Bukulmez Hulya, Dennis Adrienne T, Reese-Koc Jane, Sieg Scott F, Clagett Brian, Kleinsorge-Block Sarah, Somoza-Palacios Rodrigo, Singer Nora, Chance Mark, Highland Kristin B, Emancipator Steven N
Division of Pediatric Rheumatology, Department of Pediatrics, Metrohealth Medical Center, Case Western Reserve University, Cleveland, OH, United States.
Cellular Therapy Operations and Quality, National Center for Regenerative Medicine, School of Medicine, Case Western Reserve University, Cleveland, OH, United States.
Stem Cells. 2025 Feb 12;43(2). doi: 10.1093/stmcls/sxae078.
Mesenchymal stromal cells (MSCs) can modulate immune responses and suppress inflammation in autoimmune diseases. Although their safety has been established in clinical trials, the efficacy of MSCs is inconsistent due to variability in potency among different preparations and limited specificity in targeting mechanisms driving autoimmune diseases.
We utilized high-dimensional design of experiments methodology to identify factor combinations that modulate gene expression by MSCs to mitigate inflammation. This led to a novel MSC-based cell therapy, HXB-319. Its anti-inflammatory properties were validated in vitro by flow cytometry, RT-PCR, and mass spectrophotometry. To evaluate in vivo efficacy, we treated a diffuse alveolar hemorrhage (DAH) mouse model (C57Bl/6). Seven days post-DAH induction with pristane, mice received either MSCs or HXB-319 (2X106 cells, IP). On day 14, peritoneal lavage fluid (PLF) and lung tissue were collected for flow cytometry, histopathological examination, and mRNA.
HXB-319 increased gene expression levels of anti-inflammatory, angiogenic, and anti-fibrotic factors (eg, TSG-6, VEGF, and HGF). KEGG pathway analysis confirmed significant activation of relevant anti-inflammatory, angiogenic, and anti-fibrotic proteins, corroborating RT-PCR results. In the DAH model, HXB-319 significantly reduced lung inflammation and alveolar hemorrhage compared to MSC-treated and untreated DAH mice. HXB-319 treatment also significantly decreased neutrophils, plasmacytoid dendritic cells, and RORγT cells, increased FoxP3+ cells in PLF, and reversed alterations in mRNA encoding IL-6, IL-10, and TSG-6 in lung tissue compared to DAH mice.
HXB-319 effectively controls inflammation and prevents tissue damage in pristine-induced DAH, highlighting its therapeutic potential for autoimmune inflammatory diseases.
间充质基质细胞（MSCs）可调节免疫反应并抑制自身免疫性疾病中的炎症。尽管其安全性已在临床试验中得到证实，但由于不同制剂之间效力的差异以及驱动自身免疫性疾病的靶向机制特异性有限，MSCs的疗效并不一致。
我们利用高维实验设计方法来确定调节MSCs基因表达以减轻炎症的因子组合。这导致了一种新型的基于MSCs的细胞疗法HXB-319。其抗炎特性通过流式细胞术、逆转录-聚合酶链反应（RT-PCR）和质谱法在体外得到验证。为了评估体内疗效，我们对弥漫性肺泡出血（DAH）小鼠模型（C57Bl/6）进行了治疗。在用 pristane诱导DAH 7天后，小鼠接受MSCs或HXB-319（2×10⁶个细胞，腹腔注射）。在第14天，收集腹腔灌洗液（PLF）和肺组织用于流式细胞术、组织病理学检查和mRNA检测。
HXB-319增加了抗炎、血管生成和抗纤维化因子（如TSG-6、VEGF和HGF）的基因表达水平。京都基因与基因组百科全书（KEGG）通路分析证实了相关抗炎、血管生成和抗纤维化蛋白的显著激活，证实了RT-PCR结果。在DAH模型中，与接受MSCs治疗和未治疗的DAH小鼠相比，HXB-319显著减轻了肺部炎症和肺泡出血。与DAH小鼠相比，HXB-319治疗还显著减少了中性粒细胞、浆细胞样树突状细胞和RORγT细胞，增加了PLF中FoxP3⁺细胞，并逆转了肺组织中编码IL-6、IL-10和TSG-6的mRNA的改变。
HXB-319有效控制了pristane诱导的DAH中的炎症并预防了组织损伤，突出了其在自身免疫性炎症性疾病中的治疗潜力。
