The deubiquitinase USP40 preserves endothelial integrity by targeting the heat shock protein HSP90β
2
조회
0
다운로드
0
인용
0
참고문헌
초록
Endothelial cell (EC) barrier disruption and inflammation are the pathological hallmarks of vascular disorders and acute infectious diseases and related conditions, including the coronavirus disease 2019 (COVID-19) and sepsis. Ubiquitination plays a critical role in regulating the stability, intracellular trafficking, and enzymatic activity of proteins and is reversed by deubiquitinating enzymes (DUBs). The role of DUBs in endothelial biology is largely unknown. In this study, we report that USP40, a poorly characterized DUB, prevents EC barrier disruption through reductions in the activation of RhoA and phosphorylation of myosin light chain (MLC) and cofilin. Furthermore, USP40 reduces EC inflammation through the attenuation of NF-ĸB activation, ICAM1 expression, and leukocyte-EC adhesion. We further show that USP40 activity and expression are reduced in response to endotoxin challenge. Global depletion of USP40 and EC-targeted USP40 depletion in mice exacerbated experimental lung injury, whereas lentiviral gene transfer of USP40 protected against endotoxin-induced lung injury. Using an unbiased approach, we discovered that the protective effect of USP40 occurs through the targeting of heat shock protein 90β (HSP90β) for its deubiquitination and inactivation. Together, these data reveal a critical protective role of USP40 in vascular injury, identifying a unique mechanistic pathway that profoundly impacts endothelial function via DUBs.
Abstract
Endothelial cell (EC) barrier disruption and inflammation are the pathological hallmarks of vascular disorders and acute infectious diseases and related conditions, including the coronavirus disease 2019 (COVID-19) and sepsis. Ubiquitination plays a critical role in regulating the stability, intracellular trafficking, and enzymatic activity of proteins and is reversed by deubiquitinating enzymes (DUBs). The role of DUBs in endothelial biology is largely unknown. In this study, we report that USP40, a poorly characterized DUB, prevents EC barrier disruption through reductions in the activation of RhoA and phosphorylation of myosin light chain (MLC) and cofilin. Furthermore, USP40 reduces EC inflammation through the attenuation of NF-ĸB activation, ICAM1 expression, and leukocyte-EC adhesion. We further show that USP40 activity and expression are reduced in response to endotoxin challenge. Global depletion of USP40 and EC-targeted USP40 depletion in mice exacerbated experimental lung injury, whereas lentiviral gene transfer of USP40 protected against endotoxin-induced lung injury. Using an unbiased approach, we discovered that the protective effect of USP40 occurs through the targeting of heat shock protein 90β (HSP90β) for its deubiquitination and inactivation. Together, these data reveal a critical protective role of USP40 in vascular injury, identifying a unique mechanistic pathway that profoundly impacts endothelial function via DUBs.
참고문헌 (47)
- Rao, R. M., "Endothelial-dependent mechanisms of leukocyte recruitment to the vascular wall", Circ. Res, 101, pp.234~247 (2007)
- Gill, S. E., "Pulmonary microvascular albumin leak is associated with endothelial cell death in murine sepsis-induced lung injury in vivo", PloS One, 9, pp.e88501~ (2014)
- Goldenberg, N. M., "Broken barriers: a new take on sepsis pathogenesis", Sci. Transl. Med, 3, pp.88ps25~ (2011)
- Mackow, E. R., "Endothelial cell dysfunction in viral hemorrhage and edema", Front. Microbiol, 5, pp.733~ (2014)
- Hiyoshi, M., "Influenza A virus infection of vascular endothelial cells induces GSK-3beta-mediated beta-catenin degradation in adherens junctions, with a resultant increase in membrane permeability", Arch. Virol, 160, pp.225~234 (2015)
- Sukriti, S., "Mechanisms regulating endothelial permeability", Pulmonary Circ, 4, pp.535~551 (2014)
- Birukova, A. A., "Endothelial permeability is controlled by spatially defined cytoskeletal mechanics : atomic force microscopy force mapping of pulmonary endothelial monolayer", Nanomed. : Nanotechnol. Biol. Med, 5, pp.30~41 (2009)
- Schnoor, M., "Actin dynamics in the regulation of endothelial barrier func tions and neutrophil recruitment during endotoxemia and sepsis", Cell Mol. Life Sci, 74, pp.1985~1997 (2017)
- Lin, T., "Rho-ROCK-LIMK-cofilin pathway regulates shear stress activation of sterol regulatory element binding proteins", Circ. Res, 92, pp.1296~1304 (2003)
- Gorovoy, M., "LIM kinase 1 coordinates microtubule stability and actin polymerization in human endothelial cells", J. Biol. Chem, 280, pp.26533~26542 (2005)
- Owen-Woods, C., "Local microvascular leakage promotes trafficking of acti vated neutrophils to remote organs", J. Clin. Invest, 130, pp.2301~2318 (2020)
- Yang, L., "ICAM-1 regulates neutrophil adhesion and transcellular migration of TNF-alpha-activated vascular endothelium under flow", Blood, 106, pp.584~592 (2005)
- Kilpatrick, L. E., "Experimental approaches to evaluate leukocyte endothelial cell interactions in sepsis and inflammation", Shock, 53, pp.585~595 (2020)
- Ledebur, H. C., "Transcriptional regulation of the intercellular adhe sion molecule-1 gene by inflammatory cytokines in human endothelial cells. Essential roles of a variant NF-kappa B site and p65 homodimers", J. Biol. Chem, 270, pp.933~943 (1995)
- Kim, I., "Vascular endothelial growth factor expression of intercellular adhe sion molecule 1(ICAM-1), vascular cell adhesion molecule 1(VCAM-1), and E-selectin through nuclear factor-kappa B activation in endothelial cells", J. Biol. Chem, 276, pp.7614~7620 (2001)
- Joshi, A. D., "Heat shock protein 90 inhibitors prevent LPS-induced endo thelial barrier dysfunction by disrupting RhoA signaling", Am. J Respir. Cell Mol. Biol, 50, pp.170~179 (2014)
- Thangjam, G. S., "Hsp90 inhibition suppresses NF-kappaB transcriptional activation via Sirt-2 in human lung microvascular endothelial cells", Am. J Physiol. Lung Cell. Mol. Physiol, 310, pp.L964~L974 (2016)
- Chatterjee, A., "Heat shock protein 90 inhibitors prolong survival, attenuate inflammation, and reduce lung injury in murine sepsis", Am. J Respir. Crit. Care Med, 176, pp.667~675 (2007)
- Scroggins, B. T., "An acetylation site in the middle domain of Hsp90 regulates chaperone function", Mol. Cell, 25, pp.151~159 (2007)
- Mollapour, M., "Post-translational modifications of Hsp90 and their contributions to chaperone regulation", Biochim. Biophys. Acta, 1823, pp.648~655 (2012)
- Schnell, J. D., "Non-traditional functions of ubiquitin and ubiquitin binding proteins", J Biol. Chem, 278, pp.35857~35860 (2003)
- Mukhopadhyay, D., "Proteasome-independent functions of ubiquitin in endocytosis and signaling", Science, 315, pp.201~205 (2007)
- Kundrat, L., "Identification of residues on Hsp70 and Hsp90 ubiquiti nated by the cochaperone CHIP", J. Mol. Biol, 395, pp.587~594 (2010)
- Sarkar, A. A., "Hectd1 regulates intracellular localization and secretion of Hsp90 to control cellular behavior of the cranial mesenchyme", J. Cell Biol, 196, pp.789~800 (2012)
- Takagi, H., "USP40 gene knockdown disrupts glomerular permeability in zebrafish", Am. J. Phys. Renal Phys, 312, pp.F702~F715 (2017)
- Takahashi, S., "USP40 deubiquitinates HINT1 and stabilizes p53 in podocyte damage", Biochem. Biophys. Res. Commun, 614, pp.198~206 (2022)
- Pelletier, S., "Mouse genome engineering via CRISPR Cas9 for study of immune function", Immunity, 42, pp.18~27 (2015)
- Roy, N., "Dysregulation of lipid and glucose homeostasis in hepatocyte specific SLC25A34 knockout mice", Am. J. Pathol, 192, pp.1259~1281 (2022)
- Li, L., "TRIM21 mitigates human lung microvascular endothelial cells’ inflammatory responses to LPS", Am. J. Respir. Cell Mol. Biol, 61, pp.776~785 (2019)
- Free, R. B., "Identifying novel protein-protein interactions using co-immunoprecipitation and mass spectroscopy", Curr. Protoc. Neurosci (2009)
- Yang, N., "Cofilin phosphorylation by LIM-kinase 1 and its role in Rac mediated actin reorganization", Nature, 393, pp.809~812 (1998)
- Chen, C. C., "Selective inhi bition of E-selectin, vascular cell adhesion molecule-1, and intercellular adhesion molecule-1 expression by inhibitors of I kappa B-alpha phosphorylation", J. Immunol, 155, pp.3538~3545 (1995)
- Muller, W. A., "Mechanisms of transendothelial migration of leukocytes", Circ. Res, 105, pp.223~230 (2009)
- Kekatpure, V. D., "HDAC6 modulates Hsp90chaperone activity and regulates activation of aryl hydrocarbon receptor sig naling", J. Biol. Chem, 284, pp.7436~7445 (2009)
- Zhao, J., "F-box protein FBXL19-mediated ubiquitination and degradation of the receptor for IL-33 limits pulmonary inflammation", Nat. Immunol, 13, pp.651~658 (2012)
- Wei, J., "Regulation of the ubiquitylation and deubiquitylation of CREB binding protein modulates histone acetylation and lung inflammation", Sci. Signal, 10, pp.eaak9660~ (2017)
- Chen, B. B., "A combinatorial F box protein directed pathway controls TRAF adaptor stability to regulate inflammation", Nat. Immunol, 14, pp.470~479 (2013)
- Zhao, J., "Destabilization of lysophosphatidic acid receptor 1 reduces cytokine release and protects against lung injury", EBioMedicine, 10, pp.195~203 (2016)
- Mialki, R. K., "Overexpression of USP14protease reduces I-kappaB protein levels and increases cytokine release in lung epithelial cells", J. Biol. Chem, 288, pp.15437~15441 (2013)
- An, W., "Glucocorticoid modulatory element-binding protein 1(GMEB1)interacts with the de-ubiquitinase USP40 to stabilize CFLARL and inhibit apop tosis in human non-small cell lung cancer cells", J. Exp. Clin. Cancer Res, 38, pp.181~ (2019)
- Barabutis, N., "Hsp90 inhibitors suppress P53phosphorylation in LPS-induced endothelial inflammation", Cytokine, 113, pp.427~432 (2019)
- Thangjam, G. S., "Novel mechanism of attenuation of LPS-induced NF-kappaB activation by the heat shock protein 90 inhibitor, 17-N-allylamino-17-deme thoxygeldanamycin, in human lung microvascular endothelial cells", Am. J. Respir. Cell Mol. Biol, 50, pp.942~952 (2014)
- Uddin, M. A., "Hsp90 inhibition protects the brain microvascular endothelium against oxidative stress", Brain Disord, 1, pp.100001~ (2021)
- Lee, J. G., "Characterization of the deubiquitinating activity of USP19 and its role in endoplasmic reticulum-associated degradation", J. Biol. Chem, 289, pp.3510~3517 (2014)
- Aressy, B., "A screen for deubiquitinating enzymes involved in the G(2)", M checkpoint identifies USP50 as a regulator of HSP90-dependent Wee1 stability, 2010, pp.9, 3815~3822
- Wagner, N., "Intermediate f ilament protein nestin is expressed in developing kidney and heart and might be regulated by the Wilms’ tumor suppressor Wt1", Am. J. Physiol. Regul. Integr. Comp. Physiol, 291, pp.R779~R787 (2006)
- Kisanuki, Y. Y., "Tie2-Cre transgenic mice : a new model for endothelial cell lineage analysis in vivo", Dev. Biol, 230, pp.230~242 (2001)