MDS-MSC外泌体通过RN7SL1调控RIG-I信号通路介导克隆细胞恶性增殖的机制研究

Interactions of mesenchymal stem cells (MSC) with MDS cloned cells promote the onset of disease. How do exosomes participate in disease progression as one of the mechanisms of their interactions is unclear. In previous studies, we found that exosomes from MDS-derived MSC (MDS-MSC) expressed higher RN7SL1 than controls and MDS cloned cells expressed higher retinoic acid inducible gene-I (RIG-I) and interferon-stimulated genes (ISGs), which are the receptor and effector molecules of RN7SL1, respectively. MDS-MSC can promote tumor growth. Adding both exosomes extracted from MDS-MSC and exogenous RN7SL1 into MDS cloned cells can promote cell proliferation, and RIG-I inhibitor can block this effect. We infer that MDS-MSC may promote MDS cloned cell proliferation through RIG-ISGs signal activated by exosomes-derived RN7SL1. To test this hypothesis, we will employ MDS cell lines and primary cells to reveal the role of MSC-derived exosomes in MDS and clarify the molecular mechanism of cell proliferation induced by RN7SL1/RIG-I-ISGs signal, using the PCR, WB, FCM, RNA interference, gene arrays, CHIP, et al. We will provide evidence from the molecules and cells levels. This study will provide new ideas for MDS treatment in the aspect of microenvironment.

MSC与MDS克隆细胞的相互作用促进疾病发生，外泌体作为其相互作用的一种机制如何参与疾病的进程尚不清楚。我们前期研究发现：MDS-MSC来源的外泌体中高表达RN7SL1，而MDS克隆细胞高表达其受体RIG-I及效应分子ISGs，MDS-MSC有促瘤作用，单独MDS克隆细胞中加入MDS-MSC所提取的外泌体或外源性RN7SL1均可促进细胞增殖，RIG-I抑制剂可阻断该效应。申请人推测：MDS-MSC可能通过外泌体来源的RN7SL1激活RIG-ISGs通路介导MDS克隆细胞增殖。为验证这一假说，我们将通过MDS细胞系及原代细胞，采用PCR、WB、FCM、RNA干扰、基因芯片、CHIP等手段，从细胞及分子水平探讨MSC来源的外泌体在MDS恶性增殖中的作用，明确其介导的RN7SL1/RIG-I-ISGs信号调控MDS细胞恶性生物学行为的分子机制，为MDS治疗在微环境方面提供新思路。

MSC与MDS克隆细胞的相互作用促进疾病发生，外泌体作为其相互作用的一种机制如何参与疾病的进程尚不清楚。通过本项目我们发现MDS-MSC来源的外泌体促进MDS细胞增殖、抑制凋亡，MDS-MSC来源的外泌体中高表达RN7SL1，而MDS克隆细胞高表达其受体RIG-I及效应分子ISGs，MDS克隆细胞中加入MDS-MSC所提取的外泌体或外源性RN7SL1均可促进细胞增殖，RIG-I沉默可阻断该效应。MDS-MSC所提取的外泌体或外源性RN7SL1可激活MDS克隆细胞RIG-I-ISGs通路活化。同时，确证MDS-MSC来源的外泌体中RN7SL1表达异常的机制由NOTCH-MYC激活介导，故认为MDS-MSC中NOTCH-MYC通路激活，上调 RN7SL1表达，过表达的 RN7SL1 以外泌体的形式分泌出胞，转运至MDS克隆细胞，激活 RIG-I-ISGs信号，从而促进 MDS 克隆细胞增殖。本研究深入探讨了MSC来源的外泌体在MDS恶性增殖中的作用，明确其介导的RN7SL1/RIG-I-ISGs信号调控MDS细胞恶性生物学行为的分子机制，为MDS治疗在微环境方面提供新思路。