Bhalla Sohni Ria, Wahid Mussarat, Amartey Jason O, Zawia Amira, Riu Federica, Gao Yizhuo, Agrawal Jyoti, Lynch Amy P, Machado Maria J C, Hawtrey Tom, Kikuchi Ryosuke, Green Kathryn R, Teboul Lydia, Allen Claire, Blackley Zoe, Rajaji Keerthana, Batson Jennifer, Harper Steven J, Oltean Sebastian, Amoaku Winfried, Benest Andrew V, Morris Jonathan C, Braithwaite Bruce, Bates David O
Division of Cancer and Stem Cells, Tumour and Vascular Biology Laboratories, Centre for Cancer Sciences, School of Medicine, Biodiscovery Institute, University of Nottingham, Nottingham, United Kingdom.
School of Chemistry, University of New South Wales, Sydney, New South Wales, Australia.
Am J Physiol Heart Circ Physiol. 2025 Aug 1;329(2):H490-H504. doi: 10.1152/ajpheart.00564.2024. Epub 2025 Jun 30.
In peripheral arterial disease (PAD), antiangiogenic VEGF-Ab isoform overexpression in monocytes contributes to impaired collateralization. Serine-arginine protein-kinase-1 (SRPK1) regulates VEGF splicing. To determine whether SRPK1 controlled monocytic VEGF, impairing collateralization, we investigated SRPK1 inhibition and monocyte-specific knockout in mouse models of and in human monocytes from PAD. VEGF-Ab activity was measured by coculture of monocytes from patients with PAD with endothelial cells with SRPK1 inhibition. Mice with impaired revascularization due to soluble-frizzled-related-protein-5 knockout (), monocyte-specific Wnt5a gain-of-function (), or obese mice on a high-fat high-sucrose (HF/HS) diet were subjected to femoral artery ligation and treated with a SRPK1 inhibitor. We generated an SRPK1 conditional knockout and crossed it with monocyte-specific (LysM-Cre) driver line to specifically knockout SRPK1 in monocyte lineages. Blood flow was measured by laser speckle imaging before, and for 28 days after surgery. Monocytes from patients with PAD significantly inhibited endothelial cell migration, which was reversed by an anti-VEGF-Ab antibody. Surprisingly, migration was stimulated by SRPK1 inhibition, switching splicing from VEGF-Ab to VEGF-Aa. In , , and HF/HS mouse models of PAD, blood flow was improved by SRPK1 inhibition. Impaired revascularization in mice was rescued in : mice, which had a phenotypic shift toward M2 macrophages. Impaired blood flow recovery was also rescued in obese- mice. VEGF splicing in monocytes is regulated differently from VEGF splicing in epithelial or cancer cells suggesting that control of splicing is dependent on cell type and/or environment. SRPK1 inhibition enhances collateralization in mice, and in human in vitro models of monocyte-dependent impaired angiogenesis. A novel potential treatment for peripheral arterial disease (PAD) is described. Inhibition of SRPK1, or knockout in monocytes, induces angiogenesis by preventing splicing to antiangiogenic VEGF (VEGF-Ab) in patients and animal models. In PAD, monocyte splicing control is different from other cell types and SRPK1 inhibition by drug-like compounds can alter macrophage phenotype and reverse PAD in mice. A new cell-specific SRPK1-LoxP mouse is described.
在周围动脉疾病（PAD）中，单核细胞中抗血管生成的血管内皮生长因子-Ab（VEGF-Ab）亚型的过表达会导致侧支循环受损。丝氨酸-精氨酸蛋白激酶-1（SRPK1）调节VEGF的剪接。为了确定SRPK1是否控制单核细胞的VEGF从而损害侧支循环，我们在PAD小鼠模型以及来自PAD患者的人类单核细胞中研究了SRPK1抑制和单核细胞特异性敲除。通过对PAD患者的单核细胞与经SRPK1抑制处理的内皮细胞进行共培养来测量VEGF-Ab活性。对因可溶性卷曲相关蛋白-5敲除（）、单核细胞特异性Wnt5a功能获得（）或高脂高糖（HF/HS）饮食的肥胖小鼠导致血管重建受损的小鼠进行股动脉结扎，并给予SRPK1抑制剂治疗。我们构建了一个SRPK1条件性敲除小鼠，并将其与单核细胞特异性（LysM-Cre）驱动系杂交，以特异性敲除单核细胞谱系中的SRPK1。在手术前以及手术后28天通过激光散斑成像测量血流量。PAD患者的单核细胞显著抑制内皮细胞迁移，而抗VEGF-Ab抗体可逆转这种抑制作用。令人惊讶的是，SRPK1抑制会刺激迁移，使剪接从VEGF-Ab转变为VEGF-Aa。在PAD的、和HF/HS小鼠模型中，SRPK1抑制可改善血流量。小鼠中受损的血管重建在小鼠中得到挽救，这些小鼠表现出向M2巨噬细胞的表型转变。肥胖小鼠中受损的血流恢复也得到了挽救。单核细胞中的VEGF剪接与上皮细胞或癌细胞中的VEGF剪接受不同调节，这表明剪接的控制取决于细胞类型和/或环境。SRPK1抑制可增强小鼠以及人类单核细胞依赖性血管生成受损体外模型中的侧支循环。本文描述了一种针对周围动脉疾病（PAD）的新型潜在治疗方法。在患者和动物模型中抑制SRPK1或在单核细胞中敲除SRPK1，可通过阻止剪接为抗血管生成的VEGF（VEGF-Ab）来诱导血管生成。在PAD中，单核细胞的剪接控制与其他细胞类型不同，并且类似药物化合物对SRPK1的抑制可改变巨噬细胞表型并逆转小鼠的PAD。本文描述了一种新的细胞特异性SRPK1-LoxP小鼠。
