Kamble Prachi, Saptarshi Arati, Mudaliar Sangeeta, Kanvinde Purva, Kedar Prabhakar S
Department of Haematogenetics, ICMR-National Institute of Immunohematology, 13th Floor, NMS Building, King Edward Memorial (K.E.M.) Hospital Campus, Parel, Mumbai, 400012, India.
Department of Haematology-Oncology, B J Wadia Hospital for Children, Parel, Mumbai, 400012, Maharashtra, India.
Mol Genet Genomics. 2025 Sep 17;300(1):93. doi: 10.1007/s00438-025-02296-w.
Diamond-Blackfan anaemia (DBA) is a rare inherited disorder marked by early-onset macrocytic anaemia and erythroid hypoplasia, resulting from mutations in ribosomal protein genes. Despite growing genetic insights, data on functional validation remain limited in India; here we report a novel RPS10 mutation with functional validation and provide genotype-phenotype correlation by integrating our findings with all previously reported RPS10 variants. A clinically suspected Diamond-Blackfan anaemia (DBA) case was evaluated through haematological profiling, bone marrow examination, and erythrocyte adenosine deaminase (eADA) activity measurement. Whole exome sequencing (WES) was followed by Sanger sequencing to identify and validate a novel pathogenic variant. Gene expression of ribosomal and regulatory genes was analysed by quantitative RT-PCR, and rRNA processing analysis was carried out to assess functional impact. The proband presented with severe macrocytic anaemia, reticulocytopenia, and erythroid hypoplasia consistent with Diamond-Blackfan anaemia (DBA). Whole exome sequencing identified a novel heterozygous nonsense variant in RPS10 (c.206G > A; p.Trp69Ter), and Sanger sequencing confirmed the variant as de novo. Gene expression analysis revealed significant upregulation of TP53 and downregulation of RPS10 and GATA1, indicating ribosomal dysfunction and activation of the p53 pathway. Additionally, the rRNA processing defect validated the pathogenicity of the novel RPS10 variant. This study identifies a novel de novo nonsense variant in RPS10 associated with Diamond-Blackfan anaemia, with supporting functional evidence of haploinsufficiency and p53 pathway activation. These findings expand the mutational spectrum of RPS10 and underscore the diagnostic value of integrating genomic and functional analyses in rare haematological disorders, while also contributing to ongoing efforts to delineate genotype-phenotype correlations in DBA.
钻石黑范贫血（DBA）是一种罕见的遗传性疾病，其特征为早发性大细胞贫血和红系造血功能低下，由核糖体蛋白基因突变引起。尽管对其遗传学认识不断增加，但在印度，功能验证的数据仍然有限；在此，我们报告了一个经过功能验证的新型RPS10突变，并通过将我们的研究结果与所有先前报道的RPS10变体相结合，提供了基因型-表型相关性。通过血液学分析、骨髓检查和红细胞腺苷脱氨酶（eADA）活性测量，对一例临床疑似钻石黑范贫血（DBA）病例进行了评估。随后进行全外显子组测序（WES），接着进行桑格测序以鉴定和验证一个新型致病变体。通过定量逆转录聚合酶链反应（RT-PCR）分析核糖体和调节基因的表达，并进行rRNA加工分析以评估功能影响。先证者表现为严重的大细胞贫血、网织红细胞减少和红系造血功能低下，符合钻石黑范贫血（DBA）。全外显子组测序在RPS10中鉴定出一个新型杂合无义变体（c.206G＞A；p.Trp69Ter），桑格测序证实该变体为新发突变。基因表达分析显示TP53显著上调，RPS10和GATA1下调，表明核糖体功能障碍和p53途径激活。此外，rRNA加工缺陷验证了新型RPS10变体的致病性。本研究鉴定出一个与钻石黑范贫血相关的新型RPS10新发无义变体，有单倍体不足和p53途径激活的功能证据支持。这些发现扩展了RPS10的突变谱，强调了在罕见血液疾病中整合基因组和功能分析的诊断价值，同时也有助于正在进行的确定DBA基因型-表型相关性的工作。
