Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, 08826, Korea.
Department of Biological Sciences, Sookmyung Women's University, Seoul, Korea.
Stem Cell Res Ther. 2024 May 20;15(1):145. doi: 10.1186/s13287-024-03731-2.
Mesenchymal stem cells (MSCs) play important roles in tissue homeostasis by providing a supportive microenvironmental niche for the hematopoietic system. Cigarette smoking induces systemic abnormalities, including an impeded recovery process after hematopoietic stem cell transplantation. However, the role of cigarette smoking-mediated alterations in MSC niche function have not been investigated.
In the present study, we investigated whether exposure to cigarette smoking extract (CSE) disrupts the hematopoietic niche function of MSCs, and pathways impacted. To investigate the effects on bone marrow (BM)-derived MSCs and support of hematopoietic stem and progenitor cells (HSPCs), mice were repeatedly infused with the CSE named 3R4F, and hematopoietic stem and progenitor cells (HSPCs) supporting function was determined. The impact of 3R4F on MSCs at cellular level were screened by bulk-RNA sequencing and subsequently validated through qRT-PCR. Specific inhibitors were treated to verify the ROS or NLRP3-specific effects, and the cells were then transplanted into the animal model or subjected to coculture with HSPCs.
Both direct ex vivo and systemic in vivo MSC exposure to 3R4F resulted in impaired engraftment in a humanized mouse model. Furthermore, transcriptomic profile analysis showed significantly upregulated signaling pathways related to reactive oxygen species (ROS), inflammation, and aging in 3R4F-treated MSCs. Notably, ingenuity pathway analysis revealed the activation of NLRP3 inflammasome signaling pathway in 3R4F-treated MSCs, and pretreatment with the NLRP3 inhibitor MCC950 rescued the HSPC-supporting ability of 3R4F-treated MSCs.
In conclusion, these findings indicate that exposure to CSE reduces HSPCs supportive function of MSCs by inducing robust ROS production and subsequent NLRP3 activation.
间充质干细胞（MSCs）通过为造血系统提供支持性的微环境龛，在组织稳态中发挥重要作用。吸烟会导致全身异常，包括造血干细胞移植后恢复过程受阻。然而，吸烟介导的 MSC 生态位功能改变的作用尚未被研究。
在本研究中，我们研究了吸烟提取物（CSE）是否会破坏骨髓（BM）来源的 MSC 的造血生态位功能，以及受影响的途径。为了研究其对造血干细胞和祖细胞（HSPCs）支持功能的影响，我们对小鼠进行了重复 CSE（命名为 3R4F）输注，并确定了 HSPCs 支持功能。通过 bulk-RNA 测序筛选 CSE 对 MSC 的细胞水平影响，随后通过 qRT-PCR 进行验证。使用特异性抑制剂来验证 ROS 或 NLRP3 特异性作用，然后将细胞移植到动物模型中或与 HSPCs 共培养。
3R4F 直接在体外和系统内暴露于 MSC，导致人源化小鼠模型中的植入受损。此外，转录组谱分析显示，3R4F 处理的 MSC 中与活性氧（ROS）、炎症和衰老相关的信号通路显著上调。值得注意的是，Ingenuity 通路分析显示，NLRP3 炎性小体信号通路在 3R4F 处理的 MSC 中被激活，NLRP3 抑制剂 MCC950 的预处理挽救了 3R4F 处理的 MSC 对 HSPC 的支持能力。
总之，这些发现表明，暴露于 CSE 通过诱导强烈的 ROS 产生和随后的 NLRP3 激活，降低了 MSC 对 HSPC 的支持功能。
