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）器官，我们先前生成了一个分析平台，该平台由从不同人类样品中分离出来的缓慢生长的CRC细胞组成，以表示肿瘤异质性。在这项研究中，我们通过克隆生长测定法追踪了每个细胞的命运，发现CRC细胞显示出广泛的生长能力。克隆生长测定的进一步回合表明，CRC器官中的球体形成细胞由不同的亚群组成。产生大球体（L细胞）的细胞和产生小球体（S细胞）的细胞。源自小球体的细胞仅引起小球体，由缓慢生长的细胞（S-Pattern）组成。而源自大球体的细胞产生了小球体和大型球体，显示了双重生长表型（D-Pattern）。尽管S-Pattern球体从未产生较大的球体，但曾经隔离开来，但各种外部触发器都会过渡到D-Pattern，其中Musashi-1（MSI1）起着关键作用。我们揭示了通过使用CRISPR/CAS9系统在大球体中对MSI1的抑制，从而诱导了从D-到S-Pattern的过渡。我们还发现，S-Pattern球体对化学疗法有抵抗力，并在药物治疗后转移到D-Pattern。总之，孤立的S细胞可能是研究耐药持久性细胞（DTP）并开发DTP靶向处理的新型平台。由于过渡与耐药性有关，因此可以是治疗靶标。