Aggresomes are the product of misfolded protein aggregation, and the presence of aggresomes has been correlated with poor prognosis in cancer patients. However, the exact role of aggresomes in tumorigenesis and cancer progression remains largely unknown. Herein, the multiomics screening reveal that OTUD1 protein plays an important role in retaining ovarian cancer stem cell (OCSC) properties. Mechanistically, the elevated OTUD1 protein levels lead to the formation of OTUD1-based cytoplasmic aggresomes, which is mediated by a short peptide located in the intrinsically disordered OTUD1 N-terminal region. Furthermore, OTUD1-based aggresomes recruit ASK1 via protein-protein interactions, which in turn stabilize ASK1 in a deubiquitinase-independent manner and activate the downstream JNK signaling pathway for OCSC maintenance. Notably, the disruption of OTUD1-based aggresomes or treatment with ASK1/JNK inhibitors, including ibrutinib, an FDA-approved drug that was recently identified as an MKK7 inhibitor, effectively reduced OCSC stemness (OSCS) of OTUD1^high ovarian cancer cells. In summary, our work suggests that aggresome formation in tumor cells could function as a signaling hub and that aggresome-based therapy has translational potential for patients with OTUD1^high ovarian cancer.

聚集体是错误折叠的蛋白质聚集的产物，聚集体的存在与癌症患者的不良预后相关。然而，聚集体在肿瘤发生和癌症进展中的确切作用仍然很大程度上未知。在此，多组学筛选揭示了 OTUD1 蛋白在保留卵巢癌干细胞 (OCSC) 特性方面发挥着重要作用。从机制上讲，OTUD1 蛋白水平升高导致基于 OTUD1 的细胞质聚集体的形成，这是由位于本质上无序的 OTUD1 N 末端区域的短肽介导的。此外，基于 OTUD1 的聚集体通过蛋白质-蛋白质相互作用招募 ASK1，进而以不依赖于去泛素酶的方式稳定 ASK1，并激活下游 JNK 信号通路以维持 OCSC。值得注意的是，破坏基于 OTUD1 的聚集体或使用 ASK1/JNK 抑制剂（包括 FDA 批准的药物依鲁替尼，最近被确定为 MKK7 抑制剂）治疗，可有效降低 OTUD1^高卵巢癌细胞的 OCSC 干性（OSCS）。总之，我们的工作表明肿瘤细胞中的聚集体形成可以充当信号中枢，并且基于聚集体的治疗对于 OTUD1^高卵巢癌患者具有转化潜力。