Clonal Evolution and Prospective Isolation of Myelodysplastic Syndrome Initiating

骨髓增生异常综合征起始的克隆进化和前瞻性分离

• 批准号: 8649708
• 负责人: James Y. Chen
• 金额: $ 3.58万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-15 至 2017-01-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8649708
• 关键词: Acute Myelocytic Leukemia; Address; Aging; Allogeneic Bone Marrow Transplantation; Autologous; Autologous Stem Cell Transplantation; Azacitidine; Biological Assay; Blood; Blood Platelets; Blood Transfusion; Bone Marrow Transplantation; Cataloging; Catalogs; Cells; Chronic; Clinical; Clonal Evolution; Decitabine; Development; Disease; Dysmyelopoietic Syndromes; Effectiveness; Event; Fluorescence-Activated Cell Sorting; Future; Goals; Hematopoiesis; Hematopoietic stem cells; Immunodeficient Mouse; Incidence; Individual; Ineffective Hematopoiesis; Intervention; Knowledge; Lead; Methods; Modeling; Monoclonal Antibodies; Mouse Strains; Mus; Mutation; Neutropenia; Normal Cell; Pathogenesis; Patients; Platelet Transfusion; Prevalence; Regimen; Residual state; Resolution; Risk; SNP genotyping; Sampling; Secondary Myelodysplastic Syndrome; Secondary to; Series; Source; Stem cells; Testing; Therapeutic Intervention; Transplantation; conditioning; cytopenia; exome sequencing; improved; insight; myeloblast; novel; novel therapeutics; older patient; progenitor; prospective; public health relevance; response; stem cell therapy; therapeutic target; treatment strategy

DESCRIPTION (provided by applicant): Myelodysplastic syndrome (MDS) is a heterogeneous group of related clonal diseases with increasing incidence and prevalence, characterized by variable cytopenias due to ineffective hematopoiesis. A subset of MDS patients may also progress to secondary acute myeloid leukemia. There is considerable hope, however, that through a better understanding of MDS pathogenesis, novel therapeutics can alleviate or even correct these deficiencies. Beyond hypomethylating agents, which have variable response rates and duration of effect, no such therapy has been approved. We have previously shown that MDS is a disease of the hematopoietic stem cell (HSC). Aim 1 of this proposal describes a method to use fluorescence-activated cell sorting to deplete CD69hi and CD99hi MDS HSCs from the residual CD69lo CD99- normal HSCs to facilitate autologous stem cell transplantation as therapy. Effectiveness of separation will be functionally evaluated via transplantation into immunodeficient mouse strains, an assay we have previously shown to distinguish between normal cells and MDS cells. Combined with a promising novel non-toxic conditioning regime being developed clinically at Stanford (separately from this proposal), autologous stem cell transplantation may serve as a novel potential treatment for MDS. Aim 2 is to detect mutations in MDS at the single cell level. Through whole exome sequencing of MDS myeloblasts and progenitors, in conjunction with TaqMan SNP genotyping of individual HSC clones, we will address three fundamental questions to better characterize MDS for potential future therapeutic interventions: (1) in what order do these mutations occur? (2) do all the MDS mutations occur in the HSC pool? and (3) which mutations correlate with the conversion of a normal HSC to an MDS HSC and then to an LSC? The ordered series of mutations will provide insight into the mechanisms of disease, especially in understanding whether a stereotypical sequences of events occurs in pathogenesis and progression. Furthermore, we expect that identifying the specific mutations that occur at the transition between normal and MDS hematopoiesis will reveal critical targets for potential therapeutic interventions. Both of the aims capitalize on our knowledge of hematopoietic stem cells. These studies have the potential to offer a new application of bone marrow transplantation in the treatment of MDS and to characterize the disease mechanism to facilitate discovery of future therapies.

描述（由申请人提供）：骨髓增生综合征（MDS）是一组相关的克隆疾病的异质组，其发病率和患病率越来越高，其特征是由于无效的造血作用而导致可变的细胞质减少症。 MDS患者的一部分也可能发展为继发性急性髓样白血病。然而，有很大的希望，通过对MDS发病机理的更好理解，新型治疗剂可以减轻甚至纠正这些缺陷。除了具有可变的反应率和效果持续时间的降压剂外，尚未批准这种疗法。我们先前已经表明，MDS是造血干细胞（HSC）的疾病。该提案的目标1描述了一种使用荧光激活的细胞排序以耗尽来自残留CD69lo CD99-正常HSC的CD69HI和CD99HI MDS HSC的方法，以促进自体干细胞移植作为治疗。分离的有效性将通过移植到免疫缺陷的小鼠菌株中进行功能评估，这是我们以前已证明的测定法可以区分正常细胞和MDS细胞。结合在斯坦福大学临床上开发的有前途的新型无毒调理状态（与该提案分开），自体干细胞移植可作为MDS的新型潜在治疗方法。 AIM 2是在单细胞水平上检测MDS中的突变。通过对MD的整个外显子组测序，与Taqman SNP基因分型结合了单个HSC克隆的taqman SNP基因分型，我们将解决三个基本问题，以更好地表征MDS以进行潜在的未来治疗干预措施：（1）以什么顺序发生这些突变？ （2）所有MDS突变是否发生在HSC池中？ （3）哪些突变与正常HSC转换为MDS HSC，然后转换为LSC？一系列有序的突变将提供对疾病机制的见解，尤其是在理解事件的刻板印象序列中是否发生在发病机理和进展中。此外，我们预计确定正常造血和MDS造血之间发生的特定突变将揭示潜在的治疗干预措施的关键靶标。两个目标都利用了我们的 造血干细胞的知识。这些研究有可能在MDS治疗中提供新的骨髓移植的应用，并表征疾病机制以促进未来疗法的发现。