Interleukin 35 Delays Hindlimb Ischemia-Induced Angiogenesis Through Regulating ROS-Extracellular Matrix but Spares Later Regenerative Angiogenesis

Abstract

Interleukin (IL) 35 is a novel immunosuppressive heterodimeric cytokine in IL-12 family. Whether and how IL-35 regulates ischemia-induced angiogenesis in peripheral artery diseases are unrevealed. To fill this important knowledge gap, we used loss-of-function, gain-of-function, omics data analysis, RNA-Seq, in vivo and in vitro experiments, and we have made the following significant findings: i ) IL-35 and its receptor subunit IL-12RB2, but not IL-6ST, are induced in the muscle after hindlimb ischemia (HLI); ii ) HLI-induced angiogenesis is improved in Il12rb2−/− mice, in ApoE−/−/Il12rb2−/− mice compared to WT and ApoE−/− controls, respectively, where hyperlipidemia inhibits angiogenesis in vivo and in vitro; iii ) IL-35 cytokine injection as a gain-of-function approach delays blood perfusion recovery at day 14 after HLI; iv ) IL-35 spares regenerative angiogenesis at the late phase of HLI recovery after day 14 of HLI; v ) Transcriptome analysis of endothelial cells (ECs) at 14 days post-HLI reveals a disturbed extracellular matrix re-organization in IL-35-injected mice; vi ) IL-35 downregulates three reactive oxygen species (ROS) promoters and upregulates one ROS attenuator, which may functionally mediate IL-35 upregulation of anti-angiogenic extracellular matrix proteins in ECs; and vii ) IL-35 inhibits human microvascular EC migration and tube formation in vitro mainly through upregulating anti-angiogenic extracellular matrix-remodeling proteins. These findings provide a novel insight on the future therapeutic potential of IL-35 in suppressing ischemia/inflammation-triggered inflammatory angiogenesis at early phase but sparing regenerative angiogenesis at late phase.

Ultrasound May Suppress Tumor Growth, Inhibit Inflammation, and Establish Tolerogenesis by Remodeling Innatome via Pathways of ROS, Immune Checkpoints, Cytokines, and Trained Immunity/Tolerance

“…However, postoperative use of LIPUS after tibial fracture fixation does not accelerate radiographic healing and fails to improve functional recovery [ 123 ]. Taken together, the perspective, limitations, and drawbacks of ultrasound therapies also suggest the significance of molecular mechanistic findings presented here in order to improve our understanding on anti-inflammation therapies [ 43 , 84 , 97 , 124 ].…”

Antiangiogenic Effect of Platelet P2Y12 Inhibitor in Ischemia-Induced Angiogenesis in Mice Hindlimb

“…Accumulating data have suggested that postischemic proinflammatory cytokines play an important role in inducing angiogenesis [ 16 , 17 ]. The inflammatory action is simultaneously adjusted by anti-inflammatory cytokine production in major ischemic area.…”

Organelle Crosstalk Regulators Are Regulated in Diseases, Tumors, and Regulatory T Cells: Novel Classification of Organelle Crosstalk Regulators

“…We and others recently reported that CVD stressors and risk factors such as hyperlipidemia (3,4), hyperglycemia (5), hyperhomocysteinemia (6,7), and chronic kidney disease (8)(9)(10) promote atherosclerosis and vascular inflammation via several mechanisms. These mechanisms include innate immune activation (11) of endothelial cells (ECs) (3,(12)(13)(14)(15) promoting EC injury (16); Ly6Chigh inflammatory mouse monocyte and CD40 + human monocyte differentiation (7,(17)(18)(19); disease reprogrammed macrophages (20)(21)(22); cytokine and secretome regulation (23)(24)(25)(26)(27)(28)(29)(30); decreased/transdifferentiated CD4 + Foxp3 + regulatory T cells (Treg) (24,(31)(32)(33)(34); and impaired vascular repairability of bone marrow-derived progenitor cells (35,36). In addition, we recently proposed new models such as intracellular organelle dangers (37) and reactive oxygen species (ROS) as an integrated sensing system for metabolic homeostasis and alarming (38), which indicated that metabolic reprogramming and dysfunction trigger mitochondrial (MT) ROS (4,(39)(40)(41...…”