OTUB1通过调节糖酵解催化非依赖性地去泛素化TGFBI并介导婴儿血管瘤中的血管生成。
OTUB1 Catalytic-Independently Deubiquitinates TGFBI and Mediates the Angiogenesis in Infantile Hemangioma by Regulating Glycolysis.
作者信息
Li Mingyang, Wang Xuan, Yang Enli, Li Yiming, Geng Yiming, Chen Zhanwei, Huang Shengyun, Zhang Dongsheng, Wu Haiwei
机构信息
Department of Stomatology, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, China (M.L., X.W., E.Y., Y.L., Y.G., Z.C., S.H., D.Z., H.W.).
Department of Stomatology, Shandong Provincial Hospital, Shandong University, China (M.L., X.W., E.Y., Y.L., Y.G., Z.C., S.H., D.Z., H.W.).
出版信息
Arterioscler Thromb Vasc Biol. 2023 May;43(5):654-673. doi: 10.1161/ATVBAHA.123.319177. Epub 2023 Mar 30.
BACKGROUND
Infantile hemangioma (IH) arises as a result of dysregulation of both angiogenesis and vasculogenesis. The deubiquitylase OTUB1 (OTU domain, ubiquitin aldehyde binding 1) has been reported to play an essential role in multiple cancers; however, its function in the progression of IH and the underlying mechanisms regulating angiogenesis remain unclear.
METHODS
Transwell assays, EdU assays, and tube formation assays were performed to investigate the biological behavior of IH in vitro. IH animal models were established to estimate the progression of IH in vivo. Mass spectrometric analysis were conducted to detect the downstream of OTUB1 and ubiquitination sites of transforming growth factor beta induced (TGFBI). Half-life assays and ubiquitination test were performed to investigate the interaction between TGFBI and OTUB1. Extracellular acidification rate assays were employed to estimate the glycolysis level in IH.
RESULTS
The expression of OTUB1 was obviously increased in proliferating IH as compared to the involuting and involuted IH tissues. Through in vitro experiments, the knockdown of OTUB1 inhibited the proliferation, migration and tube formation of human hemangioma endothelial cells, while the overexpression of OTUB1 promoted the proliferation, migration and angiogenic abilities of human hemangioma endothelial cells. The knockdown of OTUB1 significantly suppressed IH progression in vivo. Furthermore, TGFBI was predicted as a functional downstream target of OTUB1 in IH by mass spectrometry. Mechanistically, OTUB1 interacted with and deubiquitylated TGFBI on the K22 and K25 residues, which was demonstrated to be independent of the catalytic activity of OTUB1. The inhibitory effects of OTUB1 knockdown on cell proliferation, migration and tube formation ability of human hemangioma endothelial cells were reversed by TGFBI overexpression. Further, we found that OTUB1 mediated glycolysis by regulating TGFBI in infantile hemangioma.
CONCLUSIONS
OTUB1 deubiquitinates TGFBI in a catalytic-independent manner and promotes angiogenesis in infantile hemangioma by regulating glycolysis. Targeting OTUB1 might be an effective therapeutic strategy for inhibiting IH progression and tumor angiogenesis.
背景
婴儿血管瘤（IH）是血管生成和血管发生失调的结果。去泛素化酶OTUB1（OTU结构域，泛素醛结合蛋白1）已被报道在多种癌症中起重要作用；然而，其在IH进展中的功能以及调节血管生成的潜在机制仍不清楚。
方法
进行Transwell实验、EdU实验和管形成实验以研究IH在体外的生物学行为。建立IH动物模型以评估IH在体内的进展。进行质谱分析以检测OTUB1的下游和转化生长因子β诱导（TGFBI）的泛素化位点。进行半衰期实验和泛素化测试以研究TGFBI与OTUB1之间的相互作用。采用细胞外酸化率实验来评估IH中的糖酵解水平。
结果
与消退期和消退后的IH组织相比，OTUB1在增殖期IH中的表达明显增加。通过体外实验，敲低OTUB1可抑制人血管瘤内皮细胞的增殖、迁移和管形成，而OTUB1的过表达则促进人血管瘤内皮细胞的增殖、迁移和血管生成能力。敲低OTUB1可显著抑制体内IH的进展。此外，通过质谱分析预测TGFBI是IH中OTUB1的功能性下游靶点。机制上，OTUB1与TGFBI在K22和K25残基上相互作用并使其去泛素化，这被证明独立于OTUB1的催化活性。TGFBI过表达可逆转OTUB1敲低对人血管瘤内皮细胞增殖、迁移和管形成能力的抑制作用。此外，我们发现OTUB1通过调节TGFBI介导婴儿血管瘤中的糖酵解。
结论
OTUB1以催化非依赖的方式使TGFBI去泛素化，并通过调节糖酵解促进婴儿血管瘤中的血管生成。靶向OTUB1可能是抑制IH进展和肿瘤血管生成的有效治疗策略。
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