12/15-lipoxygenase: Immune cell mediator linking innate immunity to hypertension

12/15-脂氧合酶：将先天免疫与高血压联系起来的免疫细胞介质

基本信息

批准号：
10318163
负责人：
WILLIAM BRYSON CAMPBELL
金额：
$ 52.58万
依托单位：
MEDICAL COLLEGE OF WISCONSIN
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-12-15 至 2023-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10318163
关键词：
ALOX15 gene Adoptive Transfer Affect Angiotensin II Anti-Inflammatory Agents Arachidonate 15-Lipoxygenase Arachidonic Acids Arteries Blood Pressure Blood Vessels Bone Marrow Cells Brain Bypass Cells Cre-LoxP Development Disease Enzymes Equilibrium Excretory function Functional disorder Genes Goals Heart Hypertrophy Human Hydroxyeicosatetraenoic Acids Hypertension Immune In Vitro Inbred SHR Rats Inflammation Inflammatory Injury Injury to Kidney Kidney Knowledge Link Mediating Mediator of activation protein Metabolic Methods Mineralocorticoids Modeling Molecular Mus Natural Immunity Norepinephrine Organ Outcome Pathway interactions Phenotype Regulation Renal function Resolution Role Sodium Source Specific qualifier value Stimulus Structure Testing Vascular remodeling Wild Type Mouse adaptive immunity blood pressure elevation constriction cytokine endothelial dysfunction hypertension control hypertensive immune activation in vivo macrophage novel strategies novel therapeutics prevent restoration vasoconstriction

项目摘要

Innate immunity and macrophages (MFs) contribute to the development and pathophysiology of hypertension. Knowledge gaps exist for the cellular and molecular mechanisms in MFs that contribute to hypertension. We identified a single gene, Alox15, that is crucial in 3 models of hypertension. Alox15 encodes 12/15-lipoxygenase (12/15-LO) that metabolizes arachidonic acid to hydroxyeicosatetraenoic acids (HETEs). An important role for MF 12/15-LO in hypertension was identified by differentially excluding MF and non-MF sources of 12/15-LO by: (1) Depletion of MFs (and thus MF 12/15-LO) in wild type (WT) mice prevents hypertension. (2) Global deletion of the Alox15 gene (Alox15-/-) prevents hypertension. (3) Transfer of 12/15-LO-containing WT MFs to these Alox15-/- mice restores hypertension. (4) Bone marrow cell-specific Alox15 deletion prevents hypertension. Also, MFs accumulate in arteries and polarize to the M1 (pro-inflammatory) phenotype, but not M2 (anti-inflammatory) phenotype, with hypertension in WT mice but not in Alox15-/- mice. Thus, 12/15-LO regulates MF M1 polarization in hypertension. These results indicate the need to understand the contributions of MF 12/15-LO to hypertension. The objective of this proposal is to define the role of 12/15-LO and its metabolites as a common pathway to hypertension through regulation of the pro- and anti-inflammatory pathways of innate immunity. The central hy- pothesis is MF 12/15-LO and its HETE metabolites regulate MF accumulation, phenotype and cytokine release that activate pro-hypertensive mechanisms in the vasculature and kidneys resulting in hypertension. (1) We will use MF-targeted deletion of Alox15 to establish the crucial role of MF 12/15-LO in regulating immune, vascular and renal pro-hypertensive mechanisms. The working hypothesis is that MF 12/15-LO stimulates MF accumu- lation, polarization and cytokine release in arteries, kidneys and brain that contribute to hypertension by activat- ing vascular and renal mechanisms to enhance constriction, inhibit dilation, promote renal injury and decrease sodium excretion.!(2) The in vivo effects of HETEs on hypertensive mechanisms are unknown. Blockade of HETE synthesis in Alox15-/- mice will be bypassed by administering exogenous HETEs to restore hypertension. The working hypothesis is that 12- and 15-HETE stimulate immune, vascular and renal mechanisms that restore hypertension in Alox15-/- mice. (3) The importance of MF polarization and its regulation by 12/15-LO in hyper- tension will be defined. To determine if Alox15 deletion prevents hypertension by redirecting MF polarization, in vivo and in vitro methods will be used to polarize M0 MFs to the M1 or M2 phenotype, and the 3 phenotypes evaluated for restoration of hypertension in MF-depleted mice. The working hypothesis is that 12/15-LO directs MF polarization to M1 phenotype, and M1, but not M2, MFs promote hypertension. The expected outcomes will advance the field by identifying a new molecular mechanism involving the 12/15-LO-HETE pathway that broadly regulates the participation of MFs and innate immunity in hypertension by specifying the M1 MF phenotype that contributes to hypertension and by mediating vascular and renal pro-hypertensive activities of MFs.

先天免疫和巨噬细胞 (MF) 有助于高血压的发生和病理生理学。对于导致高血压的 MF 的细胞和分子机制存在知识空白。我们鉴定出一个单一基因 Alox15，该基因在 3 种高血压模型中至关重要。 Alox15 编码 12/15-脂氧合酶 (12/15-LO) 将花生四烯酸代谢为羟基二十碳四烯酸 (HETE)。的重要作用高血压中的 MF 12/15-LO 是通过以下方法区别排除 MF 和非 MF 来源的 12/15-LO 来确定的： (1) 野生型 (WT) 小鼠中 MF（以及 MF 12/15-LO）的消耗可预防高血压。 (2) 全局删除 Alox15 基因 (Alox15-/-) 可以预防高血压。 (3) 将含有 12/15-LO 的 WT MF 转移至这些 Alox15-/- 小鼠恢复高血压。 (4) 骨髓细胞特异性Alox15缺失可预防高血压。还， MF 在动脉中积聚并极化为 M1（促炎）表型，但不是 M2（抗炎）表型表型，WT 小鼠有高血压，但 Alox15-/- 小鼠没有高血压。因此，12/15-LO 调节 MF M1 偏振在高血压中。这些结果表明需要了解 MF 12/15-LO 对高血压的影响。该提案的目标是定义 12/15-LO 及其代谢物作为常见途径的作用通过调节先天免疫的促炎和抗炎途径来治疗高血压。中央hy- 假设是 MF 12/15-LO 及其 HETE 代谢物调节 MF 积累、表型和细胞因子释放激活脉管系统和肾脏中的促高血压机制，导致高血压。 (1) 我们将使用 MF 靶向删除 Alox15 来确定 MF 12/15-LO 在调节免疫、血管方面的关键作用和肾脏促高血压机制。工作假设是 MF 12/15-LO 刺激 MF 累积动脉、肾脏和大脑中的极化、极化和细胞因子释放，通过激活导致高血压调节血管和肾脏机制，增强收缩、抑制扩张、促进肾损伤和减少 (2) HETEs 对高血压机制的体内作用尚不清楚。封锁通过给予外源性 HETE 来恢复高血压，可以绕过 Alox15-/- 小鼠中的 HETE 合成。工作假设是 12- 和 15-HETE 刺激免疫、血管和肾脏机制，从而恢复 Alox15-/- 小鼠的高血压。 (3) MF 极化的重要性及其在超极化中 12/15-LO 的调节张力将被定义。为了确定 Alox15 缺失是否通过改变 MF 极化来预防高血压，将使用体内和体外方法将 M0 MF 极化为 M1 或 M2 表型，以及 3 种表型评估了 MF 耗尽小鼠的高血压恢复情况。工作假设是 12/15-LO 指导 MF 极化为 M1 表型，且 M1 而非 M2，MF 会促进高血压。预期成果将通过确定涉及 12/15-LO-HETE 途径的新分子机制来推进该领域的发展，该途径广泛通过指定 M1 MF 表型来调节 MF 和先天免疫在高血压中的参与促进高血压并介导 MF 的血管和肾脏促高血压活性。