Plaque detection with the hydrocyanines

描述(申请人提供)：动脉粥样硬化和动脉粥样硬化相关并发症，如心脏病发作、中风和心绞痛，是西方世界的主要死亡原因，非常需要有效的治疗。限制动脉粥样硬化治疗的一个关键问题是无法诊断动脉粥样硬化斑块，这种斑块具有生长和破裂的高风险，也被称为“活动”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“活动性”斑块的极佳诊断标记物。这一应用的目的是开发一种新的荧光造影剂家族，称为氢花菁，它具有成像颈动脉粥样硬化斑块中的ROS和检测斑块活性所需的物理和化学性质。氢花菁是我们实验室最近开发的一类新的荧光染料，它可以检测到纳摩尔浓度的ROS，发射波长在570-810 nm之间可调，氧化后在细胞内积累。我们的方法是用氢花菁对血管损伤和动脉粥样硬化的小鼠和兔颈动脉中的ROS进行成像。这一提议的中心假设是：氢化花菁具有检测颈动脉氧化应激和检测斑块活动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，氢花菁可以在细胞培养、组织样本中以及在体内首次检测到超氧阴离子和氢氧自由基。这项建议的总体目标将通过检验我们的中心假设通过以下三个具体目标来实现：具体目标一：利用动力学同位素效应优化水蓝化合物；特定目标二：在颈动脉损伤的小鼠模型中检测ROS的产生；特定目标三：在兔动脉粥样硬化模型中检测动脉粥样硬化组织中ROS的产生；在体外，在人类冠状动脉病变的模型中检测ROS的产生本提议中的实验将确定是否可以对动脉粥样硬化斑块中的ROS进行成像，并检测小鼠和兔的斑块活动。这项提案中的实验具有创新性，因为它们将导致开发一种可以在体内成像ROS的造影剂，这是第一次。这一提议也具有重要意义，因为它将产生一种新的诊断类别，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的过度生产有关，如癌症和神经退行性疾病；因此，我们预计这项提议的结果将影响医学和生物学的几个领域。公共卫生相关性：项目描述本提案中的实验将导致开发一类新的荧光染料，称为氢花菁，它可以在体内成像活性氧物种。氢花菁的主要应用是诊断有心脏病发作和中风风险的患者。许多其他疾病，如癌症和关节炎，也可以通过对活性物质进行成像来诊断；因此，我们预计这项提议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.