PO.MCB03.02 · 分子与细胞生物学
自体结直肠癌-间皮细胞共培养模型鉴定出驱动腹膜播散的间质FGFR3依赖性
Autologous colorectal cancer-mesothelial co-culture model identifies stromal FGFR3 dependency driving peritoneal dissemination
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：结直肠癌（CRC）的腹膜播散仍是一种致命疾病，治疗选择有限。衬覆腹膜腔的间皮细胞在转移性种植和间质重塑中发挥关键作用，然而在此背景下调控肿瘤-间质相互作用的机制仍未充分了解。我们旨在建立一个生理学相关模型以阐明这些相互作用并鉴定间质易损性。
方法：我们从单个转移性CRC患者的恶性腹水中同时衍生出一种新型CRC细胞系（OMUCR-1）和癌相关间皮细胞（CAmeso）。作为对照，从一名无播散的患者中建立了正常间皮细胞（Nmeso）。表征了细胞形态、生长动力学、STR图谱和突变状态。使用划痕愈合和Matrigel实验评估CAmeso对CRC迁移和侵袭的影响。在体内，将OMUCR-1细胞单独植入或与CAmeso共移植入裸鼠。进行RNA测序以分析宿主间质基因表达，并对FGFR抑制（BGJ398）进行治疗性测试。
结果：OMUCR-1细胞表现出强大的致瘤性，并携带KRAS（G12D）和TP53（R306）突变。CAmeso在体外显著增强CRC细胞的迁移和侵袭。在异种移植中，与单独OMUCR-1相比，与CAmeso共移植产生了明显更大且富含alphaSMA阳性间质的肿瘤。批量RNA-seq揭示含CAmeso的肿瘤中鼠源Fgfr3上调。用BGJ398进行药理学FGFR阻断可缩小肿瘤体积并耗竭FGFR3阳性间质成分，表明存在间质FGFR3依赖性。
结论：我们报告了首次从同一患者同时建立CRC和间皮细胞系，提供了一个用于剖析腹膜转移过程中肿瘤-间质串扰的自体系统。我们的发现凸显间质FGFR3信号作为CRC进展的潜在介导者和治疗干预的有前景靶点。这一独特平台为晚期结直肠癌腹膜微环境的机制和转化研究提供了宝贵资源。
查看英文原文 English abstract
Background: Peritoneal dissemination of colorectal cancer (CRC) remains a fatal disease with limited therapeutic options. Mesothelial cells, which line the peritoneal cavity, play critical roles in metastatic implantation and stromal remodeling, yet the mechanisms governing tumor-stroma interactions in this setting remain poorly understood. We aimed to establish a physiologically relevant model to elucidate these interactions and identify stromal vulnerabilities.
Methods: We simultaneously derived a novel CRC cell line (OMUCR-1) and cancer-associated mesothelial cells (CAmeso) from the malignant ascites of a single patient with metastatic CRC. As a control, normal mesothelial cells (Nmeso) were established from a patient without dissemination. Cell morphology, growth kinetics, STR profile, and mutational status were characterized. The effects of CAmeso on CRC migration and invasion were evaluated using wound-healing and Matrigel assays. In vivo, OMUCR-1 cells were implanted alone or co-transplanted with CAmeso into nude mice. RNA sequencing was performed to analyze host stromal gene expression, and FGFR inhibition (BGJ398) was tested therapeutically.
Results: OMUCR-1 cells exhibited robust tumorigenicity and harbored KRAS (G12D) and TP53 (R306) mutations. CAmeso significantly enhanced CRC cell migration and invasion in vitro. In xenografts, co-transplantation with CAmeso yielded markedly larger tumors enriched in alphaSMA-positive stroma compared with OMUCR-1 alone. Bulk RNA-seq revealed upregulation of murine Fgfr3 in tumors containing CAmeso. Pharmacologic FGFR blockade with BGJ398 reduced tumor size and depleted FGFR3-positive stromal components, indicating stromal FGFR3 dependency.
Conclusions: We report the first concurrent establishment of CRC and mesothelial cell lines from the same patient, providing an autologous system for dissecting tumor-stroma crosstalk during peritoneal metastasis. Our findings highlight stromal FGFR3 signaling as a potential mediator of CRC progression and a promising target for therapeutic intervention. This unique platform offers a valuable resource for mechanistic and translational studies on the peritoneal microenvironment in advanced colorectal cancer.
利益披露 Disclosure
H. Kasashima, None..
Y. Fukui, None..
I. Omori, None..
Z. Wang, None..
N. Naito, None..
Y. Kusunoki, None..
K. Kuroda, None..
Y. Miki, None..
M. Yoshii, None..
T. Tamura, None..
M. Shibutani, None..
T. Toyokawa, None..
M. Yashiro, None..
K. Maeda, None.