从 HPV 感染的正常宫颈、癌前病变到宫颈癌，解析 HPV 持续诱导恶性转化和免疫重塑的时空奥秘：单细胞 RNA 测序和空间转录组学的整合。

BACKGROUND: The mechanism underlying cervical carcinogenesis that is mediated by persistent human papillomavirus (HPV) infection remains elusive. AIMS: Here, for the first time, we deciphered both the temporal transition and spatial distribution of cellular subsets during disease progression from normal cervix tissues to precursor lesions to cervical cancer. MATERIALS & METHODS: We generated scRNA-seq profiles and spatial transcriptomics data from nine patient samples, including two HPV-negative normal, two HPV-positive normal, two HPV-positive HSIL and three HPV-positive cancer samples. RESULTS: We not only identified three 'HPV-related epithelial clusters' that are unique to normal, high-grade squamous intraepithelial lesions (HSIL) and cervical cancer tissues but also discovered node genes that potentially regulate disease progression. Moreover, we observed the gradual transition of multiple immune cells that exhibited positive immune responses, followed by dysregulation and exhaustion, and ultimately established an immune-suppressive microenvironment during the malignant program. In addition, analysis of cellular interactions further verified that a 'homeostasis-balance-malignancy' change occurred within the cervical microenvironment during disease progression. DISCUSSION: We for the first time presented a spatiotemporal atlas that systematically described the cellular heterogeneity and spatial map along the four developmental steps of HPV-related cervical oncogenesis, including normal, HPV-positive normal, HSIL and cancer. We identified three unique HPV-related clusters, discovered critical node genes that determined the cell fate and uncovered the immune remodeling during disease escalation. CONCLUSION: Together, these findings provided novel possibilities for accurate diagnosis, precise treatment and prognosis evaluation of patients with precancer and cervical cancer.

背景：持续性人乳头瘤病毒（HPV）感染介导的宫颈癌发病机制仍不清楚。

目的：本研究首次解析了从正常宫颈组织到癌前病变再到宫颈癌的疾病进展过程中细胞亚群的时间和空间分布。

材料和方法：我们从 9 名患者样本中生成 scRNA-seq 图谱和空间转录组学数据，包括 2 例 HPV 阴性正常、2 例 HPV 阳性正常、2 例 HPV 阳性 HSIL 和 3 例 HPV 阳性癌症样本。

结果：我们不仅鉴定了三个仅存在于正常、高级别鳞状上皮内病变（HSIL）和宫颈癌组织中的“HPV 相关上皮簇”，还发现了潜在调节疾病进展的节点基因。此外，我们观察到多种免疫细胞逐渐发生阳性免疫反应，随后失调和衰竭，并最终在恶性程序中建立了免疫抑制微环境。此外，细胞相互作用的分析进一步证实，在疾病进展过程中，宫颈微环境中发生了“稳态-平衡-恶性”变化。

讨论：我们首次展示了一个时空图谱，系统地描述了 HPV 相关宫颈癌发生的四个发育阶段（正常、HPV 阳性正常、HSIL 和癌症）的细胞异质性和空间图谱。我们鉴定了三个独特的 HPV 相关簇，发现了决定细胞命运的关键节点基因，并揭示了疾病进展过程中的免疫重塑。

结论：这些发现为癌前病变和宫颈癌患者的准确诊断、精确治疗和预后评估提供了新的可能性。