Caspase-4/11 is critical for angiogenesis by repressing Notch1 signalling via inhibiting γ-secretase activity
Caspase-4/11 is critical for angiogenesis by repressing Notch1 signalling via inhibiting γ-secretase activity
复制标题
DOI：
10.1111/bph.15904
复制
发表时间：
2022-07-24
影响因子：
7.3
通讯作者：
Pang, Jinjiang
中科院分区：
文献类型：
--
作者：
Fan, Linlin;Liu, Hao;Pang, Jinjiang
背景与目的：γ分泌酶介导的Notch1激活对血管生成至关重要。GeneCards数据库预测Caspase-4 （CASP4，与小鼠同源CASP11）与γ分泌酶的催化核心-早老素-1相互作用。因此，我们研究了CASP4/11在血管生成中的作用。实验方法：在体内，我们使用Casp11野生型和敲除小鼠研究了Casp11在几种血管生成小鼠模型中的作用。在体外，我们通过在人脐静脉内皮细胞（HUVECs）中消耗或过表达CASP4来检测CASP4对内皮功能和Notch信号的影响。通过诱变和共免疫沉淀检测CASP4与早老素-1结合的功能域。关键结果：Casp11缺乏会损害成人缺血后肢、黑色素瘤异种移植和Matrigel塞的血管生成，但不会影响视网膜的发育性血管生成。骨髓移植表明，促血管生成作用依赖于来自非造血细胞的CASP11。炎症因子诱导CASP4表达，CASP4敲低可降低huvec细胞活力、增殖、迁移和小管形成。机制上，CASP4/11缺失增加了体内和体外Notch1的激活，而CASP4过表达抑制了HUVECs中的Notch1信号传导。此外，CASP4敲低可增加γ -分泌酶活性。γ -分泌酶抑制剂DAPT恢复了CASP4 siRNA对HUVECs中Notch1激活和血管生成的影响。值得注意的是，CASP4/11的催化活性是不可缺少的。CASP4通过caspase募集结构域（CARD）直接与早老素-1相互作用。结论和意义：这些发现揭示了CASP4/11在成人血管生成中的关键作用，并使其成为未来血管生成相关疾病的有希望的治疗靶点。
Background and Purpose: Notch1 activation mediated by gamma-secretase is critical for angiogenesis. GeneCards database predicted that Caspase-4 (CASP4, with murine ortholog CASP11) interacts with presenilin-1, the catalytic core of gamma-secretase. Therefore, we investigated the role of CASP4/11 in angiogenesis.Experimental Approach: In vivo, we studied the role of Casp11 in several angiogenesis mouse models using Casp11 wild-type and knockout mice. In vitro, we detected the effects of CASP4 on endothelial functions and Notch signalling by depleting or overexpressing CASP4 in human umbilical vein endothelial cells (HUVECs). The functional domain responsible for the binding of CASP4 and presenilin-1 was detected by mutagenesis and co-immunoprecipitation. KeyResultS: Casp11 deficiency impaired adult angiogenesis in ischaemic hindlimbs, melanoma xenografts and Matrigel plugs, but not the developmental angiogenesis of retina. Bone marrow transplantation revealed that the pro-angiogenic effect depended on CASP11 derived from non-haematopoietic cells. CASP4 expression was induced by inflammatory factors and CASP4 knockdown decreased cell viability, proliferation, migration and tube formation in HUVECs. Mechanistically, CASP4/11 deficiency increased Notch1 activation in vivo and in vitro, while CASP4 overexpression repressed Notch1 signalling in HUVECs. Moreover, CASP4 knockdown increased gamma-secretase activity. The gamma-Secretase inhibitor DAPT restored the effects of CASP4 siRNA on Notch1 activation and angiogenesis in HUVECs. Notably, the catalytic activity of CASP4/11 was dispensable. CASP4 directly interacted with presenilin-1 through the caspase recruitment domain (CARD).Conclusions and Implications: these findings reveal a critical role of CASP4/11 in adult angiogenesis and make this molecule a promising therapeutic target for angiogenesis-related diseases in the future.