Abstract：
Neurofibromatosis type 1 associated plexiform neurofibroma (pNF) is characterized by abundant fibroblasts and dense collagen, yet the intricate interactions between tumor-origin cells (Schwann cells) and neurofibroma-associated fibroblasts (NFAFs) remain elusive. Employing single-cell RNA sequencing on human pNF samples, we generated a comprehensive transcriptomics dataset and conducted cell-cell communication analysis to unravel the molecular dynamics between Schwann cells and NFAFs. Our focus centered on the pleiotrophin (PTN)/nucleolin (NCL) axis as a pivotal ligand-receptor pair orchestrating this interaction. Validation of PTN involvement was affirmed through coculture models and recombinant protein experiments. Functional and mechanistic investigations, employing assays such as CCK8, EdU, Western Blot, ELISA, Hydroxyproline Assay, and Human phospho-kinase array, provided critical insights. We employed siRNA or inhibitors to intercept the PTN/NCL/proline-rich Akt substrate of 40 kDa (PRAS40) axis, validating the associated molecular mechanism. Our analysis highlighted a subset of Schwann cells closely linked to collagen deposition, underscoring their significance in pNF development. The PTN/NCL axis emerged as a key mediator of the Schwann cell-NFAF interaction. Furthermore, our study demonstrated that elevated PTN levels enhanced NFAF proliferation and collagen synthesis, either independently or synergistically with TGF-β1 in vitro. Activation of the downstream molecule PRAS40 was noted in NFAFs upon PTN treatment. Crucially, by targeting NCL and PRAS40, we successfully reversed collagen synthesis within NFAFs. In conclusion, our findings unveil the pivotal role of the PTN/NCL/PRAS40 axis in driving pNF development by promoting NFAFs proliferation and function. Targeting this pathway emerges as a potential therapeutic strategy for pNF. This study contributes novel insights into the molecular mechanisms governing pNF pathogenesis.
摘要：
神经纤维瘤病1型相关丛状神经纤维瘤（pNF）的特点是丰富的成纤维细胞和致密的胶原蛋白，然而,肿瘤起源细胞(施万细胞)和神经纤维瘤相关成纤维细胞(NFAFs)之间复杂的相互作用仍然难以捉摸.对人类pNF样本进行单细胞RNA测序，我们生成了一个全面的转录组学数据集，并进行了细胞间通讯分析，以解开雪旺氏细胞和NFAFs之间的分子动力学。我们的重点集中在多效蛋白（PTN）/核仁素（NCL）轴，作为协调这种相互作用的关键配体-受体对。通过共培养模型和重组蛋白实验确认了PTN参与的验证。功能和机械调查，采用CCK8、EdU、西方印迹，ELISA,羟脯氨酸测定,和人类磷酸激酶阵列，提供了重要的见解。我们使用siRNA或抑制剂来拦截40kDa(PRAS40)轴的富PTN/NCL/脯氨酸Akt底物,验证相关的分子机制。我们的分析强调了一个与胶原蛋白沉积密切相关的雪旺氏细胞亚群，强调它们在pNF发展中的重要性。PTN/NCL轴作为施万细胞-NFAF相互作用的关键介质出现。此外，我们的研究表明，升高的PTN水平增强NFAF增殖和胶原蛋白合成，在体外独立或与TGF-β1协同作用。在PTN处理后,在NFAF中注意到下游分子PRAS40的活化。至关重要的是，通过靶向NCL和PRAS40,我们成功逆转了NFAFs内胶原蛋白的合成.总之,我们的发现揭示了PTN/NCL/PRAS40轴通过促进NFAFs增殖和功能在驱动pNF发育中的关键作用.靶向该途径成为pNF的潜在治疗策略。这项研究为控制pNF发病机理的分子机制提供了新的见解。
