Nishikubo Kou, Ohgaki Ryuichi, Liu Xingming, Okanishi Hiroki, Xu Minhui, Endou Hitoshi, Kanai Yoshikatsu
Department of Bio-system Pharmacology, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka, 565-0871, Japan.
Integrated Frontier Research for Medical Science Division, Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University, Osaka, Japan.
Cancer Cell Int. 2023 Jun 15;23(1):116. doi: 10.1186/s12935-023-02957-z.
Cytotoxic anticancer drugs widely used in cancer chemotherapy have some limitations, such as the development of side effects and drug resistance. Furthermore, monotherapy is often less effective against heterogeneous cancer tissues. Combination therapies of cytotoxic anticancer drugs with molecularly targeted drugs have been pursued to solve such fundamental problems. Nanvuranlat (JPH203 or KYT-0353), an inhibitor for L-type amino acid transporter 1 (LAT1; SLC7A5), has novel mechanisms of action to suppress the cancer cell proliferation and tumor growth by inhibiting the transport of large neutral amino acids into cancer cells. This study investigated the potential of the combined use of nanvuranlat and cytotoxic anticancer drugs.
The combination effects of cytotoxic anticancer drugs and nanvuranlat on cell growth were examined by a water-soluble tetrazolium salt assay in two-dimensional cultures of pancreatic and biliary tract cancer cell lines. To elucidate the pharmacological mechanisms underlying the combination of gemcitabine and nanvuranlat, we investigated apoptotic cell death and cell cycle by flow cytometry. The phosphorylation levels of amino acid-related signaling pathways were analyzed by Western blot. Furthermore, growth inhibition was examined in cancer cell spheroids.
All the tested seven types of cytotoxic anticancer drugs combined with nanvuranlat significantly inhibited the cell growth of pancreatic cancer MIA PaCa-2 cells compared to their single treatment. Among them, the combined effects of gemcitabine and nanvuranlat were relatively high and confirmed in multiple pancreatic and biliary tract cell lines in two-dimensional cultures. The growth inhibitory effects were suggested to be additive but not synergistic under the tested conditions. Gemcitabine generally induced cell cycle arrest at the S phase and apoptotic cell death, while nanvuranlat induced cell cycle arrest at the G0/G1 phase and affected amino acid-related mTORC1 and GAAC signaling pathways. In combination, each anticancer drug basically exerted its own pharmacological activities, although gemcitabine more strongly influenced the cell cycle than nanvuranlat. The combination effects of growth inhibition were also verified in cancer cell spheroids.
Our study demonstrates the potential of first-in-class LAT1 inhibitor nanvuranlat as a concomitant drug with cytotoxic anticancer drugs, especially gemcitabine, on pancreatic and biliary tract cancers.
广泛应用于癌症化疗的细胞毒性抗癌药物存在一些局限性，如副作用的产生和耐药性。此外，单一疗法对异质性癌组织往往效果较差。为解决这些基本问题，人们一直在探索细胞毒性抗癌药物与分子靶向药物的联合疗法。南伏拉兰特（JPH203或KYT - 0353）是一种L型氨基酸转运体1（LAT1；SLC7A5）抑制剂，具有通过抑制大中性氨基酸进入癌细胞来抑制癌细胞增殖和肿瘤生长的新作用机制。本研究探讨了南伏拉兰特与细胞毒性抗癌药物联合使用的潜力。
通过水溶性四氮唑盐试验在胰腺和胆道癌细胞系的二维培养物中检测细胞毒性抗癌药物和南伏拉兰特对细胞生长的联合作用。为阐明吉西他滨与南伏拉兰特联合使用的药理机制，我们通过流式细胞术研究了细胞凋亡和细胞周期。通过蛋白质免疫印迹法分析氨基酸相关信号通路的磷酸化水平。此外，还检测了癌细胞球体中的生长抑制情况。
与单一治疗相比，所测试的七种细胞毒性抗癌药物与南伏拉兰特联合使用均显著抑制了胰腺癌MIA PaCa - 2细胞的生长。其中，吉西他滨与南伏拉兰特的联合效果相对较高，并在二维培养的多种胰腺和胆道细胞系中得到证实。在所测试的条件下，生长抑制作用表现为相加而非协同作用。吉西他滨通常诱导细胞周期停滞在S期并导致细胞凋亡，而南伏拉兰特诱导细胞周期停滞在G0/G1期并影响与氨基酸相关的mTORC1和GAAC信号通路。联合使用时，每种抗癌药物基本上发挥其自身的药理活性，尽管吉西他滨对细胞周期的影响比南伏拉兰特更强。生长抑制的联合效果也在癌细胞球体中得到验证。
我们的研究证明了一流的LAT1抑制剂南伏拉兰特作为细胞毒性抗癌药物（尤其是吉西他滨）的伴随药物用于治疗胰腺癌和胆管癌的潜力。
