High-Throughput Screen for Small Molecules that Modulate Myocardial Damage

描述（由申请人提供）：心肌梗死（“心脏病发作”）是世界上发病率和死亡率最常见的原因之一。冠状动脉血流的突然停止在数小时内导致心肌细胞通过凋亡和坏死而大量死亡。通过血管成形术/支架植入术的及时心肌再灌注是目前最佳的治疗方法，但其有效性受到狭窄治疗时间窗的限制。在低等生物和哺乳动物中的工作已经证明，细胞死亡的很大一部分-细胞凋亡和坏死-以故意和高度调节的方式发生。这表明心肌梗死期间的细胞死亡可以在治疗上操纵。PI和其他人的大量工作已经描绘了心脏细胞死亡的多种机制。重要的是，这项工作表明，心肌梗死期间的心肌细胞死亡可以被抑制，从而减少梗死面积和保存心脏功能。我们希望通过以下目标使用化学生物学方法推动这些发现：目标1。对调节心肌细胞死亡的小分子进行高通量的基于细胞的初步筛选，以响应与心肌梗死相关的死亡刺激。我们与Sanford-Burnham研究所共同开发的初步研究证明了我们的检测方法的可行性。目标2.进一步评价在高通量筛选中影响细胞死亡的化学物质对其他死亡刺激和细胞类型的作用范围以及途径特异性。将采用更严格和特异性的细胞死亡测定。此外，还将绘制有针对性的路径。目标3.使用完整、分离、缓冲液灌注的心脏在离体心肌梗死模型中测试最终探针及其最佳相关类似物。据我们所知，这是第一个针对心肌梗死的无偏倚化学筛查。希望所产生的探针将（a）促进未来描绘连接细胞凋亡和坏死的机械联系，这是一个根本重要性的问题;和（B）为未来开发减少梗死面积和延缓其在心肌梗死期间进展的药物提供基础。 公共卫生相关性：心肌梗死（“心脏病发作”）是世界上最常见的残疾和死亡原因之一。在心肌梗死期间，大量心肌细胞死亡。本申请提出鉴定可用于（a）确定心肌细胞如何死亡和（B）产生减少这种细胞死亡的药物的化学物质。

DESCRIPTION (provided by applicant): Myocardial infarction ("heart attack") is one of the most common causes of morbidity and mortality in the world. The sudden cessation of coronary blood flow leads within hours to massive death of heart muscle cells by apoptosis and necrosis. Prompt myocardial reperfusion via angioplasty/stenting is currently the optimal treatment, but its effectiveness is limited by a narrow therapeutic time window. Work in lower organisms and mammals has demonstrated that a significant portion of cell death - both apoptosis and necrosis - occurs in a deliberate and highly regulated manner. This suggests that cell death during myocardial infarction can be therapeutically manipulated. Extensive work from the PI and others has delineated multiple mechanisms of cell death in the heart. Importantly, this work has shown that heart muscle cell death during myocardial infarction can be inhibited resulting in the reduction of infarct size and preservation of cardiac function. We wish to move these discoveries forward using a chemical biology approach through the following aims: Aim 1. To perform a high-throughput cell-based primary screen for small molecules that modulate the death of heart muscle cells in response to death stimuli relevant to myocardial infarction. Our preliminary studies, developed in conjunction with the Sanford-Burnham Institute, demonstrate the readiness of our assay. Aim 2. To further evaluate chemicals that influenced cell death in the high-throughput screen for broadness of effect against other death stimuli and cell types, and for pathway specificity. More stringent and specific cell death assays will be employed. In addition, targeted pathways will be mapped. Aim 3. To test final probe(s) and their best related analogs in an ex vivo model of myocardial infarction using intact, isolated, buffer-perfused hearts. To our knowledge, this is the first unbiased chemical screen directed at myocardial infarction. It is hoped that the probes generated will both (a) facilitate the future delineation of mechanistic linkages connecting apoptosis and necrosis, an issue of fundamental importance; and (b) provide the basis for the future development of drugs that reduce infarct size and retard its progression during myocardial infarction. PUBLIC HEALTH RELEVANCE: Myocardial infarction ("heart attack") is one of the most common causes of disability and death in the world. During myocardial infarction, large numbers of heart muscle cells die. This application proposes to identify chemicals that can be used (a) to determine how heart muscle cells die and (b) to create a medication that reduces this cell death.