TGFβ2 Regulates Human Trabecular Meshwork Cell Contractility via ERK and ROCK Pathways with Distinct Signaling Crosstalk Dependent on the Culture Substrate.
TGFβ2 Regulates Human Trabecular Meshwork Cell Contractility via ERK and ROCK Pathways with Distinct Signaling Crosstalk Dependent on the Culture Substrate.
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DOI：
10.1080/02713683.2022.2071943
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发表时间：
2022-08
影响因子：
2
通讯作者：
中科院分区：
文献类型：
--
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转化生长因子β 2（Transforming growth factor beta 2，TGFβ2）是原发性开角型青光眼小梁网（trabecular meshwork，HTM）细胞发生病理变化的主要因素。TGFβ2激活细胞外信号调节激酶（ERK）和Rho相关激酶（ROCK）信号通路，两者均影响HTM细胞行为。然而，这些信号通路究竟如何汇聚以调节HTM细胞收缩性尚不清楚。在此，我们研究了TGFβ2诱导的病理性HTM细胞收缩的分子机制，以及ERK和ROCK信号通路与不同培养基质之间的串扰。通过混合I型胶原蛋白、弹性蛋白样多肽和透明质酸（各自含有光活性官能团），随后通过UV交联来工程化水凝胶。将原代HTM细胞接种在预形成的水凝胶上以与玻璃进行比较，或包封在水凝胶内。肌动蛋白细胞骨架，细胞外基质的生产，磷酸肌球蛋白轻链（p-MLC）水平的变化，和水凝胶收缩进行了评估。HTM细胞形态和丝状（F）-肌动蛋白组织的影响，由潜在的培养基板。TGFβ2通过ERK和ROCK信号通路通过差异调节HTM细胞中的F-actin、α-smooth muscle actin、fibronectin和p-MLC增加HTM细胞的收缩性。ERK抑制，即使短至4小时，也进一步增加了水凝胶上的HTM细胞中TGFβ2诱导的p-MLC，但在玻璃上没有。这转化为HTM细胞负载水凝胶的过度收缩性。ROCK抑制具有完全相反的效果，并有效地松弛TGFβ2诱导的水凝胶。我们的数据表明，ERK信号负调控ROCK介导的HTM细胞收缩性。这些发现强调了使用组织模拟ECM底物在体外研究HTM细胞生理学和肿瘤病理生理学的至关重要性。
Transforming growth factor beta 2 (TGFβ2) is a major contributor to the pathologic changes occurring in human trabecular meshwork (HTM) cells in primary open-angle glaucoma. TGFβ2 activates extracellular-signal-regulated kinase (ERK) and Rho-associated kinase (ROCK) signaling pathways, both affecting HTM cell behavior. However, exactly how these signaling pathways converge to regulate HTM cell contractility is unclear. Here, we investigated the molecular mechanism underlying TGFβ2-induced pathologic HTM cell contractility, and the crosstalk between ERK and ROCK signaling pathways with different culture substrates. Hydrogels were engineered by mixing collagen type I, elastin-like polypeptide and hyaluronic acid, each containing photoactive functional groups, followed by UV crosslinking. Primary HTM cells were seeded atop pre-formed hydrogels for comparisons with glass, or encapsulated within the hydrogels. Changes in actin cytoskeleton, extracellular matrix production, phospho-myosin light chain (p-MLC) levels, and hydrogel contraction were assessed. HTM cell morphology and filamentous (F)-actin organization were affected by the underlying culture substrates. TGFβ2 increased HTM cell contractility via ERK and ROCK signaling pathways by differentially regulating F-actin, α-smooth muscle actin, fibronectin, and p-MLC in HTM cells. ERK inhibition, even as short as 4 h, further increased TGFβ2-induced p-MLC in HTM cells on hydrogels, but not on glass. This translated into hypercontractility of HTM cell-laden hydrogels. ROCK inhibition had precisely the opposite effects and potently relaxed the TGFβ2-induced hydrogels. Our data suggest that ERK signaling negatively regulates ROCK-mediated HTM cell contractility. These findings emphasize the critical importance of using tissue-mimetic ECM substrates for investigating HTM cell physiology and glaucomatous pathophysiology in vitro.
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影响因子：
2.9
作者：
Amano, Mutsuki;Nakayama, Masanori;Kaibuchi, Kozo
通讯作者：
Kaibuchi, Kozo
影响因子：
4.3
作者：
Hensel N;Rademacher S;Claus P
通讯作者：
Claus P
小梁网中的干细胞：现在和未来的承诺。
影响因子：
3.4
作者：
Kelley, M. J.;Rose, A. Y.;Keller, K. E.;Hessle, H.;Samples, J. R.;Acott, T. S.
通讯作者：
Acott, T. S.
影响因子：
64.8
作者：
Green JJ;Elisseeff JH
通讯作者：
Elisseeff JH
DOI：
10.1007/s004170000241
发表时间：
2001-02-01
影响因子：
2.7
作者：
Inatani, M;Tanihara, H;Honda, Y
通讯作者：
Honda, Y