Transcriptional regulatory mechanisms in B cell development and leukemogenesis

B 细胞发育和白血病发生中的转录调控机制

• 批准号: 9271811
• 负责人: ROBERT G ROEDER
• 金额: $ 56.55万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-08 至 2020-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9271811
• 关键词: Acute Lymphocytic Leukemia; Adaptive Immune System; Animals; Antibodies; B cell differentiation; B-Cell Development; B-Cell Leukemia; B-Lymphocytes; BHLH Protein; Biochemical; Biochemical Genetics; Biological Assay; Biological Process; Bone Marrow; Bone Marrow Cells; Cells; Childhood Acute Lymphocytic Leukemia; Childhood Leukemia; Chimeric Proteins; Chromatin; Chromatin Structure; Chromosomal translocation; Communication; Complement; Complex; DNA; DNA Binding Domain; Data; Development; E protein; EP300 gene; Elements; Enhancers; Event; FOXO1A gene; Fusion Protein Expression; Gene Activation; Gene Expression; Gene Targeting; Generations; Genetic; Genetic Transcription; Health; Hematopoietic stem cells; Hepatic; Histones; Human; Immune system; Knock-in; Lead; Leukemic Cell; Lymphopoiesis; Malignant Neoplasms; Mediator of activation protein; Methyltransferase; Molecular; Mus; Mutation; Oncogenic; PAX5 gene; Patients; Peptide Initiation Factors; Play; Pre-B-Cell Leukemia; Protein Family; Proteins; RNA Polymerase II; RUNX1 gene; Reagent; Recombinants; Recruitment Activity; Resolution; Role; SAGA; Series; Signal Pathway; Structure; System; TAF1 gene; TAF4 gene; TCF3 gene; Testing; Therapeutic; Transgenic Mice; Treatment Efficacy; base; cell transformation; chromatin immunoprecipitation; cofactor; design; differentiated B cell; genetic analysis; genome-wide; histone acetyltransferase; histone modification; inhibitor/antagonist; leukemia; leukemogenesis; member; mouse model; mutant; novel; peptidomimetics; promoter; protein function; public health relevance; reconstitution; transcription factor; transcriptome sequencing

DESCRIPTION (provided by applicant): Antibody-producing B cells arise from hematopoietic stem cells through a series of developmental steps that involve a hierarchical transcription factor network. The transcription factor E2A plays a central role in early B cell development through synergistic functions with other lineage-restricted transcription factors. Whereas much is known about the direct genetic targets and cooperative functions of E2A and co-regulatory factors, little is known about their actual mechanism of action through interactions with various co-activators that act on the general transcription machinery or on chromatin structure. E2A is also of significance for cancer as a result of chromosomal translocations that fuse the activation domain of E2A to the DNA- binding domain of pre-B-cell leukemia factor 1 (PBX1) or hepatic leukemia factor (HLF), generating fusion proteins (E2A-PBX1 and E2A-HLF) that result in pediatric acute lymphoblastic leukemias (ALL) through dysregulation of oncogenic target genes that are largely unknown. Since E2A and E2A fusion proteins share common activation domains (AD1, AD2 and a newly described AD3), they are likely to utilize some common mechanisms of action on respective target genes. Toward an understanding of these mechanisms, preliminary studies have demonstrated novel physical and functional interactions of AD1/2 with the KIX domain of the histone acetyltransferase/coactivator p300 and of AD3 with the TAFH domain of the TAF4 subunit of initiation factor TFIID. With the general objective of understanding the mechanism of action of E2A and E2A fusion proteins in B cell development and in ALL, respectively, our specific aims are: (i) to employ biochemically defined systems reconstituted with purified factors and either DNA templates or corresponding recombinant chromatin templates to define the mechanism of action of E2A activation domain (AD)-coactivator interactions in gene activation by E2A and E2A fusion proteins~ (ii) to determine high-resolution structures of AD3-TAFH and AD1/2-KIX interactions in order to identify mutations that block these interactions (for genetic analyses) and to design peptidomimetic inhibitors with therapeutic potential~ (iii) To investigate the biological functions of E2A-coactivator interactions in B cell development through differentiation assays with bone marrow cells from E2A-/- mice and through the generation and analysis of knockin E2A (AD1/2/3mutant), TAF4(TAFHmutant) and p300(KIXmutant) mice~ and (iv) to investigate the biological functions of E2A PBX1/HLA AD-coactivator interactions in B cell leukemogenesis (ALL), using various cell transformation and mouse models and by analysis of the effects of inhibitory peptidomimetics on patient-derived leukemic cells.

描述（由申请人提供）：产生抗体的B细胞是由造血干细胞通过一系列涉及分层转录因子的发育步骤引起的 网络。转录因子E2a通过与其他谱系限制的转录因子的协同功能在早期B细胞发育中起着核心作用。尽管对E2A和共同调节因素的直接遗传靶标和合作功能有很多了解，但很少 通过与对一般转录机制或染色质结构作用的各种共激活因子的相互作用来了解其实际作用机理。 E2A也是染色体易位的导致癌症的重要性（全部）通过失调的致癌靶基因的失调，这些基因在很大程度上未知。 由于E2A和E2A融合蛋白具有共同的激活结构域（AD1，AD2和新描述的AD3），因此它们很可能在各个靶基因上使用一些共同的作用机理。为了了解这些机制，初步研究表明，AD1/2与组蛋白乙酰基转移酶/共激活酶/共激活因子p300的Kix结构域以及AD3与启动因子TfiID因子TFIID的TAF4亚基的TAFH结构域的新型物理和功能相互作用。凭借理解E2A和E2A融合蛋白在B细胞发育中的作用机制的一般目标，分别分别，我们的具体目的分别是：（i）使用生化定义的系统重新定义的系统用纯化的因子和DNA模板或相应的重组染色体模板来定义E2A激活的动作（E2A Activation-domain-nivation-e2a激活）的动作 - E2A融合蛋白〜（ii）确定AD3-TAFH和AD1/2-KIX相互作用的高分辨率结构，以鉴定阻断这些相互作用（用于遗传分析）的突变，并设计肽抑制剂与治疗势〜（iii）的肽抑制剂，以研究通过E2A-COCOCOCTOCTORICTION INTRACTION CONTRACTION的BORNORICTION与BORNORICTION Intropartions Introvientions bone bone bone bone bone bone bone conrotions conrry con的生物学作用。 E2A-/- mice and through the generation and analysis of knockin E2A (AD1/2/3mutant), TAF4(TAFHmutant) and p300(KIXmutant) mice~ and (iv) to investigate the biological functions of E2A PBX1/HLA AD-coactivator interactions in B cell leukemogenesis (ALL), using various cell tr​​ansformation and mouse models and by analysis of the effects of对患者衍生的白血病细胞的抑制性肽仪。