使用 CRISPR/Cas9 靶向 RPS14 的斑马鱼 5q-综合征模型揭示了红细胞生成失败的 p53 非依赖性和 p53 依赖性机制。

A Zebrafish Model of 5q-Syndrome Using CRISPR/Cas9 Targeting RPS14 Reveals a p53-Independent and p53-Dependent Mechanism of Erythroid Failure.

机构信息

Department of Molecular, Cell and Developmental Biology, University of California Los Angeles, Los Angeles, CA 90095, USA.

Celgene Corporation, San Francisco 94158, USA.

出版信息

J Genet Genomics. 2016 May 20;43(5):307-18. doi: 10.1016/j.jgg.2016.03.007. Epub 2016 Apr 2.

DOI:10.1016/j.jgg.2016.03.007

PMID:27216296

Abstract

5q-syndrome is a distinct form of myelodysplastic syndrome (MDS) where a deletion on chromosome 5 is the underlying cause. MDS is characterized by bone marrow failures, including macrocytic anemia. Genetic mapping and studies using various models support the notion that ribosomal protein S14 (RPS14) is the candidate gene for the erythroid failure. Targeted disruption of RPS14 causes an increase in p53 activity and p53-mediated apoptosis, similar to what is observed with other ribosomal proteins. However, due to the higher risk for cancer development in patients with ribosome deficiency, targeting the p53 pathway is not a viable treatment option. To better understand the pathology of RPS14 deficiency in 5q-deletion, we generated a zebrafish model harboring a mutation in the RPS14 gene. This model mirrors the anemic phenotype seen in 5q-syndrome. Moreover, the anemia is due to a late-stage erythropoietic defect, where the erythropoietic defect is initially p53-independent and then becomes p53-dependent. Finally, we demonstrate the versatility of this model to test various pharmacological agents, such as RAP-011, L-leucine, and dexamethasone in order to identify molecules that can reverse the anemic phenotype.

摘要

5q 综合征是一种独特的骨髓增生异常综合征（MDS），其根本原因是 5 号染色体缺失。MDS 的特征是骨髓衰竭，包括巨细胞性贫血。遗传图谱和使用各种模型的研究支持核糖体蛋白 S14（RPS14）是红系衰竭候选基因的观点。RPS14 的靶向破坏会导致 p53 活性增加和 p53 介导的细胞凋亡，类似于其他核糖体蛋白观察到的情况。然而，由于核糖体缺陷患者癌症发展的风险较高，靶向 p53 途径不是可行的治疗选择。为了更好地了解 5q 缺失中 RPS14 缺乏的病理学，我们生成了一个携带 RPS14 基因突变的斑马鱼模型。该模型反映了 5q 综合征中观察到的贫血表型。此外，贫血是由于晚期红系发育缺陷，其中红系发育缺陷最初与 p53 无关，然后变得依赖于 p53。最后，我们证明了该模型的多功能性，可用于测试各种药理制剂，如 RAP-011、L-亮氨酸和地塞米松，以鉴定可逆转贫血表型的分子。