Role of 12-lipoxygenase and 12-HETE signaling in beta-cell dysfunction

12-脂氧合酶和 12-HETE 信号在 β 细胞功能障碍中的作用

• 批准号: 10853540
• 负责人: ROHIT N. KULKARNI
• 金额: $ 6.22万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10853540
• 关键词: Acids; Active Sites; Arachidonate 12-Lipoxygenase; Beta Cell; Biology; Cell physiology; Cells; Cessation of life; Characteristics; Diabetes Mellitus; Eicosanoids; Enzymes; Exhibits; Family; Functional disorder; G-Protein-Coupled Receptors; GPR31 receptor; Genes; Growth Factor; Human; Human Biology; Inflammation; Inflammation Mediators; Inflammatory; Insulin; Insulin Resistance; Knock-in Mouse; Knock-out; Link; Lipids; Lipoxygenase; Mediating; Molecular; Mus; Oxidative Stress; Pathway interactions; Polyunsaturated Fatty Acids; Protein Isoforms; Reagent; Research Personnel; Role; Signal Transduction; Testing; expectation; impaired glucose tolerance; in vivo; inhibitor; islet; mouse model; receptor-mediated signaling; therapeutic target; tool; type I and type II diabetes; virtual

Abstract Deficiencies in islet β-cell function and/or mass are central in the transition from impaired glucose tolerance to frank diabetes in the setting of both type 1 and type 2 diabetes. The lipoxygenases (LOXs) represent a family of enzymes that catalyzes the oxygenation of cellular poly-unsaturated fatty acids to form lipid inflammatory mediators in β-cells. The eicosanoid product of 12-LOX activity, 12-hydoxyeicosatetraenoic acid (12-HETE), imposes inflammatory and oxidative stress within β cells. A challenge in lipoxygenase biology, however, is that humans and mice express different isoforms of 12-LOX in β cells (encoded by ALOX12 and Alox15, respectively), with each isoform exhibiting different active-site characteristics. The strength of this application is the collaborative effort between Multi-PIs Drs. R. Mirmira (an expert in islet inflammation pathways) and R. Kulkarni (an expert in growth factor signaling in the islet), and coinvestigator J. Nadler (an expert in eicosanoid biology), who will collectively bring their expertise and unique reagents—including knockout and human gene knock-in mouse models that show human isoform activation, human 12-LOX-selective inhibitors, and primary human cells—to bear on the biology of 12-LOX and its inflammatory products. We hypothesize that during insulin resistance the activation of 12-LOX in β-cells promotes β cell dysfunction through receptor-mediated signaling by its downstream product 12-HETE. To test this hypothesis, we propose the following specific aims: Aim 1: Elucidate the molecular mechanisms linking β-cell insulin resistance to 12-LOX activity and cellular dysfunction. Aim 2: Determine the contribution of 12-LOX activity to β cell dysfunction in the setting of insulin resistance in vivo. Aim 3: Determine the role of the 12-HETE receptor GPR31 in mediating β-cell dysfunction downstream of 12-LOX. Until now, tools and reagents to interrogate the biology of human 12-LOX and 12-HETE did not exist. The primary impact of this proposal will be to set the stage for the expectations of therapeutically targeting the human 12-LOX pathway in insulin resistance/β-cell dysfunction, and to identify a potential new target in the 12- HETE G protein-coupled receptor GPR31.

抽象的 胰岛β细胞功能和/或质量的缺陷在从受损葡萄糖耐受性到的过渡中是核心 在类型1和2型糖尿病的情况下，Frank糖尿病。脂氧酶（LOX）代表一个家庭 催化细胞多饱和脂肪酸的氧合的酶形成脂质炎症 β细胞中的介体。 12-lox活性，12-羟基羟基乙烯酸（12-Hete），12-羟基乙烯的类类产物， 在β细胞内施加炎症和氧化应激。然而，脂氧合酶生物学的挑战是 人类和小鼠在β细胞中表达12-lox的不同同工型（由Alox12和Alox15编码， 分别），每个同工型都表现出不同的活性位点特性。该应用的优势是 多PIS DRS之间的协作工作。 R. Mirmira（胰岛注入途径的专家）和R. Kulkarni（胰岛中生长因子信号传导专家）和共依供剂J. Nadler（eicosanoid的专家 生物学），他将集体带来他们的专业知识和独特的试剂 - 包括淘汰和人类基因 显示人类同工型激活的小鼠模型，人类12-lox选择性抑制剂和主要 人类细胞 - 依靠12-Lox及其炎症产物的生物学。我们假设在 胰岛素抵抗β细胞中12-LOX的激活通过受体介导的β细胞功能障碍促进β细胞功能障碍 通过其下游产品发出信号12-Hete。为了检验这一假设，我们提出了以下特定目的： 目标1：阐明将β细胞胰岛素抵抗与12-Lox活性和 细胞功能障碍。 AIM 2：确定胰岛素环境中12-LOX活性对β细胞功能障碍的贡献 体内抗性。 AIM 3：确定12-HETE受体GPR31在介导β细胞功能障碍下游的作用 12-lox。 到目前为止，还不存在询问人类12-lox和12-Hete生物学的工具和试剂。这 该提案的主要影响是为治疗目标的期望奠定 胰岛素抵抗/β细胞功能障碍中的人类12-lox途径，并确定12--中潜在的新目标 Hete G蛋白偶联受体GPR31。

项目成果

The AAV-PCSK9 murine model of atherosclerosis and metabolic dysfunction.

• DOI: 10.1093/ehjopen/oeac028
• 发表时间: 2022-05
• 期刊: European heart journal open
• 作者: Keeter, William Coles;Carter, Nigeste M;Nadler, Jerry L;Galkina, Elena V
• 通讯作者: Galkina, Elena V

Interleukin-6 Reduces β-Cell Oxidative Stress by Linking Autophagy With the Antioxidant Response.

• DOI: 10.2337/db17-1280
• 发表时间: 2018-08
• 期刊: Diabetes
• 影响因子: 7.7
• 作者: Marasco MR;Conteh AM;Reissaus CA;Cupit JE 5th;Appleman EM;Mirmira RG;Linnemann AK
• 通讯作者: Linnemann AK

Oligomeric collagen as an encapsulation material for islet/β-cell replacement: effect of islet source, dose, implant site, and administration format.

寡聚胶原蛋白作为胰岛/β 细胞替代的封装材料：胰岛来源、剂量、植入部位和给药方式的影响。

• DOI: 10.1152/ajpendo.00066.2020
• 发表时间: 2020
• 期刊: American journal of physiology. Endocrinology and metabolism
• 作者: Stephens,ClarissaHernandez;Morrison,RachelA;McLaughlin,Madeline;Orr,Kara;Tersey,SarahA;Scott-Moncrieff,JCatharine;Mirmira,RaghavendraG;Considine,RobertV;Voytik-Harbin,Sherry
• 通讯作者: Voytik-Harbin,Sherry

Preclinical evaluation of tyrosine kinase 2 inhibitors for human beta-cell protection in type 1 diabetes.

• DOI: 10.1111/dom.14104
• 发表时间: 2020-10
• 期刊: Diabetes, obesity & metabolism
• 作者: Coomans de Brachène A;Castela A;Op de Beeck A;Mirmira RG;Marselli L;Marchetti P;Masse C;Miao W;Leit S;Evans-Molina C;Eizirik DL
• 通讯作者: Eizirik DL

PIE-FLIM Measurements of Two Different FRET-Based Biosensor Activities in the Same Living Cells.

PIE-FLIM 测量同一活细胞中两种不同的基于 FRET 的生物传感器活性。

• DOI: 10.1016/j.bpj.2020.03.003
• 发表时间: 2020
• 期刊: Biophysical journal
• 影响因子: 3.4
• 作者: Reissaus,ChristopherA;Day,KathleenH;Mirmira,RaghavendraG;Dunn,KennethW;Pavalko,FredrickM;Day,RichardN
• 通讯作者: Day,RichardN