DDO1002, an NRF2–KEAP1 inhibitor, improves hematopoietic stem cell aging and stress response

Abstract

Oxidative stress diminishes the functionality of hematopoietic stem cells (HSCs) as age advances, with heightened reactive oxygen species (ROS) levels exacerbating DNA damage, cellular senescence, and hematopoietic impairment. DDO1002, a potent inhibitor of the NRF2–KEAP1 pathway, modulates the expression of antioxidant genes. Yet, the extent to which it mitigates hematopoietic decline post-total body irradiation (TBI) or in the context of aging remains to be elucidated. Our study has elucidated the role of DDO1002 in modulating NRF2 activity, which, in turn, activates the NRF2-driven antioxidant response element (ARE) signaling cascade. This activation can diminish intracellular levels of ROS, thereby attenuating cellular senescence. In addition, DDO1002 has been demonstrated to ameliorate DNA damage and avert HSC apoptosis, underscoring its potential to mitigate hematopoietic injury precipitated by TBI. Competitive transplantation assay revealed that the administration of DDO1002 can improve the reconstitution and self-renewal capacity of HSCs in aged mice. Single-cell sequencing analysis elucidated that DDO1002 treatment attenuated intracellular inflammatory signaling pathways and mitigated ROS pathway in aged HSCs, suggesting its potential to restore the viability of these cells. Consequently, DDO1002 effectively activated the NRF2–ARE pathway, delaying cellular senescence and ameliorating impaired hematopoiesis, thereby demonstrating its potential as a therapeutic agent for age-related hematopoietic disorders.

Keywords

hematopoietic stem cell / NRF2 / aging / reactive oxygen species

References

[1] Liang Y , Van Zant G , Szilvassy SJ . Effects of aging on the homing and engraftment of murine hematopoietic stem and progenitor cells. Blood 2005; 106: 1479- 87.

[2] Ghaffari S . Oxidative stress in the regulation of normal and neoplastic hematopoiesis. Antioxid Redox Signal 2008; 10: 1923- 40.

[3] McMahon M , Thomas N , Itoh K , et al. Redox-regulated turnover of Nrf2 is determined by at least two separate protein domains, the redox-sensitive Neh2 degron and the redox-insensitive Neh6 degron. J Biol Chem 2004; 279: 31556- 67.

[4] Hayes JD , Dinkova-Kostova AT . The Nrf2 regulatory network provides an interface between redox and intermediary metabolism. Trends Biochem Sci 2014; 39: 199- 218.

[5] Itoh K , Mimura J , Yamamoto M . Discovery of the negative regulator of Nrf2, Keap1: a historical overview. Antioxid Redox Signal 2010; 13: 1665- 78.

[6] Wang H , Liu K , Geng M , et al. RXRα inhibits the NRF2-ARE signaling pathway through a direct interaction with the Neh7 domain of NRF2. Cancer Res 2013; 73: 3097- 108.

[7] Modi R , McKee N , Zhang N , et al. Stapled peptides as direct inhibitors of Nrf2-sMAF transcription factors. J Med Chem 2023; 66: 6184- 92.

[8] Kubben N , Zhang W , Wang L , et al. Repression of the antioxidant NRF2 pathway in premature aging. Cell 2016; 165: 1361- 74.

[9] Song G , Wang J , Liu J , et al. Dimethyl fumarate ameliorates erectile dysfunction in bilateral cavernous nerve injury rats by inhibiting oxidative stress and NLRP3 inflammasome-mediated pyroptosis of nerve via activation of Nrf2/HO-1 signaling pathway. Redox Biol 2023; 68: 102938.

[10] Zhang DD , Hannink M . Distinct cysteine residues in Keap1 are required for Keap1-dependent ubiquitination of Nrf2 and for stabilization of Nrf2 by chemopreventive agents and oxidative stress. Mol Cell Biol 2003; 23: 8137- 51.

[11] Cho H , Hartsock MJ , Xu Z , et al. Monomethyl fumarate promotes Nrf2-dependent neuroprotection in retinal ischemia-reperfusion. J Neuroinflammation 2015; 12: 239.

[12] Yan N , Xu Z , Qu C , et al. Dimethyl fumarate improves cognitive deficits in chronic cerebral hypoperfusion rats by alleviating inflammation, oxidative stress, and ferroptosis via NRF2/ARE/NF-κB signal pathway. Int Immunopharmacol 2021; 98: 107844.

[13] Mohamadi N , Baradaran Rahimi V , Fadaei MR , et al. A mechanistic overview of sulforaphane and its derivatives application in diabetes and its complications. Inflammopharmacol 2023; 31: 2885- 99.

[14] Reisman SA , Chertow GM , Hebbar S , et al. Bardoxolone methyl decreases megalin and activates Nrf2 in the kidney. J Am Soc Nephrol 2012; 23: 1663- 73.

[15] Rizk DV , Silva AL , Pergola PE , et al. Effects of bardoxolone methyl on magnesium in patients with Type 2 diabetes mellitus and chronic kidney disease. Cardiorenal Med 2019; 9: 316- 25.

[16] Fukutomi T , Takagi K , Mizushima T , et al. Kinetic, thermodynamic, and structural characterizations of the association between Nrf2-DLGex degron and Keap1. Mol Cell Biol 2014; 34: 832- 46.

[17] Jiang ZY , Lu M-C , Xu LL , et al. Discovery of potent Keap1-Nrf2 protein-protein interaction inhibitor based on molecular binding determinants analysis. J Med Chem 2014; 57: 2736- 45.

[18] Sun Y , Huang J , Chen Y , et al. Direct inhibition of Keap1-Nrf2 protein-protein interaction as a potential therapeutic strategy for Alzheimer’s disease. Bioorg Chem 2020; 103: 104172.

[19] Lu MC , Zhao J , Liu Y-T , et al. CPUY192018, a potent inhibitor of the Keap1-Nrf2 protein-protein interaction, alleviates renal inflammation in mice by restricting oxidative stress and NF-κB activation. Redox Biol 2019; 26: 101266.

[20] Merchant AA , Singh A , Matsui W , et al. The redox-sensitive transcription factor Nrf2 regulates murine hematopoietic stem cell survival independently of ROS levels. Blood 2011; 118: 6572- 9.

[21] Tsai JJ , Dudakov JA , Takahashi K , et al. Nrf2 regulates haematopoietic stem cell function. Nat Cell Biol 2013; 15: 309- 16.

[22] Hu L , Zhang Y , Miao W , et al. Reactive oxygen species and Nrf2: functional and transcriptional regulators of hematopoiesis. Oxid Med Cell Longev 2019; 2019: 5153268.

[23] Zhang J , Xue X , Han X , et al. Vam3 ameliorates total body irradiation-induced hematopoietic system injury partly by regulating the expression of Nrf2-targeted genes. Free Radic Biol Med 2016; 101: 455- 64.

[24] Qiu X , Brown K , Hirschey MD , et al. Calorie restriction reduces oxidative stress by SIRT3-mediated SOD2 activation. Cell Metab 2010; 12: 662- 7.

[25] Wang W , Zheng Y , Sun S , et al. A genome-wide CRISPR-based screen identifies KAT7 as a driver of cellular senescence. Sci Transl Med 2021; 13: eabd2655.

[26] Li W , Wang X , Dong Y , et al. Nicotinamide riboside intervention alleviates hematopoietic system injury of ionizing radiation-induced premature aging mice. Aging Cell 2023; 22: e13976.

[27] Han X , Zhang J , Xue X , et al. Theaflavin ameliorates ionizing radiation-induced hematopoietic injury via the NRF2 pathway. Free Radic Biol Med 2017; 113: 59- 70.

[28] Santivasi WL , Xia F . Ionizing radiation-induced DNA damage, response, and repair. Antioxid Redox Signal 2014; 21: 251- 9.

[29] Xu G , Wu H , Zhang J , et al. Metformin ameliorates ionizing irradiation-induced long-term hematopoietic stem cell injury in mice. Free Radic Biol Med 2015; 87: 15- 25.

[30] de Haan G , Lazare SS . Aging of hematopoietic stem cells. Blood 2018; 131: 479- 87.

[31] Zeng X , Li X , Li X , et al. Fecal microbiota transplantation from young mice rejuvenates aged hematopoietic stem cells by suppressing inflammation. Blood 2023; 141: 1691- 707.

[32] Zhong J , Mao X , Li H , et al. Single-cell RNA sequencing analysis reveals the relationship of bone marrow and osteopenia in STZ-induced type 1 diabetic mice. J Adv Res 2022; 41: 145- 58.

[33] Silva-Palacios A , Ostolga-Chavarría M , Zazueta C , et al. Nrf2: molecular and epigenetic regulation during aging. Ageing Res Rev 2018; 47: 31- 40.

[34] O’Connell MA , Hayes JD . The Keap1/Nrf2 pathway in health and disease: from the bench to the clinic. Biochem Soc Trans 2015; 43: 687- 9.

[35] Lu L , Dong J , Li D , et al. 3,3’-diindolylmethane mitigates total body irradiation-induced hematopoietic injury in mice. Free Radic Biol Med 2016; 99: 463- 71.

[36] Pang WW , Price EA , Sahoo D , et al. Human bone marrow hematopoietic stem cells are increased in frequency and myeloid-biased with age. Proc Natl Acad Sci USA 2011; 108: 20012- 7.

[37] Kobayashi EH , Suzuki T , Funayama R , et al. Nrf2 suppresses macrophage inflammatory response by blocking proinflammatory cytokine transcription. Nat Commun 2016; 7: 11624.

[38] El-Shitany NA , Eid BG . Icariin modulates carrageenan-induced acute inflammation through HO-1/Nrf2 and NF-kB signaling pathways. Biomed Pharmacother 2019; 120: 109567.

[39] Huang JC , Yue Z-P , Yu H-F , et al. TAZ ameliorates the microglia-mediated inflammatory response via the Nrf2-ROS-NF-κB pathway. Mol Ther Nucleic Acids 2022; 28: 435- 49.

[40] Shimizu R , Hirano I , Hasegawa A , et al. Nrf2 alleviates spaceflight-induced immunosuppression and thrombotic microangiopathy in mice. Commun Biol 2023; 6: 875.

[41] Mikolajczyk TP , Szczepaniak P , Vidler F , et al. Role of inflammatory chemokines in hypertension. Pharmacol Ther 2021; 223: 107799.

[42] Jin G , Liu Y , Xu W , et al. Tnfaip2 promotes atherogenesis by enhancing oxidative stress induced inflammation. Mol Immunol 2022; 151: 41- 51.

[43] Zhang Y , Gao Y , Jiang Y , et al. Histone demethylase KDM5B licenses macrophage-mediated inflammatory responses by repressing Nfkbia transcription. Cell Death Differ 2023; 30: 1279- 92.

[44] Sharma BR , Karki R , Rajesh Y , et al. Immune regulator IRF1 contributes to ZBP1-, AIM2-, RIPK1-, and NLRP12-PANoptosome activation and inflammatory cell death (PANoptosis). J Biol Chem 2023; 299: 105141.

[45] Lou Q , Jiang K , Xu Q , et al. The RIG-I-NRF2 axis regulates the mesenchymal stromal niche for bone marrow transplantation. Blood 2022; 139: 3204- 21.

RIGHTS & PERMISSIONS

2024 The Author(s). Published by Oxford University Press on behalf of Higher Education Press.