Jung Seung-Hyun, Lee Sung-Eun, Yun Sujin, Min Da-Eun, Shin Youngjin, Chung Yeun-Jun, Lee Sug Hyung
Departments of Biochemistry; Departments of Precision Medicine Research Center/Integrated Research Center for Genome Polymorphism; Departments of Medical Sciences.
Departments of Internal Medicine.
Haematologica. 2025 Apr 1;110(4):938-951. doi: 10.3324/haematol.2024.285598. Epub 2024 Oct 10.
Primary myelofibrosis (PMF) is a myeloid proliferative neoplasm (MPN) characterized by bone marrow fibrosis. Pre-fibrotic PMF (pre-PMF) progresses to overt PMF. Megakaryocytes play a primary role in PMF; however, the functions of megakaryocyte subsets and those of other hematopoietic cells during PMF progression remain unclear. We, therefore, analyzed bone marrow aspirates in cases of pre-PMF, overt PMF, and other MPN using single-cell RNA sequencing. We identified 14 cell types with subsets, including hematopoietic stem and progenitor cells (HSPC) and megakaryocytes. HSPC in overt PMF were megakaryocyte-biased and inflammation/fibrosis-enriched. Among megakaryocytes, the epithelial-mesenchymal transition (EMT)-enriched subset was abruptly increased in overt PMF. Megakaryocytes in non-fibrotic/non-PMF MPN were megakaryocyte differentiation-enriched, whereas those in fibrotic/non-PMF MPN were inflammation/fibrosis-enriched. Overall, the inflammation/fibrosis signatures of the HSPC, megakaryocyte, and CD14+ monocyte subsets increased from pre-PMF to overt PMF. Cytotoxic and dysfunctional scores also increased in T and NK cells. Clinically, megakaryocyte and HSPC subsets with high inflammation/fibrosis signatures were frequent in the patients with peripheral blood blasts ≥1%. Single-cell RNA-sequencing predicted higher cellular communication of megakaryocyte differentiation, inflammation/fibrosis, immunological effector/dysfunction, and tumor-associated signaling in overt PMF than in pre-PMF. However, no decisive subset emerged during PMF progression. Our study demonstrated that HSPC, monocytes, and lymphoid cells contribute to the progression of PMF, and subset specificity existed regarding inflammation/fibrosis and immunological dysfunction. PMF progression may depend on alterations of multiple cell types, and EMT-enriched megakaryocytes may be potential targets for diagnosing and treating the progression.
原发性骨髓纤维化（PMF）是一种以骨髓纤维化为特征的髓系增殖性肿瘤（MPN）。纤维化前期PMF（pre-PMF）会进展为明显的PMF。巨核细胞在PMF中起主要作用；然而，在PMF进展过程中巨核细胞亚群以及其他造血细胞的功能仍不清楚。因此，我们使用单细胞RNA测序分析了pre-PMF、明显PMF及其他MPN病例的骨髓穿刺液。我们鉴定出了14种带有亚群的细胞类型，包括造血干细胞和祖细胞（HSPC）以及巨核细胞。明显PMF中的HSPC偏向巨核细胞且富含炎症/纤维化相关成分。在巨核细胞中，富含上皮-间质转化（EMT）的亚群在明显PMF中突然增加。非纤维化/非PMF的MPN中的巨核细胞富含巨核细胞分化相关成分，而纤维化/非PMF的MPN中的巨核细胞富含炎症/纤维化相关成分。总体而言，从pre-PMF到明显PMF，HSPC、巨核细胞和CD14+单核细胞亚群的炎症/纤维化特征增加。T细胞和NK细胞的细胞毒性和功能障碍评分也增加。临床上，外周血原始细胞≥1%的患者中，具有高炎症/纤维化特征的巨核细胞和HSPC亚群很常见。单细胞RNA测序预测，与pre-PMF相比，明显PMF中巨核细胞分化、炎症/纤维化、免疫效应/功能障碍以及肿瘤相关信号的细胞间通讯更高。然而，在PMF进展过程中没有出现决定性的亚群。我们的研究表明，HSPC、单核细胞和淋巴细胞促成了PMF的进展，并且在炎症/纤维化和免疫功能障碍方面存在亚群特异性。PMF的进展可能取决于多种细胞类型的改变，富含EMT的巨核细胞可能是诊断和治疗进展的潜在靶点。
