Zubkova Ekaterina, Kalinin Alexander, Beloglazova Irina, Kurilina Ella, Menshikov Mikhail, Parfyonova Yelena, Tsokolaeva Zoya
National Medical Research Center of Cardiology Named after Academician E.I. Chazov, Moscow 121552, Russia.
Faculty of Fundamental Medicine, Lomonosov Moscow State University, Moscow 119991, Russia.
Int J Mol Sci. 2024 Dec 11;25(24):13286. doi: 10.3390/ijms252413286.
Constructing artificial tertiary lymphoid structures (TLSs) opens new avenues for advancing cancer immunotherapy and personalized medicine by creating controllable immune niches. Mesenchymal stromal cells (MSCs) offer an ideal stromal source for such constructs, given their potent immunomodulatory abilities and accessibility. In this study, we explored the potential of adipose-derived MSCs to adopt TLS-supportive phenotypes and facilitate lymphocyte organization. Single-cell RNA sequencing revealed a distinct subpopulation of MSCs expressing key fibroblastic reticular cell (FRC)-associated markers, including IL-7, PDPN, and IL-15, though lacking follicular dendritic cell (FDC) markers. TNF-α stimulation, but not LTα2β1, further enhanced FRC marker expression (IL-7, PDPN, and ICAM1). Notably, in 3D spheroid co-culture with lymphocytes, MSCs upregulated additional FRC markers, specifically CCL21. Upon implantation into adipose tissue, MSC-lymphocyte organoids maintained structural integrity and showed extensive T-cell infiltration and partial vascularization after 15 days in vivo, although organized B-cell follicles and FDC markers were still lacking. These findings highlight MSCs' intrinsic ability to adopt an FRC-like phenotype that supports T-cell and HEV organization, suggesting that further optimization, including genetic modification, may be needed to achieve an FDC phenotype and replicate the full architectural and functional complexity of TLSs.
构建人工三级淋巴结构（TLS）通过创造可控的免疫微环境，为推进癌症免疫治疗和个性化医学开辟了新途径。间充质基质细胞（MSC）由于其强大的免疫调节能力和易获取性，为此类构建物提供了理想的基质来源。在本研究中，我们探索了脂肪来源的MSC呈现支持TLS表型并促进淋巴细胞组织化的潜力。单细胞RNA测序揭示了一个独特的MSC亚群，其表达关键的成纤维网状细胞（FRC）相关标志物，包括IL-7、PDPN和IL-15，但缺乏滤泡树突状细胞（FDC）标志物。TNF-α刺激而非LTα2β1进一步增强了FRC标志物的表达（IL-7、PDPN和ICAM1）。值得注意的是，在与淋巴细胞的三维球体共培养中，MSC上调了其他FRC标志物，特别是CCL21。植入脂肪组织后，MSC-淋巴细胞类器官保持结构完整性，在体内15天后显示出广泛T细胞浸润和部分血管化，尽管仍缺乏有组织的B细胞滤泡和FDC标志物。这些发现突出了MSC具有采用支持T细胞和HEV组织化的FRC样表型的内在能力，表明可能需要进一步优化，包括基因改造，以实现FDC表型并复制TLS完整的结构和功能复杂性。
