Department of Molecular Hematology, Sanquin Research, Amsterdam, The Netherlands.
Department of Pediatric Hematology, Erasmus MC Sophia Children's Hospital, University Medical Center Rotterdam, The Netherlands; Unit of Transfusion Medicine, Sanquin Blood Supply, Amsterdam, The Netherlands.
J Thromb Haemost. 2023 Feb;21(2):359-372.e3. doi: 10.1016/j.jtha.2022.11.021. Epub 2022 Dec 22.
Inherited platelet disorders (IPDs) are a heterogeneous group of rare diseases that are caused by the defects in early megakaryopoiesis, proplatelet formation, and/or mature platelet function. Although genomic sequencing is increasingly used to identify genetic variants underlying IPD, this technique does not disclose resulting molecular changes that impact platelet function. Proteins are the functional units that shape platelet function; however, insights into how variants that cause IPDs impact platelet proteomes are limited.
The objective of this study was to profile the platelet proteomics signatures of IPDs.
We performed unbiased label-free quantitative mass spectrometry (MS)-based proteome profiling on platelets of 34 patients with IPDs with variants in 13 ISTH TIER1 genes that affect different stages of platelet development.
In line with the phenotypical heterogeneity between IPDs, proteomes were diverse between IPDs. We observed extensive proteomic alterations in patients with a GFI1B variant and for genetic variants in genes encoding proteins that impact cytoskeletal processes (MYH9, TUBB1, and WAS). Using the diversity between IPDs, we clustered protein dynamics, revealing disrupted protein-protein complexes. This analysis furthermore grouped proteins with similar cellular function and location, classifying mitochondrial protein constituents and identifying both known and putative novel alpha granule associated proteins.
With this study, we demonstrate a MS-based proteomics perspective to IPDs. By integrating the effects of IPDs that impact different aspects of platelet function, we dissected the biological contexts of protein alterations to gain further insights into the biology of platelet (dys)function.
遗传性血小板疾病(IPD)是一组异质性罕见疾病,由早期巨核细胞生成、前血小板形成和/或成熟血小板功能缺陷引起。尽管基因组测序越来越多地用于识别 IPD 的遗传变异,但该技术并不能揭示影响血小板功能的潜在分子变化。蛋白质是影响血小板功能的功能单位;然而,关于导致 IPD 的变异如何影响血小板蛋白质组的见解有限。
本研究旨在分析 IPD 的血小板蛋白质组特征。
我们对 34 名 IPD 患者的血小板进行了无偏标签自由定量质谱(MS)基于蛋白质组学的分析,这些患者的变异位于影响血小板发育不同阶段的 13 个 ISTH TIER1 基因中。
与 IPD 之间的表型异质性一致,IPD 的蛋白质组之间存在多样性。我们观察到具有 GFI1B 变异的患者和编码影响细胞骨架过程的蛋白质(MYH9、TUBB1 和 WAS)的基因中的遗传变异的患者存在广泛的蛋白质组改变。利用 IPD 之间的多样性,我们对蛋白质动力学进行聚类,揭示了破坏的蛋白质-蛋白质复合物。这种分析还将具有相似细胞功能和位置的蛋白质进行分类,对线粒体蛋白质成分进行分类,并确定已知和潜在的新的 alpha 颗粒相关蛋白质。
通过这项研究,我们展示了 IPD 的 MS 基于蛋白质组学的观点。通过整合影响血小板功能不同方面的 IPD 的影响,我们剖析了蛋白质改变的生物学背景,以深入了解血小板(功能障碍)的生物学。