Novel p97 / VCP inhibitor induces endoplasmic reticulum stress and apoptosis in both bortezomib‐sensitive and ‐resistant multiple myeloma cells
Novel p97 / VCP inhibitor induces endoplasmic reticulum stress and apoptosis in both bortezomib‐sensitive and ‐resistant multiple myeloma cells
复制标题
新型 p97 / VCP 抑制剂可诱导硼替佐米敏感和耐药的多发性骨髓瘤细胞内质网应激和细胞凋亡
DOI：
10.1111/cas.14154
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发表时间：
2019
期刊：
影响因子：
5.7
通讯作者：
Okuno Yutaka
中科院分区：
文献类型：
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作者：
Nishimura Nao;Radwan Mohamed O.;Amano Masayuki;Endo Shinya;Fujii Eri;Hayashi Hironori;Ueno Shikiko;Ueno Niina;Tatetsu Hiro;Hata Hiroyuki;Okamoto Yoshinari;Otsuka Masami;Mitsuya Hiroaki;Matsuoka Masao;Okuno Yutaka
P97/VCP是一种内质网(ER)相关蛋白，属于AAA(与多种细胞活动相关的ATPase)ATPase家族。它具有多种细胞功能，包括内质网相关蛋白的降解、自噬和侵袭体的形成。最近的研究表明，p97/VCP在包括多发性骨髓瘤(MM)在内的几种癌症亚型中的作用及其作为治疗靶点的潜力。我们进行了基于细胞的化合物筛选，以开发在骨髓瘤细胞中具有细胞毒活性的新的小分子化合物。在约2,000个化合物中，OSSL_325096对MM细胞具有较强的抗增殖活性(IC_(50)，100-500nmol/L)。OSSL_325096可诱导骨髓瘤细胞系的凋亡，包括对硼替佐米耐药的骨髓瘤细胞系和从患者体内纯化的原代骨髓瘤细胞。在用OSSL325096处理的MM细胞系中，观察到多泛素化蛋白PERK、CHOP和IREα的积累，这表明它诱导了MM细胞的内质网应激。OSSl_325096与已知的p97/VCP抑制剂DBeQ具有相似的化学结构。P97/VCP基因敲除可诱导骨髓瘤细胞发生凋亡，并伴随多泛素化蛋白的积聚。OSSL_325096对骨髓瘤细胞株的IC50(0.1%~0.8%μ摩尔/L)低于苯乙醇组(2~5%μ摩尔/L)。蛋白质药物结合模拟表明OSSL325096可能与p97/VCP的D2区的三磷酸腺苷结合位点结合。在无细胞ATPase实验中，OSSL325096呈剂量依赖性抑制p97/VCPATPase活性。最后，OSSl_325096在体内抑制了皮下骨髓瘤细胞瘤的生长。本研究结果提示OSSL325096具有抗骨髓瘤作用，至少部分是通过抑制p97/vcp发挥作用的。
p97/VCP is an endoplasmic reticulum (ER)‐associated protein that belongs to the AAA (ATPases associated with diverse cellular activities) ATPase family. It has a variety of cellular functions including ER‐associated protein degradation, autophagy, and aggresome formation. Recent studies have shown emerging roles of p97/VCP and its potential as a therapeutic target in several cancer subtypes including multiple myeloma (MM). We conducted a cell‐based compound screen to exploit novel small compounds that have cytotoxic activity in myeloma cells. Among approximately 2000 compounds, OSSL_325096 showed relatively strong antiproliferative activity in MM cell lines (IC50, 100‐500 nmol/L). OSSL_325096 induced apoptosis in myeloma cell lines, including a bortezomib‐resistant cell line and primary myeloma cells purified from patients. Accumulation of poly‐ubiquitinated proteins, PERK, CHOP, and IREα, was observed in MM cell lines treated with OSSL_325096, suggesting that it induces ER stress in MM cells. OSSL_325096 has a similar chemical structure to DBeQ, a known p97/VCP inhibitor. Knockdown of the gene encoding p97/VCP induced apoptosis in myeloma cells, accompanied by accumulation of poly‐ubiquitinated protein. IC50of OSSL_325096 to myeloma cell lines were found to be lower (0.1‐0.8 μmol/L) than those of DBeQ (2‐5 μmol/L). In silico protein–drug‐binding simulation suggested possible binding of OSSL_325096 to the ATP binding site in the D2 domain of p97/VCP. In cell‐free ATPase assays, OSSL_325096 showed dose‐dependent inhibition of p97/VCP ATPase activity. Finally, OSSL_325096 inhibited the growth of subcutaneous myeloma cell tumors in vivo. The present data suggest that OSSL_325096 exerts anti‐myeloma activity, at least in part through p97/VCP inhibition.