结直肠癌（CRC）是全球癌症相关死亡的主要原因之一，其进展由结直肠癌干细胞（CR-CSCs）驱动，这些细胞受内源性和微环境信号调节。白细胞介素（IL）-30已被证明对CSC的生存和肿瘤进展至关重要。它是否参与CRC肿瘤发生并影响临床行为尚不清楚。通过蛋白质印迹、免疫电镜、流式细胞术、细胞存活和球形形成测定，确定了干和非干CRC细胞中IL30的产生和功能。通过CRISPR/Cas9介导的IL30基因删除，RNA-Seq和将IL30转染或删除的CR-CSCs植入NSG小鼠，研究了IL30在CRC肿瘤发生中的作用。生物信息学和CRC标本的免疫病理学突出了临床意义。我们证明了CR-CSC和CRC细胞都表达膜锚定的IL30，该蛋白质通过WNT5A和RAB33A调节其自我更新，通过STAT3上调CXCR4主要上调其增殖和迁移，在IL30基因删除的情况下，这些途径以及WNT和RAS通路都被抑制。IL30基因的删除降低了蛋白酶，如MMP2和MMP13，趋化因子受体，主要是CCR7、CCR3和CXCR4，以及生长和炎症介质，包括ANGPT2、CXCL10、EPO、IGF1和EGF的表达。这些因素促进了IL30驱动的CR-CSC和CRC细胞扩张，这些扩张可以通过选择性阻断来消除。删除IL30基因的CR-CSC显示出较低的肿瘤生成能力，并且在80%的小鼠中形成生长缓慢、转移性较低的肿瘤，它们的存活时间比对照组长得多。对“结直肠腺癌TCGA自然2012”收藏进行了生物信息学和CIBERSORTx分析，对临床CRC标本中IL30表达进行了形态学评估，发现CRC和浸润性白细胞中缺乏IL30与总体生存时间延长相关。IL30是一种新的CRC驱动因素，因为它的失活，可以禁用致癌途径和多个内源性循环，从而抑制CR-CSC的肿瘤生成能力和转移能力。CRISPR/Cas9介导的靶向IL30的开发可能有助于改善CRC的现有治疗局面。©作者（或其雇主）2023年。在CC BY-NC下允许重复使用。无商业再利用。由BMJ出版。

Progression of colorectal cancer (CRC), a leading cause of cancer-related death worldwide, is driven by colorectal cancer stem cells (CR-CSCs), which are regulated by endogenous and microenvironmental signals. Interleukin (IL)-30 has proven to be crucial for CSC viability and tumor progression. Whether it is involved in CRC tumorigenesis and impacts clinical behavior is unknown.IL30 production and functions, in stem and non-stem CRC cells, were determined by western blot, immunoelectron microscopy, flow cytometry, cell viability and sphere formation assays. CRISPR/Cas9-mediated deletion of the IL30 gene, RNA-Seq and implantation of IL30 gene transfected or deleted CR-CSCs in NSG mice allowed to investigate IL30's role in CRC oncogenesis. Bioinformatics and immunopathology of CRC samples highlighted the clinical implications.We demonstrated that both CR-CSCs and CRC cells express membrane-anchored IL30 that regulates their self-renewal, via WNT5A and RAB33A, and/or proliferation and migration, primarily by upregulating CXCR4 via STAT3, which are suppressed by IL30 gene deletion, along with WNT and RAS pathways. Deletion of IL30 gene downregulates the expression of proteases, such as MMP2 and MMP13, chemokine receptors, mostly CCR7, CCR3 and CXCR4, and growth and inflammatory mediators, including ANGPT2, CXCL10, EPO, IGF1 and EGF. These factors contribute to IL30-driven CR-CSC and CRC cell expansion, which is abrogated by their selective blockade. IL30 gene deleted CR-CSCs displayed reduced tumorigenicity and gave rise to slow-growing and low metastatic tumors in 80% of mice, which survived much longer than controls. Bioinformatics and CIBERSORTx of the 'Colorectal Adenocarcinoma TCGA Nature 2012' collection, and morphometric assessment of IL30 expression in clinical CRC samples revealed that the lack of IL30 in CRC and infiltrating leucocytes correlates with prolonged overall survival.IL30 is a new CRC driver, since its inactivation, which disables oncogenic pathways and multiple autocrine loops, inhibits CR-CSC tumorigenicity and metastatic ability. The development of CRISPR/Cas9-mediated targeting of IL30 could improve the current therapeutic landscape of CRC.© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.