Crenolanib Regulates ERK and AKT/mTOR Signaling Pathways in RAS/BRAF-Mutated Colorectal Cancer Cells and Organoids
Crenolanib Regulates ERK and AKT/mTOR Signaling Pathways in RAS/BRAF-Mutated Colorectal Cancer Cells and Organoids
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DOI：
10.1158/1541-7786.mcr-20-0600
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发表时间：
2021-05-01
影响因子：
5.2
通讯作者：
Eguchi, Hidetoshi
中科院分区：
文献类型：
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作者：
Fujino, Shiki;Miyoshi, Norikatsu;Eguchi, Hidetoshi
最近开发的分子靶向治疗，如EGFR抑制剂，显著改善了癌症患者的预后。然而，患有KRAS和BRAF突变的患者目前并没有从这些治疗中受益。在这里，我们的目标是研究Crenolanib作为一种新的分子靶向治疗结直肠癌的潜在作用。我们使用多个结直肠癌细胞株来研究crenolanib的生长抑制作用及其与其他细胞毒剂的联合作用。患者衍生器官(PDO)的原代培养是一种反映临床结直肠癌异质性的模型，用来进一步验证crenolanib的效果。与西妥昔单抗不同，crenolanib显著抑制BRAF突变的HT29细胞和KRAS突变的HCT116细胞的ERK和AKT/mTOR通路，并具有相应的生长抑制作用。Crenolanib与其他细胞毒药物联合治疗时，可观察到相加或协同作用。此外，crenolanib抑制干细胞标记物的表达，如OCT4、NANOG和SOX2。这些观察结果在7个带有KRAS突变的PDO和2个没有KRAS/BRAF突变的PDO中得到证实，crenolanib抑制了所有PDO的生长，而无论它们的KRAS突变状态如何。此外，crenolanib可阻断PDGF和转化生长因子β诱导的PDO中OCT4阳性细胞的增加。总之，这些发现提示Crenolanib可能对结直肠癌患者，特别是KRAS/BRAF突变患者具有临床实用价值。
Recently developed molecularly targeted therapies such as EGFR inhibitors have notably improved the prognosis of patients with cancer. However, patients with KRAS and BRAF mutations do not currently benefit from these therapies. Here, we aimed to examine potential effects of crenolanib as a new molecularly targeted therapy in colorectal cancer. We used multiple colorectal cancer cell lines to investigate the growth-inhibitory effect of crenolanib and its effect in combination with other cytotoxic agents. Primary cultures of patient-derived organoids (PDO), a model that reflects the heterogeneity of clinical colorectal cancer, were used to further validate the effects of crenolanib. Unlike cetuximab, crenolanib remarkably suppressed ERK and AKT/mTOR pathways in HT29 cells with BRAF mutation and in HCT116 cells with KRAS mutation with corresponding growth-suppressing effects. Additive or synergistic effects were observed in treatments with combination of crenolanib and other cytotoxic drugs. Moreover, crenolanib suppressed the expression of stem cell markers, such as OCT4, NANOG, and SOX2. These observations were substantiated in seven PDOs with KRAS mutation and two PDOs without KRAS/BRAF mutations, with crenolanib suppressing the growth of all PDOs regardless of their KRAS mutation status. Furthermore, crenolanib abrogated PDGF- and TGF beta-induced increase of OCT4-positive cells in PDOs. Together, these findings suggest that crenolanib may have clinical utility for patients with colorectal cancer, especially patients with KRAS/BRAF mutations.Implications: These findings indicate that crenolanib can be a useful target agent for patients with colorectal cancer, especially patients with KRAS/BRAF mutations.