Cohesin Mutations in Acute Myelogenous Leukemia

急性髓系白血病的粘连蛋白突变

• 批准号: 10224847
• 负责人: Sridhar Rao
• 金额: $ 37.67万
• 依托单位: VERSITI WISCONSIN, INC.
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-19 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10224847
• 关键词: Acute Myelocytic Leukemia; Address; Allogeneic Bone Marrow Transplantation; Area; Bone Marrow; Bone Marrow Transplantation; Cancer Biology; Cells; ChIP-seq; Chemicals; Clinical; Clinical Trials; Complex; Coupled; Data; Development; Disease; Distal; Drosophila genus; Epigenetic Process; Gene Expression; Gene Mutation; Gene Silencing; Genes; Genomics; Goals; Hematopoietic; Hematopoietic stem cells; Histones; Human; In Vitro; Induced Mutation; Lysine; Malignant Bone Neoplasm; Malignant Neoplasms; Mammals; Mediating; Modeling; Molecular Conformation; Mus; Mutate; Mutation; NPM1 gene; Pathogenesis; Patients; Pilot Projects; Play; Polycomb; Publications; Recurrence; Regulatory Element; Research; Role; Secondary to; Site; Somatic Mutation; Supportive care; Survival Rate; Testing; Therapeutic Agents; Untranslated RNA; Vertebrates; Work; base; chemotherapy; chromatin immunoprecipitation; cohesin; derepression; epigenetic silencing; experimental study; genetic signature; genome editing; genome-wide; histone methyltransferase; improved; in vivo; inhibitor/antagonist; innovation; leukemia; leukemogenesis; mouse model; next generation sequencing; novel; novel therapeutics; prevent; recruit; self-renewal; targeted treatment; transcription factor; transcriptome

Abstract: Acute Myelogenous Leukemia (AML) is a bone marrow-derived malignancy and remains a clinical challenge, with long-term survival of approximately 50%. Improvements in AML therapy have predominantly come through better supportive care and the improvements in allogeneic bone marrow transplantation, rather than improved chemotherapy approaches. This plateauing in AML survival points to a need for better, targeted therapies. Part of the difficulty with developing novel treatments to AML is that it is a heterogeneous disease, with well over 250 different mutated genes, although any one patient has on average 13 somatic mutations. Thus, developing a “unifying genetic signature” that drives AML remains elusive. To understand how mutations found in AML contribute to leukemia development, we have chosen to focus on mutations in genes encoding the cohesin complex. Cohesin mutations occur in approximately 10-20% of patients, but its role in leukemogenesis is unknown. Our preliminary data demonstrate that cohesin loss causes increased self- renewal in wild-type murine bone marrow, a hallmark of leukemia development. Interestingly, this increased self-renewal is augmented in Npm1+/- after cohesin loss. While Npm1 mutations are the most common single gene mutation in AML, Npm1-mutated mice rarely develop AML, implying additional mutations are required. In both wild-type (WT) and Npm1-mutated bone marrow, cohesin loss causes epigenetic derepression of the self- renewal associated transcription factor HoxA9. Given that the Polycomb Repressive Complex 2 (PRC2), which mediates trimethylation of Histone 3 on Lysine 27 (H3K27me3), normally silences HoxA9 epigenetically we hypothesize that cohesin haploinsufficiency promotes AML by derepressing PRC2 target genes. The goal of this proposal is to use a combination of genome-wide approaches and genomic editing to determine the mechanism by which cohesin mutations induce abnormal self-renewal and promote leukemia development. In Aim 1 we will use chromatin immunoprecipitation coupled with next-generation sequencing (ChIP-seq) to identify changes in PRC2 genomic localization after cohesin loss. In Aim 2, we will use circular chromosomal conformational capture (4C) to identify which distal cis-regulatory elements (CREs) are normally recruited to regulate HoxA9 expression, and the changes in distal CREs after cohesin loss. In Aim 3 we will further characterize both in vitro and in vivo the effects of cohesin depletion on WT and Npm1cA/+ bone marrow. Also in Aim 3, we will test the role of the histone methyltransferase Dot1l loss/inhibition in preventing HoxA9 expression and cohesin-depletion associated self-renewal. Collectively, the long-term goal of our studies is to develop a novel mouse model of AML based upon loss of cohesin in Npm1cA AML, a combination of mutations that represents a sizeable fraction of patients with AML. In addition, we will use this model to address mechanistic question regarding how altered targeting of epigenetic complexes can cause disordered gene expression and leukemogenesis.

摘要：急性骨髓性白血病（AML）是一种骨髓源性恶性肿瘤，目前仍是临床上的常见病 长期存活率约为50%。AML治疗的改善主要是 通过更好的支持性护理和异基因骨髓移植的改进， 而不是改进化疗方法。AML生存率的这种稳定表明需要更好的、有针对性的治疗。 治疗开发AML新疗法的部分困难在于它是一种异质性疾病， 有超过250种不同的突变基因，尽管任何一个病人平均有13个体细胞突变。 因此，开发一种驱动AML的“统一基因签名”仍然是难以捉摸的。为了了解突变是如何 在AML中发现的导致白血病发展的基因突变，我们选择专注于编码 cohesin complex.在大约10-20%的患者中发生粘着蛋白突变，但其在 白血病发生是未知的。我们的初步数据表明，内聚蛋白的缺失会导致自我调节能力的增加， 野生型小鼠骨髓中的更新，白血病发展的标志。有趣的是，这增加了 自我更新在Npm 1 +/-中在粘着蛋白损失后增强。虽然Npm 1突变是最常见的单一突变， 在AML中，Npm 1基因突变的小鼠很少发生AML，这意味着需要额外的突变。在 无论是野生型（WT）和Npm 1突变的骨髓，粘连蛋白的损失导致表观遗传的自我去抑制， 更新相关转录因子HoxA 9。鉴于Polycomb抑制复合物2（PRC 2）， 其介导组蛋白3在赖氨酸27（H3 K27 me 3）上的三甲基化，通常使HoxA 9表观遗传学沉默 我们推测，粘附素单倍不足通过去抑制PRC 2靶基因促进AML。 该提案的目标是使用全基因组方法和基因组编辑的组合， 确定粘附素突变诱导异常自我更新和促进白血病的机制 发展在目标1中，我们将使用染色质免疫沉淀结合下一代测序技术， （ChIP-seq）来鉴定粘附素损失后PRC 2基因组定位的变化。在目标2中，我们将使用循环 染色体构象捕获（4C），以确定哪些远端顺式调控元件（克雷斯）通常 募集来调节HoxA 9表达，以及在粘附素丧失后远端克雷斯的变化。在目标3中， 进一步在体外和体内表征内聚蛋白耗竭对WT和Npm 1cA/+骨髓的影响。 同样在目标3中，我们将测试组蛋白甲基转移酶Dot 1 l丢失/抑制在阻止HoxA 9 表达和粘着蛋白耗竭相关的自我更新。总的来说，我们研究的长期目标是 基于Npm 1cA AML中粘附素的丢失，开发了一种新的AML小鼠模型， 这代表了相当大一部分AML患者。此外，我们还将使用该模型来解决 关于表观遗传复合体靶向改变如何导致基因紊乱的机制问题 表达和白血病发生。

项目成果

Nanog Expression in Embryonic Stem Cells - An Ideal Model System to Dissect Enhancer Function.

• DOI: 10.1002/bies.201700086
• 发表时间: 2017-12
• 期刊: BioEssays : news and reviews in molecular, cellular and developmental biology
• 作者: Blinka S;Rao S
• 通讯作者: Rao S

DOT1L inhibitors block abnormal self-renewal induced by cohesin loss.

• DOI: 10.1038/s41598-021-86646-9
• 发表时间: 2021-03-31
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Heimbruch KE;Fisher JB;Stelloh CT;Phillips E;Reimer MH Jr;Wargolet AJ;Meyer AE;Pulakanti K;Viny AD;Loppnow JJ;Levine RL;Pulikkan JA;Zhu N;Rao S
• 通讯作者: Rao S

GATA4 Controls Epithelial Morphogenesis in the Developing Stomach to Promote Establishment of Glandular Columnar Epithelium.

• DOI: 10.1016/j.jcmgh.2021.05.021
• 发表时间: 2021
• 期刊: Cellular and molecular gastroenterology and hepatology
• 影响因子: 7.2
• 作者: DeLaForest A;Kohlnhofer BM;Franklin OD;Stavniichuk R;Thompson CA;Pulakanti K;Rao S;Battle MA
• 通讯作者: Battle MA

Cohesin loss and MLL-AF9 are not synthetic lethal in murine hematopoietic stem and progenitor cells.

粘连蛋白丢失和 MLL-AF9 在小鼠造血干细胞和祖细胞中并不具有合成致死性。

• DOI: 10.21203/rs.3.rs-3894962/v1
• 发表时间: 2024
• 期刊: Research square
• 作者: Meyer,Alison;Stelloh,Cary;Zhu,Nan;Rao,Sridhar
• 通讯作者: Rao,Sridhar

Rap1A, Rap1B, and β-Adrenergic Signaling in Autologous HCT: A Randomized Controlled Trial of Propranolol.

自体 HCT 中的 Rap1A、Rap1B 和 β-肾上腺素信号传导：普萘洛尔的随机对照试验。

• 发表时间: 2022
• 期刊: The Yale journal of biology and medicine
• 作者: Johnson,AlexanderK;Lorimer,EllenL;Szabo,Aniko;Wu,Ruizhe;Shah,NiravN;D'Souza,Anita;Chhabra,Saurabh;Hamadani,Mehdi;Dhakal,Binod;Hari,Parameswaran;Rao,Sridhar;Carlson,Karen;Williams,CarolL;Knight,JenniferM
• 通讯作者: Knight,JenniferM