The cell membrane repair protein MG53 modulates transcription factor NF-κB signaling to control kidney fibrosis

Abstract

Kidney fibrosis is associated with the progression of acute kidney injury to chronic kidney disease. MG53, a cell membrane repair protein, has been shown to protect against injury to kidney epithelial cells and acute kidney injury. Here, we evaluated the role of MG53 in modulation of kidney fibrosis in aging mice and in mice with unilateral ureteral obstruction (UUO) a known model of progressive kidney fibrosis. Mice with ablation of MG53 developed more interstitial fibrosis with age than MG53-intact mice of the same age. Similarly, in the absence of MG53, kidney fibrosis was exaggerated compared to mice with intact MG53 in the obstructed kidney compared to the contralateral unobstructed kidney or the kidneys of sham operated mice. The ureteral obstructed kidneys from MG53 deficient mice also showed significantly more inflammation than ureteral obstructed kidneys from MG53 intact mice. In vitro experiments demonstrated that MG53 could enter the nuclei of proximal tubular epithelial cells and directly interact with the p65 component of transcription factor NF-κB, providing a possible explanation of enhanced inflammation in the absence of MG53. To test this, enhanced MG53 expression through engineered cells or direct recombinant protein delivery was given to mice subject to UUO. This reduced NF-κB activation and inflammation and attenuated kidney fibrosis. Thus, MG53 may have a therapeutic role in treating chronic kidney inflammation and thereby provide protection against fibrosis that leads to the chronic kidney disease phenotype. Kidney fibrosis is associated with the progression of acute kidney injury to chronic kidney disease. MG53, a cell membrane repair protein, has been shown to protect against injury to kidney epithelial cells and acute kidney injury. Here, we evaluated the role of MG53 in modulation of kidney fibrosis in aging mice and in mice with unilateral ureteral obstruction (UUO) a known model of progressive kidney fibrosis. Mice with ablation of MG53 developed more interstitial fibrosis with age than MG53-intact mice of the same age. Similarly, in the absence of MG53, kidney fibrosis was exaggerated compared to mice with intact MG53 in the obstructed kidney compared to the contralateral unobstructed kidney or the kidneys of sham operated mice. The ureteral obstructed kidneys from MG53 deficient mice also showed significantly more inflammation than ureteral obstructed kidneys from MG53 intact mice. In vitro experiments demonstrated that MG53 could enter the nuclei of proximal tubular epithelial cells and directly interact with the p65 component of transcription factor NF-κB, providing a possible explanation of enhanced inflammation in the absence of MG53. To test this, enhanced MG53 expression through engineered cells or direct recombinant protein delivery was given to mice subject to UUO. This reduced NF-κB activation and inflammation and attenuated kidney fibrosis. Thus, MG53 may have a therapeutic role in treating chronic kidney inflammation and thereby provide protection against fibrosis that leads to the chronic kidney disease phenotype. Translational StatementChronic inflammation leads to fibrotic remodeling that may also underlie the transition from acute kidney injury to chronic kidney disease. Activation of the proinflammatory transcription factor nuclear factor κB (NF-κB) is involved in the pathogenesis of kidney inflammation. Here, we provide evidence that MG53, a previously identified cell membrane repair protein, directly interacts with NF-κB and reduces its transcriptional activity. Concomitantly, exogenously administered MG53 can decrease fibrotic remodeling of the inflamed kidney. These findings point to a protective interplay between MG53 and NF-κB to attenuate the development of inflammation-mediated kidney fibrosis. Pharmacologic administration of MG53 might be a promising approach for the treatment of progressive kidney fibrosis. Chronic inflammation leads to fibrotic remodeling that may also underlie the transition from acute kidney injury to chronic kidney disease. Activation of the proinflammatory transcription factor nuclear factor κB (NF-κB) is involved in the pathogenesis of kidney inflammation. Here, we provide evidence that MG53, a previously identified cell membrane repair protein, directly interacts with NF-κB and reduces its transcriptional activity. Concomitantly, exogenously administered MG53 can decrease fibrotic remodeling of the inflamed kidney. These findings point to a protective interplay between MG53 and NF-κB to attenuate the development of inflammation-mediated kidney fibrosis. Pharmacologic administration of MG53 might be a promising approach for the treatment of progressive kidney fibrosis. Kidney dysfunction, which can be classified as acute kidney injury (AKI) or chronic kidney disease (CKD), has grown into an epidemic in older populations. In 2017, the prevalence of CKD reached 14.5% of people aged 65 and over, resulting in Medicare expenditures of over $84 billion for treatment.1Saran R. Robinson B. Abbott K.C. et al.US Renal Data System 2018 annual data report: epidemiology of kidney disease in the United States.Am J Kidney Dis. 2019; 73: A7-A8Abstract Full Text Full Text PDF PubMed Scopus (413) Google Scholar,2Saran R. Robinson B. Abbott K.C. et al.US Renal Data System 2019 Annual Data Report: epidemiology of kidney disease in the United States.Am J Kidney Dis. 2020; 75: A6-A7Abstract Full Text Full Text PDF PubMed Scopus (298) Google Scholar AKI and CKD are more common in older individuals, mainly due to increased susceptibility to injury plus a diminished ability to repair the aging kidney. Clinical studies have shown that CKD is a major risk factor for AKI, and, conversely, episodes of AKI can accelerate the progression of CKD toward end-stage kidney disease. Currently, the treatment of AKI is mainly supportive, and the treatment of established CKD is focused on slowing progression through inhibition of the renin-angiotensin-aldosterone system and now possibly through sodium-glucose cotransporter-2 inhibition.3Alicic R.Z. Johnson E.J. Tuttle K.R. SGLT2 inhibition for the prevention and treatment of diabetic kidney disease: a review.Am J Kidney Dis. 2018; 72: 267-277Abstract Full Text Full Text PDF PubMed Scopus (62) Google Scholar,4Hocher B. Tsuprykov O. Diabetic nephropathy: renoprotective effects of GLP1R agonists and SGLT2 inhibitors.Nat Rev Nephrol. 2017; 13: 728-730Crossref PubMed Scopus (18) Google Scholar Depending on the severity of injury, proximal tubular cells, the major target of acute injury, may undergo changes in cell cycle progression,5Yang L. Besschetnova T.Y. Brooks C.R. et al.Epithelial cell cycle arrest in G2/M mediates kidney fibrosis after injury.Nat Med. 2010; 16: 535-543Crossref PubMed Scopus (807) Google Scholar,6Canaud G. Brooks C.R. Kishi S. et al.Cyclin G1 and TASCC regulate kidney epithelial cell G2-M arrest and fibrotic maladaptive repair.Sci Transl Med. 2019; 11: 476Crossref Scopus (51) Google Scholar metabolism,7Kang H.M. Ahn S.H. Choi P. et al.Defective fatty acid oxidation in renal tubular epithelial cells has a key role in kidney fibrosis development.Nat Med. 2015; 21: 37-46Crossref PubMed Scopus (589) Google Scholar secretion of proinflammatory and profibrotic cytokines, or partial epithelial-mesenchymal transition,8Lovisa S. LeBleu V.S. Tampe B. et al.Epithelial-to-mesenchymal transition induces cell cycle arrest and parenchymal damage in renal fibrosis.Nat Med. 2015; 21: 998-1009Crossref PubMed Scopus (514) Google Scholar which will determine whether remodeling/repair will be successful or maladaptive and result in kidney fibrosis, a hallmark of CKD.9Humphreys B.D. Mechanisms of renal fibrosis.Annu Rev Physiol. 2018; 80: 309-326Crossref PubMed Scopus (282) Google Scholar,10Liu B.C. Tang T.T. Lv L.L. et al.Renal tubule injury: a driving force toward chronic kidney disease.Kidney Int. 2018; 93: 568-579Abstract Full Text Full Text PDF PubMed Scopus (245) Google Scholar This remodeling generally occurs on a background of kidney inflammation, diminished vascular supply, and the production of extracellular matrix (ECM) proteins and includes loss of epithelial cells and accumulation of collagens, α-smooth muscle actin, and fibronectin; it also has a high correlation with deterioration of kidney function. Chronic inflammation leads to fibrotic remodeling that may also underlie the transition from AKI to CKD.11Zhang H. Sun S.C. NF-kappaB in inflammation and renal diseases.Cell Biosci. 2015; 5: 63Crossref PubMed Scopus (143) Google Scholar, 12Song N. Thaiss F. Guo L. NFκB and Kidney Injury.Front Immunol. 2019; 10: 815Crossref PubMed Scopus (39) Google Scholar, 13Andrade-Oliveira V. Foresto-Neto O. Watanabe I.K. et al.Inflammation in renal diseases: new and old players.Front Pharmacol. 2019; 10: 1192Crossref PubMed Scopus (84) Google Scholar Cumulative research suggests the involvement of the nuclear factor κB (NF-κB) transcription factor in the pathogenesis of kidney inflammation caused by infection, injury, or transplantation.11Zhang H. Sun S.C. NF-kappaB in inflammation and renal diseases.Cell Biosci. 2015; 5: 63Crossref PubMed Scopus (143) Google Scholar,12Song N. Thaiss F. Guo L. NFκB and Kidney Injury.Front Immunol. 2019; 10: 815Crossref PubMed Scopus (39) Google Scholar,14Reid S. Scholey J. Recent approaches to targeting canonical NFκB signalling in the early inflammatory response to renal IRI.J Am Soc Nephrol. 2021; 32: 2117-2124Crossref PubMed Scopus (3) Google Scholar Activation of the canonical NF-κB pathway starts with activation of the inhibitor of NF-κB (IκB) kinase, which leads to phosphorylation and degradation of IκBα and the nuclear translocation of NF-κB heterodimers.15Hoffmann A. Levchenko A. Scott M.L. et al.The IkappaB-NF-kappaB signaling module: temporal control and selective gene activation.Science. 2002; 298: 1241-1245Crossref PubMed Scopus (1436) Google Scholar Activation of NF-κB signaling in kidney epithelial cells and infiltrating immune cells can be elicited by pathophysiological triggers such as exposure to lipopolysaccharides (LPSs) or ischemia-reperfusion injury.16Marko L. Vigolo E. Hinze C. et al.Tubular epithelial NF-kappaB activity regulates ischemic AKI.J Am Soc Nephrol. 2016; 27: 2658-2669Crossref PubMed Scopus (94) Google Scholar Molecular profiling studies reveal nfkb1 as a major driver of kidney fibrosis.17Wu H. Lai C.F. Chang-Panesso M. et al.Proximal tubule translational profiling during kidney fibrosis reveals proinflammatory and long noncoding RNA expression patterns with sexual dimorphism.J Am Soc Nephrol. 2020; 31: 23-38Crossref PubMed Scopus (25) Google Scholar In addition to kidney tubular cells, innate immune cells such as macrophages and dendritic cells also contribute to kidney injury, inflammation, and fibrotic remodeling.13Andrade-Oliveira V. Foresto-Neto O. Watanabe I.K. et al.Inflammation in renal diseases: new and old players.Front Pharmacol. 2019; 10: 1192Crossref PubMed Scopus (84) Google Scholar,18Singbartl K. Formeck C.L. Kellum J.A. Kidney-immune system crosstalk in AKI.Semin Nephrol. 2019; 39: 96-106Abstract Full Text Full Text PDF PubMed Scopus (59) Google Scholar, 19Hato T. Dagher P.C. How the innate immune system senses trouble and causes trouble.Clin J Am Soc Nephrol. 2015; 10: 1459-1469Crossref PubMed Scopus (76) Google Scholar, 20Komada T. Muruve D.A. The role of inflammasomes in kidney disease.Nat Rev Nephrol. 2019; 15: 501-520Crossref PubMed Scopus (90) Google Scholar Growing evidence suggests that muscle-derived secretory factors (i.e., myokines) modulate systemic physiology via tissue crosstalk to influence the progression of kidney diseases.21Peng H. Wang Q. Lou T. et al.Myokine mediated muscle-kidney crosstalk suppresses metabolic reprogramming and fibrosis in damaged kidneys.Nat Commun. 2017; 8: 1493Crossref PubMed Scopus (71) Google Scholar MG53 (also named TRIM72) is a muscle-enriched tripartite protein with a in cell membrane C. H. N. et of cell membrane repair 11: PubMed Scopus Google C. H. N. et regulates membrane and in muscle Full Text Full Text PDF PubMed Scopus (84) Google Scholar proteins have from the of immune signaling to tissue S. in and 2017; Full Text Full Text PDF PubMed Scopus Google Scholar membrane repair is involved in acute injury to C. N. et repair in are to between MG53, and Full Text Full Text PDF PubMed Scopus Google Scholar P. H. P. et cell membrane repair against acute kidney Transl Med. 2015; Scopus Google Scholar J. H. et of recombinant MG53 protein in a model of and 2015; 80: Full Text Full Text PDF PubMed Scopus Google Scholar K. P. et of acute injury by targeting cell membrane Commun. 5: PubMed Scopus (71) Google Scholar F. T. Wang et al.The protein and therapeutic of cells in 2019; 10: PubMed Scopus Google B. H. et through to protect ischemic 2016; PubMed Scopus Google Scholar and H. P. et of and by MG53 cell membrane 2015; Full Text Full Text PDF PubMed Scopus Google Scholar studies identified a of MG53 expression in the which is a key component in AKI and mice deficient in MG53 more to P. H. P. et cell membrane repair against acute kidney Transl Med. 2015; Scopus Google Scholar also demonstrated that of MG53 in the tissue injury repair and Wang Q. et of MG53 in the tissue metabolic Commun. 2019; 10: PubMed Scopus Google Scholar The of and MG53 in kidney fibrosis during progressive CKD are an role of MG53 in tissue protection through the inhibition of inflammation, via modulation of NF-κB has MG53 and by the F. P. et and via pathway in vitro and in 2019; PubMed Scopus Google Scholar and MG53 regulates expression to control remodeling during Wang G. C. et a of and a role in remodeling in 2019; PubMed Scopus Google Scholar In we demonstrated a of MG53 in a maladaptive response during a infection, by via of activation and inhibition of signaling in M. et suppresses and inflammation via of Commun. 2020; 11: PubMed Scopus Google Scholar a of infection, MG53 was shown to and by directly by and through of the and prevention of of the et MG53 protein mice from J Med. 2021; PubMed Scopus Google Scholar chronic treatment of MG53 was to of inflammation in the aged H. et suppresses NFκB activation to 2021; PubMed Scopus Google Scholar These studies MG53 can modulate NF-κB signaling during inflammation whether occurs in the of kidney inflammation has been that MG53 regulates kidney inflammation by the transcriptional activity of NF-κB and in can attenuate the kidney fibrosis that occurs as a of chronic inflammation. This was to experiments with to the of for the and of and by the and or of the C. H. N. et of cell membrane repair 11: PubMed Scopus Google Scholar to the role of MG53 in the modulation of and unilateral obstruction kidney fibrosis. mice from kidney injury mice to to kidney fibrosis. Mice via to a of The was by the with to obstruction as a model of renal interstitial fibrosis and Int. 75: Full Text Full Text PDF PubMed Scopus Google Scholar mice an To the of on kidney injury, of mice via after and for with on and control of mice to the same Mice on after kidneys in with and tissue from the kidney and the contralateral kidney for and In and mice or sham and on after the MG53 was given to mice a R. A. M. et kidney fibrosis in the mice 2017; PubMed Scopus Google Scholar macrophages with for The cells a of into mice after Mice of cells on and after the cell mice into a that and a that in in for and from the kidney and contralateral kidney for and protein Data are as was the test and of for more than by the or test of was Full cell and tubular epithelial cell and cell infection, NF-κB p65 nuclear translocation and NF-κB tissue and production are in the compared the of aging on kidney and fibrosis in and mice. that of mice with and MG53 significantly older mice a significantly than on we that mice more kidney fibrosis compared with the the age of and through the age of and was to the of MG53 ablation on the of and macrophages in and kidneys and from and mice of significantly more inflammatory cells in kidneys than kidneys in mice. for α-smooth muscle and extracellular matrix proteins in of kidney fibrosis, showed enhanced to in the and of kidneys from mice compared with mice Kidney MG53 by significantly increased after The kidneys exaggerated fibrosis, with a than kidneys was also significantly increased in kidneys Kidney by and macrophages was significantly increased after in and mice was increased in and The activity of NF-κB was in the The of p65 was significantly increased in kidneys from and NF-κB as phosphorylation of was in the kidney than the kidney compared with the RNA of factor and in the obstructed Tang J. et al.The response to repair and progression of fibrotic disease in the obstructed 2021; PubMed Scopus (3) Google Scholar of gene expression in sham and in the kidney from and To the of MG53 in the kidney renal tissue more protein than the nuclear The in the between and mice in the nuclear of mice MG53 was in the as a a of MG53 was the nuclear as a nuclear To we also MG53 in muscle studies in which MG53 to and the membrane as a of membrane C. H. N. et of cell membrane repair 11: PubMed Scopus Google C. H. N. et regulates membrane and in muscle Full Text Full Text PDF PubMed Scopus (84) Google C. N. et repair in are to between MG53, and Full Text Full Text PDF PubMed Scopus Google Scholar also nuclear MG53 from To tissue we the In proximal tubular epithelial cells, p65 was mainly in the to nuclei in the absence of MG53 cells with for proinflammatory secretion was significantly in cells than cells data MG53 may interact with p65 to regulate inflammatory a direct between p65 and MG53 was by cells with and demonstrated a of and and of p65 and MG53 whether MG53 NF-κB transcriptional activity after by the activity from a transcriptional cell that was with or The cells a expression by a in with of NF-κB transcriptional response In the absence of was activity in the cells MG53 MG53 NF-κB transcriptional activity by NF-κB p65 to the treatment in the kidney proximal tubule P. A. Wang B. et al.The protein mediates NF-kappaB Am Soc Nephrol. 2010; 21: PubMed Scopus Google Scholar To whether treatment with MG53 could attenuate NF-κB p65 nuclear translocation we MG53 in cells via or through treatment with recombinant MG53 a after MG53 was by MG53 was a high in cells, to In and a of of by was by the to p65 activation was to NF-κB p65 mainly in the and of MG53 and after p65 to the nuclei in cells MG53. In nuclear translocation of p65 by given to cells given in cells MG53 and To whether treatment with MG53 could attenuate fibrosis, MG53 was increased in the model via engineered macrophages engineered via of of expression was by of with a cell after and of of engineered cells into the model was with cells administered through after Mice into a that and a that in mice for a of after Mice with engineered by significantly reduced kidney fibrosis compared with cells or with engineered cells fibrotic and a the of system by the of the that treatment MG53 expression in the kidney kidneys from mice and MG53 a than or given and The reduced was with of kidneys MG53 by and by kidney proximal tubule cells could in and decrease kidney inflammation. with and expression was increased significantly in kidneys from mice with MG53 of profibrotic could be to the of a from cells into mice. kidneys from mice with and given significantly and of and than mice treatment whether a as the engineered cell data the in of N. N. P. et MG53 protein therapeutic cell membrane repair in treatment of Transl Med. Scopus Google Scholar Mice chronic administration of to P. H. P. et cell membrane repair against acute kidney Transl Med. 2015; Scopus Google H. et suppresses NFκB activation to 2021; PubMed Scopus Google Scholar the administration shown in was mice given or for the and on and contralateral kidneys and expression with the increased in contralateral kidneys compared with sham kidney with the in the kidneys of was reduced by IκBα increased by and by and was in kidneys after treatment fibrosis was reduced in kidneys RNA expression of and was in mice with or MG53 is a muscle-enriched protein identified as a component of membrane repair C. H. N. et of cell membrane repair 11: PubMed Scopus Google P. H. P. et cell membrane repair against acute kidney Transl Med. 2015; Scopus Google N. N. P. et MG53 protein therapeutic cell membrane repair in treatment of Transl Med. Scopus Google N. J. et targeting membrane repair and contribute to inflammatory 2020; Google Scholar the of MG53 are to and MG53 can be into the as a Wang Q. et of MG53 in the tissue metabolic Commun. 2019; 10: PubMed Scopus Google Scholar have previously shown MG53 can control inflammation during tissue damage in J. H. et of recombinant MG53 protein in a model of and 2015; 80: Full Text Full Text PDF PubMed Scopus Google F. T. Wang et al.The protein and therapeutic of cells in 2019; 10: PubMed Scopus Google F. P. et and via pathway in vitro and in 2019; PubMed Scopus Google M. et suppresses and inflammation via of Commun. 2020; 11: PubMed Scopus Google Scholar, et MG53 protein mice from J Med. 2021; PubMed Scopus Google Scholar, H. et suppresses NFκB activation to 2021; PubMed Scopus Google Scholar The the of protection to the kidney. that MG53 can attenuate inflammation and kidney fibrosis by activation of the proinflammatory and profibrotic transcription factor This is on the of MG53 in mice in a in kidney as the mice aged that was by an in kidney fibrosis and of the kidney by and in a kidney injury the obstructed kidneys of mice more fibrosis and inflammation than and production by proximal tubular epithelial cells was attenuated in the of treatment with or of MG53 in a proximal tubular epithelial cell diminished NF-κB activation by the nuclear translocation of the p65 component of NF-κB was to directly to and phosphorylation of a in the activation of NF-κB, was in the obstructed kidneys of mice with recombinant MG53 or to MG53 in the inhibitor of NF-κB was increased in the obstructed kidneys of mice with recombinant MG53. of MG53 in mice was to the effects of aging on the This suggests that the of MG53 a of protection during Similarly, the loss of MG53 AKI due to obstruction The kidney MG53, possibly activity from muscle MG53 may be to the kidney during a damaged kidney and R. et in renal injury and 2016; PubMed Scopus Google L. Q. J. et on injury, and the Int. 2017; Full Text Full Text PDF PubMed Scopus Google Scholar that could to MG53. could be to attenuate kidney M. of the Rev Nephrol. 2019; 15: PubMed Scopus Google Scholar kidney cells of MG53 P. H. P. et cell membrane repair against acute kidney Transl Med. 2015; Scopus Google Scholar and, during acute or chronic injury, MG53 production could be NF-κB is a transcriptional driver of inflammation and fibrosis in kidney injury and S. Scholey J. Recent approaches to targeting canonical NFκB signalling in the early inflammatory response to renal IRI.J Am Soc Nephrol. 2021; 32: 2117-2124Crossref PubMed Scopus (3) Google H. Lai C.F. Chang-Panesso M. et al.Proximal tubule translational profiling during kidney fibrosis reveals proinflammatory and long noncoding RNA expression patterns with sexual dimorphism.J Am Soc Nephrol. 2020; 31: 23-38Crossref PubMed Scopus (25) Google et al.The inflammatory transcription factors and transcriptional changes in the 2015; PubMed Scopus Google M. et in NF-kappaB signaling control the of gene PubMed Scopus Google Scholar to whether the effects of MG53 could be mediated via NF-κB the of MG53 in the nuclear of kidney the for transcriptional of MG53 is In a model MG53, nuclear MG53 the expression of the transcriptional and F. R. et of MG53 induces diabetic through transcriptional activation of 2015; PubMed Scopus Google Scholar we demonstrated NF-κB activation to through proinflammatory cytokines, and in aged H. et suppresses NFκB activation to 2021; PubMed Scopus Google Scholar The of MG53 p65 MG53 nuclear it nuclear a by nuclear protein J. et for nuclear signaling and the the 2019; PubMed Scopus Google Scholar The direct between MG53 and p65 was as the or of MG53 on NF-κB MG53 to the of NF-κB the to be The of to be to decrease fibrotic remodeling through the of inflammation. Kidney fibrosis is a of P. matrix in kidney more than a 2019; PubMed Scopus Google Scholar MG53 to have effects on the protein increased in kidneys deficient in MG53 and in mice with MG53. The expression of proteins in the and absence of MG53 is more to previously demonstrated that regulates signaling and suppresses the of proteins in an in vitro H. P. et of and by MG53 cell membrane 2015; Full Text Full Text PDF PubMed Scopus Google Scholar In and in kidneys from deficient These decrease in to MG53 through the administration of recombinant MG53 was was This may be a of the MG53 in given the as to recombinant MG53. protein and could the in UUO. research is to the role of MG53 in In has demonstrated that a may regulate inflammation in a the kidney and, in the of kidney injury, may to attenuate and control inflammation. a model of the role of MG53 in patterns such as and patterns such as and ischemic the NF-κB is in the kidney. MG53 the kidney by the phosphorylation and activation of NF-κB and p65 nuclear activation of proinflammatory MG53 may we more protection may be MG53 which is on the ability of MG53 to injury in UUO. is to may be has an in which for treatment of on the of MG53 are by and The the This was by of and and and This was also by an from The and an from the for Translational The is the of the and the of the or the and the and the and data and the and and and the and the with

Topics & Concepts