Role of HOTTIP/beta-catenin-HOXA9/PRMT1 axis in hematopoietic and leukemic stem cells

HOTTIP/β-catenin-HOXA9/PRMT1 轴在造血干细胞和白血病干细胞中的作用

• 批准号: 10537540
• 负责人: So Eric
• 金额: $ 52.03万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10537540
• 关键词: Acute Myelocytic Leukemia; Adult; Binding; Biochemical; Biological; Bone marrow failure; Cell Line; Cells; Complement; Complex; DNA Damage; DNA Repair; Degenerative Disorder; Development; Disease; Enhancers; Epigenetic Process; Future; Genetic; Genetic Transcription; Genome; Goals; HOXA9 gene; HOXA9 protein; Hematological Disease; Hematopoiesis; Hematopoietic; Hematopoietic stem cells; Homeostasis; Human; Individual; Knock-in Mouse; Lead; Link; Malignant - descriptor; Malignant Neoplasms; Mediating; Mediator of activation protein; Molecular; Molecular Analysis; Mus; NURF; Natural regeneration; Oncogenic; Pathogenesis; Pathologic; Pathway interactions; Patients; Pharmacology; Play; Population; Premature aging syndrome; Property; Protein-Arginine N-Methyltransferase; Proteins; RUNX1 gene; Regulation; Repair Complex; Reporting; Role; Stat5 protein; Techniques; Testing; Therapeutic; Tissues; Transcriptional Regulation; Translating; Untranslated RNA; Validation; WNT Signaling Pathway; acute myeloid leukemia cell; beta catenin; cancer stem cell; cohesin; cohesion; design; gain of function; gene regulatory network; hematopoietic gene; hematopoietic stem cell self-renewal; insight; leukemic stem cell; loss of function; malignant phenotype; novel; programs; promoter; protein function; repair function; response; self-renewal; small molecule inhibitor; stem cell self renewal; stem cells; success; therapeutic target; therapeutically effective; transcription factor

Abstract: Self-renewal is an indispensable property that allows stem cells to regenerate and maintain the homeostasis of functionally diverse cell populations. Dergulation of self-renewal in hematopoietic stem cell (HSC) directly links to a number of hematopoietic degenerative disorders including bone marrow failure and various blood diseases, whereas its aberrant activation is a defining and indispensable feature of leukemia stem cells (LSCs) that sustain the malignant phenotypes. We and others have shown that self-renewal of acute myeloid leukemia (AML) stem cells heavily rely on the key component of canonical Wnt signaling pathway, β-catenin and the homeobox protein, HOXA9 that are largely dispensable for adult HSCs. Although these findings reveal a contrasting functional requirement and a potential therapeutic opportunity for AML, development of small molecule inhibitors against oncogenic transcription factors has so far met with very little success largely due to our lack of understanding about the molecular regulations of these proteins in mediating hematopoietic self-renewal, which has significantly hindered progress in developing effective therapeutic strategies to treat the resultant diseases. Intriguingly, we have recently revealed coregulation of both canonical Wnt pathway and posterior HOXA loci including HOXA9 by the long non-coding RNA (lncRNA), HOTTIP. Strikingly, we also report a novel crosstalk between β-catenin and HOXA9 that can override their individual requirement in both mouse and human AML of HSC origins. In search for crucial mediators for β-catenin and Hoxa9 functions, we further identified that protein arginine methyltransferase 1 (PRMT1), which also implicates in DNA damage and repair (DDR), can functionally replace HOXA9 or β-catenin in AML stem cell originated from HSCs. PRMT1 together with DDR complex are unbiasedly isolated along with CTCF/cohesin complex, hematopoietic transcription factors (TFs) and nucleosome remodeling factors as HOTTIP interacting partners by ChIRP-MS in AML cells. These findings not only discover a novel HOTTIP/β-catenin-HOXA9/PRMT1 axis critical for mediating hematopoietic self-renewal, but also lead to central hypothesis that HOTTIP/β-catenin-HOXA9/PRMT1 axis coordinates hematopoietic self-renewal and characterization of the functions of individual components and their crosstalk along the axis regulates hematopoietic specific transcription networks and DDR pathways to modulate hematopoietic self-renewal in the disease setting. In this proposal, we will 1 decipher cooperative action of HOTTIP and hematopoietic TFs in regulating HOXA9 and β-catenin axis; 2) dissect the molecular functions and regulation of β-catenin- HOXA9/Prmt1 axis during normal and malignant hematopoiesis. Success completion of proposed studies not only will establish the molecular principles, but also facilitate the design of specific therapeutics in modulating self-renewal activities in normal and malignant stem cells, which can be potentially translated into patient benefits.

抽象的： 自我更新是干细胞再生和维持体内平衡不可或缺的特性。 功能多样的细胞群。造血干细胞（HSC）自我更新的失调直接联系 一些造血退行性疾病，包括骨髓衰竭和各种血液疾病， 而其异常激活是白血病干细胞 (LSC) 的一个决定性且不可或缺的特征，它维持着 恶性表型。我们和其他人已经证明，急性髓系白血病（AML）的自我更新 细胞严重依赖经典 Wnt 信号通路的关键成分、β-连环蛋白和同源盒蛋白， HOXA9 对于成人 HSC 来说基本上是可有可无的。尽管这些发现揭示了对比的功能 AML 的需求和潜在的治疗机会，开发针对 AML 的小分子抑制剂 迄今为止，致癌转录因子几乎没有取得成功，这主要是由于我们缺乏了解 关于这些蛋白质在介导造血自我更新中的分子调控，这对 阻碍了开发有效治疗策略来治疗由此产生的疾病的进展。有趣的是，我们 最近揭示了经典 Wnt 通路和后 HOXA 基因座（包括 HOXA9）的共调控 由长非编码 RNA (lncRNA) HOTTIP 组成。引人注目的是，我们还报告了 β-连环蛋白之间的一种新的串扰 HOXA9 可以超越 HSC 来源的小鼠和人类 AML 的个体要求。在 在寻找 β-catenin 和 Hoxa9 功能的关键介质时，我们进一步确定了蛋白质精氨酸 甲基转移酶 1 (PRMT1) 也参与 DNA 损伤和修复 (DDR)，可以在功能上替代 AML干细胞中的HOXA9或β-catenin源自HSC。 PRMT1 与 DDR 复合体是无偏的 与 CTCF/粘连蛋白复合物、造血转录因子 (TF) 和核小体一起分离 通过 ChIRP-MS 在 AML 细胞中将重塑因子视为 HOTTIP 相互作用伙伴。这些发现不仅发现 一种新型 HOTTIP/β-catenin-HOXA9/PRMT1 轴对于介导造血自我更新至关重要，但也导致 HOTTIP/β-catenin-HOXA9/PRMT1 轴协调造血自我更新和 各个组件的功能表征及其沿轴的串扰调节 造血特异性转录网络和 DDR 通路调节造血自我更新 疾病设置。在本提案中，我们将 1 破译 HOTTIP 与造血 TF 的合作作用 调节 HOXA9 和 β-catenin 轴； 2）剖析β-catenin的分子功能和调控 正常和恶性造血过程中的 HOXA9/Prmt1 轴。未成功完成拟议研究 不仅会建立分子原理，而且还有助于设计调节特定疗法 正常和恶性干细胞的自我更新活动，有可能转化为患者 好处。