ULK-mediated autophagy of α-globin in ß-thalassemia

α-地中海贫血中 ULK 介导的 α-珠蛋白自噬

基本信息

批准号：
10649565
负责人：
Mitchell J Weiss
金额：
$ 65.26万
依托单位：
ST. JUDE CHILDREN'S RESEARCH HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10649565
关键词：
5&apos -AMP-activated protein kinase Ablation Adult Affect Alleles Anemia Apoptosis Attenuated Autophagocytosis Biological Markers Biology CD34 gene Cell Line Cells Chronic Complex Data Deformity Disease Dose Drug Metabolic Detoxication Elements Erythroblasts Erythrocyte Transfusion Erythrocytes Erythroid Erythroid Cells Erythropoiesis FRAP1 gene Functional disorder Gene Mutation Genetic Goals Hematological Disease Hematopoietic Stem Cell Transplantation Hematopoietic stem cells Hemoglobin Hemoglobinopathies Hemolysis Human Immunologic Deficiency Syndromes Impairment In Vitro Iron Iron Chelating Agents Iron Chelation Iron Overload Life Mediating Messenger RNA MicroRNAs Mus Newborn Infant Oxygen Pathology Pathway interactions Patient Selection Patients Phosphotransferases Pre-Clinical Model Process Production Proteins Quality Control Regulation Reporting Repression STK11 gene Severities Sickle Cell Trait Sirolimus Testing Therapeutic Therapeutic Effect Translating Transplantation alpha Globin beta Globin beta Thalassemia bone cofactor cytotoxic derepression hepcidin improved in vivo inhibitor innovation insight low and middle-income countries mimetics new therapeutic target novel therapeutic intervention novel therapeutics pharmacologic prevent protein aggregation protein degradation proteostasis response

项目摘要

PROJECT SUMMARY Our long-term goal is to better define the proteostasis pathways that coordinate erythropoiesis, a specialized process distinguished by massive hemoglobin synthesis and the elimination of most other proteins. This application investigates how protein quality control modulates ß-thalassemia, a common hemoglobinopathy caused by HBB gene mutations that impair the production of the ß-globin subunit of adult hemoglobin (HbA, α2ß2). Consequently, free α-globin forms cytotoxic precipitates that cause maturation arrest and apoptosis of erythroid precursors (ineffective erythropoiesis) and hemolysis, leading to anemia, bone deformities and iron overload. Current therapies, including red blood cell transfusion, iron chelation and hematopoietic stem cell transplantation for selected patients, are effective but not uniformly available, particularly in low/middle income countries where the disease is most prevalent. Hence, new therapies are needed. Our preliminary data demonstrate that the Unc-51–like autophagy activating kinase 1 (ULK1) mediates the autophagy of free α-globin in ß-thalassemia. In general, ULK1 is inhibited by the mammalian target of rapamycin complex 1 (mTORC1) kinase and stimulated by AMP-activated protein kinase (AMPK). Administration of the mTORC1 inhibitor rapamycin to HbbTh3/+ mice, a validated preclinical model for ß-thalassemia, stimulated the autophagy of free α- globin to reduce ineffective erythropoiesis and hemolysis in an ULK1-dependent fashion. Our data support the central hypothesis that mTORC1 inhibition or AMPK activation can alleviate the pathophysiology of ß- thalassemia by stimulating ULK1-mediated autophagic clearance of free α-globin. We will test this by: Aim 1, optimizing the pharmacological inhibition of mTORC1 for ULK1 activation in HbbTh3/+ mice and by defining the regulatory circuitry of α-globin autophagy in mouse and human ß-thalassemic erythroblasts; Aim 2, determining whether elemental iron, a known activator of mTORC1, suppresses ULK1-mediated clearance of α-globin in ß- thalassemic erythroblasts and whether this deleterious effect can be prevented by iron restriction; and Aim 3, elucidating the genetic interactions between ß-thalassemia and miR-451, an abundantly expressed erythroid microRNA that we showed to inhibit the LKB1 kinase and its substrate AMPK. In support of Aim 3, disruption of the bi-cistronic miR-144/451 locus in HbbTh3/+ mice caused a reduction in α-globin precipitates and ß-thalassemia pathologies. Overall, our studies promise to elucidate the biology of proteostasis networks that maintain balanced hemoglobin synthesis through targeted protein degradation and validate mTORC1, AMPK and ULK1 as “druggable” targets for novel ß-thalassemia therapies.

项目概要我们的长期目标是更好地定义协调红细胞生成的蛋白质稳态途径，这是一种专门的该过程以大量血红蛋白合成和大多数其他蛋白质的消除为特征。应用研究蛋白质质量控制如何调节 β 地中海贫血（一种常见的血红蛋白病）由 HBB 基因突变引起，该突变损害成人血红蛋白 (HbA、 α2ß2) 经检查，游离 α-珠蛋白形成细胞毒性沉淀物，导致成熟停滞和细胞凋亡。红细胞前体细胞（无效红细胞生成）和溶血，导致贫血、骨骼畸形和铁目前的治疗方法包括红细胞输注、铁螯合和造血干细胞。对选定的患者进行移植是有效的，但并非普遍可用，特别是在低/中等收入国家因此，我们需要新的治疗方法。证明 Unc-51 样自噬激活激酶 1 (ULK1) 介导游离 α-珠蛋白的自噬在β-地中海贫血中，ULK1 通常受到哺乳动物雷帕霉素复合物 1 (mTORC1) 的抑制。 mTORC1抑制剂的给药雷帕霉素对 HbbTh3/+ 小鼠（经过验证的 β-地中海贫血临床前模型）刺激游离 α- 的自噬珠蛋白以 ULK1 依赖性方式减少无效红细胞生成和溶血。我们的数据支持这一点。中心假设是 mTORC1 抑制或 AMPK 激活可以减轻 ß- 的病理生理学通过刺激 ULK1 介导的游离 α-珠蛋白的自噬清除来治疗地中海贫血我们将通过以下方式进行测试：目标 1，优化 HbbTh3/+ 小鼠中 mTORC1 对 ULK1 激活的药理学抑制作用，并定义小鼠和人类 β-地中海贫血红细胞中 α-珠蛋白自噬的调节回路，确定；元素铁（一种已知的 mTORC1 激活剂）是否会抑制 β- 中 ULK1 介导的 α-球蛋白清除地中海贫血性成红细胞以及是否可以通过限制铁来预防这种有害作用；以及目标 3，阐明 ß-地中海贫血和 miR-451（一种丰富表达的红系细胞）之间的遗传相互作用我们证明 microRNA 可以抑制 LKB1 激酶及其底物 AMPK 支持目标 3：破坏 HbbTh3/+ 小鼠中的双顺反子 miR-144/451 位点导致 α-珠蛋白沉淀和 β-地中海贫血减少总的来说，我们的研究有望阐明维持平衡的蛋白质稳态网络的生物学。通过靶向蛋白质降解合成血红蛋白，并验证 mTORC1、AMPK 和 ULK1 新型β-地中海贫血疗法的“可药物”靶标。