Genetic Basis of Polycythemia Vera

真性红细胞增多症(PV)是最常见的骨髓增生性疾病。PV被认为是由单个造血干细胞的躯体变化引起的，但PV的诊断和治疗都是 有争议的是它的分子缺陷或缺陷还没有被表征。 我们认为JAK2V617F不是导致PV克隆增殖的原因，而是先于其他体细胞和胚系突变(S)。 因此，我们假设，除了JAK2 V617F突变外，完整的PV表型的发展还需要额外的遗传事件；鉴定这些事件是本应用的主要目标。这些研究将需要结合和整合来自几种基因组学方法的数据。 基于这些考虑，我们计划追求三个具体目标来实现我们的目标： SA 1.通过整合这些互补方法鉴定引起克隆性造血的JAK2前体细胞突变： Sa 1a.通过家族性共分离基因组区域和PV表型来确定位置候选。 散发性PV患者个体的克隆细胞和多克隆细胞的比较。 Sa 1c.使用三重家族方法鉴定基因组结构的共有区域以确定易患遗传单倍型。 SA 1D。候选基因组区域将通过全外显子基因组测序和全基因组测序进行详细评估，因为这项技术正在迅速发展，其成本正在变得负担得起。我们大学正在获取太平洋生物科学平台，该平台能够通过该项目的联合研究员Jorde博士带头的努力，每次运行分析10kb的序列。 Sa 2.寻找非常规遗传损伤，即miRNA。在与Croce博士的合作中，我们将重点关注6号染色体区域，作为PV发生的潜在生殖系或获得性贡献者。 SA 3.确定聚乙二醇化干扰素a治疗的PV患者的序贯基因组变化 降低JAK2 V617F等位基因负荷，恢复多克隆造血。

Polycythemia Vera ( PV) is the most common myeloproliterative disorder. PV is believed to arise from a somatic change of a single hematopoietic stem cell, but both the diagnosis and therapy of PV are controversial as its molecular defect or defects have not been characterized. We conclude that JAK2 V617F is not the cause of clonal proliferation of PV but is preceded by other somatic and germ line mutation(s). Thus we hypothesize that besides the JAK2 V617F mutation, additional genetic events are needed for the development of the full PV phenotype; identification of these is the principal goal of this application. These studies will require the combination and integration of data from several genomics approaches. Based on these considerations we plan to pursue three Specific Aims to accomplish our goal: SA 1. Identification of pre-JAK2 somatic mutations causing clonal hematopoiesis by integration of these complementary approaches: SA 1a. Identification of positional candidates through familial co-segregation of genomic regions with the PV phenotype. SA 1 b. Comparison of clonal and polyclonal cells from individual patients with sporadic PV. SA 1c. Identification of shared regions of genome architecture using the Trio family approach for determination of a predisposing inherited haplotype. SA 1d. The candidate genomic regions will be evaluated in detail by whole exonic genome sequencing and entire whole genome sequencing, as this technology is rapidly advancing and its costs are becoming affordable. Our University is acquiring Pacific biosciences platform capable of analyzing 10kb sequence per run by effort spearheaded by this project co-investigator Dr. Jorde. SA 2. Search for nonconventional genetic lesions; i.e. miRNA. In collaboration with Dr. Croce, we will focus on the region of chromosome 6 as a potential germ-line or acquired contributor to the genesis of PV. SA 3. Determine the sequential genomic changes in PV treated by pegylated interferon a in those patients with decreasing JAK2 V617F allelic burden and return of polyclonal hematopoiesis.