CYP1B1, NF-kB , and Regulation of Angiogenesis

CYP1B1、NF-kB 和血管生成的调节

• 批准号: 8007269
• 负责人: Tammy Lynn Palenski
• 金额: $ 3.13万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-30 至 2013-08-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8007269
• 关键词: Adult; Age related macular degeneration; Angiogenesis Inhibitors; Blindness; Blood Vessels; Blood capillaries; CYP1B1 gene; Cell Adhesion; Cells; Cytochrome P450; Development; Diabetic Retinopathy; Disease; Down-Regulation; Embryonic Development; Endothelial Cells; Exhibits; Genes; Homeostasis; In Vitro; Luciferases; Morphogenesis; Mus; NF-kappa B; Oxidative Stress; Oxygen; Pathogenesis; Pattern; Phenotype; Play; Process; Production; Regulation; Retinal; Retinal Diseases; Retinopathy of Prematurity; Role; Smooth Muscle Myocytes; Transgenic Mice; Up-Regulation; Wound Healing; angiogenesis; attenuation; capillary; density; effective therapy; insight; matrigel; migration; neovascular; neovascularization; novel; postnatal; public health relevance; response; thrombospondin 2

DESCRIPTION (provided by applicant): Angiogenesis, the process of new blood vessel formation from pre-existing capillaries, is very tightly regulated and normally does not occur except during embryonic development and reparative processes in the adult. However, dysregulation of angiogenesis is associated with pathogenesis many diseases including retinopathy of prematurity, diabetic retinopathy and age-related macular degeneration. This is generally accomplished by down-regulation of negative and up-regulation of positive regulators of angiogenesis. Therefore, a better understanding of the regulatory processes that govern normal angiogenesis is key to understanding how abnormal angiogenesis occurs during disease. The recent discovery of specific cytochrome P450 expression (CYPs) in the vascular smooth muscle cells and endothelial cells (EC), and their contribution to vascular function suggest critical roles for these genes in vascular homeostasis. We recently showed that CYP1B1 plays an essential role in retinal vascular development and neovascularization during oxygen induced ischemic retinopathy (OIR). CYP1B1 deficient mice exhibited reduced retinal vascular density and failed to elicit a neovascular response during OIR. We also showed retinal EC prepared from CYP1B1-/- mice are less migratory and fail to undergo capillary morphogenesis in Matrigel. These cells also expressed significant amounts of an endogenous inhibitor of angiogenesis, thrombospondin-2 (TSP2), a gene shown to be up-regulated in response to oxidative stress. My hypothesis is that the increased oxidative stress in the absence of CYP1B1 results in sustained NF-:B activation and promotes increased production of TSP2. In Aim 1, we will determine the expression patterns of NF-:B and TSP2 during postnatal retinal vascular development and neovascularization during OIR using NF-:B-GFP/luciferase and TSP2-GFP transgenic mice. We will investigate whether NF-:B and TSP2 expression are altered in the absence of CYP1B1. In Aim 2, we will determine whether sustained activation of NF-:B in wild type retinal EC in vitro is sufficient to recapitulate the CYP1B1 null phenotype, attenuation of retinal EC adhesion, migration and capillary morphogenesis, and induce TSP2 expression. Understanding how CYP1B1 expression regulates retinal EC phenotype will provide new insight into novel mechanisms that regulate angiogenesis and aid in the development of more effective treatments. PUBLIC HEALTH RELEVANCE: Angiogenesis, the formation of new blood vessels from preexisting capillaries, occurs during development and in many reparative processes in adults, such as wound healing. Dysregulation of this process is associated with pathogenesis of a number of diseases including retinopathy of prematurity, diabetic retinopathy, and age-related macular degeneration, the major causes of blindness. A better understanding of how angiogenesis is regulated will aid in development of new and more effective therapies for these blinding diseases, as well as other diseases with a neovascular component.

描述（由申请人提供）：血管生成，即从预先存在的毛细血管形成新血管的过程，受到非常严格的调控，通常不会发生，除非在胚胎发育和成人修复过程中。然而，血管生成失调与许多疾病的发病机制相关，包括早产儿视网膜病变、糖尿病视网膜病变和年龄相关性黄斑变性。这通常通过下调血管生成的负调节剂和上调血管生成的正调节剂来实现。因此，更好地了解控制正常血管生成的调节过程是了解疾病期间异常血管生成如何发生的关键。细胞色素P450在血管平滑肌细胞和内皮细胞中的特异性表达及其对血管功能的影响提示了这些基因在血管稳态中的重要作用。我们最近发现，CYP 1B 1在氧诱导缺血性视网膜病变（OIR）的视网膜血管发育和新生血管形成中起重要作用。CYP 1B 1缺陷小鼠表现出视网膜血管密度降低，在OIR过程中未能引起新生血管反应。我们还表明，从CYP 1B 1-/-小鼠制备的视网膜EC迁移性较低，在Matrigel中未能进行毛细血管形态发生。这些细胞还表达了大量的内源性血管生成抑制剂，血小板反应蛋白-2（TSP 2），一种在氧化应激反应中被上调的基因。我的假设是，在缺乏CYP 1B 1的情况下，氧化应激的增加导致持续的NF-：B激活，并促进TSP 2的产生增加。在目的1中，我们将使用NF-：B-GFP/荧光素酶和TSP 2-GFP转基因小鼠确定出生后视网膜血管发育和OIR期间新生血管形成期间NF-：B和TSP 2的表达模式。我们将研究NF-：B和TSP 2表达是否在CYP 1B 1缺失的情况下改变。在目的2中，我们将确定体外野生型视网膜EC中NF-：B的持续激活是否足以重现CYP 1B 1无效表型，减弱视网膜EC粘附、迁移和毛细血管形态发生，并诱导TSP 2表达。了解CYP 1B 1表达如何调节视网膜EC表型将为调节血管生成的新机制提供新的见解，并有助于开发更有效的治疗方法。 公共卫生关系：血管生成，即从先前存在的毛细血管形成新血管，发生在发育期间和成人的许多修复过程中，如伤口愈合。这一过程的失调与许多疾病的发病机制有关，包括早产儿视网膜病变、糖尿病视网膜病变和年龄相关性黄斑变性，这些疾病是失明的主要原因。更好地了解血管生成是如何调节的，将有助于开发新的和更有效的治疗这些致盲性疾病，以及其他疾病的新生血管成分。