Matrix stiffness primes lymphatic tube formation directed by vascular endothelial growth factor-C.
Matrix stiffness primes lymphatic tube formation directed by vascular endothelial growth factor-C.
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DOI：
10.1096/fj.202002426rr
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发表时间：
2021-05
期刊：
影响因子：
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通讯作者：
Hanjaya-Putra D
中科院分区：
文献类型：
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作者：
Alderfer L;Russo E;Archilla A;Coe B;Hanjaya-Putra D
淋巴系统功能障碍与多种疾病表型相关。功能失调淋巴管的修复被认为是一种创新的方法，可以拯救疾病状态下的健康表型，包括神经系统疾病、代谢综合征和心血管疾病。与血管系统相比，我们对控制淋巴管形态发生的分子调控知之甚少。利用合成透明质酸（HA）水凝胶作为化学和机械可调节的系统来保护淋巴内皮细胞（LECs）表型，我们证明了低基质弹性在高浓度血管内皮生长因子- c （VEGF-C）的指导下促进淋巴索样结构（CLS）的形成。底物硬度降低导致关键淋巴标志物上调，包括PROX-1、淋巴管内皮透明质酸受体1 （LYVE-1）和VEGFR-3。因此，较高水平的VEGFR-3可以用VEGF-C刺激LEC，这是激活基质金属蛋白酶（MMPs）和促进LEC迁移所必需的。这两个步骤都是在体外建立CLS形成的关键。随着底物弹性的降低，我们观察到MMP表达增加，细胞伸长增加，以及细胞内液泡的形成，这些液泡可以进一步合并成凝聚液泡。RNAi研究表明，MMP-14是CLS形成的必要条件，LECs通过YAP/TAZ机械传感器感知基质刚度，从而激活其下游靶基因。总之，我们表明，通过调节基质刚度和VEGF-C浓度，CLS形成的信号通路可以在合成基质中调节，从而产生淋巴网络，这将有助于淋巴生物学的研究和未来的组织再生方法。
Dysfunction of the lymphatic system is associated with a wide range of disease phenotypes. The restoration of dysfunctional lymphatic vessels has been hypothesized as an innovative method to rescue healthy phenotypes in diseased states including neurological conditions, metabolic syndromes, and cardiovascular disease. Compared to the vascular system, little is known about the molecular regulation that controls lymphatic tube morphogenesis. Using synthetic hyaluronic acid (HA) hydrogels as a chemically and mechanically tunable system to preserve lymphatic endothelial cell (LECs) phenotypes, we demonstrate that low matrix elasticity primes lymphatic cord-like structure (CLS) formation directed by a high concentration of vascular endothelial growth factor-C (VEGF-C). Decreasing the substrate stiffness results in the upregulation of key lymphatic markers, including PROX-1, lymphatic vessel endothelial hyaluronan receptor 1 (LYVE-1), and VEGFR-3. Consequently, higher levels of VEGFR-3 enable stimulation of LECs with VEGF-C which is required to both activate matrix metalloproteinases (MMPs) and to facilitate LEC migration. Both of these steps are critical in establishing CLS formation in vitro. With decreases in substrate elasticity, we observe increased MMP expression and increased cellular elongation, as well as formation of intracellular vacuoles, which can further merge into coalescent vacuoles. RNAi studies demonstrate that MMP-14 is required to enable CLS formation and that LECs sense matrix stiffness through YAP/TAZ mechanosensors leading to the activation of their downstream target genes. Collectively, we show that by tuning both the matrix stiffness and VEGF-C concentration, the signaling pathways of CLS formation can be regulated in a synthetic matrix, resulting in lymphatic networks which will be useful for the study of lymphatic biology and future approaches in tissue regeneration.