TY - JOUR
T1 - An ERK5-NRF2 Axis Mediates Senescence-Associated Stemness and Atherosclerosis
AU - Abe, Jun-Ichi
AU - Imanishi, Masaki
AU - Li, Shengyu
AU - Zhang, Aijun
AU - Ko, Kyung Ae
AU - Samanthapudi, Venkata S K
AU - Lee, Ling-Ling
AU - Bojorges, Angelica Paniagua
AU - Gi, Young Jin
AU - Hobbs, Brian P
AU - Deswal, Anita
AU - Herrmann, Joerg
AU - Lin, Steven H
AU - Chini, Eduardo N
AU - Shen, Ying H
AU - Schadler, Keri L
AU - Nguyen, Thi-Hong-Minh
AU - Gupte, Anisha A
AU - Reyes-Gibby, Cielito
AU - Yeung, Sai-Ching J
AU - Abe, Rei J.
AU - Olmsted-Davis, Elizabeth A
AU - Krishnan, Sunil
AU - Dantzer, Robert
AU - Palaskas, Nicolas L
AU - Cooke, John P
AU - Pownall, Henry J
AU - Yoshimoto, Momoko
AU - Fujiwara, Keigi
AU - Hamilton, Dale J
AU - Burks, Jared K
AU - Wang, Guangyu
AU - Le, Nhat-Tu
AU - Kotla, Sivareddy
N1 - Funding Information:
This work was partially supported by grants from the National Institutes of Health to J.-i. Abe (HL-149303 and AI-156921), J.P. Cooke (HL-149303), and N.-T. Le (HL-134740 and HL-149303) and from the Cancer Prevention and Research Institute of Texas (CPRIT) to J.-i. Abe and K.L. Schadler (RP190256). This work is also partially supported by the University of Texas MD Anderson Cancer Center Institutional Research Grant Program to S. Kotla. This research was performed in the Flow Cytometry and Cellular Imaging Core Facility, which is supported, in part, by the National Institutes of Health through MD Anderson’s Cancer Center Support Grant CA016672, the NCI’s Research Specialist 1 R50 CA243707-01A1, and a Shared Instrumentation Award from CPRIT, RP121010.
Publisher Copyright:
© 2023 Lippincott Williams and Wilkins. All rights reserved.
PY - 2023/6/23
Y1 - 2023/6/23
N2 - BACKGROUND: ERK5 (extracellular signal-regulated kinase 5) is a dual kinase transcription factor containing an N-terminal kinase domain and a C-terminal transcriptional activation domain. Many ERK5 kinase inhibitors have been developed and tested to treat cancer and inflammatory diseases. However, recent data have raised questions about the role of the catalytic activity of ERK5 in proliferation and inflammation. We aimed to investigate how ERK5 reprograms myeloid cells to the proinflammatory senescent phenotype, subsequently leading to atherosclerosis.METHODS: A ERK5 S496A (dephosphorylation mimic) knock in (KI) mouse model was generated using CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/clustered regularly interspaced short palindromic repeat-associated 9), and atherosclerosis was characterized by hypercholesterolemia induction. The plaque phenotyping in homozygous ERK5 S496A KI and wild type (WT) mice was studied using imaging mass cytometry. Bone marrow-derived macrophages were isolated from hypercholesterolemic mice and characterized using RNA sequencing and functional in vitro approaches, including senescence, mitochondria reactive oxygen species, and inflammation assays, as well as by metabolic extracellular flux analysis.RESULTS: We show that atherosclerosis was inhibited in ERK5 S496A KI mice. Furthermore, ERK5 S496 phosphorylation mediates both senescence-associated secretory phenotype and senescence-associated stemness by upregulating AHR (aryl hydrocarbon receptor) in plaque and bone marrow-derived macrophages isolated from hypercholesterolemic mice. We also discovered that ERK5 S496 phosphorylation could induce NRF2 (NFE2-related factor 2) SUMOylation at a novel K518 site to inhibit NRF2 transcriptional activity without altering ERK5 catalytic activity and mediates oxidized LDL (low-density lipoprotein)-induced senescence-associated secretory phenotype. Specific ERK5 kinase inhibitors (AX15836 and XMD8-92) also inhibited ERK5 S496 phosphorylation, suggesting the involvement of ERK5 S496 phosphorylation in the anti-inflammatory effects of these ERK5 kinase inhibitors.CONCLUSIONS: We discovered a novel mechanism by which the macrophage ERK5-NRF2 axis develops a unique senescence-associated secretory phenotype/stemness phenotype by upregulating AHR to engender atherogenesis. The finding of senescence-associated stemness phenotype provides a molecular explanation to resolve the paradox of senescence in proliferative plaque by permitting myeloid cells to escape the senescence-induced cell cycle arrest during atherosclerosis formation.
AB - BACKGROUND: ERK5 (extracellular signal-regulated kinase 5) is a dual kinase transcription factor containing an N-terminal kinase domain and a C-terminal transcriptional activation domain. Many ERK5 kinase inhibitors have been developed and tested to treat cancer and inflammatory diseases. However, recent data have raised questions about the role of the catalytic activity of ERK5 in proliferation and inflammation. We aimed to investigate how ERK5 reprograms myeloid cells to the proinflammatory senescent phenotype, subsequently leading to atherosclerosis.METHODS: A ERK5 S496A (dephosphorylation mimic) knock in (KI) mouse model was generated using CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/clustered regularly interspaced short palindromic repeat-associated 9), and atherosclerosis was characterized by hypercholesterolemia induction. The plaque phenotyping in homozygous ERK5 S496A KI and wild type (WT) mice was studied using imaging mass cytometry. Bone marrow-derived macrophages were isolated from hypercholesterolemic mice and characterized using RNA sequencing and functional in vitro approaches, including senescence, mitochondria reactive oxygen species, and inflammation assays, as well as by metabolic extracellular flux analysis.RESULTS: We show that atherosclerosis was inhibited in ERK5 S496A KI mice. Furthermore, ERK5 S496 phosphorylation mediates both senescence-associated secretory phenotype and senescence-associated stemness by upregulating AHR (aryl hydrocarbon receptor) in plaque and bone marrow-derived macrophages isolated from hypercholesterolemic mice. We also discovered that ERK5 S496 phosphorylation could induce NRF2 (NFE2-related factor 2) SUMOylation at a novel K518 site to inhibit NRF2 transcriptional activity without altering ERK5 catalytic activity and mediates oxidized LDL (low-density lipoprotein)-induced senescence-associated secretory phenotype. Specific ERK5 kinase inhibitors (AX15836 and XMD8-92) also inhibited ERK5 S496 phosphorylation, suggesting the involvement of ERK5 S496 phosphorylation in the anti-inflammatory effects of these ERK5 kinase inhibitors.CONCLUSIONS: We discovered a novel mechanism by which the macrophage ERK5-NRF2 axis develops a unique senescence-associated secretory phenotype/stemness phenotype by upregulating AHR to engender atherogenesis. The finding of senescence-associated stemness phenotype provides a molecular explanation to resolve the paradox of senescence in proliferative plaque by permitting myeloid cells to escape the senescence-induced cell cycle arrest during atherosclerosis formation.
KW - Animals
KW - Atherosclerosis/metabolism
KW - Inflammation
KW - Mice
KW - Mitogen-Activated Protein Kinase 7/genetics
KW - NF-E2-Related Factor 2/genetics
KW - Plaque, Atherosclerotic
KW - atherosclerosis
KW - sumoylation
KW - antioxidants
KW - NF-E2-related factor 2 (NRF2)
KW - senescence-associated secretory phenotype
KW - receptors, aryl hydrocarbon (AHR)
KW - mitogen-activated protein kinase 7 (ERK5)
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U2 - 10.1161/CIRCRESAHA.122.322017
DO - 10.1161/CIRCRESAHA.122.322017
M3 - Article
C2 - 37264926
SN - 0009-7330
VL - 133
SP - 25
EP - 44
JO - Circulation Research
JF - Circulation Research
IS - 1
ER -