Modelling erythropoiesis in congenital dyserythropoietic anaemia type I (CDA-I) for diagnosis and molecular dissection.

Congenital Dyserythropoietic Anaemia type 1 (CDA-I) is an inherited anaemia arising primarily from mutations in C15orf41 and CDAN1, however the molecular mechanisms that cause the disease remain to be fully elucidated. We use an in vitro culture system to study multiple stages of erythropoiesis from CD34+ progenitors of patients with CDA-I. Applying a number of techniques, including ATAC-seq, we show that differentiation of CDA-I patient erythroblasts closely matches that of healthy donors during the early and intermediate stages of erythroid differentiation and maturation. However, a defect in terminal erythropoiesis can be observed in the CDA-I patient derived erythroblasts, resulting in a reduction in the number of enucleated erythroid cells. Overall design: 32 ATAC-seq samples of cultured erythroblasts from Cogenital DyserythropoieticAnemia Type 1 (CDA-I) CD34 progenitors from 7 Donors, with up to 2 technical replicates for each timepoint during culture (Day7, Day10, Day11, Day13)

1型先天性红细胞生成异常性贫血（Congenital Dyserythropoietic Anaemia type 1, CDA-I）是一类遗传性贫血，其主要致病诱因是C15orf41与CDAN1基因发生突变，但目前该疾病的完整分子致病机制仍有待完全阐明。本研究采用体外培养体系，对CDA-I患者CD34+造血祖细胞（CD34+ progenitors）的红细胞生成多个阶段进行研究。通过运用包括ATAC-seq（转座酶可及性测序）在内的多种实验技术，我们发现：在红系分化与成熟的早期及中期阶段，CDA-I患者来源的成红细胞分化模式与健康供体高度相似；然而，CDA-I患者来源的成红细胞在终末红细胞生成阶段存在明显缺陷，最终导致去核红系细胞数量减少。

实验整体设计：本研究共包含32例ATAC-seq测序样本，均取自7名1型先天性红细胞生成异常性贫血（CDA-I）患者的CD34+造血祖细胞体外培养的成红细胞；在细胞培养过程中分别于第7天、第10天、第11天、第13天四个时间点取样，每个时间点最多设置2次技术重复。