Functional characterization of CUX1 regulators for the treatment of myelodysplastic syndromes

CUX1 调节剂治疗骨髓增生异常综合征的功能特征

• 批准号: 10544145
• 负责人: Madhavi Dushyanthi Senagolage
• 金额: $ 7.43万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10544145
• 关键词: Alleles; Anemia; Biological; Biology; Biometry; Blood; Blood Cells; Bone Marrow; CD34 gene; CRISPR/Cas technology; Cell Line; Cell Lineage; Cells; Chromosome 7; Chromosome abnormality; Clinical; Communication; Cytogenetics; Data; Defect; Deficiency Diseases; Development; Differentiation Therapy; Disease; Disease model; Dysmyelopoietic Syndromes; Dysplasia; Erythroid; Foundations; Genes; Glycogen Synthase Kinase 3; Goals; Grant; Hematopoiesis; Hematopoietic; Hematopoietic stem cells; Hemorrhage; Homeobox; Human; Impairment; Ineffective Hematopoiesis; Infection; Mediating; Megakaryocytes; Mentors; Modeling; Molecular; Monosomy 7; Morbidity - disease rate; Mus; Mutation; Mutation Analysis; Myeloid Cells; Myeloproliferative disease; Online Systems; Pathway interactions; Patients; Pharmaceutical Preparations; Phosphotransferases; Post-Translational Protein Processing; Prognosis; Protein Isoforms; Proteins; Regulation; Regulatory Pathway; Research; Residual state; Risk; Signs and Symptoms; Site; Testing; Therapeutic; Training; Transcript; Tumor Suppressor Proteins; Work; Writing; arm; career; chromosome 7 loss; chromosome 7q loss; computerized tools; cytopenia; efficacy evaluation; efficacy testing; erythroid differentiation; genome-wide; granulocyte; high risk; improved outcome; in vivo; inhibitor; innovation; knock-down; leukemia; leukemic transformation; loss of function; mouse model; novel; patient retention; pharmacologic; progenitor; restoration; stem cell homeostasis; therapeutic effectiveness; therapeutic evaluation; transcription factor

PROJECT SUMMARY/ABSTRACT The myelodysplastic syndromes (MDS) are a heterogeneous group of disorders characterized by cytopenias and dysplasia in one or more of the myeloid cell lines due to ineffective hematopoiesis. Patients present with signs and symptoms of anemia, accompanied by infections and bleeding leading to serious morbidity. Loss of chromosome 7 (-7) or the long-arm of 7 [del(7q)] is commonly seen in high-risk MDS patients. Treatment options are limited and high-risk patients have less than one-year survival. Cut-like homeobox 1 (CUX1), a transcription factor encoded in the commonly deleted segment of 7q22, is expressed at haploinsufficient transcript and protein levels in -7/del(7q) myeloid diseases. Knockdown of CUX1 causes a spontaneous hematopoietic disorder with many features of human MDS, including erythroid dysplasia, and fatal anemia, indicating that loss of this single gene is sufficient to cause disease. Remarkably, CUX1 re-expression reverses MDS in mice, which provides the premise for the current proposal, that restoration of CUX1 is a viable therapeutic strategy for MDS patients. However, there exists a major gap in our understanding of the mechanisms by which CUX1 levels are regulated. Preliminary studies indicate that Glycogen synthase kinase- 3 (GSK3) inhibition increases cellular CUX1 protein levels. The central hypothesis is that pharmacological induction of CUX1 levels will restore hematopoietic stem cell homeostasis and reverse impaired differentiation in CUX1 deficient myeloid disease models. The central hypothesis will be tested by pursuing two specific aims: 1) define the cellular and molecular mechanisms by which CUX1 levels are regulated, and 2) drive normal erythroid differentiation of -7/del(7q) myelodysplasia through therapeutic restoration of CUX1. Under the first aim, GSK3 mediated post-translational regulatory mechanism of CUX1 levels will be determined using site- directed mutational analysis in immortalized human blood cell lines. Additionally, a genome-wide CRISPR/Cas9 loss of function screen will be utilized to identify novel regulators of CUX1. For the second aim, CD34+ human -7/del(7q) MDS cells and CUX1-deficient mouse models will be used to determine the efficacy of GSK3 isoform selective inhibitors in restoring CUX1 protein and reversing erythroid differentiation defects. The proposed research is expected to reveal druggable and regulatory pathways of CUX1 to restore its levels as a way of reversing anemia and dysplasia in -7/del(7q) MDS patients.

项目概要/摘要 骨髓增生异常综合征 (MDS) 是一组以血细胞减少为特征的异质性疾病 以及由于无效造血作用导致的一种或多种骨髓细胞系的发育不良。患者就诊时患有 贫血的体征和症状，伴有感染和出血，导致严重发病。损失 7 号染色体 (-7) 或 7 号长臂 [del(7q)] 常见于高危 MDS 患者。治疗 选择有限，高危患者的生存期不到一年。切割样同源框 1 (CUX1) 编码在 7q22 常见缺失片段中的转录因子，在单倍体不足处表达 -7/del(7q) 骨髓疾病中的转录物和蛋白质水平。 CUX1 的敲低会导致自发的 具有人类 MDS 的许多特征的造血障碍，包括红细胞发育不良和致命性贫血， 表明这个单一基因的丢失就足以引起疾病。值得注意的是，CUX1 重新表达逆转 小鼠 MDS，这为当前提议提供了前提，即恢复 CUX1 是可行的 MDS 患者的治疗策略。然而，我们对这个问题的理解却存在着很大的差距。 CUX1 水平的调节机制。初步研究表明糖原合酶激酶- 3 (GSK3) 抑制会增加细胞 CUX1 蛋白水平。中心假设是药理学 诱导 CUX1 水平将恢复造血干细胞稳态并逆转受损的分化 在 CUX1 缺陷性骨髓疾病模型中。中心假设将通过追求两个具体目标来检验： 1) 定义调节 CUX1 水平的细胞和分子机制，以及 2) 驱动正常 通过 CUX1 的治疗性恢复实现 -7/del(7q) 骨髓增生异常的红系分化。在第一个下 目的，GSK3介导的CUX1水平的翻译后调节机制将使用位点确定 永生化人类血细胞系的定向突变分析。此外，全基因组 CRISPR/Cas9 功能丧失筛选将用于鉴定 CUX1 的新型调节因子。为了第二个目标， CD34+人-7/del(7q) MDS细胞和CUX1缺陷小鼠模型将用于确定疗效 GSK3 亚型选择性抑制剂在恢复 CUX1 蛋白和逆转红细胞分化缺陷方面的作用。 拟议的研究预计将揭示 CUX1 恢复其水平的药物和调节途径 作为逆转 -7/del(7q) MDS 患者贫血和发育不良的一种方法。