PO.ET03.06 · 实验与分子治疗
The anticancer effect of TROP2-targeted antibody-drug conjugates (ADC) is potentially attenuated by dysregulated cholesterol metabolism in cancer cells
作者与单位
摘要 Abstract
Background: Trop-2 is a widely expressed glycoprotein on a variety of different tumors. Datopotamab deruxtecan (Dato-DXd) is a Trop-2-targeted ADC that has demonstrated anticancer efficacy in patients with EGFR -mutated non-small cell lung cancer (NSCLC). However, their efficacy is inconsistent. In cancer cells, cholesterol synthesis is promoted, and cholesterol localizes to the cell membrane, playing a crucial role in the endocytosis of various receptors on the cell membrane. The current study aimed to explore the mechanisms underlying the limited efficacy of Dato-DXd using EGFR -mutated NSCLC cells.
Materials and methods: We established a Dato-DXd-resistant PC9DR cell line by long-term exposure of EGFR -mutated NSCLC PC9 cells to Dato-DXd. A colony formation assay was performed to evaluate anticancer effects. Trop-2 expression on the cell membrane was measured immunologically. Intracellular uptake of Dato-DXd was measured using a pH-Rhod method. Microarray data were analyzed using Transcriptome Analysis Console Software to examine differentially expressed genes and related-pathways.
Result: According to the colony formation assay, 1µg/mL Dato-DXd inhibited 89% colony formation in PC9 cells, but inhibited 5% colony formation in PC9DR cells. In PC9DR cells, cell surface Trop-2 expression was greater than PC9 cells (434,338 vs 400,722 molecules per cell, p = 0.0012). However, intracellular uptake of Dato-DXd was significantly reduced in PC9DR cells compared to PC9 cells (two-way ANOVA, p < 0.0001). These results suggested that reduced internalization of Trop-2 protein on the cell membrane is hypothetically associated with resistance to Dato-DXd. Second, pathway analysis revealed that the cholesterol metabolism pathway showed the most significant change in PC9DR cells compared to PC9 cells. Specifically, compared to PC9 cells, PC9DR cells showed a marked decrease in the expression of cholesterol synthesis-related enzymes, including HMGCR, MVD, and SQLE, while conversely exhibiting increased expression of the cholesterol transporter ABCA1 (ANOVA p<0.001, greater than two-fold change). Therefore, we hypothesized that impaired cholesterol metabolism cause resistance to Dato-DXd. Finally, we exposed the Dato-DXd-sensitive cells including PC9 and HCC827 to the HMGCoA reductase inhibitor, 1µM mevastatin, and evaluated anticancer effect of 1µg/mL Dato-DXd using a colony formation assay. In PC9 cells, Dato-DXd potently inhibited colony formation (relative colony formation area, Control; 100%, Dato-DXd; 3.6%). However, PC9 cells treated with mevastatin, Dato-DXd minimally inhibited colony formation (Control; 100%, Dato-DXd; 77.4%). Similar trend was observed in HCC827 cells.
Conclusion: These results suggest that impaired cholesterol metabolism may cause resistance to Dato-DXd in EGFR -mutated NSCLC.
利益披露 Disclosure
K. Yonesaka,
Daiichi-Sankyo Patent.
Y. Kawanaka, None.
T. Kurosaki,
Boehringer Ingelheim ).
Chugai Pharmaceutical Co., Ltd. ).
S. Watanabe, None..
J. Tanizaki, None..
K. Sakai, None..
T. Teramura, None.
K. Nishio,
Nippon Boehringer Ingelheim ).
West Japan Oncology Group ).
Nichirei Biosciences Inc. ).
Eli Lilly Japan ).
Hitachi ).
Sysmex ).
Otsuka Pharmaceutical ).
Thoracic Oncology Research Group ).
Okayama University ).
Japan Breast Cancer Research Group ).
H. Hayashi,
Boehringer Ingelheim ).
IQVIA Service JAPAN KK ).
Eisai Co., Ltd. ).
SYNEOS HEALTH CLINICAL KK ).
Shionogi & Co. Ltd. ).
EP-CRSU Co., Ltd ).
Nippon Kayaku Co.,Ltd. ).
EPS Corporation ).
GlaxoSmithKline KK ).
Otsuka Pharmaceutical ).
Chugai ).
MSD KK ).
Bristol Myers Squibb Company ).
Amgen Inc. ).
SRL Mediserch Inc. ).
Chugai Pharmaceutical Co. Ltd. ).
PRA Health Sciences Inc. ).
Nippon BOehringer Ingelheim CO.,Ltd. ).
CMIC CO., Ltd ).
Janssen Pharmaceutical KK ).