Transcriptome profiling of c-Kit+ hematopoietic stem and progenitor cells enriched from 10-week-old wild-type and Rps19R67?/R67? mutant mice.

This RNA sequencing experiment is part of the study "Preclinical animal model of Diamond-Blackfan anemia with single amino acid mutation of ribosomal protein Rps19". A mouse model with arginine 67 mutation of ribosomal protein Rps19 develops features characteristic of human Diamond-Blackfan anemia, a rare bone marrow failure syndrome. These include hematologic dysfunctions, early onset growth delay, intrinsic anemia, severe craniofacial, skeletal, urogenital, cardiovascular, and cerebral abnormalities leading to premature lethality during the adolescence of the mouse. This model exhibits cell intrinsic activation of the Trp53 signaling pathway in hematopoietic stem cells (HSCs) leading to reduced erythroid lineage development that may be rescued after inactivation of the tumor suppressor Trp53. Using preliminary RNA sequencing study we identify a set of non-canonical components of the p53 signaling pathway which with high likelihood mediate the wide range of pathologies associated with DBA, the experiment if followed up by single cell transcriptome analysis of bone marrow hematopoietic progenitors and RNA sequencing of E14.5 fetal liver from wild-type control and Rps19R67?/R67?, Rps19R67?/R67? Trp53-/- and Trp53-/- mutant embryos.

本RNA测序实验隶属于研究《携带核糖体蛋白Rps19单氨基酸突变的戴蒙德布莱克凡贫血（Diamond-Blackfan anemia, DBA）临床前动物模型》。本研究构建的核糖体蛋白Rps19第67位精氨酸突变小鼠模型，可重现人类罕见骨髓衰竭综合征——戴蒙德布莱克凡贫血的典型病理特征，具体表现为血液系统功能异常、早发性生长迟缓、先天性贫血，以及严重的颅面部、骨骼、泌尿生殖系统、心血管系统与中枢神经系统异常，最终导致小鼠青春期提前死亡。该模型的造血干细胞（Hematopoietic Stem Cells, HSCs）中存在细胞自主性的Trp53信号通路激活，可引发红细胞系发育受阻，而抑癌基因Trp53失活后可逆转该表型。本研究通过前期RNA测序实验，鉴定出一组非经典的p53信号通路组分，它们极有可能介导了DBA相关的多种病理损伤。本后续实验将对野生型对照、Rps19R67?/R67?、Rps19R67?/R67? Trp53-/-以及Trp53-/-突变胚胎的骨髓造血祖细胞开展单细胞转录组分析，并对其E14.5胎肝进行RNA测序。