RUNX-fusion Target Genes in Normal and Leukemic Hematopoiesis

正常和白血病造血中的 RUNX 融合靶基因

• 批准号: 7229966
• 负责人: JAMES C MULLOY
• 金额: $ 14.57万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7229966
• 关键词: AML1-ETO fusion protein; Accounting; Acute; Appendix; Biological Models; Biology; CBFbeta-MYH11 fusion protein; CBFp-MYH11; CD34 gene; Cell physiology; Cells; Cellular biology; Childhood; Chimeric Proteins; Chromosome Breakage; Chromosome abnormality; Clinical; Complementary DNA; Core-Binding Factor; DNA Sequence Rearrangement; Family; Fusion Protein Expression; Gene Expression; Gene Targeting; Genes; Hematologic Neoplasms; Hematopoiesis; Hematopoietic; Human; In Vitro; Lymphoblastic Leukemia; Malignant Neoplasms; Manuscripts; Myelogenous; Myeloid Leukemia; Oncogenes; Patients; Play; Process; Property; RUNX1 gene; Recurrence; Role; Sampling; Screening procedure; Stem cells; Subfamily lentivirinae; System; Transgenes; density; fusion gene; in vivo; insight; knock-down; leukemia; research study; self-renewal; small hairpin RNA; stem; t(8; 21)(q22; q22); transcription factor

DESCRIPTION (provided by applicant): RUNX1/CBFbeta is a key regulator of normal hematopoiesis that is frequently involved in leukemia. Rearrangements involving the CBF family account for approximately 25% of de novo myeloid leukemias and 22% of pediatric lymphoblastic leukemias. The overall objective of the proposed study is to understand gene expression that is determined by leukemia-associated translocation fusion proteins of the RUNX1/CBFbeta transcription factor, and to evaluate the role of these regulated genes in normal and leukemic stem cell self-renewal, survival and differentiation. The fusion proteins we will focus on are the t(8;21) encoding AML1-ETO, t(3;21) encoding AML1-EVI1, and the inv16/t(16;16) encoding CBFbeta- MYH11. These will be expressed in human HSPC by lentiviral transduction. Controls will include a lentivirus lacking a transgene as well as a lentivirus expressing RUNX1. Differentially expressed target genes will be validated using human HSPC expressing either AML1-ETO or CBFp-MYH11 that have acquired enhanced self-renewal properties due to the oncogene expression. Clinical samples from AML patients expressing a RUNX-fusion oncogene will also be analyzed for expression of the identified genes (Aim 1). The specific contribution of identified target genes to HSPC function will be evaluated by over-expressing genes that are repressed by the fusion proteins, and conversely by knocking-down those genes that become upregulated upon fusion protein expression, using lentiviral transduction of the human HSPC that express these oncogenes and that self-renew in vitro. Primary human HSPC will be similarly transduced to determine the contribution of the targeted gene to normal stem cell function (Aim 2). Understanding the modulation of the self-renewal process by these fusion proteins could give insight into the normal mechanisms of self-renewal employed by the stem cell.

描述（由申请人提供）：RUNX1/CBFbeta 是正常造血的关键调节因子，经常与白血病有关。涉及 CBF 家族的重排约占新发髓细胞白血病的 25% 和儿童淋巴细胞白血病的 22%。本研究的总体目标是了解由 RUNX1/CBFbeta 转录因子的白血病相关易位融合蛋白决定的基因表达，并评估这些调节基因在正常和白血病干细胞自我更新、存活和分化中的作用。我们将重点关注的融合蛋白是编码 AML1-ETO 的 t(8;21)、编码 AML1-EVI1 的 t(3;21) 和编码 CBFbeta-MYH11 的 inv16/t(16;16)。这些将通过慢病毒转导在人 HSPC 中表达。对照将包括缺乏转基因的慢病毒以及表达 RUNX1 的慢病毒。将使用表达 AML1-ETO 或 CBFp-MYH11 的人类 HSPC 来验证差异表达的靶基因，这些基因由于癌基因的表达而获得了增强的自我更新特性。还将分析来自表达 RUNX 融合癌基因的 AML 患者的临床样本，以确定已识别基因的表达（目标 1）。所鉴定的靶基因对 HSPC 功能的具体贡献将通过被融合蛋白抑制的基因过表达来评估，反之，通过使用表达这些癌基因并在体外自我更新的人 HSPC 的慢病毒转导来敲低融合蛋白表达上调的那些基因来评估。原代人 HSPC 将进行类似的转导，以确定目标基因对正常干细胞功能的贡献（目标 2）。了解这些融合蛋白对自我更新过程的调节可以帮助我们深入了解干细胞自我更新的正常机制。