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体细胞变化引起的，但PV的诊断和治疗都非常困难。 由于其分子缺陷或缺陷尚未被表征，因此存在争议。 我们得出结论，JAK2 V617F 不是 PV 克隆增殖的原因，而是先于其他体细胞和种系突变。 因此，我们假设除了 JAK2 V617F 突变外，完整 PV 表型的发展还需要其他遗传事件；识别这些是本应用程序的主要目标。这些研究需要组合和整合来自多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来鉴定导致克隆造血的前 JAK2 体细胞突变： SA 1a。通过基因组区域与 PV 表型的家族共分离来识别候选位置。 SA 1 b．散发性真性红细胞增多症个体患者的克隆细胞和多克隆细胞的比较。 SA 1c。使用 Trio 家族方法鉴定基因组结构的共享区域，以确定易遗传的单倍型。 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，其成本也变得可以承受。在该项目联合研究员 Jorde 博士的带领下，我们大学正在收购能够每次运行分析 10kb 序列的 Pacific 生物科学平台。 SA 2.寻找非常规遗传病变；即 miRNA。我们将与 Croce 博士合作，重点研究 6 号染色体区域作为 PV 发生的潜在种系或后天贡献者。 SA 3. 确定这些患者接受聚乙二醇化干扰素 a 治疗后 PV 的连续基因组变化 随着 JAK2 V617F 等位基因负担的减少和多克隆造血的恢复。

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.