谱系特异性诱导多能干细胞衍生平滑肌细胞建模预测整合素 α-V 拮抗作用可减少马凡综合征小鼠的主动脉根部动脉瘤形成。
Lineage-Specific Induced Pluripotent Stem Cell-Derived Smooth Muscle Cell Modeling Predicts Integrin Alpha-V Antagonism Reduces Aortic Root Aneurysm Formation in Marfan Syndrome Mice.
机构信息
Department of Cardiothoracic Surgery (K.N., A.R.D., N.Y., A.J.P., S.K., O.M., C.G., B.M., M.L., W.H., M.P.F.).
Department of Comparative Medicine (K.M.C.), Stanford University School of Medicine, CA.
出版信息
Arterioscler Thromb Vasc Biol. 2023 Jul;43(7):1134-1153. doi: 10.1161/ATVBAHA.122.318448. Epub 2023 Apr 20.
BACKGROUND
The role of increased smooth muscle cell (SMC) integrin αv signaling in Marfan syndrome (MFS) aortic aneurysm remains unclear. Herein, we examine the mechanism and potential efficacy of integrin αv blockade as a therapeutic strategy to reduce aneurysm progression in MFS.
METHODS
Induced pluripotent stem cells (iPSCs) were differentiated into aortic SMCs of the second heart field (SHF) and neural crest (NC) lineages, enabling in vitro modeling of MFS thoracic aortic aneurysms. The pathological role of integrin αv during aneurysm formation was confirmed by blockade of integrin αv with GLPG0187 in MFS mice.
RESULTS
iPSC-derived MFS SHF SMCs overexpress integrin αv relative to MFS NC and healthy control SHF cells. Furthermore, integrin αv downstream targets (FAK [focal adhesion kinase]/Akt/mTORC1 [mechanistic target of rapamycin complex 1]) were activated, especially in MFS SHF. Treatment of MFS SHF SMCs with GLPG0187 reduced p-FAK/p-Akt/mTORC1 activity back to control SHF levels. Functionally, MFS SHF SMCs had increased proliferation and migration compared to MFS NC SMCs and control SMCs, which normalized with GLPG0187 treatment. In the MFS mouse model, integrin αv, p-Akt, and downstream targets of mTORC1 proteins were elevated in the aortic root/ascending segment compared to littermate wild-type control. Mice treated with GLPG0187 (age 6-14 weeks) had reduced aneurysm growth, elastin fragmentation, and reduction of the FAK/Akt/mTORC1 pathway. GLPG0187 treatment reduced the amount and severity of SMC modulation assessed by single-cell RNA sequencing.
CONCLUSIONS
The integrin αv-FAK-Akt signaling pathway is activated in iPSC SMCs from MFS patients, specifically from the SHF lineage. Mechanistically, this signaling pathway promotes SMC proliferation and migration in vitro. As biological proof of concept, GLPG0187 treatment slowed aneurysm growth and p-Akt signaling in mice. Integrin αv blockade via GLPG0187 may be a promising therapeutic approach to inhibit MFS aneurysmal growth.
背景
平滑肌细胞（SMC）整合素 αv 信号增强在马凡综合征（MFS）主动脉瘤中的作用尚不清楚。在此，我们研究整合素 αv 阻断作为一种治疗策略的机制和潜在疗效，以减少 MFS 中的动脉瘤进展。
方法
诱导多能干细胞（iPSC）分化为第二心区（SHF）和神经嵴（NC）谱系的主动脉 SMC，能够体外模拟 MFS 胸主动脉瘤。通过在 MFS 小鼠中用 GLPG0187 阻断整合素 αv ，证实了整合素 αv 在动脉瘤形成过程中的病理作用。
结果
iPSC 衍生的 MFS SHF SMC 相对于 MFS NC 和健康对照 SHF 细胞过表达整合素 αv。此外，整合素 αv 下游靶标（粘着斑激酶（FAK）/Akt/mTORC1 [雷帕霉素靶蛋白复合物 1]）被激活，尤其是在 MFS SHF 中。用 GLPG0187 处理 MFS SHF SMC 可使 p-FAK/p-Akt/mTORC1 活性恢复至对照 SHF 水平。功能上，与 MFS NC SMC 和对照 SMC 相比，MFS SHF SMC 的增殖和迁移增加，而用 GLPG0187 处理则正常化。在 MFS 小鼠模型中，与同窝野生型对照相比，主动脉根部/升段中的整合素 αv、p-Akt 和 mTORC1 下游靶蛋白升高。用 GLPG0187 （6-14 周龄）治疗的小鼠的动脉瘤生长、弹性蛋白片段化和 FAK/Akt/mTORC1 途径减少。GLPG0187 治疗减少了通过单细胞 RNA 测序评估的 SMC 调节的数量和严重程度。
结论
整合素 αv-FAK-Akt 信号通路在来自 MFS 患者的 iPSC SMC 中被激活，特别是来自 SHF 谱系的 SMC。从机制上讲，这种信号通路促进了体外 SMC 的增殖和迁移。作为生物学概念验证，GLPG0187 治疗减缓了小鼠的动脉瘤生长和 p-Akt 信号。通过 GLPG0187 阻断整合素 αv 可能是抑制 MFS 动脉瘤生长的一种很有前途的治疗方法。
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