Single-cell transcriptome sequencing

  • 文章类型: Journal Article
    “组学”通常涉及使用高通量分析方法在特定水平上探索生物系统的整个组成的结构和功能,以探测和分析大量数据,包括基因组学,转录组学,蛋白质组学,和代谢组学,在其他类型中。基因组学表征和量化生物体的所有基因,研究它们的相互关系及其对生物体的影响。然而,常规转录组测序技术靶向群体细胞,他们的结果仅反映了群体细胞中基因的平均表达水平,因为它们无法揭示单个细胞之间的基因表达异质性和空间异质性,从而掩盖了不同细胞之间的表达特异性。单细胞转录组测序和空间转录组测序技术分析植物或动物组织中单个细胞的转录组,能够理解每个细胞的代谢产物和表达的基因。因此,可以进行相应组织的统计分析,为了实现细胞分类,进化生长,以及生理和病理分析。本文综述了植物单细胞和空间转录组学的研究进展,以及它们在植物中的应用和挑战。此外,提出了单细胞和空间转录组学的发展前景。
    \"Omics\" typically involves exploration of the structure and function of the entire composition of a biological system at a specific level using high-throughput analytical methods to probe and analyze large amounts of data, including genomics, transcriptomics, proteomics, and metabolomics, among other types. Genomics characterizes and quantifies all genes of an organism collectively, studying their interrelationships and their impacts on the organism. However, conventional transcriptomic sequencing techniques target population cells, and their results only reflect the average expression levels of genes in population cells, as they are unable to reveal the gene expression heterogeneity and spatial heterogeneity among individual cells, thus masking the expression specificity between different cells. Single-cell transcriptomic sequencing and spatial transcriptomic sequencing techniques analyze the transcriptome of individual cells in plant or animal tissues, enabling the understanding of each cell\'s metabolites and expressed genes. Consequently, statistical analysis of the corresponding tissues can be performed, with the purpose of achieving cell classification, evolutionary growth, and physiological and pathological analyses. This article provides an overview of the research progress in plant single-cell and spatial transcriptomics, as well as their applications and challenges in plants. Furthermore, prospects for the development of single-cell and spatial transcriptomics are proposed.
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  • 文章类型: Journal Article
    结直肠癌(CRC)患者的预后由于转移而仍然比预期差。复发,对化疗有抵抗力.结直肠癌干细胞(CRCSCs)在肿瘤转移中起着至关重要的作用,复发,和化疗抵抗。然而,目前尚无基于CRCSCs相关基因的预后标志物可用于临床.在这项研究中,采用单细胞转录组测序技术在CRC微环境中区分癌症干细胞(CSC),并在单细胞水平分析其特性.随后,来自TCGA和GEO数据库的数据被用于建立CRCSCs相关基因的预后风险模型,并验证其诊断性能.此外,功能富集,免疫反应,并对风险模型中相关基因的化疗药物敏感性进行研究。最后,风险模型中的关键基因RPS17被鉴定为潜在的预后标志物和治疗靶点,用于进一步的综合研究.我们的发现为CRC的预后治疗提供了新的见解,并为系统和全面地了解CRC的发展提供了新的观点。
    The prognosis for patients with colorectal cancer (CRC) remains worse than expected due to metastasis, recurrence, and resistance to chemotherapy. Colorectal cancer stem cells (CRCSCs) play a vital role in tumor metastasis, recurrence, and chemotherapy resistance. However, there are currently no prognostic markers based on CRCSCs-related genes available for clinical use. In this study, single-cell transcriptome sequencing was employed to distinguish cancer stem cells (CSCs) in the CRC microenvironment and analyze their properties at the single-cell level. Subsequently, data from TCGA and GEO databases were utilized to develop a prognostic risk model for CRCSCs-related genes and validate its diagnostic performance. Additionally, functional enrichment, immune response, and chemotherapeutic drug sensitivity of the relevant genes in the risk model were investigated. Lastly, the key gene RPS17 in the risk model was identified as a potential prognostic marker and therapeutic target for further comprehensive studies. Our findings provide new insights into the prognostic treatment of CRC and offer novel perspectives for a systematic and comprehensive understanding of CRC development.
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  • 文章类型: Journal Article
    本研究旨在探讨睾丸组织形态学的差异,基因表达,和性不成熟(1岁)和性成熟(10岁)的蒙古马之间的标记基因。我们研究的目的是提供对雄性蒙古马的生殖生理的见解,并确定性成熟的潜在标记。我们应用的方法包括使用单细胞测序技术对睾丸细胞进行转录组学分析。我们的结果表明,两个年龄组之间的组织形态和基因表达模式存在显着差异。具体来说,确定了25个细胞簇和10种细胞类型,包括精原和体细胞。差异基因表达分析强调了与性未成熟马的细胞基础结构和性成熟马的精子发生有关的不同模式。还鉴定了每个阶段特有的标记基因,包括APOA1,AMH,TAC3,INHA,SPARC,和SOX9在性不成熟阶段,和PRM1,PRM2,LOC100051500,PRSS37,HMGB4和H1-9为性成熟阶段。这些发现有助于更深入地了解蒙古马的睾丸发育和精子发生,并在马生殖生物学和育种计划中具有潜在的应用。总之,这项研究为蒙古马性成熟的分子机制提供了有价值的见解。
    This study aimed to investigate differences in testicular tissue morphology, gene expression, and marker genes between sexually immature (1-year-old) and sexually mature (10-year-old) Mongolian horses. The purposes of our research were to provide insights into the reproductive physiology of male Mongolian horses and to identify potential markers for sexual maturity. The methods we applied included the transcriptomic profiling of testicular cells using single-cell sequencing techniques. Our results revealed significant differences in tissue morphology and gene expression patterns between the two age groups. Specifically, 25 cell clusters and 10 cell types were identified, including spermatogonial and somatic cells. Differential gene expression analysis highlighted distinct patterns related to cellular infrastructure in sexually immature horses and spermatogenesis in sexually mature horses. Marker genes specific to each stage were also identified, including APOA1, AMH, TAC3, INHA, SPARC, and SOX9 for the sexually immature stage, and PRM1, PRM2, LOC100051500, PRSS37, HMGB4, and H1-9 for the sexually mature stage. These findings contribute to a deeper understanding of testicular development and spermatogenesis in Mongolian horses and have potential applications in equine reproductive biology and breeding programs. In conclusion, this study provides valuable insights into the molecular mechanisms underlying sexual maturity in Mongolian horses.
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  • 文章类型: Journal Article
    胶质母细胞瘤(GBM),神经肿瘤领域内普遍存在的恶性肿瘤,一直是一个相当仔细审查的主题。巨噬细胞,作为主要的免疫学成分,深刻渗透到GBM的微环境中。然而,在单细胞水平上阐明控制巨噬细胞参与GBM的复杂免疫机制的研究仍然非常有限。
    我们采用单细胞分析进行了全面调查,旨在重新定义GBM肿瘤核心和外周区域复杂的细胞景观。我们的分析重点扩展到对巨噬细胞的深入研究,阐明它们在氧化应激背景下的作用,细胞间信息交换,以及有关GBM及其分类亚群的细胞轨迹。我们致力于鉴定与巨噬细胞复杂相关的GBM预后基因。利用实验研究探讨MANBA在GBM背景下的相关性。
    我们的研究阐明了巨噬细胞在GBM微环境中各种亚群之间的复杂相互作用中的核心作用。值得注意的是,与其他亚群中的GBM对应物相比,我们观察到巨噬细胞内的氧化应激反应强度明显.此外,巨噬细胞协调了错综复杂的蜂窝通信网络,由SPP1-CD44轴促进,内部和邻近的亚群。这些发现共同表明巨噬细胞极化从M1到M2表型的可能性,有助于肿瘤微环境内的免疫抑制。此外,我们的探索发现了与巨噬细胞密切相关的GBM预后基因,最著名的是MANBA和TCF12。值得注意的是,MANBA似乎通过对M1极化的巨噬细胞施加抑制作用来参与神经免疫功能的调节,从而促进肿瘤进展。为了支持这些断言,实验验证明确肯定了MANBA对GBM的推广影响,通过其抑制细胞增殖的能力得以阐明,侵入性,和转移潜力。
    这些启示代表了揭示控制巨噬细胞与GBM环境内不同亚群之间相互作用的复杂免疫机制的关键一步。此外,它们为开发创新的GBM预后模型奠定了基础,曼巴在震中,并强调了在持续追求这种强大的恶性肿瘤的增强治疗方式中,新的免疫治疗靶标的潜力。
    Glioblastoma (GBM), a prevalent malignant neoplasm within the neuro-oncological domain, has been a subject of considerable scrutiny. Macrophages, serving as the principal immunological constituents, profoundly infiltrate the microenvironment of GBM. However, investigations elucidating the intricate immunological mechanisms governing macrophage involvement in GBM at the single-cell level remain notably limited.
    We conducted a comprehensive investigation employing single-cell analysis, aiming to redefine the intricate cellular landscape within both the core and peripheral regions of GBM tumors. Our analytical focus extended to the profound study of macrophages, elucidating their roles within the context of oxidative stress, intercellular information exchange, and cellular trajectories concerning GBM and its assorted subpopulations. We pursued the identification of GBM prognostic genes intricately associated with macrophages. Utilizing experimental research to investigate the relevance of MANBA in the context of GBM.
    Our investigations have illuminated the central role of macrophages in the intricate interplay among various subpopulations within the GBM microenvironment. Notably, we observed a pronounced intensity of oxidative stress responses within macrophages when compared to their GBM counterparts in other subpopulations. Moreover, macrophages orchestrated intricate cellular communication networks, facilitated by the SPP1-CD44 axis, both internally and with neighboring subpopulations. These findings collectively suggest the potential for macrophage polarization from an M1 to an M2 phenotype, contributing to immune suppression within the tumor microenvironment. Furthermore, our exploration unearthed GBM prognostic genes closely associated with macrophages, most notably MANBA and TCF12. Remarkably, MANBA appears to participate in the modulation of neuroimmune functionality by exerting inhibitory effects on M1-polarized macrophages, thereby fostering tumor progression. To bolster these assertions, experimental validations unequivocally affirmed the promotional impact of MANBA on GBM, elucidated through its capacity to curb cell proliferation, invasiveness, and metastatic potential.
    These revelations represent a pivotal step towards unraveling the intricate immunological mechanisms governing the interactions between macrophages and diverse subpopulations within the GBM milieu. Furthermore, they lay the foundation for the development of an innovative GBM prognostic model, with MANBA at its epicenter, and underscore the potential for novel immunotherapeutic targets in the ongoing pursuit of enhanced treatment modalities for this formidable malignancy.
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  • 文章类型: Journal Article
    单细胞RNA测序(scRNA-Seq)为研究不同细胞类型与生物体之间的相互作用提供了新的视角和思路。通过将scRNA-seq与新的计算框架或特定技术集成,可以开发更好的阿尔茨海默病(AD)治疗方法。
    单细胞测序数据集GSE158234从GEO数据库获得。预处理,质量控制,降维聚类,并对其进行注释以识别细胞类型。RNA-seq分析数据集GSE238013用于确定不同样品中特定细胞亚群的组分。OMIM中包含的一组基因,基因卡,CTD,和DisGeNET数据库被选择为高度可信的AD相关基因。然后,使用KO组中显著表达的基因作为hub基因创建ROC曲线以预测诊断价值。将上述基因映射到Coremine医学数据库,以预测未来的治疗中药,并建立了“中药-成分-靶标”网络,以筛选潜在的治疗靶标。最后一步是进行孟德尔随机化研究,以确定全基因组关联研究中关键基因IL1B和AD之间的因果关系。
    使用scRNA-seq数据集,发现了五个独特的细胞簇。使用标记基因将这些簇进一步细分为四种不同的细胞类型。与WT组相比,KO组显示出更实质性的巨噬细胞差异亚组。通过使用可用的数据集和PPI网络分析,发现了54个常见基因。使用MCODE方法确定了四个集群,相关分析表明,4个基因簇中的7个基因与巨噬细胞呈显著负相关。四组中的六个基因呈显著正相关。五种基因在WT和KO组中具有不同的表达水平。String数据库用于鉴定四个基因(IL10,CX3CR1,IL1B,和IL6),最终被选为ADhub基因。筛选鉴定出干预AD转化过程的潜在中药,包括丹参,人参,灵芝,甘草,黄连,黄芩,除了有希望的治疗目标,例如PTGS1、PTGS2和RXRA。最后,表明IL1B与AD中的免疫细胞浸润直接相关。在方差倒数加权中,我们发现IL1B与AD的高风险相关,OR为1.003(95%CI=1.001-1.006,p=0.038)。
    我们的研究结合了网络药理学和scRNA-seq计算框架,以揭示相关的枢纽基因和潜在的中药治疗AD的潜在目标。这些发现可能有助于理解AD基因背后的分子过程以及治疗该疾病的新型药物的开发。
    UNASSIGNED: Single-cell RNA sequencing (scRNA-Seq) provides new perspectives and ideas to investigate the interactions between different cell types and organisms. By integrating scRNA-seq with new computational frameworks or specific technologies, better Alzheimer\'s disease (AD) treatments may be developed.
    UNASSIGNED: The single-cell sequencing dataset GSE158234 was obtained from the GEO database. Preprocessing, quality control, dimensionality-reducing clustering, and annotation to identify cell types were performed on it. RNA-seq profiling dataset GSE238013 was used to determine the components of specific cell subpopulations in diverse samples. A set of genes included in the OMIM, Genecards, CTD, and DisGeNET databases were selected as highly plausible AD-related genes. Then, ROC curves were created to predict the diagnostic value using the significantly expressed genes in the KO group as hub genes. The genes mentioned above were mapped to the Coremine Medical database to forecast prospective therapeutic Chinese medicines, and a \"Chinese medicine-ingredient-target\" network was constructed to screen for potential therapeutic targets. The last step was to undertake Mendelian randomization research to determine the causal link between the critical gene IL1B and AD in the genome-wide association study.
    UNASSIGNED: Using the scRNA-seq dataset, five unique cell clusters were discovered. These clusters were further subdivided into four distinct cell types using marker genes. The KO group showed a more substantial differential subgroup of macrophages than the WT group. By using the available datasets and PPI network analysis, 54 common genes were discovered. Four clusters were identified using the MCODE approach, and correlation analysis showed that seven genes in those four clusters had a significantly negative correlation with macrophages. Six genes in four sets had a significantly positive correlation. Five genes had different levels of expression in the WT and KO groups. The String database was used to identify the regulatory relationships between the four genes (IL10, CX3CR1, IL1B, and IL6) that were finally selected as AD hub genes. Screening identified potential traditional Chinese medicine to intervene in the transformation process of AD, including Radix Salviae, ginseng, Ganoderma, licorice, Coptidis Rhizoma, and Scutellariae Radix, in addition to promising therapeutic targets, such as PTGS1, PTGS2, and RXRA. Finally, it was shown that IL1B directly correlated with immune cell infiltration in AD. In inverse variance weighting, we found that IL1B was associated with a higher risk of AD, with an OR of 1.003 (95% CI = 1.001-1.006, p = 0.038).
    UNASSIGNED: Our research combined network pharmacology and the scRNA-seq computational framework to uncover pertinent hub genes and prospective traditional Chinese medicine potential therapeutic targets for AD. These discoveries may aid in understanding the molecular processes behind AD genes and the development of novel medications to treat the condition.
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  • 文章类型: Journal Article
    鄂尔多斯细毛羊是中国优质品种,生产优质天然纺织品和羊毛、羊肉等原料。然而,与澳大利亚美利奴羊相比,在羊毛纤维细度和羊毛产量方面仍有差距。毛囊是控制羊毛纤维类型的主要器官,次级毛囊的形态变化对决定羊毛质量至关重要。然而,鄂尔多斯细毛羊毛囊形态发生的过程和分子机制尚不清楚。因此,分析毛囊形成过程的分子机制对提高鄂尔多斯细毛羊纤维直径和产毛量具有重要意义。差异表达基因,APOD,POSTN,与初级毛囊和次级毛囊相关的KRT5和KRT15,是从真皮乳头中提取的。基于伪时间分析,成功构建了鄂尔多斯细毛羊真皮谱系细胞和表皮谱系细胞的分化轨迹,为鄂尔多斯细毛羊的育种研究提供理论依据。
    The Ordos fine-wool sheep is a high-quality breed in China that produces superior natural textiles and raw materials such as wool and lamb meat. However, compared to the Australian Merino sheep, there is still a gap in terms of the wool fiber fineness and wool yield. The hair follicle is the main organ that controls the type of wool fiber, and the morphological changes in the secondary hair follicle are crucial in determining wool quality. However, the process and molecular mechanisms of hair follicle morphogenesis in Ordos fine-wool sheep are not yet clear. Therefore, analyzing the molecular mechanisms underlying the process of follicle formation is of great significance for improving the fiber diameter and wool production of Ordos fine-wool sheep. The differential expressed genes, APOD, POSTN, KRT5, and KRT15, which related to primary hair follicles and secondary hair follicles, were extracted from the dermal papillae. Based on pseudo-time analysis, the differentiation trajectories of dermal lineage cells and epidermal lineage cells in the Ordos fine-wool sheep were successfully constructed, providing a theoretical basis for breeding research in Ordos fine-wool sheep.
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  • 文章类型: Case Reports
    我们使用10倍基因组学单细胞转录组测序技术来揭示恶性胸腔积液(MPE)患者小细胞肺癌(SCLC)的肿瘤免疫微环境特征。最终获得8008个高质量细胞用于后续的生物信息学分析,将其分成10个细胞簇,进一步鉴定为B细胞,T细胞,骨髓细胞,NK细胞,和癌细胞。这些SCLC相关基因如NOTCH1,MYC,TSC22D1,SOX4,BLNK,YBX3,VIM,CD8A,CD8B,在T细胞和B细胞分化过程中,KLF6有不同程度的表达。T之间不同的配体和受体,B和肿瘤细胞几乎通过MHCII相互作用,IL-16,半乳糖凝集素,和APP信号通路。
    We used 10 × genomics single-cell transcriptome sequencing technology to reveal the tumor immune microenvironment characteristics of small cell lung cancer (SCLC) in a patient with malignant pleural effusion (MPE). A total of 8008 high-quality cells were finally obtained for subsequent bioinformatic analysis, which were divided into 10 cell clusters further identified as B cells, T cells, myeloid cells, NK cells, and cancer cells. Such SCLC related genes as NOTCH1, MYC, TSC22D1, SOX4, BLNK, YBX3, VIM, CD8A, CD8B, and KLF6 were expressed in different degrees during differentiation of T and B cells. Different ligands and receptors between T, B and tumor cells almost interact through MHC II, IL-16, galectin, and APP signaling pathway.
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  • 文章类型: Journal Article
    乳腺癌转移是一个复杂的,多步骤过程,在原发性和转移性乳腺癌之间具有高度的细胞异质性,以及肿瘤微环境中转移性癌细胞和其他细胞之间更复杂的相互作用。高分辨率单细胞转录组测序技术可以实时可视化肿瘤微环境中恶性和非恶性细胞的异质性,特别是结合空间转录组分析,可以直接比较不同阶段的转移样本之间的变化。因此,本研究以单细胞分析为第一视角,深入探讨与乳腺癌转移相关的特殊或罕见细胞亚群,系统地总结了它们的功能,分子特征,以及相应的治疗策略,这将有助于准确识别,理解,和目标肿瘤转移相关的驱动事件,为乳腺癌转移机理研究提供研究基础,为其个性化精准治疗提供新线索。
    Breast cancer metastasis is a complex, multi-step process, with high cellular heterogeneity between primary and metastatic breast cancer, and more complex interactions between metastatic cancer cells and other cells in the tumor microenvironment. High-resolution single-cell transcriptome sequencing technology can visualize the heterogeneity of malignant and non-malignant cells in the tumor microenvironment in real time, especially combined with spatial transcriptome analysis, which can directly compare changes between different stages of metastatic samples. Therefore, this study takes single-cell analysis as the first perspective to deeply explore special or rare cell subpopulations related to breast cancer metastasis, systematically summarizes their functions, molecular features, and corresponding treatment strategies, which will contribute to accurately identify, understand, and target tumor metastasis-related driving events, provide a research basis for the mechanistic study of breast cancer metastasis, and provide new clues for its personalized precision treatment.
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  • 文章类型: Editorial
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  • 文章类型: Journal Article
    干扰素-γ(IFN-γ),通常被称为II型干扰素,是协调肿瘤免疫过程的关键细胞因子,在肿瘤免疫治疗研究中受到了广泛关注。先前的研究已经讨论了IFN-γ在特定肿瘤或疾病中的作用和机制。但IFN-γ在泛癌症中的相关作用仍不确定。
    下载TCGA和GTExRNA表达数据和临床数据。此外,我们通过使用生物信息学方法分析了IFN-γ在肿瘤中的作用,其中包括分析IFN-γ在不同肿瘤中的表达和表达之间的相关性,预后,功能状态,TMB,MSI,免疫细胞浸润,和潮汐。我们还开发了一个用于网络拓扑分析的PPI网络,将集线器基因鉴定为程度大于IFN-γ水平的基因。
    IFN-γ在TCGA中的大多数肿瘤类型中差异表达并预测不同的生存状态。此外,IFN-γ表达与T细胞浸润等因素密切相关,免疫检查点,免疫激活基因,免疫抑制基因,趋化因子,和趋化因子受体,以及肿瘤纯度,功能状态,和预后价值。此外,预后,CNV,和治疗反应均与IFN-γ相关基因表达相关。特别是,IFN-γ相关基因STAT1在UCEC中表现出最大的SNV百分比和最大的SNPs百分比。在多种肿瘤类型中发现IFN-γ相关基因的表达水平升高,这与20种不同类型药物的药物敏感性呈正相关。
    IFN-γ是肿瘤免疫治疗反应的良好指标,可能会限制肿瘤进展,为免疫治疗的未来发展提供了一种新的方法。
    Interferon-gamma (IFN-γ), commonly referred to as type II interferon, is a crucial cytokine that coordinates the tumor immune process and has received considerable attention in tumor immunotherapy research. Previous studies have discussed the role and mechanisms associated with IFN-γ in specific tumors or diseases, but the relevant role of IFN-γ in pan-cancer remains uncertain.
    TCGA and GTEx RNA expression data and clinical data were downloaded. Additionally, we analyzed the role of IFN-γ on tumors by using a bioinformatic approach, which included the analysis of the correlation between IFN-γ in different tumors and expression, prognosis, functional status, TMB, MSI, immune cell infiltration, and TIDE. We also developed a PPI network for topological analysis of the network, identifying hub genes as those having a degree greater than IFN-γ levels.
    IFN-γ was differentially expressed and predicted different survival statuses in a majority of tumor types in TCGA. Additionally, IFN-γ expression was strongly linked to factors like infiltration of T cells, immune checkpoints, immune-activating genes, immunosuppressive genes, chemokines, and chemokine receptors, as well as tumor purity, functional statuses, and prognostic value. Also, prognosis, CNV, and treatment response were all substantially correlated with IFN-γ-related gene expression. Particularly, the IFN-γ-related gene STAT1 exhibited the greatest percentage of SNVs and the largest percentage of SNPs in UCEC. Elevated expression levels of IFN-γ-related genes were found in a wide variety of tumor types, and this was shown to be positively linked to drug sensitivity for 20 different types of drugs.
    IFN-γ is a good indicator of response to tumor immunotherapy and is likely to limit tumor progression, offering a novel approach for immunotherapy\'s future development.
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