美国威尔康奈尔医学中心Dan A. Landau和加拿大大学健康网络Federico Gaiti课题组合作，利用单细胞多组学揭示了剪接畸变对人造血克隆细胞类型的特异性影响。相关论文发表在2023年8月14日出版的《细胞-干细胞》杂志上。

为了克服技术限制，研究人员将转录组基因分型（GoT）与长读长单细胞转录组学和蛋白质基因组学相结合，实现对转录组、表面蛋白、体细胞突变和RNA剪接（GoT-剪接）进行单细胞分析。研究人员将GoT-Splice应用于骨髓增生异常综合征（MDS）患者的造血祖细胞，这些患者的核心剪接因子SF3B1发生突变。SF3B1^mut细胞在巨核细胞-红系中富集，并且SF3B1^mut红系祖细胞会发生扩增。

研究发现不同祖细胞群体会使用不同的3'剪接位点以及红系分化过程中会发生阶段特异性异常剪接。对SF3B1突变克隆造血样本的分析显示，SF3B1^mut细胞中的红系偏倚和细胞类型特异性3′剪接位点的使用先于MDS发生。总的来说，GoT-Splice揭示了细胞类型特异性体细胞突变对RNA剪接的影响，并直接在人类样本中阐释了从早期克隆生长到明显肿瘤形成的过程。

据悉，RNA剪接因子在克隆血液疾病中发生反复突变，但尚不清楚剪接失调对造血的影响。

附：英文原文

Title: Single-cell multi-omics defines the cell-type-specific impact of splicing aberrations in human hematopoietic clonal outgrowths

Author: Mariela Cortés-López, Paulina Chamely, Allegra G. Hawkins, Robert F. Stanley, Ariel D. Swett, Saravanan Ganesan, Tarek H. Mouhieddine, Xiaoguang Dai, Lloyd Kluegel, Celine Chen, Kiran Batta, Nili Furer, Rahul S. Vedula, John Beaulaurier, Alexander W. Drong, Scott Hickey, Neville Dusaj, Gavriel Mullokandov, Adam M. Stasiw, Jiayu Su, Ronan Chaligné, Sissel Juul, Eoghan Harrington, David A. Knowles, Catherine J. Potenski, Daniel H. Wiseman, Amos Tanay, Liran Shlush, Robert C. Lindsley, Irene M. Ghobrial, Justin Taylor, Omar Abdel-Wahab, Federico Gaiti, Dan A. Landau

Issue&Volume: 2023-08-14

Abstract: RNA splicing factors are recurrently mutated in clonal blood disorders, but the impactof dysregulated splicing in hematopoiesis remains unclear. To overcome technical limitations,we integrated genotyping of transcriptomes (GoT) with long-read single-cell transcriptomicsand proteogenomics for single-cell profiling of transcriptomes, surface proteins,somatic mutations, and RNA splicing (GoT-Splice). We applied GoT-Splice to hematopoieticprogenitors from myelodysplastic syndrome (MDS) patients with mutations in the coresplicing factor SF3B1. SF3B1mut cells were enriched in the megakaryocytic-erythroid lineage, with expansion of SF3B1mut erythroid progenitor cells. We uncovered distinct cryptic 3′ splice site usage indifferent progenitor populations and stage-specific aberrant splicing during erythroiddifferentiation. Profiling SF3B1-mutated clonal hematopoiesis samples revealed that erythroid bias and cell-type-specificcryptic 3′ splice site usage in SF3B1mut cells precede overt MDS. Collectively, GoT-Splice defines the cell-type-specificimpact of somatic mutations on RNA splicing, from early clonal outgrowths to overtneoplasia, directly in human samples.

DOI: 10.1016/j.stem.2023.07.012

Source: https://www.cell.com/cell-stem-cell/fulltext/S1934-5909(23)00257-6