A Single-cell Perturbation Landscape of Colonic Stem Cell Polarisation
A Single-cell Perturbation Landscape of Colonic Stem Cell Polarisation
复制标题
DOI：
10.1101/2023.02.15.528008
复制
发表时间：
2023-02
期刊：
影响因子：
--
通讯作者：
X. Qin;Ferran Cardoso Rodriguez;J. Sufi;P. Vlckova;J. Claus;C. Tape
中科院分区：
文献类型：
--
作者：
X. Qin;Ferran Cardoso Rodriguez;J. Sufi;P. Vlckova;J. Claus;C. Tape
癌细胞受致癌突变和微环境信号的调控，但这些过程通常被单独研究。为了在功能上映射细胞内在和细胞外在线索如何共同调节结直肠癌（CRC）中的细胞命运，我们对1，071个结肠类器官培养物进行了系统的单细胞分析，这些培养物受1）CRC致癌突变，2）微环境成纤维细胞和巨噬细胞，3）基质配体和4）信号传导抑制剂的调节。多重单细胞分析揭示了由癌基因和基质配体的组合所决定的逐步上皮分化景观，从成纤维细胞诱导的LRIG 1+再生结肠干细胞（revCSC）到癌基因驱动的LRIG 1+过度增殖CSC（proCSC）。从revCSC到proCSC的转变通过减少WNT 3 A和TGF-β驱动的雅普信号传导和增加KRASG 12 D或基质EGF/上皮调节蛋白激活的MAPK/PI 3 K通量来调节。我们发现APC缺失和KRASG 12 D协同限制了对revCSC的访问，并破坏了基质-上皮通讯-在proCSC命运中捕获上皮细胞。这些结果表明，致癌突变占主导地位的稳态分化，阻碍细胞命运可塑性的细胞外源性调节。突出了由致癌和微环境线索调节的结肠干细胞极化的1，071条件单细胞转换图。成纤维细胞通过TGF-β1和雅普信号传导使WT结肠上皮朝向Clu+ revCSC极化。APC缺失和KRASG 12 D通过MAPK和PI 3 K驱动Birc 5+、Lrig 1+和Ephb 2 + proCSC命运。致癌突变破坏上皮可塑性的基质调节，使细胞陷入proCSC命运。
Cancer cells are regulated by oncogenic mutations and microenvironmental signals, yet these processes are often studied separately. To functionally map how cell-intrinsic and cell-extrinsic cues co-regulate cell-fate in colorectal cancer (CRC), we performed a systematic single-cell analysis of 1,071 colonic organoid cultures regulated by 1) CRC oncogenic mutations, 2) microenvironmental fibroblasts and macrophages, 3) stromal ligands, and 4) signalling inhibitors. Multiplexed single-cell analysis revealed a stepwise epithelial differentiation landscape dictated by combinations of oncogenes and stromal ligands, spanning from fibroblast-induced Clusterin (CLU)+ revival colonic stem cells (revCSC) to oncogene-driven LRIG1+ hyper-proliferative CSC (proCSC). The transition from revCSC to proCSC is regulated by decreasing WNT3A and TGF-β-driven YAP signalling and increasing KRASG12D or stromal EGF/Epiregulin-activated MAPK/PI3K flux. We find APC-loss and KRASG12D collaboratively limit access to revCSC and disrupt stromal-epithelial communication – trapping epithelia in the proCSC fate. These results reveal that oncogenic mutations dominate homeostatic differentiation by obstructing cell-extrinsic regulation of cell-fate plasticity. Highlights 1,071-condition single-cell transition map of colonic stem cell polarisation regulated by oncogenic and mircoenvironmental cues. Fibroblasts polarise WT colonic epithelia towards Clu+ revCSC via TGF-β1 and YAP signalling. APC-loss and KRASG12D drive a Birc5+, Lrig1+, and Ephb2+ proCSC fate via MAPK and PI3K. Oncogenic mutations disrupt stromal regulation of epithelial plasticity, trapping cells in the proCSC fate.