INVOLVEMENT OF THE EVI-2 LOCUS IN MYELOID LEUKEMIA

嗜亲性病毒整合位点2（Evi-2）被鉴定为常见的 来源于BXH-2小鼠的骨髓肿瘤中的病毒整合位点。 Evi-2基因座的表征显示， 发生在编码跨膜蛋白的2个新基因（Evi-2a和 Evi-2b）。 Evi-2定位于小鼠11号染色体和人11号染色体， 染色体17q11.2。 通过使用Evi-2， 克隆并显示Evi-2a、Evi-2b和第三个基因（Omgp） 位于一个内含子内 有趣的是， 患有NF 1的患者发生青少年期的频率更高， 骨髓性白血病，这表明，无论是什么基因参与了 小鼠骨髓性白血病，可能是与人类骨髓性白血病相关的相同基因。 白血病 因此，通过使用病毒整合的共同位点， 鉴定了一个重要的基因组区域， 人类疾病 我们的长期目标是描述在这些细胞中发现的新基因。 Evi-2基因座，并确定其在正常发育中的作用， 白血病 初步分析表明这不是 NF 1基因的表达，但基因（或基因）的改变 导致骨髓性白血病的NF 1内含子中 BXH-2小鼠 将被老化以分离额外的骨髓肿瘤和整合在 将鉴定Evi-2基因座。 这些肿瘤将在 RNA和蛋白质水平检测表达的改变， Evi-2基因座内的基因 此外，我们建议 表征Evi-2a和Evi-2b基因及其产物，以确定 它们在正常发育和骨髓性白血病发生中的作用。 充分 将对两种基因的长度mRNA进行测序， 剪接模式和外显子的描绘。 这些蛋白质将 在表达载体中产生并用于产生多克隆抗血清 来研究蛋白质的细胞定位。 调查 蛋白质的功能将随着鉴定而沿着启动 相关蛋白质。 此外，发展概况的 mRNA和蛋白质的表达，通过原位RNA杂交 免疫组化染色，将确定，以获得 深入了解这些基因在正常小鼠发育中的作用。 的 这些基因产物过表达和表达缺失的影响 以及过表达在细胞中的作用。 转基因动物 这些研究将阐明 Evi-2基因座及其在髓系肿瘤中作用

Ecotropic viral integration site-2 (Evi-2) was identified as a common site of viral integration in myeloid tumors derived from BXH-2 mice. Characterization of the Evi-2 locus revealed that the viral integrations occurred within 2 novel genes encoding transmembrane proteins (Evi-2a and Evi-2b). Evi-2 was localized to mouse chromosome 11 and to human chromosome 17q11.2. Through the use of Evi-2, the gene for NF1 was cloned and it was revealed that Evi-2a, Evi-2b and a third gene (Omgp) were localized within an intron. Interestingly, it has been observed that patients with NF1 have a higher frequency of developing juvenile myelogenous leukemia, suggesting that whatever gene is involved in the murine myeloid leukemia, could be the same gene involved in human myeloid leukemia. Thus, through the use of a common site of viral integration an important genomic region was identified that is involved in murine and human disease. Our long term goal is to characterize the novel genes found within the Evi-2 locus and to determine their role in normal development and leukemia. Preliminary analysis indicates that it is not the alteration of expression of the NF1 gene, but the alteration of the gene (or genes) in the NF1 intron that contributes to the myeloid leukemia. BXH-2 mice will be aged to isolate additional myeloid tumors and integrations within the Evi-2 locus will be identified. These tumors will be analyzed at both the RNA and protein level to detect alterations in the expression of the genes within the Evi-2 locus. Additionally, we propose to characterize the Evi-2a and Evi-2b genes and their products to determine their role in normal development and myeloid leukemogenesis. The full length mRNA of both genes will be sequenced with the characterization of splicing patterns and delineation of exons. The proteins will be produced in expression vectors and used to generate polyclonal antisera to study the cellular localization of the proteins. Investigation of the function of the proteins will be initiated along with the identification of associated proteins. Additionally, a developmental profile of the expression of both mRNA and protein, through in situ RNA hybridization and immunohistochemical staining, will be determined in order to gain insights into the role these genes play in normal mouse development. The effect of overexpression and loss of expression of these gene products in vitro will be assessed as well as the effect of overexpression in transgenic animals. These studies will elucidate the normal function of the Evi-2 locus and its influence in myeloid neoplasia.