The Role of MMSET in the Pathogenesis and Progression of Lymphoid Malignancy

MMSET 在淋巴恶性肿瘤发病机制和进展中的作用

• 批准号: 9759647
• 负责人: Jonathan D. Licht
• 金额: $ 33.84万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-11 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9759647
• 关键词: Acute Lymphocytic Leukemia; Affect; Alleles; Animal Disease Models; B-Cell Development; B-Lymphocytes; Binding; Biological; CRISPR/Cas technology; Cell Line; Cell model; Child; Childhood Acute Lymphocytic Leukemia; Chromatin; Chronic Lymphocytic Leukemia; DNA Repair; Data; Development; Disease; Disease Progression; EZH2 gene; Epithelial; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genome; Grant; Growth; Hematology; Histone-Lysine N-Methyltransferase; Histones; Immunoglobulins; In Vitro; Knock-in; Knock-in Mouse; Lead; Link; Lysine; MYC gene; Malignant Neoplasms; Malignant lymphoid neoplasm; Malignant neoplasm of lung; Malignant neoplasm of prostate; Mantle Cell Lymphoma; Mediating; Mesenchymal; Modeling; Molecular; Multiple Myeloma; Mutate; Mutation; Neuroblastoma; Oncogenes; Oncogenic; Pathogenesis; Pathway interactions; Point Mutation; Property; Prostate; Proteins; Recurrence; Relapse; Role; SET Domain; Transferase; WHSC1 gene; base; c-myc Genes; cancer cell; cancer therapy; cell growth; cell motility; cell transformation; chromatin modification; gain of function mutation; genetically modified cells; genome sequencing; histone methylation; in vivo; inhibitor/antagonist; molecular sequence database; mouse model; mutant; novel therapeutics; overexpression; prostate cancer cell; public health relevance; response; therapeutic target; tumor

 DESCRIPTION (provided by applicant): We have studied the MMSET (also known as WHSC1 or NSD2) protein a histone lysine methyltransferase identified by its rearrangement with the immunoglobulin locus aberrant overexpression in multiple myeloma (MM). Overexpression of MMSET leads to dramatic shifts in chromatin modification and gene expression and stimulates cell growth. MMSET overexpression has also been linked to aberrant cell growth and invasive properties in prostate, lung cancer and neuroblastoma. More recently we and others described a recurrent point mutation (E1099K) in the SET domain of MMSET in acute lymphoblastic leukemia (ALL). The importance of this mutation is growing as genome sequencing showed that this mutation in present in ~10% of cases of mantle cell lymphoma (MCL) and is found in ~10-20% of cases of relapsed pediatric ALL. Collectively these data indicate that MMSET and the pathways it regulates represent therapeutic targets in MM and other malignancies. Our overarching hypothesis is that overexpression or gain of function mutations of MMSET alter gene regulation leading to the pathogenesis of MM and to the progression of ALL. While some target genes may be activated in common across these tumors, our preliminary data suggests that MMSET mutation dysregulates a unique set of genes in ALL. By constructing genetically engineered cell line and mouse models of models of the action of the E1099K mutation we will ascertain the detailed molecular mechanisms, by which MMSETE1099K affects gene expression, identify critical target genes and pathways and uncover new therapeutic opportunities for cancer treatment. Our specific aims will be: Aim 1: Evaluate the Biological Activity of a Recurrent Point Mutation of MMSET in Malignancy. We will determine the mechanism of action of this protein and how it alters histone methylation levels. Using newly prepared CRISPR/CAS9 edited cell lines we will determine how mutant MMSET modulates cell growth and response to therapy. Aim 2: Define the Genetic Targets and Pathways Affected by Oncogenic Mutations of MMSET. Preliminary data suggest that mutant MMSET activates a different set of genes in ALL compared to those affected in MM. Using genetically edited cell lines, we will determine how the mutation alters chromatin configuration across the genome and identify critical genes and pathways of its oncogenic action. Aim 3: Evaluate the Ability of Mutant MMSET to Collaborate with Known MM and ALL Oncogenes. We will cross a newly created knockin mouse expressing MMSETE1099K with established mouse models of ALL and MM to determine how this disease allele may drive pathogenesis and progression of disease.

 描述(申请人提供)：我们研究了MMSET(也称为WHSC1或NSD2)蛋白，一种组蛋白赖氨酸甲基转移酶，通过其与免疫球蛋白基因座的重排而在多发性骨髓瘤(MM)中异常过度表达。MMSET的过表达导致染色质修饰和基因表达的戏剧性变化，并刺激细胞生长。MMSET的过度表达也与前列腺癌、肺癌和神经母细胞瘤的细胞异常生长和侵袭性有关。最近，我们和其他人描述了急性淋巴细胞白血病(ALL)中MMSET SET结构域的一个复发点突变(E1099K)。随着基因组测序显示，该突变存在于约10%的套细胞淋巴瘤(MCL)和约10%-20%的复发性儿童ALL中，该突变的重要性与日俱增。总的来说，这些数据表明，MMSET及其调节的通路代表了MM和其他恶性肿瘤的治疗靶点。我们的主要假设是MMSET功能突变的过度表达或获得改变了导致MM发病和ALL进展的基因调控。虽然某些靶基因可能在这些肿瘤中被共同激活，但我们的初步数据表明，MMSET突变在所有肿瘤中都扰乱了一组独特的基因。通过构建E1099K突变作用的基因工程细胞系和小鼠模型，我们将确定MMSETE1099K影响基因表达的详细分子机制，识别关键的靶基因和途径，并发现癌症治疗的新治疗机会。我们的具体目标是：目标1：评估MMSET复发点突变在恶性肿瘤中的生物学活性。我们将确定这种蛋白质的作用机制，以及它如何改变组蛋白甲基化水平。使用新准备的CRISPR/Cas9编辑的细胞系，我们将确定突变的MMSET如何调节细胞生长和对治疗的反应。目的2：明确MMSET癌基因突变影响的遗传靶点和途径。初步数据表明，突变的MMSET激活了一组与MM中受影响的基因不同的基因。使用经过基因编辑的细胞系，我们将确定突变如何改变整个基因组的染色质配置，并确定其致癌作用的关键基因和途径。目的3：评价突变型MMSET与已知MM及所有癌基因协同作用的能力。我们将新创造的表达MMSETE1099K的敲门小鼠与已建立的ALL和MM小鼠模型进行杂交，以确定这种疾病等位基因如何推动疾病的发病和进展。