Huang Hua, Ma Jiaze, Kang An, Guo Tianwei, Sun Wei, Xu Yan, Ji Lijiang
Department of Anorectal Surgery, Changshu Hospital Affiliated to Nanjing University of Chinese Medicine, Changshu, Jiangsu, China.
No. 1 Clinical Medical College, Nanjing University of Chinese Medicine, Nanjing, Jiangsu, China.
Front Immunol. 2025 May 13;16:1534768. doi: 10.3389/fimmu.2025.1534768. eCollection 2025.
Ulcerative colitis (UC) is a chronic inflammatory bowel disease marked by dysregulated immune responses, resulting in sustained inflammation and ulceration of the colonic and rectal mucosa. To elucidate the cellular subtypes and gene expression profiles implicated in the pathogenesis of UC, we utilized single-cell and spatial transcriptomic analyses.
We conducted an analysis of single-cell data to identify cell types involved in the pathogenesis of UC. Employing machine learning methodologies, we screened for key genes implicated in UC and validated these findings through spatial transcriptomics. Additionally, immunohistochemistry was performed on UC lesion samples to investigate the expression patterns of the identified key genes. In an animal model, we utilized immunofluorescence and western blotting to validate the expression of these genes in the affected intestinal segments.
Our investigation identified specific monocyte subtypes associated with UC through a comprehensive analysis involving cell communication, Least Absolute Shrinkage and Selection Operator (LASSO), and Support Vector Machine (SVM) methodologies. Notably, two genes, G protein subunit gamma 5 () and tissue inhibitor of metalloproteinase 1 (), were identified as key regulators of UC development. Spatial transcriptomic indicated a downregulation of expression in UC, whereas expression was upregulated. Furthermore, a significant correlation was detected between and T cell exhaustion-related genes such as genes related to T cell exhaustion, including T cell immunoreceptor with Ig and ITIM domains () and cytotoxic T-lymphocyte-associated protein 4 (). Immunohistochemical analysis of UC lesion samples revealed diminished expression levels of and elevated expression levels of . A dextran sulfate sodium (DSS)-induced colitis mouse model was developed, demonstrating that the protein expression levels of in the colonic tissue of model mice were significantly decreased compared to controls w)ile the expression levels of were increased ( < 0.01). Furthermore, immunofluorescence staining indicated co-localization of with the macrophage marker F4/80 in monocytes.
Our research delineated distinct monocyte subtypes correlated with UC and identified two pivotal genes, and , that contribute to the disease's pathogenesis. These insights offer a significant theoretical basis for enhancing the clinical diagnosis and therapeutic strategies for patients with UC.
溃疡性结肠炎（UC）是一种慢性炎症性肠病，其特征是免疫反应失调，导致结肠和直肠黏膜持续炎症和溃疡。为了阐明与UC发病机制相关的细胞亚型和基因表达谱，我们采用了单细胞和空间转录组分析。
我们对单细胞数据进行分析，以确定参与UC发病机制的细胞类型。运用机器学习方法，我们筛选出与UC相关的关键基因，并通过空间转录组学验证这些发现。此外，对UC病变样本进行免疫组织化学分析，以研究已鉴定关键基因的表达模式。在动物模型中，我们利用免疫荧光和蛋白质印迹法验证这些基因在受影响肠段中的表达。
我们的研究通过涉及细胞通讯、最小绝对收缩和选择算子（LASSO）以及支持向量机（SVM）方法的综合分析，确定了与UC相关的特定单核细胞亚型。值得注意的是，两个基因，G蛋白亚基γ5（）和金属蛋白酶组织抑制剂1（），被确定为UC发展的关键调节因子。空间转录组学表明UC中表达下调，而表达上调。此外，在与T细胞耗竭相关的基因之间检测到显著相关性，例如与T细胞耗竭相关的基因，包括具有Ig和ITIM结构域的T细胞免疫受体（）和细胞毒性T淋巴细胞相关蛋白4（）。UC病变样本的免疫组织化学分析显示表达水平降低，而表达水平升高。建立了葡聚糖硫酸钠（DSS）诱导的结肠炎小鼠模型，表明模型小鼠结肠组织中的蛋白表达水平与对照组相比显著降低，而的表达水平升高（<0.01）。此外，免疫荧光染色表明在单核细胞中与巨噬细胞标志物F4/80共定位。
我们的研究描绘了与UC相关的不同单核细胞亚型，并确定了两个关键基因，和，它们有助于疾病的发病机制。这些见解为加强UC患者的临床诊断和治疗策略提供了重要的理论基础。
