Therapeutic Strategies for Anemia in 3q26 Rearranged Leukemia

项目总结/摘要 根据最新的NIH癌症统计评论，白血病，一种血细胞癌症，是第九大癌症。 常见的癌症类型。急性髓细胞白血病（AML）是一种侵袭性白血病，具有高致死性 （约75%的患者在诊断后5年死亡），其特征是贫血和过度增殖。 骨髓（BM）中异常的髓样祖细胞。染色体3q 26区带的重排 预示着生存率的进一步降低，并导致癌基因嗜亲性病毒整合的过度表达 站点1（EVI 1）。3q 26重排AML的严重性，对EVI 1在AML中的作用缺乏深入的了解， 白血病和治疗策略的不足引起了我们实验室和其他人的兴趣，以研究EVI 1相关的 白血病发生虽然以前的研究组使用病毒转导的BM移植来过表达EVI 1，但我们发现， 是第一个通过建立一个诱导型基因组来概括小鼠3q 26重排效应的实验室。 EVI 1-过表达模型，这为我们提供了新的见解EVI 1的机制， 诱发白血病我们使用我们的体内和体外模型得出结论，EVI 1引起骨髓扩张， 阻断红细胞生成和淋巴细胞生成。作为对分子机制的深入了解，我们以前 证明EVI 1与GACAAGATA结合，GACAAGATA与EVI 1的主调节因子的结合位点重叠。 红细胞生成加塔-1。此外，我们的数据表明，EVI 1上调先前发表的加塔-1， 阻断剂PU.1，我们发现EVI 1与PU.1编码基因（Spi-1）上游的增强子结合。因此，在本发明中， 我们假设EVI 1通过两种机制阻止红细胞分化：1）直接与加塔竞争- 1）包含EVI 1/加塔-1重叠基序的关键基因组结合位点;与Spi-1增强子结合， 上调PU.1，其抑制GATA 1功能。我们将调查这两个假设的机制，使用 尖端技术，包括ChIP-seq，ATAC-seq和CRISPR，在我的赞助商的培训下， 合作者为了将提出的机制见解转化为临床环境和治疗 我们将使用体内模型进行CRISPR文库筛选，以识别逆转 与EVI 1过表达相关的红细胞生成阻断。 总之，这项研究将侧重于调查红细胞生成障碍和由此导致的贫血， 解释了与3q 26重排白血病相关的致命性增加，并将揭示新的治疗方法 逆转白血病相关贫血的策略。

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.