Diabetic kidney disease (DKD) is the leading cause of end-stage kidney disease, and podocyte injury is one of the major contributors to DKD. As a crucial transcriptional factor that regulates cellular response to oxidative stress, nuclear factor erythroid 2-related factor 2 (Nrf2) is an attractive therapeutic target for DKD. In this study, we evaluated the therapeutic potential of DDO-1039, a novel small-molecule Nrf2 activator developed with protein–protein interaction strategy, on podocyte injury in DKD.
Results:
DDO-1039 treatment significantly increased Nrf2 protein level and Nrf2 nuclear translocation, thereby upregulating Nrf2 target genes [heme oxygenase 1, NAD(P)H quinone dehydrogenase 1, glutamate-cysteine ligase modifier, and tyrosine-protein kinase receptor] both in vitro and in vivo. DDO-1039 attenuated glomerular sclerosis and podocyte injury in the high-fat diet/streptozotocin-induced (HFD/STZ) diabetic mice and db/db diabetic mice. It also significantly improved hyperglycemia in both diabetic mice and mitigated proteinuria in HFD/STZ mice. Meanwhile, DDO-1039 attenuated oxidative stress and inflammation as well as apoptosis in vivo and in podocytes stimulated with palmitic acid and high glucose. Interestingly, we identified podocyte protective factor Tyro3 as a novel Nrf2-regulated gene. In addition, podocyte ferroptosis is reduced via activation of glutathione peroxidase 4 by the novel Nrf2 activator.
Innovation and conclusion:
DDO-1039 activates the Nrf2-based cytoprotective system to mitigate podocyte injury in the context of diabetes, suggesting the potential of DDO-1039 in the treatment of DKD. Antioxid. Redox Signal. 42, 787–806.
AlicicRZ, RooneyMT, TuttleKR. Diabetic kidney disease: Challenges, progress, and possibilities. Clin J Am Soc Nephrol, 2017; 12(12):2032–2045; doi: 10.2215/CJN.11491116
3.
AssadyS, WannerN, SkoreckiKL, et al.New insights into podocyte biology in glomerular health and disease. J Am Soc Nephrol, 2017; 28(6):1707–1715; doi: 10.1681/asn.2017010027
4.
BelavgeniA, MeyerC, StumpfJ, et al.Ferroptosis and necroptosis in the kidney. Cell Chem Biol, 2020; 27(4):448–462; doi: 10.1016/j.chembiol.2020.03.016
5.
BenzingT, SalantD. Insights into glomerular filtration and albuminuria. N Engl J Med, 2021; 384(15):1437–1446; doi: 10.1056/NEJMra1808786
ChenJ, OuZ, GaoT, et al.Ginkgolide B alleviates oxidative stress and ferroptosis by inhibiting GPX4 ubiquitination to improve diabetic nephropathy. Biomed Pharmacother, 2022; 156:113953; doi: 10.1016/j.biopha.2022.113953
8.
TanaseDM, GosavEM, AntonMI, et al.Oxidative stress and NRF2/KEAP1/ARE pathway in Diabetic Kidney Disease (DKD): New perspectives. Biomolecules, 2022, 12, 9; doi: 10.3390/biom12091227
9.
DE ZeeuwD, AkizawaT, AudhyaP, et al.BEACON Trial Investigators. Bardoxolone methyl in type 2 diabetes and stage 4 chronic kidney disease. N Engl J Med, 2013; 369(26):2492–2503; doi: 10.1056/NEJMoa1306033
10.
DengHF, YueLX, WangNN, et al.Mitochondrial iron overload-mediated inhibition of Nrf2-HO-1/GPX4 assisted ALI-induced nephrotoxicity. Front Pharmacol, 2020; 11:624529; doi: 10.3389/fphar.2020.624529
11.
DodsonM, Castro-PortuguezR, ZhangDD. NRF2 plays a critical role in mitigating lipid peroxidation and ferroptosis. Redox Biol, 2019a;23:101107; doi: 10.1016/j.redox.2019.101107
12.
DodsonM, DE LA VegaMR, CholaniansAB, et al.Modulating NRF2 in disease: Timing is everything. Annu Rev Pharmacol Toxicol, 2019b;59:555–575; doi: 10.1146/annurev-pharmtox-010818-021856
13.
FengJ, XieL, YuX, et al.Perilipin 5 ameliorates high-glucose-induced podocyte injury via Akt/GSK-3beta/Nrf2-mediated suppression of apoptosis, oxidative stress, and inflammation. Biochem Biophys Res Commun, 2021; 544:22–30; doi: 10.1016/j.bbrc.2021.01.069
14.
FuC, WuY, LiuS, et al.Rehmannioside A improves cognitive impairment and alleviates ferroptosis via activating PI3K/AKT/Nrf2 and SLC7A11/GPX4 signaling pathway after ischemia. J Ethnopharmacol, 2022; 289:115021; doi: 10.1016/j.jep.2022.115021
15.
GaoZ, ZhangZ, GuD, et al.Hemin mitigates contrast-induced nephropathy by inhibiting ferroptosis via HO-1/Nrf2/GPX4 pathway. Clin Exp Pharmacol Physiol, 2022; 49(8):858–870; doi: 10.1111/1440-1681.13673
16.
GargP. A review of podocyte biology. Am J Nephrol, 2018; 47 (Suppl 1):3–13; doi: 10.1159/000481633
17.
HeM, LiY, WangL, et al.MYDGF attenuates podocyte injury and proteinuria by activating Akt/BAD signal pathway in Mice with diabetic kidney disease. Diabetologia, 2020; 63(9):1916–1931; doi: 10.1007/s00125-020-05197-2
18.
HerremaH, GuanD, ChoiJW, et al.FKBP11 rewires UPR signaling to promote glucose homeostasis in type 2 diabetes and obesity. Cell Metab, 2022; 34(7):1004–1022 e8; doi: 10.1016/j.cmet.2022.06.007
19.
ItoM, TanakaT, NangakuM. Nuclear factor erythroid 2-related factor 2 as a treatment target of kidney diseases. Curr Opin Nephrol Hypertens, 2020; 29(1):128–135; doi: 10.1097/MNH.0000000000000556
20.
JiangA, SongA, ZhangC. Modes of podocyte death in diabetic kidney disease: An update. J Nephrol, 2022; 35(6):1571–1584; doi: 10.1007/s40620-022-01269-1
21.
Hu A#J, Ning Ma AWGB, Fan AX, et al.Leonurine hydrochloride alleviates ferroptosis in cisplatin-induced acute kidney injury by activating the Nrf2 signaling pathway. Br J Pharmacol, 2022; 179:3991–4009; doi: 10.1002/bph.15834
22.
JinJ, WangY, ZhengD, et al.A novel identified circular RNA, mmu_mmu_circRNA_0000309, involves in germacrone-mediated improvement of diabetic nephropathy through regulating ferroptosis by targeting miR-188-3p/GPX4 signaling axis. Antioxid Redox Signal, 2022; 36(10–12):740–759; doi: 10.1089/ars.2021.0063
23.
KandaH, YamawakiK. Bardoxolone methyl: Drug development for diabetic kidney disease. Clin Exp Nephrol, 2020; 24(10):857–864; doi: 10.1007/s10157-020-01917-5
KimS, KangSW, JooJ, et al.Characterization of ferroptosis in kidney tubular cell death under diabetic conditions. Cell Death Dis, 2021; 12(2):160; doi: 10.1038/s41419-021-03452-x
26.
LiS, ZhengL, ZhangJ, et al.Inhibition of ferroptosis by up-regulating Nrf2 delayed the progression of diabetic nephropathy. Free Radic Biol Med, 2021; 162:435–449; doi: 10.1016/j.freeradbiomed.2020.10.323
27.
LiuX-Q, JiangL, LiY-Y, et al.Wogonin protects glomerular podocytes by targeting Bcl-2-mediated autophagy and apoptosis in diabetic kidney disease. Acta Pharmacol Sin, 2022a;43(1):96–110; doi: 10.1038/s41401-021-00721-5
28.
LiuY, UrunoA, SaitoR, et al.Nrf2 deficiency deteriorates diabetic kidney disease in Akita model Mice. Redox Biol, 2022b;58:102525; doi: 10.1016/j.redox.2022.102525
29.
LuM-C, JiJ-A, JiangY-L, et al.An inhibitor of the Keap1-Nrf2 protein-protein interaction protects NCM460 colonic cells and alleviates experimental colitis. Sci Rep, 2016; 6:26585; doi: 10.1038/srep26585
30.
LuMC, ShaoHL, LiuT, et al.Discovery of 2-oxy-2-phenylacetic acid substituted naphthalene sulfonamide derivatives as potent KEAP1-NRF2 protein-protein interaction inhibitors for inflammatory conditions. Eur J Med Chem, 2020; 207:112734; doi: 10.1016/j.ejmech.2020.112734
31.
LuMC, ZhaoJ, LiuYT, et al.CPUY192018, a potent inhibitor of the Keap1-Nrf2 protein-protein interaction, alleviates renal inflammation in Mice by restricting oxidative stress and NF-kappaB activation. Redox Biol, 2019; 26:101266; doi: 10.1016/j.redox.2019.101266
32.
LytvynY, BjornstadP, VAN RaalteDH, et al.The new biology of diabetic kidney disease-mechanisms and therapeutic implications. Endocr Rev, 2020; 41(2):202–231; doi: 10.1210/endrev/bnz010
33.
MaJ, LiC, LiuT, et al.Identification of markers for diagnosis and treatment of diabetic kidney disease based on the ferroptosis and immune. Oxid Med Cell Longev, 2022; 2022:9957172; doi: 10.1155/2022/9957172
34.
MathiesonPW. The podocyte as a target for therapies–new and old. Nat Rev Nephrol, 2011; 8(1):52–56; doi: 10.1038/nrneph.2011.171
35.
MatobaK, TakedaY, NagaiY, et al.Unraveling the role of inflammation in the pathogenesis of diabetic kidney disease. Int J Mol Sci, 2019; 20(14); doi: 10.3390/ijms20143393
36.
MohandesS, DokeT, HuH, et al.Molecular pathways that drive diabetic kidney disease. J Clin Invest, 2023; 133(4); doi: 10.1172/JCI165654
37.
MohanyM, AhmedMM, Al-RejaieSS. Molecular mechanistic pathways targeted by natural antioxidants in the prevention and treatment of chronic kidney disease. Antioxidants (Basel), 2021; 11(1); doi: 10.3390/antiox11010015
38.
Moin A SaleemMJOH, ReiserJ, CowardRJ, et al.A conditionally immortalized human podocyte cell line demonstrating nephrin and podocin expression. J Am Soc Nephrol, 2002; 13(3):630–638; doi: 10.1681/ASN.V133630
39.
MolinariP, CaldiroliL, DozioE, et al.AGEs and sRAGE variations at different timepoints in patients with chronic kidney disease. Antioxidants (Basel), 2021; 10(12); doi: 10.3390/antiox10121994
40.
NagataM. Podocyte injury and its consequences. Kidney Int, 2016; 89(6):1221–1230; doi: 10.1016/j.kint.2016.01.012
41.
NezuM, SuzukiN. Roles of Nrf2 in protecting the kidney from oxidative damage. Int J Mol Sci, 2020; 21(8); doi: 10.3390/ijms21082951
42.
NezuM, SuzukiN, YamamotoM. Targeting the KEAP1-NRF2 system to prevent kidney disease progression. Am J Nephrol, 2017; 45(6):473–483; doi: 10.1159/000475890
43.
NguyenD, PingF, MuW, et al.Macrophage accumulation in human progressive diabetic nephropathy. Nephrology (Carlton), 2006; 11(3):226–231; doi: 10.1111/j.1440-1797.2006.00576.x
44.
OstergaardJA, CooperME, Jandeleit-DahmKAM. Targeting oxidative stress and anti-oxidant defence in diabetic kidney disease. J Nephrol, 2020; 33(5):917–929; doi: 10.1007/s40620-020-00749-6
45.
PiaoH-L, LiuH, XiaT, et al.KEAP1 promotes anti-tumor immunity by inhibiting PD-L1 expression in NSCLC. Cell Death Di, 2023; doi: 10.21203/rs.3.rs-3158334/v1
46.
PodgorskiP, KoniecznyA, LisL, et al.Glomerular podocytes in diabetic renal disease. Adv Clin Exp Med, 2019; 28(12):1711–1715; doi: 10.17219/acem/104534
47.
QiuW, AnS, WangT, et al.Melatonin suppresses ferroptosis via activation of the Nrf2/HO-1 signaling pathway in the mouse model of sepsis-induced acute kidney injury. Int Immunopharmacol, 2022; 112:109162; doi: 10.1016/j.intimp.2022.109162
48.
RajeshK, ThimmulappaRalph J, MalhotraD, et al.Preclinical evaluation of targeting the Nrf2 pathway by triterpenoids (CDDO-Im and CDDO-Me) for protection from LPSInduced inflammatory response and reactive oxygen species in human peripheral blood mononuclear cells and neutrophils. Antioxid Redox Signal, 2007; 9:7
49.
RuizS, PergolaPE, ZagerRA, et al.Targeting the transcription factor Nrf2 to ameliorate oxidative stress and inflammation in chronic kidney disease. Kidney Int, 2013; 83(6):1029–1041; doi: 10.1038/ki.2012.439
50.
RushBM, BondiCD, StockerSD, et al.Genetic or pharmacologic Nrf2 activation increases proteinuria in chronic kidney disease in mice. Kidney Int, 2021; 99(1):102–116; doi: 10.1016/j.kint.2020.07.036
51.
SakashitaM, TanakaT, InagiR. Metabolic changes and oxidative stress in diabetic kidney disease. Antioxidants (Basel), 2021; 10(7); doi: 10.3390/antiox10071143
52.
SeibtTM, PronethB, ConradM. Role of GPX4 in ferroptosis and its pharmacological implication. Free Radic Biol Med, 2019; 133:144–152; doi: 10.1016/j.freeradbiomed.2018.09.014
ShebanD, MerblY. EMSY stabilization in KEAP1-mutant lung cancer disrupts genome stability and type I interferon signaling. Cell Death Differ, 2023; 30(5):1397–1399; doi: 10.1038/s41418-023-01150-z
55.
StockwellBR, Friedmann AngeliJP, BayirH, et al.Ferroptosis: A regulated cell death nexus linking metabolism, redox biology, and disease. Cell, 2017; 171(2):273–285; doi: 10.1016/j.cell.2017.09.021
TuttleKR, AgarwalR, AlpersCE, et al.Molecular mechanisms and therapeutic targets for diabetic kidney disease. Kidney Int, 2022; 102(2):248–260; doi: 10.1016/j.kint.2022.05.012
58.
UlasovAV, RosenkranzAA, GeorgievGP, et al.Nrf2/Keap1/ARE signaling: Towards specific regulation. Life Sci, 2022; 291:120111; doi: 10.1016/j.lfs.2021.120111
59.
UrunoA, FurusawaY, YagishitaY, et al.The keap1-Nrf2 system prevents onset of diabetes mellitus. Mol Cell Biol, 2013; 33(15):2996–3010; doi: 10.1128/mcb.00225-13
60.
WangYH, ChangDY, ZhaoMH, et al.Glutathione peroxidase 4 is a predictor of diabetic kidney disease progression in type 2 diabetes mellitus. Oxid Med Cell Longev, 2022d;2022:2948248; doi: 10.1155/2022/2948248
61.
WangC, ChenS, GuoH, et al.Forsythoside a mitigates alzheimer’s-like pathology by inhibiting ferroptosis-mediated neuroinflammation via Nrf2/GPX4 axis activation. Int J Biol Sci, 2022a;18(5):2075–2090; doi: 10.7150/ijbs.69714
62.
WangWJ, JiangX, GaoCC, et al.Salusinbeta participates in high glucoseinduced HK2 cell ferroptosis in a Nrf2dependent manner. Mol Med Rep, 2021:24; doi: 10.3892/mmr.2021.12313
63.
WangX, LiuQ, KongD, et al.Down-regulation of SETD6 protects podocyte against high glucose and palmitic acid-induced apoptosis, and mitochondrial dysfunction via activating Nrf2-Keap1 signaling pathway in diabetic nephropathy. J Mol Histol, 2020; 51(5):549–558; doi: 10.1007/s10735-020-09904-6
64.
WangY, YanS, LiuX, et al.PRMT4 promotes ferroptosis to aggravate doxorubicin-induced cardiomyopathy via inhibition of the Nrf2/GPX4 pathway. Cell Death Differ, 2022c;29(10):1982–1995; doi: 10.1038/s41418-022-00990-5
65.
WangJ, ZhuQ, WangY, et al.Irisin protects against sepsis-associated encephalopathy by suppressing ferroptosis via activation of the Nrf2/GPX4 signal axis. Free Radic Biol Med, 2022b;187:171–184; doi: 10.1016/j.freeradbiomed.2022.05.023
66.
WuY, ChenY. Research progress on ferroptosis in diabetic kidney disease. Front Endocrinol (Lausanne), 2022; 13:945976; doi: 10.3389/fendo.2022.945976
67.
WuWY, WangZX, LiTS, et al.SSBP1 drives high fructose-induced glomerular podocyte ferroptosis via activating DNA-PK/p53 pathway. Redox Biol, 2022; 52:102303; doi: 10.1016/j.redox.2022.102303
68.
WuY, ZhaoY, YangHZ, et al.HMGB1 regulates ferroptosis through Nrf2 pathway in mesangial cells in response to high glucose. Biosci Rep, 2021; 41(2); doi: 10.1042/BSR20202924
69.
XiaoL, XuX, ZhangF, et al.The mitochondria-targeted antioxidant MitoQ ameliorated tubular injury mediated by mitophagy in diabetic kidney disease via Nrf2/PINK1. Redox Biol, 2017; 11:297–311; doi: 10.1016/j.redox.2016.12.022
70.
XingL, FangJ, ZhuB, et al.Astragaloside IV protects against podocyte apoptosis by inhibiting oxidative stress via activating PPARgamma-Klotho-FoxO1 axis in diabetic nephropathy. Life Sci, 2021a;269:119068; doi: 10.1016/j.lfs.2021.119068
71.
XingL, GuoH, MengS, et al.Klotho ameliorates diabetic nephropathy by activating Nrf2 signaling pathway in podocytes. Biochem Biophys Res Commun, 2021b;534:450–456; doi: 10.1016/j.bbrc.2020.11.061
72.
XuC, LiuX, ZhaiX, et al.CDDO-Me ameliorates podocyte injury through anti-oxidative stress and regulation of actin cytoskeleton in adriamycin nephropathy. Biomed Pharmacother, 2023; 167:115617; doi: 10.1016/j.biopha.2023.115617
73.
YagishitaY, UrunoA, FukutomiT, et al.Nrf2 improves leptin and insulin resistance provoked by hypothalamic oxidative stress. Cell Rep, 2017; 18(8):2030–2044; doi: 10.1016/j.celrep.2017.01.064
74.
ZhangQ, HuY, HuJ-E, et al.Sp1-mediated upregulation of Prdx6 expression prevents podocyte injury in diabetic nephropathy via mitigation of oxidative stress and ferroptosis. Life Sci, 2021; 278:119529; doi: 10.1016/j.lfs.2021.119529
75.
ZhangQ, HuY, HuJE, et al.Solasonine alleviates high glucose-induced podocyte injury through increasing Nrf2-medicated inhibition of NLRP3 activation. Drug Dev Res, 2022a;83(7):1697–1706; doi: 10.1002/ddr.21988
76.
ZhangL, JiangS, ShiJ, et al.TYRO3 protects podocyte via JNK/c-jun-P53 pathway. Arch Biochem Biophys, 2023; 739:109578; doi: 10.1016/j.abb.2023.109578
77.
ZhangY, WuQ, LiuJ, et al.Sulforaphane alleviates high fat diet-induced insulin resistance via AMPK/Nrf2/GPx4 axis. Biomed Pharmacother, 2022b;152:113273; doi: 10.1016/j.biopha.2022.113273
78.
ZhengH, WhitmanSA, WuW, et al.Therapeutic potential of Nrf2 activators in streptozotocin-induced diabetic nephropathy. Diabetes, 2011; 60(11):3055–3066; doi: 10.2337/db11-0807
79.
ZhongF, CaiH, FuJ, et al.TYRO3 agonist as therapy for glomerular disease. JCI Insight, 2023; 8(1); doi: 10.1172/jci.insight.165207
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