Plaque detection with the hydrocyanines

描述（由申请人提供）：动脉粥样硬化及其相关并发症，如心脏病发作、中风、心绞痛，是西方世界的主要死亡原因，迫切需要有效的治疗。限制动脉粥样硬化治疗的一个关键问题是无法诊断具有生长和破裂高风险的动脉粥样硬化斑块，也称为“活性”斑块。活性氧（ROS）的过量产生是动脉粥样硬化斑块生长和破裂所必需的，使ROS成为“活性”斑块的良好诊断标志物。本应用程序的目的是开发一种新的荧光造影剂家族，称为氢氰胺，其具有成像动脉粥样硬化斑块，颈动脉中的ROS所需的物理和化学性质，并检测斑块活性。氢菁染料是我们实验室最近开发的一种新型荧光染料，可以检测纳米摩尔浓度的活性氧，发射波长在570-810 nm范围内可调，并且在氧化后在细胞内积累。我们的方法是用氢花青素成像血管损伤和动脉粥样硬化小鼠和兔子颈动脉中的ROS。该提议的中心假设是：氢氰化物具有检测颈动脉氧化应激和检测斑块活性所需的物理和化学性质。这一假设是基于我们的初步数据，这些数据表明氢花青素可以在细胞培养、组织样品和体内首次检测超氧化物和氢氧根。本提案的总体目标将通过以下三个具体目标来检验我们的中心假设来实现；专项目的1：利用动力学同位素效应优化氢花青素。专项目的2：在小鼠颈动脉损伤模型中检测ROS的产生。专项目的3：在兔动脉粥样硬化模型中检测动脉粥样硬化组织体内和人患病冠状动脉离体中ROS的产生。本课题的实验将确定氢花青素是否可以成像动脉粥样硬化斑块中的ROS，并检测小鼠和兔斑块活性。本提案中的实验具有创新性，因为它们将首次导致能够在体内成像ROS的造影剂的开发。这一提议意义重大，因为它将产生一种新的诊断方法，有可能诊断出有心脏病发作和中风风险的患者。此外，许多其他疾病也与ROS的过量产生有关，如癌症和神经退行性疾病；因此，我们预计这项提案的结果将影响医学和生物学的几个领域。本提案中的实验将导致开发一类新的荧光染料，称为氢菁，它可以对体内的活性氧进行成像。氢氰胺的主要应用是诊断有心脏病发作和中风危险的病人。许多其他疾病，如癌症和关节炎也可以通过成像反应性物质来诊断；因此，我们预计这项提案的结果将影响医学和生物学的几个领域。

DESCRIPTION (provided by applicant): Atherosclerosis and atherosclerosis related complications such as heart attack, stroke, and angina, are the leading cause of death in the western world, and effective treatments are greatly needed. A key issue limiting the treatment of atherosclerosis is an inability to diagnose atherosclerotic plaques that have a high risk of growing and rupturing, also known as "active" plaques. The overproduction of reactive oxygen species (ROS) is necessary for the growth and rupture of atherosclerotic plaques, making ROS an excellent diagnostic marker for "active" plaques. The objective of this application is to develop a new family of fluorescent contrast agents, termed the hydrocyanines, which have the physical and chemical properties needed to image ROS in atherosclerotic plaques, in the carotid artery, and detect plaque activity. The hydrocyanines are a new family of fluorescent dyes, recently developed in our laboratory, which can detect ROS at nanomolar concentrations, have tunable emission wavelengths ranging from 570-810 nm, and accumulate within cells after oxidation. Our approach is to image ROS in the carotid artery of mice and rabbits, suffering from vascular injury and atherosclerosis, with the hydrocyanines. The central hypothesis of this proposal is that: The hydrocyanines have the physical and chemical properties needed to detect oxidative stress in the carotid artery and detect plaque activity. This hypothesis is based on our preliminary data, which demonstrates that the hydrocyanines can detect superoxide and the hydroxide radical in cell culture, in tissue samples and for the first time in vivo. The overall objective of this proposal will be accomplished by testing our central hypothesis through the following three Specific Aims; Specific Aim I: Optimize the hydrocyanines using the kinetic isotope effect Specific Aim II: Detect ROS production in a murine model of carotid artery injury Specific Aim III: Detect ROS production by atherosclerotic tissues in vivo in a rabbit model of atherosclerosis and ex-vivo in diseased human coronary arteries The experiments in this proposal will determine if the hydrocyanines can image ROS in atherosclerotic plaques and detect plaque activity in mice and rabbits. The experiments in this proposal are innovative because they will lead to the development of a contrast agent that can image ROS, in vivo, for the first time. This proposal is also significant because it will generate a new class of diagnostics, which have the potential to diagnose patients at risk of developing heart attacks and strokes. Furthermore, numerous other diseases are also associated with an overproduction of ROS, such as cancer and neurodegenerative disease; we therefore anticipate that the results of this proposal will impact several areas of medicine and biology. PUBLIC HEALTH RELEVANCE: Project Narrative The experiments in this proposal will lead to the development of a new class of fluorescent dyes, termed the hydrocyanines, which can image reactive oxygen species in vivo. The major applications of the hydrocyanines are to diagnose patients at risk of developing heart attacks and strokes. Numerous other diseases such as cancer and arthritis can also be diagnosed by imaging reactive species; we therefore anticipate that the results of this proposal will impact several areas of medicine and biology.