Ogawa Keita, Chiba Tetsuhiro, Nakamura Masato, Arai Jun, Zhang Jiaqi, Ma Yaojia, Qiang N A, Ao Junjie, Yumita Sae, Ishino Takamasa, Kan Motoyasu, Iwanaga Terunao, Nakagawa Miyuki, Fujiwara Kisako, Sakuma Takafumi, Kanzaki Hiroaki, Koroki Keisuke, Kusakabe Yuko, Kobayashi Kazufumi, Kanogawa Naoya, Kiyono Soichiro, Kondo Takayuki, Nakagawa Ryo, Ogasawara Sadahisa, Muroyama Ryosuke, Nakamoto Shingo, Kanda Tatsuo, Maruyama Hitoshi, Kato Jun, Matsumoto Shoji, Arai Takayoshi, Motohashi Shinichiro, Kato Naoya
Department of Gastroenterology, Graduate School of Medicine, Chiba University, Chiba, Japan.
Department of Gastroenterology, Graduate School of Medicine, Chiba University, Chiba, Japan;
Anticancer Res. 2023 Mar;43(3):1043-1052. doi: 10.21873/anticanres.16249.
BACKGROUND/AIM: MHC-class I-related chain A (MICA) functions as a ligand for natural killer group D, an activating receptor on natural killer (NK) cells, and its expression correlates with the carcinogenesis and progression of hepatocellular carcinoma (HCC). Although membranous MICA (mMICA) activates NK cells, soluble forms of MICA (sMICA), shed by cleaving enzymes, such as A disintegrin and metalloprotease (ADAM) 9, suppress NK cells. Therefore, the prevention of MICA shedding through the inhibition of ADAM9 has the potential to activate cancer immunity. Although we have discovered several ADAM inhibitors, many did not sufficiently activate NK cells without being cytotoxic, and, thus, new ADAM9 inhibitor candidates are needed.
To identify possible compounds for drug development, chemical library screening (a total of 741 compounds) was conducted using a fluorescence assay. Compounds with reduced fluorescence intensity were used as hit compounds in a subsequent analysis. Their impact on sMICA and mMICA in HCC cell lines was assessed using ELISA and flow cytometry, respectively. The cytotoxicity of NK cells was also evaluated by co-culturing NK cells with HCC cells.
CCL347, a symmetrical compound with five benzene rings, was identified as a hit compound. CCL347 significantly reduced sMICA levels in the culture medium supernatant with negligible cytotoxicity. Although mMICA was also reduced, CCL347 successfully enhanced NK cell cytotoxicity in co-cultures of NK cells and HCC cells.
CCL347 has potential as a novel therapeutic drug for HCC.
背景/目的：MHC I类相关链A（MICA）作为自然杀伤细胞D组（自然杀伤（NK）细胞上的一种激活受体）的配体发挥作用，其表达与肝细胞癌（HCC）的发生和进展相关。尽管膜结合型MICA（mMICA）可激活NK细胞，但由诸如解聚素和金属蛋白酶（ADAM）9等裂解酶所释放的可溶性MICA（sMICA）形式会抑制NK细胞。因此，通过抑制ADAM9来防止MICA脱落具有激活癌症免疫的潜力。尽管我们已经发现了几种ADAM抑制剂，但许多抑制剂在无细胞毒性的情况下并不能充分激活NK细胞，因此，需要新的ADAM9抑制剂候选物。
为了鉴定可能用于药物开发的化合物，使用荧光测定法进行了化学文库筛选（总共741种化合物）。荧光强度降低的化合物在后续分析中用作命中化合物。分别使用酶联免疫吸附测定（ELISA）和流式细胞术评估它们对HCC细胞系中sMICA和mMICA的影响。还通过将NK细胞与HCC细胞共培养来评估NK细胞的细胞毒性。
CCL347，一种具有五个苯环的对称化合物，被鉴定为命中化合物。CCL347显著降低了培养基上清液中的sMICA水平，且细胞毒性可忽略不计。尽管mMICA也有所降低，但CCL347在NK细胞与HCC细胞的共培养中成功增强了NK细胞的细胞毒性。
CCL347有潜力成为一种用于HCC的新型治疗药物。
