-Nicolas, Filippo Rusconi

Ion mobility mass spectrometry has become popular in proteomics lately, in particular because the Bruker timsTOF instruments have found significant adoption in proteomics facilities. The Bruker's implementation of the ion mobility dimension generates massive amounts of mass spectrometric data that require carefully designed software both to extract meaningful information and to perform processing tasks at reasonable speed. In a historical move, the Bruker company decided to harness the skills of the scientific software development community by releasing to the public the timsTOF data file format specification. As a proteomics facility that has been developing Free Open Source Software (FOSS) solutions since decades, we took advantage of this opportunity to implement the very first FOSS proteomics complete solution to natively read the timsTOF data, low-level process them, and explore them in an integrated quantitative proteomics software environment. We dubbed our software i2MassChroQ because it implements a (peptide)identification-(protein)inference-mass-chromatogram-quantification processing workflow. The software benchmarking results reported in this paper show that i2MassChroQ performed better than competing software on two critical characteristics: (1) feature extraction capability and (2) protein quantitative dynamic range. Altogether, i2MassChroQ yielded better quantified protein numbers, both in a technical replicate MS runs setting and in a differential protein abundance analysis setting.

离子迁移质谱法最近在蛋白质组学领域大受欢迎,特别是因为布鲁克公司的 timsTOF 仪器已在蛋白质组学设备中得到广泛应用。布鲁克公司实施的离子迁移量产生了大量的质谱数据,需要精心设计的软件来提取有意义的信息,并以合理的速度执行处理任务。作为一项具有历史意义的举措,布鲁克公司决定利用科学软件开发社区的技能,向公众发布 timsTOF 数据文件格式规范。作为一家几十年来一直致力于开发自由开放源码软件 (FOSS) 解决方案的蛋白质组学机构,我们利用这个机会实施了首个 FOSS 蛋白质组学完整解决方案,在一个集成的定量蛋白质组学软件环境中读取 timsTOF 数据、对其进行底层处理和探索。我们将我们的软件命名为 i2MassChroQ,因为它实现了(肽)识别-(蛋白质)推断-质谱-色谱-定量处理的工作流程。本文中报告的软件基准测试结果表明,i2MassChroQ 在两个关键特性上优于同类软件:(1) 特征提取能力和 (2) 蛋白质定量动态范围。总之,i2MassChroQ 在技术重复质谱运行设置和差异蛋白质丰度分析设置中都能获得更好的量化蛋白质数量。
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引用次数: 0
Extracellular Matrix Protein Signatures of the Outer and Inner Zones of the Rat Adrenal Cortex. 大鼠肾上腺皮质外区和内区的细胞外基质蛋白特征
IF 3.8 2区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-17 DOI: 10.1021/acs.jproteome.4c00071
Jean Lucas Kremer, Veronica Feijoli Santiago, Fernanda Bongiovani Rodrigues, Thais Barabba Auricino, Danilo Henriques de Oliveira Freitas, Giuseppe Palmisano, Claudimara Ferini Pacicco Lotfi

This study analyzes the extracellular matrix (ECM) signatures of the outer (OF = capsule + subcapsular + zona glomerulosa cells) and inner fractions (IF = zona fasciculata cells) of the rat adrenal cortex, which comprise two distinct microenvironment niches. Proteomic profiles of decellularized OF and IF samples, male and female rats, identified 252 proteins, with 32 classified as ECM-component and ECM-related. Among these, 25 proteins were differentially regulated: 17 more abundant in OF, including Col1a1, Col1a2, Col6a1, Col6a2, Col6a3, Col12a1, Col14a1, Lama5, Lamb2, Lamc1, Eln, Emilin, Fbln5, Fbn1, Fbn2, Nid1, and Ltbp4, and eight more abundant in IF, including Col4a1, Col4a2, Lama2, Lama4, Lamb1, Fn1, Hspg2, and Ecm1. Eln, Tnc, and Nid2 were abundant in the female OF, while Lama2, Lama5, Lamb2, and Lamc1 were more abundant in the male IF. The complex protein signature of the OF suggests areas of tissue stress, stiffness, and regulatory proteins for growth factor signaling. The higher concentrations of Col4a1 and Col4a2 and their role in steroidogenesis should be further investigated in IF. These findings could significantly enhance our understanding of adrenal cortex functionality and its implications for human health and disease. Key findings were validated, and data are available in ProteomeXchange (PXD046828).

本研究分析了大鼠肾上腺皮质外层(OF = 囊 + 囊下 + 肾小球细胞)和内层(IF = 筋膜细胞)的细胞外基质(ECM)特征,这两个部分构成了两个不同的微环境龛位。对雄性和雌性大鼠的脱细胞 OF 和 IF 样品进行蛋白质组学分析,发现了 252 种蛋白质,其中 32 种归类为 ECM 成分和 ECM 相关蛋白。在这些蛋白质中,有 25 种蛋白质受到不同程度的调控:17 个蛋白质在 OF 中含量较高,包括 Col1a1、Col1a2、Col6a1、Col6a2、Col6a3、Col12a1、Col14a1、Lama5、Lamb2、Lamc1、Eln、Emilin、Fbln5、Fbn1、Fbn2、Nid1 和 Ltbp4,以及在 IF 中含量较高的 8 个物种,包括 Col4a1、Col4a2、Lama2、Lama4、Lamb1、Fn1、Hspg2 和 Ecm1。Eln、Tnc和Nid2在雌性OF中含量较高,而Lama2、Lama5、Lamb2和Lamc1在雄性IF中含量较高。OF 蛋白质特征复杂,表明存在组织应力、僵硬和生长因子信号转导调节蛋白的区域。Col4a1和Col4a2的浓度较高,它们在类固醇生成中的作用应在IF中进一步研究。这些发现将大大提高我们对肾上腺皮质功能及其对人类健康和疾病影响的认识。主要发现已得到验证,数据可在 ProteomeXchange (PXD046828) 上查阅。
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引用次数: 0
Integrated MICROFASP Method with CZE-Based Fractionation Technique and NanoRPLC-ESI-MS/MS for a Comprehensive Proteomics Analysis of a Submicrogram Sample. 将 MICROFASP 方法与基于 CZE 的分馏技术和 NanoRPLC-ESI-MS/MS 集成在一起,对亚微克样品进行全面的蛋白质组学分析。
IF 3.8 2区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-16 DOI: 10.1021/acs.jproteome.4c00396
Gang Lu, Guojin Ying, Yu He, Yang Li, Zhenbin Zhang

We report a loss-less two-dimensional (2D) separation platform that integrated capillary zone electrophoresis (CZE) fractionation and nanoRPLC-ESI-MS/MS for a comprehensive proteomics analysis of a submicrogram sample. Protein digest was injected into the linear polyacrylamide-coated capillary, followed by CZE separation. The schemes for collecting the fractions were carefully optimized to maximize the protein coverage. The peptide fractions were directly eluted into the autosampler insert vials, followed by the nanoRPLC-ESI-MS/MS analysis without lyophilization and redissolution, thus dramatically minimizing sample loss and potential contamination. The integrated platform generated 30,845 unique peptides and 5231 protein groups from 500 ng of a HeLa protein digest within 11.5 h (90 min CZE fractionation plus 10 h LC-MS analysis). Finally, the developed platform was used to analyze the protein digest prepared by the MICROFASP method with 1 μg of cell lysate as the starting material. Three thousand seven hundred ninety-six (N = 2, RSD = 4.95%) protein groups and 20,577 (N = 2, RSD = 7.89%) peptides were identified from only 200 ng of the resulted tryptic digest within 5.5 h. The results indicated that the combination of the MICROFASP method and the developed CZE/nanoRPLC-MS/MS 2D separation platform enabled comprehensive proteome profiling of a submicrogram biological sample. Data are available via ProteomeXchange with the identifier PXD052735.

我们报告了一种无损耗二维(2D)分离平台,该平台集成了毛细管区带电泳(CZE)分馏和 nanoRPLC-ESI-MS/MS 技术,可对亚微克样品进行全面的蛋白质组学分析。将蛋白质消化液注入线性聚丙烯酰胺涂层毛细管,然后进行 CZE 分离。收集馏分的方案经过精心优化,以最大限度地提高蛋白质的覆盖率。肽馏分直接洗脱到自动进样器插入瓶中,然后进行 nanoRPLC-ESI-MS/MS 分析,无需冻干和再溶解,从而大大减少了样品损失和潜在污染。该集成平台在 11.5 小时内(90 分钟 CZE 分馏加 10 小时 LC-MS 分析)从 500 纳克 HeLa 蛋白消化液中生成了 30,845 个独特肽段和 5231 个蛋白质基团。最后,利用所开发的平台分析了以 1 μg 细胞裂解液为起始材料、采用 MICROFASP 方法制备的蛋白质消化液。结果表明,结合 MICROFASP 方法和所开发的 CZE/nanoRPLC-MS/MS 二维分离平台,可以对亚微克生物样本进行全面的蛋白质组分析。数据可通过 ProteomeXchange 获取,标识符为 PXD052735。
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引用次数: 0
Immunoaffinity Intact-Mass Spectrometry for the Detection of Endogenous Concentrations of the Acetylated Protein Tumor Biomarker Neuron Specific Enolase. 用于检测乙酰化蛋白质肿瘤生物标志物神经元特异性烯醇化酶内源性浓度的免疫亲和性完整质谱法
IF 3.8 2区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-16 DOI: 10.1021/acs.jproteome.4c00391
Sebastian A H van den Wildenberg, Sylvia A A M Genet, Maarten A C Broeren, Joost L J van Dongen, Luc Brunsveld, Volkher Scharnhorst, Daan van de Kerkhof

Intact-mass spectrometry has huge potential for clinical application, as it enables both quantitative and qualitative analysis of intact proteins and possibly unlocks additional pathophysiological information via, e.g., detection of specific post-translational modifications (PTMs). Such valuable and clinically useful selectivity is typically lost during conventional bottom-up mass spectrometry. We demonstrate an innovative immunoprecipitation protein enrichment assay coupled to ultrahigh performance liquid chromatography quadrupole time-of-flight high resolution mass spectrometry (UPLC-QToF-HRMS) for the fast and simple identification of the protein tumor marker Neuron Specific Enolase Gamma (NSEγ) at low endogenous concentrations in human serum. Additionally, using the combination of immunoaffinity purification with intact mass spectrometry, the presence of NSEγ in an acetylated form in human serum was detected. This highlights the unique potential of immunoaffinity intact mass spectrometry in clinical diagnostics.

完整质谱具有巨大的临床应用潜力,因为它可以对完整蛋白质进行定量和定性分析,并可能通过检测特定的翻译后修饰(PTMs)等方式获取更多的病理生理信息。传统的自下而上质谱法通常会丧失这种宝贵的临床实用选择性。我们展示了一种创新的免疫沉淀蛋白质富集测定与超高效液相色谱四极杆飞行时间高分辨质谱(UPLC-QToF-HRMS)相结合的方法,用于快速、简单地鉴定人血清中低浓度内源性蛋白质肿瘤标志物神经元特异性酵解酶γ(NSEγ)。此外,利用免疫亲和纯化与完整质谱的结合,还检测到人血清中存在乙酰化形式的 NSEγ。这凸显了免疫亲和完整质谱法在临床诊断中的独特潜力。
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引用次数: 0
Metabolic and Microbial Dysregulation in Preterm Infants with Neonatal Respiratory Distress Syndrome: An Early Developmental Perspective. 患有新生儿呼吸窘迫综合征的早产儿的代谢和微生物失调:早期发育视角。
IF 3.8 2区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-16 DOI: 10.1021/acs.jproteome.4c00114
Yanxu Bi, Wenyi Yu, Wenjie Bian, Mengtong Jin, Yukun He, Jinglei Wang, Xiaofeng Miao, Tiantian Guo, Xiaojun Ma, Pihua Gong, Ran Li, Jiangli Xi, Shuming Guo, Zhancheng Gao

Neonatal respiratory distress syndrome (NRDS) is one of the most severe respiratory disorders in preterm infants (PTIs) due to immature lung development. To delineate the serum metabolic alterations and gut microbiota variations in NRDS and assess their implications on neonatal development, we enrolled 13 NRDS neonates and 12 PTIs and collected fecal and serum specimens after birth. Longitudinal fecal sampling was conducted weekly for a month in NRDS neonates. NRDS neonates were characterized by notably reduced gestational ages and birth weights and a higher rate of asphyxia at birth relative to PTIs. Early postnatal disturbances in tryptophan metabolism were evident in the NRDS group, concomitant with elevated relative abundance of Haemophilus, Fusicatenibacter, and Vibrio. Integrative multiomics analyses revealed an inverse relationship between tryptophan concentrations and Blautia abundance. At one-week old, NRDS neonates exhibited cortisol regulation anomalies and augmented hepatic catabolism. Sequential microbial profiling revealed distinct gut microbiota evolution in NRDS subjects, characterized by a general reduction in potentially pathogenic bacteria. The acute perinatal stress of NRDS leads to mitochondrial compromise, hormonal imbalance, and delayed gut microbiota evolution. Despite the short duration of NRDS, its impact on neonatal development is significant and requires extended attention.

新生儿呼吸窘迫综合征(NRDS)是早产儿(PTIs)中最严重的呼吸系统疾病之一,原因是肺部发育不成熟。为了明确 NRDS 中的血清代谢改变和肠道微生物群变化并评估其对新生儿发育的影响,我们招募了 13 名 NRDS 新生儿和 12 名 PTI,并在出生后收集了粪便和血清标本。在为期一个月的时间里,我们每周对 NRDS 新生儿进行一次纵向粪便采样。与 PTI 相比,NRDS 新生儿的胎龄和出生体重明显降低,出生时窒息率较高。NRDS 组新生儿出生后早期色氨酸代谢紊乱明显,同时嗜血杆菌、镰刀菌和弧菌的相对丰度升高。多组学综合分析表明,色氨酸浓度与布劳氏菌丰度之间存在反比关系。一周大时,NRDS新生儿表现出皮质醇调节异常和肝脏分解代谢增强。连续微生物图谱分析显示,NRDS受试者的肠道微生物群发生了独特的演变,其特点是潜在致病菌普遍减少。NRDS 的围产期急性应激导致线粒体受损、激素失衡和肠道微生物群进化延迟。尽管 NRDS 的持续时间很短,但它对新生儿发育的影响却很大,需要引起更多关注。
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引用次数: 0
期刊
Journal of Proteome Research
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