Identification of novel y-globin corepressors and advanced inhibitor development

摘要： 这项研究的最终目标是为人类提供安全、有效、可及和持久的疾病修饰 镰状细胞性贫血（SCD）和β-地中海贫血（CA），将改善多器官病理生理学并减少 早逝。过去40年基础和临床研究无可争议的数据支持 前提是诱导胎儿 γ-珠蛋白 (HBG) 基因抑制对人类患者具有治疗作用。的 该提案的两个具体目标侧重于转录的操纵和特定靶向 在发育过程中抑制 HBG 基因的调节机制。我们的首要目标是致力于发现 以及在成人红细胞中引发 γ-珠蛋白抑制活性的新型表观遗传修饰酶的验证 祖细胞，我们描述了几种有前途的新候选酶，我们建议对其进行研究 潜在的药物靶向。我们的第二个目标是完善基于结构的表观遗传修饰抑制剂 LSD1 (KDM1a) 酶（我们在概念验证研究中展示）可以构成 治疗这些疾病的治疗干预。通过结构辅助设计耦合 迭代酶抑制和基于细胞的 HbF 诱导测定，我们目前正在完善和测试第四种 和第五代抑制剂（由密歇根大学设计）。这些化合物已经进化到 指出它们对 LSD1 具有高度特异性（IC50 < 90 nM），是可逆的，不表现出 行为或心脏副作用，可以口服给药。我们已经开发出LSD1抑制剂 在临床前动物研究中取得了部分成功，在这里我们建议开发新化合物 对目标的不良影响最小。我们建议合作跟进这些临床前研究 与项目 2 的研究人员合作，通过分析这些新型抑制剂中最好的在狒狒身上的效果，如果 这些与项目 3 合作在人体临床试验中取得了成功。对患者的预计影响 患有 β-球蛋白病的一个重要原因是，这些提议的 HbF 诱导疗法将足够有效 对抗这些疾病的毁灭性并发症，如中风和急性胸部综合症， 可访问性和安全性参数将允许普遍应用以及终身使用。

Abstract: The ultimate goal of this research is to provide safe, effective, accessible and durable disease modification for sickle cell anemia (SCD) and β-thalassaemia (CA) that will improve multi-organ pathophysiology and reduce early death. Incontrovertible data from both basic and clinical research studies over the past 40 years support the premise that inducing fetal γ-globin (HBG) gene repression would be therapeutic in human patients. The two specific aims of this proposal focus on the manipulation and specific targeting of the transcriptional regulatory machinery that represses HBG genes during development. Our first aim is devoted to the discovery and validation of new epigenetic modifying enzymes that elicit γ-globin repressive activity in adult erythroid progenitor cells, and we describe several promising new candidate enzymes that we propose to investigate for potential drug targeting. Our second aim is to refine structure-based inhibitors of the epigenetic modifying enzyme LSD1 (KDM1a), which (we showed in a proof of concept study) could constitute an ideal target for therapeutic intervention for the treatment of these diseases. Through structure-aided design coupled to iterative enzyme inhibition and cell-based HbF induction assays, we are currently refining and testing fourth and fifth generation inhibitors (designed at the University of Michigan). These compounds have evolved to the point that they are proposed to be highly specific for LSD1 (with IC50s < 90 nM), are reversible, exhibit no behavioral or cardiac side effects and can be orally administered. We have already developed LSD1 inhibitors that were partially successful in preclinical animal studies, and here we propose to develop novel compounds with minimal undesirable on target effects. We propose to follow up these preclinical studies, in collaboration with investigators in project 2, by analysis of the effects of the best of these novel inhibitors in baboons, and if those are successful, in collaboration with project 3, in human clinical trials. The projected impact for patients suffering from β-globinopathies is that these proposed HbF-inducing therapies will be sufficiently efficacious to counter the devastating complications of these diseases such as stroke and acute chest syndrome, and with accessibility and safety parameters that will permit universal application as well as life-long use.