PO.TB07.01 · 肿瘤生物学
Netrin-1/UNC5B-TFF3轴调节转移性结直肠癌中癌症干细胞的自我更新和化疗耐药
Netrin-1/UNC5B-TFF3 axis modulates cancer stem cells self-renewal and chemoresistance in metastatic colorectal cancer
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
目的：转移性结直肠癌（mCRC）因癌症干细胞（CSC）驱动的复发和化疗耐药而结局不佳。我们研究了依赖性受体配体netrin-1及其受体UNC5B在CSC自我更新中的作用，并评估了使用抗netrin-1抗体NP137对netrin-1的治疗性抑制。
实验方法：用重组netrin-1、NP137或对照处理来源于结直肠癌肝转移的患者来源类器官（PDO）。通过极限稀释实验量化自我更新。用CRISPR/Cas9沉默UNC5B以确定该Netrin-1受体的作用。单细胞RNA测序阐明受NP137影响的信号通路。在PDO异种移植小鼠中测试NP137单用或与FOLFOX化疗联用的体内疗效，并对一名参加NP137临床试验的mCRC患者的配对肿瘤活检进行RNA-seq分析。
结果：Netrin-1增强了CSC的自我更新和存活，这些效应被NP137或UNC5B敲除所消除。NP137处理触发了CSC凋亡，该效应可被caspase抑制所逆转。单细胞转录组学揭示UNC5B阳性细胞分泌三叶因子3（TFF3），其以旁分泌方式维持干性基因表达（LGR5、SOX4、SMOC2、PROM1）。NP137在PDO和一名接受治疗的患者肿瘤中均抑制了TFF3和干性转录本。阻断TFF3二聚化模拟了NP137的活性，确认TFF3是一个关键的下游效应因子。FOLFOX暴露上调了netrin-1和UNC5B，联合疗法（FOLFOX + NP137）在小鼠中显著减少了自我更新和肿瘤生长，同时降低了肿瘤内TFF3。
结论：Netrin-1通过UNC5B依赖性、TFF3介导的旁分泌存活机制维持mCRC CSC的自我更新。用NP137药理学抑制netrin-1可诱导CSC凋亡并增强化疗疗效，确认netrin-1/UNC5B/TFF3轴为克服转移性结直肠癌中干性驱动耐药的有前景的治疗靶点。
查看英文原文 English abstract
Purpose: Metastatic colorectal cancer (mCRC) exhibits poor outcomes due to recurrence and resistance to chemotherapy driven by cancer stem cells (CSCs). We investigated the role of the dependence receptor ligand netrin-1 and its receptor UNC5B in CSC self-renewal and evaluated therapeutic inhibition of netrin-1 using the anti-netrin-1 antibody NP137.
Experimental Procedures: Patient-derived organoids (PDOs) from colorectal liver metastases were treated with recombinant netrin-1, NP137, or controls. Self-renewal was quantified by extreme limiting dilution assays. UNC5B was silenced by CRISPR/Cas9 to determine receptor the implication of this Netrin-1 receptor. Single-cell RNA-sequencing elucidated signaling pathways affected by NP137. In vivo efficacy of NP137 alone or combined with FOLFOX chemotherapy was tested in PDO-xenografted mice, and paired tumor biopsies from an mCRC patient enrolled in an NP137 clinical trial were analyzed by RNA-seq.
Results: Netrin-1 enhanced CSC self-renewal and survival, effects abolished by NP137 or UNC5B knockout. NP137 treatment triggered CSC apoptosis, an effect reversed by caspase inhibition. Single-cell transcriptomics revealed that UNC5B-positive cells secreted trefoil factor 3 (TFF3), which acted paracrinally to maintain stemness gene expression (LGR5, SOX4, SMOC2, PROM1). NP137 suppressed TFF3 and stemness transcripts in both PDOs and a treated patient's tumor. Blocking TFF3 dimerization phenocopied NP137 activity, confirming TFF3 as a critical downstream effector. FOLFOX exposure upregulated netrin-1 and UNC5B, and combination therapy (FOLFOX + NP137) significantly reduced self-renewal and tumor growth in mice while decreasing intratumoral TFF3.
Conclusions: Netrin-1 sustains mCRC CSC self-renewal through an UNC5B-dependent, TFF3-mediated paracrine survival mechanism. Pharmacologic inhibition of netrin-1 with NP137 induces CSC apoptosis and enhances chemotherapy efficacy, identifying the netrin-1/UNC5B/TFF3 axis as a promising therapeutic target to overcome stemness-driven resistance in metastatic colorectal cancer.
利益披露 Disclosure
M. Brisset, None..
K. Radkova, None..
A. Paradisi, None..
L. Stephan, None..
R. Wagner, None..
F. Luiggi, None..
A. Heriot, None..
C. Behrenbruch, None..
T. Vu, None.