Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

Keynote webinar | Spotlight on medication adherence

LIVE: Thursday 27th June 2024, 18:00-19:30 (CEST)
Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.
ADVANCED SEARCH
Log in
Top
Arthritis Research & Therapy
Published in: Arthritis Research & Therapy 1/2019

Open Access 01-12-2019 | Systemic Lupus Erythematosus | Research article

Reversible SAHH inhibitor protects against glomerulonephritis in lupus-prone mice by downregulating renal α-actinin-4 expression and stabilizing integrin-cytoskeleton linkage

Authors: Shijun He, Xing Liu, Zemin Lin, Yuting Liu, Lei Gu, Hu Zhou, Wei Tang, Jianping Zuo

Published in: Arthritis Research & Therapy | Issue 1/2019

Login to get access

Abstract

Background

Glomerulonephritis is one of the major complications and causes of death in systemic lupus erythematosus (SLE) and is characterized by glomerulosclerosis, interstitial fibrosis, and tubular atrophy, along with severe persistent proteinuria. DZ2002 is a reversible S-adenosyl-l-homocysteine hydrolase (SAHH) inhibitor with potent therapeutic activity against lupus nephritis in mice. However, the molecular events underlying the renal protective effects of DZ2002 remained unclear. This study is designed to uncover the molecular mechanisms of DZ2002 on glomerulonephritis of lupus-prone mice.

Methods

We conducted a twice-daily treatment of DZ2002 on the lupus-prone NZB/WF1 mice, and the progression of lupus nephritis and alteration of renal function were monitored. The LC-MS-based label-free quantitative (LFQ) proteomic approach was applied to analyze the kidney tissue samples from the normal C57BL/6 mice and the NZB/WF1 mice treated with DZ2002 or vehicle. KEGG pathway enrichment and direct protein-protein interaction (PPI) network analyses were used to map the pathways in which the significantly changed proteins (SCPs) are involved. The selected proteins from proteomic analysis were validated by Western blot analysis and immunohistochemistry in the kidney tissues.

Results

The twice-daily regimen of DZ2002 administration significantly ameliorated the lupus nephritis and improved the renal function in NZB/WF1 mice. A total of 3275 proteins were quantified, of which 253 proteins were significantly changed across normal C57BL/6 mice and the NZB/WF1 mice treated with DZ2002 or vehicle. Pathway analysis revealed that 13 SCPs were involved in tight junction and focal adhesion process. Further protein expression validation demonstrated that DZ2002-treated NZB/WF1 mice exhibited downregulation of α-actinin-4 and integrin-linked kinase (ILK), as well as the restoration of β1-integrin activation in the kidney tissues compared with the vehicle-treated ones.

Conclusions

Our study demonstrated the first evidence for the molecular mechanism of SAHH inhibitor on glomerulonephritis in SLE via the modulation of α-actinin-4 expression and focal adhesion-associated signaling proteins in the kidney.
Appendix
Available only for authorised users
Literature
1.
Teramoto K, Negoro N, Kitamoto K, Iwai T, Iwao H, Okamura M, et al. Microarray analysis of glomerular gene expression in murine lupus nephritis. J Pharmacol Sci. 2008;106(1):56–67.PubMedCrossRef
2.
Devarapu S. Update on immune mechanisms in systemic lupus and lupus nephritis 2016. 116.
3.
Kang YS, Li Y, Dai C, Kiss LP, Wu C, Liu Y. Inhibition of integrin-linked kinase blocks podocyte epithelial-mesenchymal transition and ameliorates proteinuria. Kidney Int. 2010;78(4):363–73.PubMedPubMedCentralCrossRef
4.
Durvasula RV, Shankland SJ. Podocyte injury and targeting therapy: an update. Curr Opin Nephrol Hypertens. 2006;15(1):1–7.PubMedCrossRef
5.
6.
Smoyer WE, Mundel P, Gupta A, Welsh MJ. Podocyte alpha-actinin induction precedes foot process effacement in experimental nephrotic syndrome. Am J Physiol. 1997;273(1 Pt 2):F150.PubMed
7.
Alberts B, Bray D, Lewis J, Raff M, Roberts K, Watson JD. The cytoskeleton. In: molecular biology of the cell. Genetics. 2002;25(5):547–54.
8.
Shirato I, Sakai T, Kimura K, Tomino Y, Kriz W. Cytoskeletal changes in podocytes associated with foot process effacement in Masugi nephritis. Am J Pathol. 1996;148(4):1283–96.PubMedPubMedCentral
9.
Goode NP, Shires M, Khan TN, Mooney AF. Expression of α-actinin-4 in acquired human nephrotic syndrome: a quantitative immunoelectron microscopy study. Nephrol Dial Transplant. 2004;19(4):844–51.PubMedCrossRef
10.
Weins A, Schlondorff JS, Nakamura F, Denker BM, Hartwig JH, Stossel TP, et al. Disease-associated mutant α-actinin-4 reveals a mechanism for regulating its F-actin-binding affinity. Proc Natl Acad Sci U S A. 2007;104(41):16080–5.PubMedPubMedCentralCrossRef
11.
Kaplan JM, Kim SH, North KN, Rennke H, Correia LA, Tong HQ, et al. Mutations in ACTN4, encoding alpha-actinin-4, cause familial focal segmental glomerulosclerosis. Nat Genet. 2000;24(3):251–6.PubMedCrossRef
12.
Kreidberg JA, Symons JM. Integrins in kidney development, function, and disease. Am J Physiol Renal Physiol. 2000;279(2):F233.PubMedCrossRef
13.
Mulrooney JP, Hong T, Grabel LB. Serine 785 phosphorylation of the beta1 cytoplasmic domain modulates beta1A-integrin-dependent functions. J Cell Sci. 2001;114(13):2525–33.PubMed
14.
Wickström SA, Lange A, Montanez E, Fässler R. The ILK/PINCH/parvin complex: the kinase is dead, long live the pseudokinase! EMBO J. 2014;29(2):281–91.CrossRef
15.
Kretzler M, Teixeira VPC, Unschuld PG, Cohen CD, Wanke R, Edenhofer I, et al. Integrin linked kinase as a candidate downstream effector in proteinuria. FASEB J. 2001;15(10):1843–5.PubMedCrossRef
16.
Chiang PK, Gordon RK, Tal J, Zeng GC, Doctor BP, Pardhasaradhi K, et al. S-Adenosylmethionine and methylation. FASEB J. 1996;10(4):471–80.PubMedCrossRef
17.
Tardif V, Manenkova Y, Berger M, Hoebe K, Zuo JP, Yuan C, et al. Critical role of transmethylation in TLR signaling and systemic lupus erythematosus. Clin Immunol. 2013;147(2):133–43.PubMedPubMedCentralCrossRef
18.
He SJ, Lin ZM, Wu YW, Bai BX, Yang XQ, He PL, et al. Therapeutic effects of DZ2002, a reversible SAHH inhibitor, on lupus-prone NZBxNZW F1 mice via interference with TLR-mediated APC response. Acta Pharmacol Sin. 2014;35(2):219–29.PubMedCrossRef
19.
Lin ZM, Ma M, Li H, Qi Q, Liu YT, Yan YX, et al. Topical administration of reversible SAHH inhibitor ameliorates imiquimod-induced psoriasis-like skin lesions in mice via suppression of TNF-α/IFN-γ-induced inflammatory response in keratinocytes and T cell-derived IL-17. Pharmacol Res. 2018;129:443–52.PubMedCrossRef
20.
Witzmann FA, Grant RA. Pharmacoproteomics in drug development. Pharmacogenomics J. 2003;3(2):69–76.PubMedCrossRef
21.
Hueber W, Robinson WH. Genomics and proteomics: applications in autoimmune diseases. Pharmacogenomics Personalized Med. 2009;2:39–48.CrossRef
22.
Pinna S, Pasella S, Deiana M, Baralla A, Mannu A, Masala AGE, et al. Proteomic analysis of human plasma and peripheral blood mononuclear cells in systemic lupus erythematosus patients. J Immunol Methods. 2017;446:37–46.PubMedCrossRef
23.
Ferreira TAR, de Andrade HM, de Padua PM, Carvalho MDG, Pires SDF, Oliveira IHR, et al. Identification of potential biomarkers for systemic lupus erythematosus diagnosis using two-dimensional differential gel electrophoresis (2D-DIGE) and mass spectrometry. Autoimmunity. 2017;50(4):247–56.PubMedCrossRef
24.
Nicolaou O, Kousios A, Hadjisavvas A, Lauwerys B, Sokratous K, Kyriacou K. Biomarkers of systemic lupus erythematosus identified using mass spectrometry-based proteomics: a systematic review. J Cell Mol Med. 2017;21(5):993–1012.PubMedCrossRef
25.
Aggarwal A, Gupta R, Negi VS, Rajasekhar L, Misra R, Singh P, et al. Urinary haptoglobin, alpha-1 anti-chymotrypsin and retinol binding protein identified by proteomics as potential biomarkers for lupus nephritis. Clin Exp Immunol. 2017;188(2):254–62.PubMedPubMedCentralCrossRef
26.
Abrao AL, Falcao DP, de Amorim RF, Bezerra AC, Pombeiro GA, Guimaraes LJ, et al. Salivary proteomics: a new adjuvant approach to the early diagnosis of familial juvenile systemic lupus erythematosus. Med Hypotheses. 2016;89:97–100.PubMedCrossRef
27.
Alaiya A, Assad L, Alkhafaji D, Shinwari Z, Almana H, Shoukri M, et al. Proteomic analysis of class IV lupus nephritis. Nephrol Dial Transplant. 2015;30(1):62–70.PubMedCrossRef
28.
Bahjat FR, Pine PR, Reitsma A, Cassafer G, Baluom M, Grillo S, et al. An orally bioavailable spleen tyrosine kinase inhibitor delays disease progression and prolongs survival in murine lupus. Arthritis Rheum. 2008;58(5):1433–44.PubMedCrossRef
29.
Hou LF, He SJ, Li X, Yang Y, He PL, Zhou Y, et al. Oral administration of artemisinin analog SM934 ameliorates lupus syndromes in MRL/lpr mice by inhibiting Th1 and Th17 cell responses. Arthritis Rheum. 2011;63(8):2445–55.PubMedCrossRef
30.
Monahan E, Yamazaki K. An Improved Urine Collection Technique for Laboratory Mice: The Bladder Massage Method. Lab Animal. 1993;22(6):38-39.
31.
Wisniewski JR. Proteomic sample preparation from formalin fixed and paraffin embedded tissue. Journal of visualized experiments: JoVE. 2013;(79):50589.
32.
Wisniewski JR, Zougman A, Nagaraj N, Mann M. Universal sample preparation method for proteome analysis. Nat Methods. 2009;6(5):359–62.PubMedCrossRef
33.
Liu X, Xu Y, Meng Q, Zheng Q, Wu J, Wang C, et al. Proteomic analysis of minute amount of colonic biopsies by enteroscopy sampling. Biochem Biophys Res Commun. 2016;476(4):286–92.PubMedCrossRef
34.
Chen ZG, Liu X, Wang W, Geng F, Gao J, Gan CL, et al. Dissociative role for dorsal hippocampus in mediating heroin self-administration and relapse through CDK5 and RhoB signaling revealed by proteomic analysis. Addict Biol. 2017;22(6):1731–42.PubMedCrossRef
35.
Cox J, Mann M. MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification. Nat Biotechnol. 2008;26(12):1367–72.PubMedCrossRef
36.
Cox J, Neuhauser N, Michalski A, Scheltema RA, Olsen JV, Mann M. Andromeda: a peptide search engine integrated into the MaxQuant environment. J Proteome Res. 2011;10(4):1794–805.PubMedCrossRef
37.
Cox J, Hein MY, Luber CA, Paron I, Nagaraj N, Mann M. Accurate proteome-wide label-free quantification by delayed normalization and maximal peptide ratio extraction, termed MaxLFQ. Mol Cell Proteomics. 2014;13(9):2513–26.PubMedPubMedCentralCrossRef
38.
Vizcaino JA, Csordas A, del-Toro N, Dianes JA, Griss J, Lavidas I, et al. 2016 update of the PRIDE database and its related tools. Nucleic Acids Res. 2016;44(D1):D447–56.PubMedCrossRef
39.
40.
Yang Y, Guo L, Blattner SM, Mundel P, Kretzler M, Wu C. Formation and phosphorylation of the PINCH-1-integrin linked kinase-alpha-parvin complex are important for regulation of renal glomerular podocyte adhesion, architecture, and survival. J Am Soc Nephrol. 2005;16(7):1966–76.PubMedCrossRef
41.
Dai C, Stolz DB, Bastacky SI, St-Arnaud R, Wu C, Dedhar S, et al. Essential role of integrin-linked kinase in podocyte biology: bridging the integrin and slit diaphragm signaling. J Am Soc Nephrol. 2006;17(8):2164–75.PubMedCrossRef
42.
Tassiulas IO, Boumpas DT. Clinical features and treatment of systemic lupus erythematosus; 2008.
43.
Wang Y, Hu Q, Madri JA, Rollins SA, Chodera A, Matis LA. Amelioration of lupus-like autoimmune disease in NZB/W F1 mice after treatment with a blocking monoclonal antibody specific for complement component C5. Proc Natl Acad Sci U S A. 1996;93(16):8563–8.PubMedPubMedCentralCrossRef
44.
45.
Perico L, Conti S, Benigni A, Remuzzi G. Podocyte-actin dynamics in health and disease. Nat Rev Nephrol. 2016;12(11):692–710.PubMedCrossRef
46.
Asanuma K, Yanagida-Asanuma E, Faul C, Tomino Y, Kim K, Mundel P. Synaptopodin orchestrates actin organization and cell motility via regulation of RhoA signalling. Nat Cell Biol. 2006;8(5):485–91.PubMedCrossRef
47.
Faul C, Donnelly M, Merschergomez S, Chang YH, Franz S, Delfgaauw J, et al. The actin cytoskeleton of kidney podocytes is a direct target of the antiproteinuric effect of cyclosporine A. Nat Med. 2008;14(9):931–8.PubMedPubMedCentralCrossRef
48.
Yao J, Le TC, Kos CH, Henderson JM, Allen PG, Denker BM, et al. Alpha-actinin-4-mediated FSGS: an inherited kidney disease caused by an aggregated and rapidly degraded cytoskeletal protein. PLoS Biol. 2004;2(6):e167.PubMedPubMedCentralCrossRef
49.
Lennon R, Randles MJ, Humphries MJ. The importance of podocyte adhesion for a healthy glomerulus. Front Endocrinol (Lausanne). 2014;5:160.CrossRef
50.
Yen CF, Wang HS, Lee CL, Liao SK. Roles of integrin-linked kinase in cell signaling and its perspectives as a therapeutic target. Gynecol Minim Invasive Ther. 2014;3(3):67–72.CrossRef
51.
Ghatak S, Morgner J, Wickström SA. ILK: a pseudokinase with a unique function in the integrin-actin linkage. Biochem Soc Trans. 2013;41(4):995–1001.PubMedCrossRef
52.
Fukuda K, Gupta S, Chen K, Wu C, Qin J. The pseudo-active site of ILK is essential for its binding to α-parvin and localization to focal adhesions. Mol Cell. 2009;36(5):819.PubMedPubMedCentralCrossRef
53.
Brunton VG, MacPherson IRJ, Frame MC. Cell adhesion receptors, tyrosine kinases and actin modulators: a complex three-way circuitry. Biochimica et Biophysica Acta (BBA)-Molecular Cell Research. 2004;1692(2-3):121-144.
54.
Goto H, Wakui H, Komatsuda A, Ohtani H, Imai H, Sawada K, et al. Renal alpha-actinin-4: purification and puromycin aminonucleoside-binding property. Nephron Exp Nephrol. 2003;93(1):e27–35.PubMedCrossRef
55.
Guan N, Ding J, Deng J, Zhang J, Yang J. Key molecular events in puromycin aminonucleoside nephrosis rats. Pathol Int. 2004;54(9):703–11.PubMedCrossRef
56.
Deocharan B, Qing X, Lichauco J, Putterman C. α-Actinin is a cross-reactive renal target for pathogenic anti-DNA antibodies. J Immunol. 2002;168(6):3072–8.PubMedCrossRef
57.
Mostoslavsky G, Fischel R, Yachimovich N, Yarkoni Y, Rosenmann E, Monestier M, et al. Lupus anti-DNA autoantibodies cross-react with a glomerular structural protein: a case for tissue injury by molecular mimicry. Eur J Immunol. 2001;31(4):1221–7.PubMedCrossRef
58.
Terman JR, Kashina A. Post-translational modification and regulation of actin. Curr Opin Cell Biol. 2013;25(1):30–8.PubMedCrossRef
Metadata
Title
Reversible SAHH inhibitor protects against glomerulonephritis in lupus-prone mice by downregulating renal α-actinin-4 expression and stabilizing integrin-cytoskeleton linkage
Authors
Shijun He
Xing Liu
Zemin Lin
Yuting Liu
Lei Gu
Hu Zhou
Wei Tang
Jianping Zuo
Publication date
01-12-2019
Publisher
BioMed Central
Published in
Arthritis Research & Therapy / Issue 1/2019
Electronic ISSN: 1478-6362
DOI
https://doi.org/10.1186/s13075-019-1820-3

Other articles of this Issue 1/2019

Arthritis Research & Therapy 1/2019 Go to the issue

Research article

High-throughput sequencing of CD4+ T cell repertoire reveals disease-specific signatures in IgG4-related disease

Research article

11β-HSD1 plays a critical role in trabecular bone loss associated with systemic glucocorticoid therapy

Letter

Response to: “A somatization comorbidity phenotype impacts response to therapy in rheumatoid arthritis: post hoc results from the certolizumab pegol phase 4 PREDICT trial”

Research article

Antifibrotic effects of 2-carba cyclic phosphatidic acid (2ccPA) in systemic sclerosis: contribution to the novel treatment

Research article

Effect of disease-modifying anti-rheumatic drugs on bone structure and strength in psoriatic arthritis patients

Research article

Microbiome dysbiosis is associated with disease duration and increased inflammatory gene expression in systemic sclerosis skin
Live Webinar | 27-06-2024 | 18:00 (CEST)

Keynote webinar | Spotlight on medication adherence

Reserve your place

Live: Thursday 27th June 2024, 18:00-19:30 (CEST)

WHO estimates that half of all patients worldwide are non-adherent to their prescribed medication. The consequences of poor adherence can be catastrophic, on both the individual and population level.

Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.

Prof. Kevin Dolgin
Prof. Florian Limbourg
Prof. Anoop Chauhan
Developed by: Springer Medicine
Obesity Clinical Trial Summary

At a glance: The STEP trials

Read more

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Highlights from the ACC 2024 Congress

Year in Review: Pediatric cardiology

Pediatric Cardiology Video

Watch Dr. Anne Marie Valente present the last year's highlights in pediatric and congenital heart disease in the official ACC.24 Year in Review session.

Year in Review: Pulmonary vascular disease

Cardiopulmonary Comorbidity Video

The last year's highlights in pulmonary vascular disease are presented by Dr. Jane Leopold in this official video from ACC.24.

Year in Review: Valvular heart disease

Valvular Heart Disease Video

Watch Prof. William Zoghbi present the last year's highlights in valvular heart disease from the official ACC.24 Year in Review session.

Year in Review: Heart failure and cardiomyopathies

Heart Failure Video

Watch this official video from ACC.24. Dr. Biykem Bozkurt discusses last year's major advances in heart failure and cardiomyopathies.

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Heart Failure Video

Watch this official video from ACC.24. Dr. Biykem Bozkurt discusses last year's major advances in heart failure and cardiomyopathies.

Watch now

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

16-04-2024 Type 2 Diabetes News

Incentivised to exercise

11-04-2024 Lifestyle Modifications News

New intervention for STEMI-related cardiogenic shock

10-04-2024 Myocardial Infarction News

» View more highlights on the ACC 2024 congress hub page

Image Credits