Functional single-cell analysis of the stem cell plasticity associated with drug-tolerant persister cells in colorectal cancer

与结直肠癌耐药持久细胞相关的干细胞可塑性的功能单细胞分析

基本信息

批准号：
22K15539
负责人：
COPPO Roberto
金额：
$ 2.91万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Early-Career Scientists
财政年份：
2022
资助国家：
日本
起止时间：
2022-04-01 至 2023-03-31
项目状态：
已结题

项目摘要

Using patient-derived colorectal cancer (CRC) organoids, we previously generated an analysis platform consisting of slow-growing CRC cells isolated from different human samples to represent the tumor heterogeneity. In this study, we tracked the fate of each cell through a clonogenic growth assay and found that the CRC cells showed a wide range of growth ability. Further rounds of the clonogenic growth assay revealed that the spheroid forming cells in CRC organoids consisted of distinct subpopulations; the cells generating large spheroids (L-cells) and the cells generating small spheroids (S-cells). The cells derived from the small spheroids gave rise to only small spheroids, consisting of slow-growing cells (S-pattern). While the cells derived from the large spheroids gave rise to both small and large spheroids, showing a dual-growing phenotype (D-pattern). Although the S-pattern spheroids never gave rise to large spheroids once isolated, transition to the D-pattern occurred by various extrinsic triggers, in which Musashi-1 (MSI1) played a key role. We revealed that the suppression of MSI1 in large spheroids, by using the CRISPR/Cas9 system, induced a transition from the D- to the S-pattern. We also found that the S-pattern spheroids were resistant to chemotherapy and transited to the D-pattern upon drug treatment. In conclusion, the isolated S-cells could be a novel platform for investigating drug-tolerant persister cells (DTPs) and developing the DTP targeting treatment. As the transition is linked to the drug resistance, it can be a therapeutic target.

使用患者来源的结直肠癌 (CRC) 类器官，我们之前生成了一个分析平台，该平台由从不同人类样本中分离出的缓慢生长的 CRC 细胞组成，以代表肿瘤异质性。在这项研究中，我们通过克隆生长测定追踪了每个细胞的命运，发现CRC细胞表现出广泛的生长能力。进一步的克隆生长测定表明，CRC 类器官中的球状形成细胞由不同的亚群组成；产生大球体的细胞（L 细胞）和产生小球体的细胞（S 细胞）。源自小球体的细胞仅产生由生长缓慢的细胞组成的小球体（S型）。而源自大球体的细胞则产生小球体和大球体，显示出双重生长表型（D型）。虽然 S 型球体一旦分离就不会产生大球体，但各种外在触发因素会导致向 D 型的转变，其中 Musashi-1 (MSI1) 发挥了关键作用。我们发现，通过使用 CRISPR/Cas9 系统抑制大球体中的 MSI1，诱导了从 D 型到 S 型的转变。我们还发现 S 型球体对化疗有抵抗力，并在药物治疗后转变为 D 型。总之，分离的 S 细胞可能成为研究耐药持久细胞 (DTP) 和开发 DTP 靶向治疗的新平台。由于这种转变与耐药性有关，因此它可以成为治疗靶点。