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Hepatitis B small surface protein hijacking Bip is initial and essential to promote lipid synthesis. 乙型肝炎小体表面蛋白劫持 Bip 是促进脂质合成的初始和必要条件。
IF 2.8 2区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-11-21 DOI: 10.1016/j.jprot.2024.105358
Tao Zuo, Sha Jing, Peiru Chen, Tao Zhang, Yihao Wang, Yanchang Li, Lei Chang, Xingyu Rong, Na Li, Zhenwen Zhao, Chao Zhao, Ping Xu

To date, the molecular pathogenic mechanisms between HBsAg and liver metabolic disorders have not been fully understood. To explore the overall effects of HBsAg on liver tissues from HBV transgenic mice, proteome, interactome, and signal pathway analysis were employed to uncover the underlying mechanisms. Bioinformatics analysis of 191 differentially expressed proteins suggested that HBV upregulated the expression of multiple enzymes involved in lipid synthesis, and small HBs (SHBs) caused lipid accumulation in cells. Further studies showed that SHBs bound to binding immunoglobulin protein (Bip), which normally functions in cell homeostasis against the unfolded protein response (UPR) signaling via occupying inositol-requiring enzyme 1 (IRE1). Hijacking Bip by SHBs alleviated the inhibition of post-endoplasmic reticulum (ER) signaling and sequential activation of the IRE1 downstream transcription factors involved in lipid synthesis, such as spliced X-box binding protein 1 (sXBP1) and sterol regulatory element-binding protein 1 (SREBP1), leading to lipid metabolism disorder. The restoration of Bip can alleviate ER stress, and block the sequential post-ER signaling caused by SHBs. This study revealed a new pathway through which SHBs promote lipid disorder, and suggests that Bip may serve as a novel target for intervention in HBV related liver diseases. SIGNIFICANCE: In this study, we found a new pathway promoting the lipid disorder by SHBs through quantitative proteomics studies, and Bip may serve as a novel target for intervention in HBV related liver diseases. These findings highlight a novel role of SHBs in regulating cell lipid metabolism and provide an insight into the relationship between HBV infection and liver fatty disorders, which may serve as a potential therapeutic target for intervention of HBV related liver diseases.

迄今为止,HBsAg 与肝脏代谢紊乱之间的分子致病机制尚未完全明了。为了探索 HBsAg 对 HBV 转基因小鼠肝脏组织的整体影响,我们采用了蛋白质组、相互作用组和信号通路分析来揭示其潜在机制。对 191 个差异表达蛋白的生物信息学分析表明,HBV 上调了多种参与脂质合成的酶的表达,小 HBs(SHBs)导致细胞内脂质积累。进一步的研究表明,SHBs 与结合免疫球蛋白(Bip)结合,Bip 通常通过占据肌醇需要酶 1(IRE1),在细胞平衡中发挥对抗未折叠蛋白反应(UPR)信号的功能。SHBs 劫持 Bip 可减轻内质网(ER)后信号传导的抑制以及 IRE1 下游转录因子(如剪接 X-box 结合蛋白 1(sXBP1)和固醇调节元件结合蛋白 1(SREBP1))参与脂质合成的顺序激活,从而导致脂质代谢紊乱。Bip的恢复可以缓解ER应激,阻断SHBs引起的ER后信号传导。这项研究揭示了 SHBs 促进脂质代谢紊乱的新途径,并提示 Bip 可作为干预 HBV 相关肝病的新靶点。意义:在这项研究中,我们通过定量蛋白质组学研究发现了SHBs促进脂质紊乱的新途径,Bip可能成为干预HBV相关肝病的新靶点。这些发现凸显了 SHBs 在调控细胞脂质代谢中的新作用,并为 HBV 感染与肝脏脂肪紊乱之间的关系提供了新的视角,可能成为干预 HBV 相关肝病的潜在治疗靶点。
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引用次数: 0
Validation of urine p-cresol glucuronide as renal cell carcinoma non-invasive biomarker 尿液中对甲酚葡萄糖醛酸苷作为肾细胞癌非侵入性生物标记物的验证。
IF 2.8 2区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-11-17 DOI: 10.1016/j.jprot.2024.105357
Julia Oto , Raquel Herranz , Patricia Verger , Marta Roca , Emma Plana , Manuel Miralles , Manuel Martínez-Sarmiento , César D. Vera-Donoso , Pilar Medina
Renal cell carcinoma (RCC) stands among the most lethal urological malignancies. Most RCCs are incidentally diagnosed as initial symptoms are unspecific. Novel, minimally-invasive diagnostic and prognostic methods for RCC are needed, ideally in urine.
Using UPLC-Q-ToF MS untargeted metabolomic analysis in urine, we previously revealed p-cresol glucuronide as potential RCC diagnostic marker. Additionally, urine samples one-year post-nephrectomy revealed isobutyryl-l-carnitine and L-proline betaine as potential RCC prognostic markers. Our present aim was to validate these differences in an independent cohort of RCC patients and healthy controls to strengthen their value as non-invasive biomarkers.
In an independent cohort of 69 RCC patients and 52 controls we validated an increase in p-cresol glucuronide in urine from patients at diagnosis compared to controls (P = 0.0043). It remained increased one-year post-nephrectomy (P = 0.0288). The value of p-cresol glucuronide for RCC diagnosis was assessed with ROC curves analysis (AUC = 0.66, 95 % Confidence Interval 0.56–0.76). The role of isobutyryl-l-carnitine and L-proline betaine as prognostic markers could not be validated and will require a larger cohort.
Our findings confirm the value of p-cresol glucuronide in urine as diagnostic marker for RCC in an independent cohort. This non-invasive method holds promise for enhancing patient care by reducing the need for potentially risky diagnostic procedures. Further metaproteomics-oriented approaches towards the tyrosine oxidation pathway and microbiota metagenomics studies may promote a holistic management of RCC.

Significance

Current imaging techniques available to diagnose and monitor renal cell carcinoma (RCC) are harmful for the patient given the high-radiation dose, and unspecific in low-grade tumors. Thus, novel non-invasive biomarkers with diagnostic and prognostic capabilities are of utmost importance. Herein, we have validated urine p-cresol glucuronide as diagnostic marker for RCC. This novel non-invasive biomarker could improve accurate assessments of tumor behavior, while enhancing patient outcomes by reducing discomfort and detrimental side effects.
肾细胞癌(RCC)是最致命的泌尿系统恶性肿瘤之一。大多数 RCC 都是偶然诊断出来的,因为最初的症状没有特异性。我们需要新型、微创的 RCC 诊断和预后方法,最好是在尿液中进行诊断。利用 UPLC-Q-ToF MS 对尿液进行非靶向代谢组分析,我们发现对甲酚葡萄糖醛酸苷是潜在的 RCC 诊断标志物。此外,肾切除术后一年的尿液样本显示异丁酰-l-肉碱和 L-脯氨酸甜菜碱是潜在的 RCC 预后标志物。我们目前的目的是在一组独立的 RCC 患者和健康对照组中验证这些差异,以加强它们作为非侵入性生物标志物的价值。在一个由 69 名 RCC 患者和 52 名对照组组成的独立队列中,我们验证了与对照组相比,诊断时患者尿液中对甲酚葡萄糖醛酸的含量有所增加(P = 0.0043)。肾切除术后一年,该数值仍在增加(P = 0.0288)。通过 ROC 曲线分析评估了对甲酚葡萄糖醛酸对诊断 RCC 的价值(AUC = 0.66,95 % 置信区间为 0.56-0.76)。异丁酰基-l-肉碱和L-脯氨酸甜菜碱作为预后标志物的作用尚未得到验证,需要更大规模的队列研究。我们的研究结果证实了尿液中的对甲酚葡萄糖醛酸苷作为RCC诊断标志物在一个独立队列中的价值。这种非侵入性方法减少了对潜在风险诊断程序的需求,有望加强对患者的护理。针对酪氨酸氧化途径的进一步元蛋白质组学方法和微生物群元组学研究可促进对 RCC 的整体管理。意义:目前用于诊断和监测肾细胞癌(RCC)的成像技术对患者有害,因为辐射剂量大,而且对低级别肿瘤没有特异性。因此,具有诊断和预后能力的新型非侵入性生物标志物至关重要。在此,我们验证了尿液对甲酚葡萄糖醛酸苷可作为 RCC 的诊断标志物。这种新型非侵入性生物标记物可以提高对肿瘤行为的准确评估,同时通过减少不适感和有害副作用来改善患者的预后。
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引用次数: 0
An integrated proteomic and phosphoproteomic landscape of chronic kidney disease 慢性肾脏病的综合蛋白质组和磷酸蛋白质组图谱。
IF 2.8 2区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-11-14 DOI: 10.1016/j.jprot.2024.105355
Linxiao Sun , Cheng Wang , Zhongjing Zhou , Qiangqiang Li
The prevalence of chronic kidney disease (CKD) is gradually rising worldwide. Patients often remain asymptomatic for an extended period, leaving them unaware of their condition, which can lead to progressing to end-stage renal disease and cause significant economic burden. Improved understanding of CKD pathogenesis can enhance early detection and facilitate advances in drug development. Here, we performed proteomic and phosphoproteomic analyses of the mouse unilateral ureteral obstruction model to explore the molecular mechanisms of chronic kidney injury. 474 significantly differentially expressed proteins and 96 significantly differentially expressed phosphoproteins were screened, respectively. Chronic kidney injury involves complex metabolic pathways such as citrate cycle and hematopoietic system in proteome, and mitochondrial oxidative phosphorylation suppression is a notable alteration. The phosphoproteomic analysis revealed a significant upregulation in epithelial mesenchymal transition and P53 pathways, with a corresponding increase in the phosphorylation of Jun at serine 73. Utilizing HK2 cells, we observed that the reduction oxidative phosphorylation was consistently associated with an augmentation in oxidative stress, which subsequently activated Jun and induced apoptosis. Proteins that act as hubs in these pathways may be candidate targets for CKD intervention. These findings contribute significantly to the current understanding of CKD and provide valuable insights for future studies.

Significance

Chronic kidney disease (CKD) incidence rising annually with varied etiologies, kidney often irreversibly fibrotic, the treatment options are limited and often ineffective due to deficient understanding of renal fibrosis mechanisms. Despite the extensive efforts and numerous omics studies conducted on renal fibrosis, to date, no study has been undertaken to investigate the role of phosphorylated proteins in UUO models. Previously, we performed a comprehensive transcriptome and proteome analysis based on the CKD model, but the potential alterations in the phosphoproteome were not addressed. Here, an integrated proteomic and phosphoproteomic landscape of CKD was completed, which was the the first phosphoproteomic profiles of UUO model. Phosphoproteomic profile suggests that the epithelial mesenchymal transition and P53 pathways is significantly activated in mouse models of kidney injury, and the core protein Jun played a key role in CKD. And a preliminary correlation between P-Jun and oxidative phosphorylation was found base on HK2 cells. Our work contributes to a deeper understanding of the disease characteristics and molecular mechanisms of CKD. Identifying potential CKD targets from proteome and phosphoproteome may provide valuable insights for early diagnosis and treatment of CKD.
慢性肾脏病(CKD)的发病率在全球范围内逐渐上升。患者往往长期无症状,对自己的病情毫无察觉,这可能导致病情恶化为终末期肾病,并造成巨大的经济负担。加深对慢性肾功能衰竭发病机制的了解可以提高早期发现率,促进药物开发。在此,我们对小鼠单侧输尿管梗阻模型进行了蛋白质组学和磷酸化蛋白质组学分析,以探索慢性肾损伤的分子机制。分别筛选出了 474 个明显差异表达的蛋白质和 96 个明显差异表达的磷酸蛋白。慢性肾损伤在蛋白质组中涉及复杂的代谢途径,如柠檬酸循环和造血系统,线粒体氧化磷酸化抑制是一个明显的改变。磷酸化蛋白质组分析显示,上皮间质转化和 P53 通路显著上调,Jun 在丝氨酸 73 处的磷酸化也相应增加。利用 HK2 细胞,我们观察到氧化磷酸化的减少始终与氧化应激的增加有关,氧化应激随后激活 Jun 并诱导细胞凋亡。在这些通路中起枢纽作用的蛋白质可能是干预 CKD 的候选靶标。这些发现极大地促进了目前对 CKD 的了解,并为今后的研究提供了宝贵的见解。意义:慢性肾脏病(CKD)的发病率每年都在上升,其病因多种多样,肾脏经常发生不可逆的纤维化,由于对肾脏纤维化机制的认识不足,治疗方案有限且往往无效。尽管对肾脏纤维化进行了大量的努力和omics研究,但迄今为止,还没有研究调查磷酸化蛋白在UUO模型中的作用。此前,我们基于 CKD 模型进行了全面的转录组和蛋白质组分析,但并未涉及磷酸化蛋白质组的潜在改变。在此,我们完成了 CKD 的综合蛋白质组和磷酸蛋白组图谱,这是 UUO 模型的首个磷酸蛋白组图谱。磷酸蛋白组图谱表明,上皮间充质转化和P53通路在小鼠肾损伤模型中被显著激活,核心蛋白Jun在CKD中起着关键作用。在HK2细胞的基础上,还初步发现了P-Jun与氧化磷酸化之间的相关性。我们的工作有助于加深对 CKD 疾病特征和分子机制的理解。从蛋白质组和磷酸化蛋白质组中识别潜在的 CKD 靶点可能为 CKD 的早期诊断和治疗提供有价值的见解。
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引用次数: 0
Serum proteomics identifies biomarkers for predicting non-survivors in elderly COVID-19 patients 血清蛋白质组学确定了预测 COVID-19 老年患者非存活者的生物标志物。
IF 2.8 2区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-11-14 DOI: 10.1016/j.jprot.2024.105356
Lin Wang , Wenmin Tian , Sen Wang , Yuhong Liu , Hongli Wang , Junjie Xiao , Zhongkuo Yu , Lixin Xie , Yang Chen
In December 2022, China ceased the zero-COVID-19 policy, resulting in an increase in hospitalizations and deaths due to COVID-19, particularly among the elderly population. Predicting non-survivors aims to identify high-risk patients and enable targeted interventions to improve survival rates. Additionally, understanding factors affecting prognosis provides essential insights for further research and optimization of treatment strategies. We applied 4D-DIA mass spectrometry for serum proteome analysis and provided a comprehensive characterization of disease features in elderly patients within the Chinese population. Our study elucidated that immune disorders, lung damage, and cardiovascular disorders are predominant causes of death in these patients. Compared to clinical indices, proteomic analysis is more sensitive in tracing these disorders. We also provided a prediction panel for survival outcomes of elderly patients using levels of CXCL10, CXCL16 and IL1RA, which were validated by ELISA. These biomarkers will help improve predictive efficacy for survival outcomes in elderly patients.
2022 年 12 月,中国停止了 COVID-19 零政策,导致 COVID-19 引起的住院和死亡人数增加,尤其是在老年人群中。预测非存活患者的目的是识别高危患者,并采取有针对性的干预措施以提高存活率。此外,了解影响预后的因素还能为进一步研究和优化治疗策略提供重要启示。我们应用 4D-DIA 质谱技术进行血清蛋白质组分析,全面描述了中国老年患者的疾病特征。我们的研究阐明,免疫紊乱、肺损伤和心血管疾病是这些患者的主要死因。与临床指标相比,蛋白质组分析在追踪这些疾病方面更为敏感。我们还利用 CXCL10、CXCL16 和 IL1RA 的水平为老年患者的生存结果提供了一个预测面板,并通过 ELISA 进行了验证。这些生物标志物将有助于提高对老年患者生存结果的预测效力。
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引用次数: 0
Decellularized extracellular matrix from bovine ovarian tissue maintains the protein composition of the native matrisome 牛卵巢组织脱细胞细胞外基质保持了原生基质的蛋白质组成。
IF 2.8 2区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-11-07 DOI: 10.1016/j.jprot.2024.105347
Cecibel M. León-Félix , Emna Ouni , Gaëtan Herinckx , Didier Vertommen , Christiani A. Amorim , Carolina M. Lucci
Recent approaches of regenerative reproductive medicine investigate the decellularized extracellular matrix to develop a transplantable engineered ovary (TEO). However, a full proteomic analysis is not usually performed after the decellularization process to evaluate the preservation of the extracellular matrix (ECM). In this study, the ECM of the bovine ovarian cortex was analyzed before and after decellularization using mass spectrometry and bioinformatics. A total of 155 matrisome proteins were identified in the native ECM of the bovine ovarian cortex, with 145 matrisome proteins detected in the decellularized ECM. After decellularization, only 10 matrisome proteins were lost, and notably, none belonged to the category of reproductive biological processes. Histology and histochemistry analyses were employed to assess the general morphology of both native and decellularized ECM, allowing for the identification of the most abundant ECM proteins. Moreover, our study highlighted collagen type VI alpha 3 and heparan sulfate proteoglycan 2 as the most abundant components in the bovine ovarian ECM, mirroring the composition observed in the human ovary. These findings enhance our understanding of the composition of both native and decellularized ECM, with the potential implications for the development of a TEO.

Significance

The significance of the present study lies on the possibility of advancing towards developing a bioengineered ovary, which is the ultimate strategy to regain fertility in women. The results demonstrate that the decellularized extracellular matrix of the bovine ovary maintains the protein composition of the native matrisome, using a recently described decellularization protocol. The decellularized matrix may serve as scaffolding for seeding ovarian stromal cells and follicles to create a bioengineered ovary, and as closer its composition is to the native matrix the better. Also, comparing the bovine ovarian matrisome, which was described for the first time here, with the human ovarian matrisome, we could see a great similarity, suggesting that the bovine ovary decellularized matrix may serve as a model for developing a human bioengineered ovary.
最近的再生生殖医学方法研究了脱细胞细胞外基质,以开发可移植工程卵巢(TEO)。然而,通常不会在脱细胞过程后进行全面的蛋白质组分析,以评估细胞外基质(ECM)的保存情况。本研究利用质谱法和生物信息学分析了脱细胞前后牛卵巢皮质的 ECM。在牛卵巢皮质的原生 ECM 中共鉴定出 155 个 matrisome 蛋白,在脱细胞 ECM 中检测到 145 个 matrisome 蛋白。脱细胞后,只有 10 个 matrisome 蛋白丢失,值得注意的是,没有一个属于生殖生物过程类别。通过组织学和组织化学分析评估了原生和脱细胞 ECM 的总体形态,从而确定了最丰富的 ECM 蛋白。此外,我们的研究突出表明,VI 型胶原蛋白α 3 和硫酸肝素蛋白多糖 2 是牛卵巢 ECM 中含量最高的成分,这与在人类卵巢中观察到的成分一致。这些发现加深了我们对原生和脱细胞 ECM 组成的了解,对 TEO 的开发具有潜在影响。意义:本研究的意义在于有可能推进生物工程卵巢的开发,这是恢复女性生育能力的最终策略。研究结果表明,采用最近描述的脱细胞方案,脱细胞的牛卵巢细胞外基质保持了原生基质的蛋白质组成。脱细胞基质可作为播种卵巢基质细胞和卵泡的支架,以创建生物工程卵巢,其成分越接近原生基质越好。此外,我们还将这里首次描述的牛卵巢基质组与人类卵巢基质组进行了比较,发现两者非常相似,这表明牛卵巢脱细胞基质可作为开发人类生物工程卵巢的模型。
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引用次数: 0
DIA proteomic and PRM validation through human granulose cells profiles screen suitable biomarkers for polycystic ovary syndrome patients 通过人类粒细胞图谱进行 DIA 蛋白质组学和 PRM 验证,筛选出适合多囊卵巢综合征患者的生物标记物。
IF 2.8 2区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-30 DOI: 10.1016/j.jprot.2024.105332
Faying Liu , Lifeng Tian , Ying Zhang , Wei Deng , Xiaoyun Xu , Yang Zou , Ruifang An
The aim of this study is to identify differentially expressed proteins (DEPs) in granulose cells (GCs) from women with or withoutpolycystic ovary syndrome (PCOS) via data independent acquisition (DIA) proteomic analysis.A total of 63 women were recruited for this study, 34 PCOS patients as experimental group (P), and 29 women without PCOS as Normal group (NP). DIA-based proteomic analysis was performed to identify DEPs in GCs between the P and NP samples. Certain typical DEPs were further validated by Parallel reaction monitoring (PRM), and correlation analysis was performed between these DEPs and the clinical characteristics.Cell vitality was measured by CCK-8 assay. DIA analysis revealed 174 significantly DEPs, of which 7 were upregulated and 167 downregulated. Bioinformatics analysis was performed to analysis the significantly DEPs. The PRM experiment confirmed TOP2A and SPHKAP were upregulated significantly in P by comparing to NP, while GM2A, MRPS16, APOA2 and FGF2 were downregulated significantly. Most notably, Correlation analysis revealed that TOP2A, SPHKAP, MRPS16 and FGF2were positively correlated with TG, AMH and Age, but negatively correlated with Menarche age, DBIL, FT3, Basal serum FSH and LH.Meanwhile, CCK-8 assay has shown that downregulation of FGF2 could weaken cell viability. Finally, a panel of DEPs were identified in the GCs of patients with PCOS, of which certain significant DEPs might play essential roles in the pathogenesis of PCOS, could be regarded as candidate biomarkers for PCOS.
本研究旨在通过数据独立采集(DIA)蛋白质组学分析,鉴定患有或未患有多囊卵巢综合征(PCOS)的女性颗粒细胞(GCs)中的差异表达蛋白质(DEPs)。本研究共招募了 63 名女性,其中 34 名 PCOS 患者为实验组(P),29 名未患有 PCOS 的女性为正常组(NP)。通过基于 DIA 的蛋白质组学分析,确定了 P 组和 NP 组样本 GC 中的 DEPs。平行反应监测(PRM)进一步验证了某些典型的DEPs,并对这些DEPs与临床特征进行了相关性分析。DIA 分析发现了 174 个明显的 DEPs,其中 7 个上调,167 个下调。生物信息学分析对明显的 DEPs 进行了分析。PRM实验证实,与NP相比,TOP2A和SPHKAP在P中明显上调,而GM2A、MRPS16、APOA2和FGF2则明显下调。最值得注意的是,相关性分析表明,TOP2A、SPHKAP、MRPS16 和 FGF2 与 TG、AMH 和年龄呈正相关,但与初潮年龄、DBIL、FT3、基础血清 FSH 和 LH 呈负相关。最后,研究人员在多囊卵巢综合征患者的GC中鉴定出了一组DEPs,其中某些重要的DEPs可能在多囊卵巢综合征的发病机制中起着至关重要的作用,可被视为多囊卵巢综合征的候选生物标志物。
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引用次数: 0
Untargeted metabolomics of 3xTg-AD neurotoxic astrocytes 3xTg-AD神经毒性星形胶质细胞的非靶向代谢组学研究
IF 2.8 2区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-23 DOI: 10.1016/j.jprot.2024.105336
Diego Carvalho , Pablo Diaz-Amarilla , Mathew R. Smith , María Daniela Santi , Marcela Martinez-Busi , Young-Mi Go , Dean P. Jones , Pablo Duarte , Eduardo Savio , Juan A. Abin-Carriquiry , Florencia Arredondo
Alzheimer's disease (AD) is the most common form of dementia, affecting approximately 47 M people worldwide. Histological features and genetic risk factors, among other evidence, supported the amyloid hypothesis of the disease. This neuronocentric paradigm is currently undergoing a shift, considering evidence of the role of other cell types, such as microglia and astrocytes, in disease progression. Previously, we described a particular astrocyte subtype obtained from the 3xTg-AD murine model that displays neurotoxic properties in vitro. We continue here our exploratory analysis through the lens of metabolomics to identify potentially altered metabolites and biological pathways.
Cell extracts from neurotoxic and control astrocytes were compared using high-resolution mass spectrometry-based metabolomics. Around 12 % of metabolic features demonstrated significant differences between neurotoxic and control astrocytes, including alterations in the key metabolite glutamate. Consistent with our previous transcriptomic study, the present results illustrate many homeostatic and regulatory functions of metabolites, suggesting that neurotoxic 3xTg-AD astrocytes exhibit alterations in the Krebs cycle as well as the prostaglandin pathway.
This is the first metabolomic study performed in 3xTg-AD neurotoxic astrocytes. These results provide insight into metabolic alterations potentially associated with neurotoxicity and pathology progression in the 3xTg-AD mouse model and strengthen the therapeutic potential of astrocytes in AD.

Biological significance

Our study is the first high-resolution metabolomic characterization of the novel neurotoxic 3xTg-AD astrocytes. We propose key metabolites and pathway alterations, as well as possible associations with gene expression alterations in the model. Our results are in line with recent hypotheses beyond the amyloid cascade, considering the involvement of several stress response cascades during the development of Alzheimer's disease. This work could inspire other researchers to initiate similar studies in related models. Furthermore, this work illustrates a powerful workflow for metabolite annotation and selection that can be implemented in other studies.
阿尔茨海默病(AD)是最常见的痴呆症,全球约有 4700 万人患病。组织学特征和遗传风险因素等证据支持该病的淀粉样蛋白假说。考虑到有证据表明小胶质细胞和星形胶质细胞等其他细胞类型在疾病进展中的作用,这种以神经元为中心的范式目前正在发生转变。此前,我们描述了从 3xTg-AD 小鼠模型中获得的一种特殊星形胶质细胞亚型,这种亚型在体外显示出神经毒性特性。在此,我们继续通过代谢组学的视角进行探索性分析,以确定可能发生改变的代谢物和生物通路。我们使用基于 HRMS 的代谢组学方法比较了神经毒性星形胶质细胞和对照星形胶质细胞的细胞提取物。约有 12% 的代谢特征在神经毒性星形胶质细胞和对照组星形胶质细胞之间表现出显著差异,其中包括关键代谢物谷氨酸的改变。与我们之前的转录组学研究一致,本研究结果说明了代谢物的许多平衡和调节功能,表明神经毒性 3xTg-AD 星形胶质细胞表现出克雷布斯循环和前列腺素途径的改变。这是首次对 3xTg-AD 神经毒性星形胶质细胞进行代谢组学研究。这些结果有助于深入了解与 3xTg-AD 小鼠模型的神经毒性和病理进展可能相关的代谢改变,并增强了星形胶质细胞治疗 AD 的潜力。生物学意义:我们的研究首次对新型神经毒性 3xTg-AD 星形胶质细胞进行了高分辨率代谢组学表征。我们提出了该模型中关键代谢物和通路的改变,以及与基因表达改变可能存在的关联。我们的研究结果与最近提出的淀粉样蛋白级联之外的假说一致,认为阿尔茨海默病的发展过程中涉及多个应激反应级联。这项工作可以激励其他研究人员在相关模型中开展类似研究。此外,这项工作还展示了一个强大的代谢物注释和筛选工作流程,可在其他研究中实施。
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引用次数: 0
Serum proteomic analysis identified ITIH4 as a potential novel biomarker for feline infectious peritonitis 血清蛋白质组分析发现 ITIH4 是猫传染性腹膜炎的潜在新型生物标记物。
IF 2.8 2区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-23 DOI: 10.1016/j.jprot.2024.105338
Yuzhou Jiao , Mengfang Yang , Lingying Fang , Yuanyuan Yan , Zhen Fu , Mengxia Li , Lisha Li , Zirui Liu , Xiaoshuai Hu , Benyuan Wu , Yuejun Shi , Chao Kang , Zhou Shen , Guiqing Peng
Feline infectious peritonitis (FIP) is a fatal feline disease. At present, the reference standard for FIP diagnosis is immunohistochemistry (IHC) of organs, but this method involves high time-related costs, invasive sampling procedures and professional requirements. Serological detection is a common auxiliary method for diagnosing diseases. As a result, we assessed the changes in the serum proteome of FIP patients with the aim of identifying novel specific serum biomarkers that could aid in the clinical diagnosis of FIP. Pre- and postinfection groups were compared and 92 differentially expressed proteins (DEPs) were identified. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses of the DEPs revealed that the enriched GO terms and KEGG pathways among the DEPs were immune activation, peptidase regulator activity and the complement and coagulation cascade pathways. The level of peptidase regulator interalpha-trypsin inhibitor heavy chain 4 (ITIH4) in cat serum was significantly correlated with FIP. The areas under the ROC curve (AUCs) of full-length ITIH4 (f-ITIH4) and cleaved ITIH4 (c-ITIH4) expression were 0.922 and 1.000, respectively, which allowed the discrimination of FIP cats from healthy cats. These results suggest that ITIH4 may be a potential serum biomarker for detecting early FIP.

Significance

FIP causes fatal disease in cats of almost all ages, and there is currently no effective vaccine or treatment for FIP. Therefore, early diagnosis is extremely important for disease prevention and control. The results of the model and clinical samples revealed that ITIH4 was significantly increased in the serum of FIP cats. This study is the first to propose ITIH4 as a diagnostic biomarker in cats with FIP and our results suggest that serum ITIH4 levels might identify cats with FIP during the early stage.
猫传染性腹膜炎(FIP)是一种致命的猫科动物疾病。目前,诊断 FIP 的参考标准是器官免疫组化(IHC),但这种方法时间成本高、取样过程具有侵入性且对专业人员要求高。血清学检测是诊断疾病的常用辅助方法。因此,我们对 FIP 患者血清蛋白质组的变化进行了评估,目的是找出有助于 FIP 临床诊断的新型特异性血清生物标记物。我们比较了感染前组和感染后组,并确定了 92 个差异表达蛋白 (DEP)。对差异表达蛋白进行的基因本体(GO)和京都基因组百科全书(KEGG)通路富集分析表明,差异表达蛋白中富集的GO术语和KEGG通路包括免疫激活、肽酶调节剂活性以及补体和凝血级联通路。猫血清中肽酶调节剂α-胰蛋白酶抑制剂重链 4(ITIH4)的水平与 FIP 显著相关。全长 ITIH4(f-ITIH4)和裂解 ITIH4(c-ITIH4)表达的 ROC 曲线下面积(AUCs)分别为 0.922 和 1.000,可将 FIP 猫与健康猫区分开来。这些结果表明 ITIH4 可能是检测早期 FIP 的潜在血清生物标记物。意义:FIP 几乎会导致所有年龄段的猫患上致命疾病,目前还没有有效的疫苗或治疗方法。因此,早期诊断对疾病的预防和控制极为重要。模型和临床样本的结果显示,FIP 猫血清中 ITIH4 明显增加。本研究首次提出将 ITIH4 作为 FIP 猫的诊断生物标志物,我们的结果表明血清 ITIH4 水平可在早期阶段识别 FIP 猫。
{"title":"Serum proteomic analysis identified ITIH4 as a potential novel biomarker for feline infectious peritonitis","authors":"Yuzhou Jiao ,&nbsp;Mengfang Yang ,&nbsp;Lingying Fang ,&nbsp;Yuanyuan Yan ,&nbsp;Zhen Fu ,&nbsp;Mengxia Li ,&nbsp;Lisha Li ,&nbsp;Zirui Liu ,&nbsp;Xiaoshuai Hu ,&nbsp;Benyuan Wu ,&nbsp;Yuejun Shi ,&nbsp;Chao Kang ,&nbsp;Zhou Shen ,&nbsp;Guiqing Peng","doi":"10.1016/j.jprot.2024.105338","DOIUrl":"10.1016/j.jprot.2024.105338","url":null,"abstract":"<div><div>Feline infectious peritonitis (FIP) is a fatal feline disease. At present, the reference standard for FIP diagnosis is immunohistochemistry (IHC) of organs, but this method involves high time-related costs, invasive sampling procedures and professional requirements. Serological detection is a common auxiliary method for diagnosing diseases. As a result, we assessed the changes in the serum proteome of FIP patients with the aim of identifying novel specific serum biomarkers that could aid in the clinical diagnosis of FIP. Pre- and postinfection groups were compared and 92 differentially expressed proteins (DEPs) were identified. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses of the DEPs revealed that the enriched GO terms and KEGG pathways among the DEPs were immune activation, peptidase regulator activity and the complement and coagulation cascade pathways. The level of peptidase regulator interalpha-trypsin inhibitor heavy chain 4 (ITIH4) in cat serum was significantly correlated with FIP. The areas under the ROC curve (AUCs) of full-length ITIH4 (f-ITIH4) and cleaved ITIH4 (c-ITIH4) expression were 0.922 and 1.000, respectively, which allowed the discrimination of FIP cats from healthy cats. These results suggest that ITIH4 may be a potential serum biomarker for detecting early FIP.</div></div><div><h3>Significance</h3><div>FIP causes fatal disease in cats of almost all ages, and there is currently no effective vaccine or treatment for FIP. Therefore, early diagnosis is extremely important for disease prevention and control. The results of the model and clinical samples revealed that ITIH4 was significantly increased in the serum of FIP cats. This study is the first to propose ITIH4 as a diagnostic biomarker in cats with FIP and our results suggest that serum ITIH4 levels might identify cats with FIP during the early stage.</div></div>","PeriodicalId":16891,"journal":{"name":"Journal of proteomics","volume":"310 ","pages":"Article 105338"},"PeriodicalIF":2.8,"publicationDate":"2024-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142502654","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Proteomic analysis of the human amniotic mesenchymal stromal cell secretome by integrated approaches via filter-aided sample preparation 通过过滤辅助样品制备综合方法对人类羊膜间充质基质细胞分泌组进行蛋白质组分析。
IF 2.8 2区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-22 DOI: 10.1016/j.jprot.2024.105339
Alexandra Muntiu , Andrea Papait , Federica Vincenzoni , Diana Valeria Rossetti , Pietro Romele , Anna Cargnoni , Antonietta Silini , Ornella Parolini , Claudia Desiderio
The immunomodulatory, anti-inflammatory and regenerative properties of the human amniotic mesenchymal stromal cells (hAMSCs) secretome are acknowledged but the understanding of the specific bioactive components remains incomplete. To address these limitations, the present investigation aimed to profile the proteins and peptides content of the hAMSC secretome through sample pretreatment and fractionation on 10 kDa molecular cut-off FASP (Filter Aided Sample Preparation) device and LC-MS analysis. The filter retained protein fraction underwent trypsin digestion, while the unretained was collected unchanged for intact small proteins and peptides analysis. This combined approach (C-FASP) collects in a single step two complementary fractions, advantageously saving sample volume and time of analysis. The bottom-up analysis of the C-FASP proteins fraction >10 kDa confirmed our previous findings, establishing a set of proteins consistently characterizing the hAMSC secretome. The analysis of the fraction <10 kDa, never been investigated to our knowledge, identified peptide fragments of thymosin beta 4 and beta 10, collagen alpha 1 chains I and III, alpha-enolase, and glyceraldehyde-3-phosphate dehydrogenase, involved in wound healing, anti-inflammatory response, tissue repair and regeneration, key biological activities of the secretome. C-FASP provided a comprehensive molecular profile of the hAMSC secretome offering new insights for enhanced therapeutic applications in regenerative medicine.

Significance

In this investigation we originally present the comprehensive proteomic investigation of the human amniotic mesenchymal stromal cell secretome by combining the analysis of the proteome and of the peptidome following sample pretreatment and fractionation by Filter Aided Sample Preparation (FASP) with 10 kDa molecular cut-off in coupling with LC-MS analysis. The proteome fraction retained by FASP filter was analyzed after enzymatic digestion, while the unretained fraction, below 10 kDa molecular mass, was analyzed unchanged in its intact form. This dual approach provides novel insights, previously unexplored, into the molecular components potentially responsible for the immunomodulatory and anti-inflammatory properties of the hAMSC secretome. These findings could significantly enhance the therapeutic potential of hAMSCs in regenerative medicine.
人羊膜间充质基质细胞(hAMSCs)分泌组的免疫调节、抗炎和再生特性已得到公认,但对其特定生物活性成分的了解仍不全面。为了解决这些局限性,本研究旨在通过样品预处理、在 10 kDa 分子截断的 FASP(过滤辅助样品制备)装置上分馏以及 LC-MS 分析,对人羊膜间充质干细胞分泌组的蛋白质和肽含量进行分析。过滤保留的蛋白质部分经过胰蛋白酶消化,而未保留的则原样收集,用于完整的小蛋白质和肽分析。这种组合方法(C-FASP)可在一个步骤中收集两个互补的馏分,从而节省样品量和分析时间。对 >10 kDa 的 C-FASP 蛋白馏分进行的自下而上分析证实了我们之前的研究结果,建立了一组能一致描述 hAMSC 分泌组特征的蛋白质。对
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引用次数: 0
Intraspecific venom variation in the Iberian asp viper (Vipera aspis zinnikeri) across natural and intensive agricultural habitats 伊比利亚蝰蛇(Vipera aspis zinnikeri)在自然栖息地和集约化农业栖息地的种内毒液变异。
IF 2.8 2区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-10-22 DOI: 10.1016/j.jprot.2024.105337
Jon Buldain , Rui Vitorino , Tânia Lima , Ignazio Avella , Óscar Zuazo , Fernando Martínez-Freiría
Snake venom composition varies at different levels. To date, comparative venom studies have seldom taken into account the role of habitat type in the occurrence of snake venom variation. Here we investigated the presence of venom variation across different populations of the Iberian asp viper (Vipera aspis zinnikeri) inhabiting two contrasting habitats: natural vs. intensive agricultural. We used shotgun proteomics to describe the protein composition of the venoms of six adults from two distinct localities. Furthermore, to test whether local conditions and habitat can alter venom composition in this taxon, we compared the SDS-PAGE profiles of 40 adult venoms from six populations, three in natural habitats and three in intensive agricultural environments. The venoms were composed of 21 toxin families, of which five (CTL, PLA2, VEGF, svSP, and svMP) comprised 69–82 % of each proteome. The relative abundances of toxin families varied considerably at inter- and intra-population levels. Linear regression performed on non-metric multidimensional scaling values showed a significant effect of locality of origin and habitat type on the differences detected between individual SDS-PAGE venom profiles. Our results suggest the presence of regional variation in V. a. zinnikeri venom, potentially reinforcing the role of local pressures in shaping snake venom composition.

Significance

This work provides the first proteomic characterization of the venom of the Iberian asp viper, Vipera aspis zinnikeri, obtained by means of shotgun proteomics. The statistical analysis of 40 individual SDS-PAGE venom profiles highlights that venom variation in this taxon can be associated with geographical origin and habitat type of the area where each viper was collected. Our results suggest the presence of regional variation in V. a. zinnikeri venom, reinforcing the role that local pressures may play as drivers of snake venom variation.
蛇毒成分在不同层次上存在差异。迄今为止,毒液比较研究很少考虑栖息地类型在蛇毒变异中的作用。在这里,我们研究了伊比利亚蝰蛇(Vipera aspis zinnikeri)在两种截然不同的栖息地(自然栖息地和集约化农业栖息地)中不同种群的毒液变异情况。我们使用霰弹枪蛋白质组学描述了来自两个不同地区的六条成年蝰蛇毒液的蛋白质组成。此外,为了检验当地条件和栖息地是否会改变该类群的毒液组成,我们比较了来自六个种群(三个在自然栖息地,三个在集约农业环境)的 40 个成体毒液的 SDS-PAGE 图谱。这些毒液由 21 个毒素家族组成,其中 5 个毒素家族(CTL、PLA2、VEGF、svSP 和 svMP)占每个蛋白质组的 69-82%。毒素家族的相对丰度在种群间和种群内差异很大。对非度量多维缩放值进行的线性回归表明,原产地和栖息地类型对个体 SDS-PAGE 毒液图谱之间的差异有显著影响。我们的研究结果表明,V. a. zinnikeri 毒液存在区域差异,这可能加强了当地压力在塑造蛇毒成分方面的作用。意义:这项研究首次通过霰弹枪蛋白质组学方法对伊比利亚蝰蛇(Vipera aspis zinnikeri)的毒液进行了蛋白质组学表征。对 40 个 SDS-PAGE 毒液图谱的统计分析结果表明,该类群的毒液差异与地理起源和每条蝰蛇采集地的栖息地类型有关。我们的研究结果表明,V. a. zinnikeri 毒液存在区域性差异,这进一步证实了地方压力可能是蛇毒变异的驱动因素。
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引用次数: 0
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Journal of proteomics
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