Therapeutic Strategies for Anemia in 3q26 Rearranged Leukemia

3q26重排白血病贫血的治疗策略

• 批准号: 9754589
• 负责人: Edward Ayoub
• 金额: $ 0.28万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9754589
• 关键词: 3q26; ATAC-seq; Acute Myelocytic Leukemia; Address; Alleles; Anemia; Attenuated; Binding; Binding Proteins; Binding Sites; Blood Cells; Bone Marrow; Bone Marrow Transplantation; CRISPR library; Cells; ChIP-seq; Chromatin; Chromatin Remodeling Factor; Chromosomal Rearrangement; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; DNA Binding; Data; Diagnosis; EVI1 gene; Elements; Enhancers; Erythroblasts; Erythroid; Erythroid Cells; Erythropoiesis; Erythropoietin; Fellowship; GATA1 gene; Genes; Genetic Screening; Genome; Genomics; Goals; Grant; Growth; Hematopoietic Neoplasms; Hematopoietic stem cells; Knock-out; Lead; Lymphopoiesis; Mediating; Modeling; Molecular; Mus; Myelogenous; Myeloid Progenitor Cells; Myelopoiesis; Oncogenes; Patients; Pattern; Publishing; Purines; Relapse; Reporting; Research; Role; Severities; Site; Survival Rate; System; Techniques; Testing; Tetracyclines; Therapeutic; Training; Translating; United States National Institutes of Health; Up-Regulation; Upstream Enhancer; Viral; cancer statistics; cancer type; clinical application; erythroid differentiation; experimental study; hematopoietic differentiation; improved; in vitro Model; in vivo; in vivo Model; insight; interest; knock-down; lambda Spi-1; leukemia; leukemogenesis; mouse model; new therapeutic target; novel therapeutic intervention; overexpression; progenitor; recruit; screening; small hairpin RNA; therapeutic target; transcription factor

Project Summary/ Abstract According to the most recent NIH Cancer Statistics Review, leukemia, a cancer of blood cells, is the ninth most common type of cancer. Acute myeloid leukemia (AML) is an aggressive form of leukemia with high lethality (~75% of patients die 5 years after being diagnosed) characterized by anemia, and excessive proliferation of abnormal myeloid progenitor cells in the bone marrow (BM). Rearrangements of the chromosomal band 3q26 portend further reduction in survival, and lead to the overexpression of the oncogene Ecotropic Viral Integration Site 1 (EVI1). The severity of 3q26 rearranged AML, the lack of in-depth understanding of the role of EVI1 in leukemia, and the inadequate therapeutic strategies interested our lab and others to investigate EVI1 associated leukemogenesis. While previous groups used transplantation of BM virally transduced to overexpress EVI1, we are the first lab to recapitulate the effects of the 3q26 rearrangements in the mouse by establishing an inducible EVI1-overexpression model, which has provided us with new insights into the mechanisms by which EVI1 induces leukemia. We concluded using our in vivo and in vitro models that EVI1 causes myeloid expansion and blocks both erythropoiesis and lymphopoiesis. As an insight to the molecular mechanism, we previously documented that EVI1 binds to GACAAGATA, which overlaps with the binding site of the master regulator of erythropoiesis GATA-1. Additionally, our data indicate that EVI1 upregulates a previously published GATA-1 blocker, PU.1, and we showed that EVI1 binds to an enhancer upstream of PU.1 encoding gene (Spi-1). Thus, we hypothesize that EVI1 blocks erythroid differentiation by two mechanisms: 1) directly competing with GATA- 1 for key genomic binding sites harboring EVI1/GATA-1 overlap motifs and 2)!binding to Spi-1 enhancer and upregulating PU.1, which suppresses GATA1 function. We will investigate both hypothesized mechanisms using cutting edge techniques including ChIP-seq, ATAC-seq, and CRISPR under the training of my sponsor and collaborator. In order to translate the proposed mechanistic insights into clinical settings and therapeutic strategies, we will perform CRISPR library screening using an in vivo model to identify genes that reverse erythropoiesis blockage associated with EVI1-overexpression. In summary, this fellowship will focus on investigating erythropoiesis blockage and resulting anemia that might explain the increased lethality associated with 3q26 rearranged leukemia, and It will unveil new therapeutic strategies that reverse the leukemia-associated anemia.

项目概要/摘要 根据最新的 NIH 癌症统计评论，白血病（一种血细胞癌症）在最常见的癌症中排名第九。 常见的癌症类型。急性髓系白血病 (AML) 是一种具有高致死率的侵袭性白血病 （约 75% 的患者在诊断后 5 年内死亡）以贫血和细胞过度增殖为特征 骨髓（BM）中异常的骨髓祖细胞。染色体带 3q26 的重排 预示着生存率进一步下降，并导致致癌基因生态性病毒整合的过度表达 站点 1 (EVI1)。 3q26重排AML的严重性，缺乏对EVI1在AML中的作用的深入了解 白血病，以及不充分的治疗策略使我们的实验室和其他人感兴趣，以研究 EVI1 相关性 白血病发生。虽然之前的小组使用病毒转导的 BM 移植来过度表达 EVI1，但我们 是第一个通过建立诱导型来重现 3q26 重排对小鼠的影响的实验室 EVI1过度表达模型，为我们提供了关于EVI1的机制的新见解 诱发白血病。我们使用我们的体内和体外模型得出结论，EVI1 会导致骨髓扩张和 阻断红细胞生成和淋巴细胞生成。为了深入了解分子机制，我们之前 文献报道 EVI1 与 GACAAGATA 结合，GACAAGATA 与 GACAAGATA 的主调节因子的结合位点重叠 红细胞生成GATA-1。此外，我们的数据表明 EVI1 上调先前发表的 GATA-1 阻断剂 PU.1，我们发现 EVI1 与 PU.1 编码基因 (Spi-1) 上游的增强子结合。因此， 我们假设 EVI1 通过两种机制阻止红细胞分化：1）直接与 GATA-竞争 1 为含有 EVI1/GATA-1 重叠基序的关键基因组结合位点，2)! 与 Spi-1 增强子结合 上调 PU.1，抑制 GATA1 功能。我们将使用以下方法研究这两种假设的机制 在我的赞助商的培训下，包括 ChIP-seq、ATAC-seq 和 CRISPR 在内的尖端技术 合作者。为了将所提出的机制见解转化为临床环境和治疗 策略，我们将使用体内模型进行 CRISPR 文库筛选，以识别逆转的基因 与 EVI1 过度表达相关的红细胞生成受阻。 总之，该奖学金将重点研究红细胞生成受阻和由此导致的贫血，这可能会导致贫血。 解释与 3q26 重排白血病相关的致死率增加，并将推出新的治疗方法 逆转白血病相关贫血的策略。