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Role of inflammation in initiation and progression of clonal hematopoiesis

炎症在克隆造血的起始和进展中的作用

基本信息

批准号：
18F18408
负责人：
滝澤仁
金额：
$ 1.41万
依托单位：
Kumamoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for JSPS Fellows
财政年份：
2018
资助国家：
日本
起止时间：
2018-10-12 至 2021-03-31
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-18F18408/
关键词：
Clonal Hematopoiesis Lineage Tracing Leukemia Inflammation clonal hematopoiesis inflammation

项目摘要

Many labs around the world has been working to develop novel methods for lineage tracing employing different mechanisms. But, these methods cannot provide the information on clone size of the dys-regulated and expanded clones, which led us to design this study to develop a method to determine both clonality and clone size simultaneously.I designed, developed and evaluated the novel clonal tracing method in cell line transfected with sleeping beauty transposon vectors and analysis of data revealed that the method can detect around 500 clones (both expanded and non-expanded) with their respective clone size. Distribution of integration site analysis showed random and stochastic distribution of transposon integration in all the chromosomes. I also evaluated the method with cell number and with as low as 10000 cells, the method work well and can detect enough clones with their respective clone size. To apply this method in vivo, novel strains of mice were needed. So, I also generated two novel strains of mice- Rosa26-Hyper sleeping beauty (HSB), which contains transposase enzyme and polyIR mice, which contains multiple copy of IR transposon in silent locus of chromosome.Upon successful evaluation of the method developed in this study in mouse model, it will provide useful tools not only in the field of hematology but also in other biomedical research fields to trace lineage to single cell resolution. The knowledge obtained from this study is expected to provide contribution towards establishment of preventive methods for pre-leukemia initiation and progression.

世界各地的许多实验室一直在努力开发使用不同机制进行血统追踪的新方法。但是，这些方法不能提供关于差异调节克隆和扩增克隆的克隆大小的信息，这使得我们设计了一种同时确定克隆和克隆大小的方法。我设计、开发并评估了一种新的克隆示踪方法，该方法在转染睡美人转座子载体的细胞系中进行了检测，数据分析表明，该方法可以检测到大约500个克隆(包括扩增的和非扩增的)。整合位点的分布分析表明，转座子整合在所有染色体上都是随机分布的。我还用细胞数对该方法进行了评估，在低至10000个细胞的情况下，该方法工作良好，可以检测到足够多的克隆和各自的克隆大小。为了在体内应用这种方法，需要新的小鼠品系。因此，我还培育了两个新品系的小鼠--含有转座酶的ROSA26-超级睡美人(ROSA26-Hyper Slear Beauty，HSB)和含有多拷贝IR转座子的PolyIR小鼠。在小鼠模型中成功评估本研究的方法后，不仅将为血液学领域提供有用的工具，也将为其他生物医学研究领域提供有用的工具，以追溯谱系到单细胞分辨率。从这项研究中获得的知识有望为白血病前期发病和进展的预防方法的建立做出贡献。