间充质基质细胞的代谢组学和细胞因子分析鉴定了预测 T 细胞抑制的标志物。

Metabolomics and cytokine profiling of mesenchymal stromal cells identify markers predictive of T-cell suppression.

机构信息

School of Chemical, Materials, and Biomedical Engineering, University of Georgia, Athens, Georgia, USA; Regenerative Bioscience Center, University of Georgia, Athens, Georgia, USA.

Complex Carbohydrate Research Center and Institute of Bioinformatics, University of Georgia, Athens, Georgia, USA.

出版信息

Cytotherapy. 2022 Feb;24(2):137-148. doi: 10.1016/j.jcyt.2021.08.002. Epub 2021 Oct 22.

DOI:10.1016/j.jcyt.2021.08.002

PMID:34696960

Abstract

BACKGROUND AIMS

Mesenchymal stromal cells (MSCs) have shown great promise in the field of regenerative medicine, as many studies have shown that MSCs possess immunomodulatory function. Despite this promise, no MSC therapies have been licensed by the Food and Drug Administration. This lack of successful clinical translation is due in part to MSC heterogeneity and a lack of critical quality attributes. Although MSC indoleamine 2,3-dioxygnease (IDO) activity has been shown to correlate with MSC function, multiple predictive markers may be needed to better predict MSC function.

METHODS

Three MSC lines (two bone marrow-derived, one induced pluripotent stem cell-derived) were expanded to three passages. At the time of harvest for each passage, cell pellets were collected for nuclear magnetic resonance (NMR) and ultra-performance liquid chromatography mass spectrometry (MS), and media were collected for cytokine profiling. Harvested cells were also cryopreserved for assessing function using T-cell proliferation and IDO activity assays. Linear regression was performed on functional data against NMR, MS and cytokines to reduce the number of important features, and partial least squares regression (PLSR) was used to obtain predictive markers of T-cell suppression based on variable importance in projection scores.

RESULTS

Significant functional heterogeneity (in terms of T-cell suppression and IDO activity) was observed between the three MSC lines, as were donor-dependent differences based on passage. Omics characterization revealed distinct differences between cell lines using principal component analysis. Cell lines separated along principal component one based on tissue source (bone marrow-derived versus induced pluripotent stem cell-derived) for NMR, MS and cytokine profiles. PLSR modeling of important features predicted MSC functional capacity with NMR (R = 0.86), MS (R = 0.83), cytokines (R = 0.70) and a combination of all features (R = 0.88).

CONCLUSIONS

The work described here provides a platform for identifying markers for predicting MSC functional capacity using PLSR modeling that could be used as release criteria and guide future manufacturing strategies for MSCs and other cell therapies.

摘要

背景目的

间充质基质细胞（MSCs）在再生医学领域显示出巨大的应用潜力，因为许多研究表明 MSCs 具有免疫调节功能。尽管有这种前景，但食品和药物管理局尚未批准任何 MSC 疗法。这种临床转化的失败部分是由于 MSC 的异质性和缺乏关键质量属性。尽管 MSC 吲哚胺 2,3-双加氧酶（IDO）活性已被证明与 MSC 功能相关，但可能需要多种预测性标志物来更好地预测 MSC 功能。

方法

将三条 MSC 系（两条骨髓源性，一条诱导多能干细胞源性）扩增至三代。在每一代的收获时，收集细胞沉淀进行核磁共振（NMR）和超高效液相色谱质谱联用（MS），并收集培养基进行细胞因子谱分析。收获的细胞也被冷冻保存，以通过 T 细胞增殖和 IDO 活性测定来评估其功能。对功能数据与 NMR、MS 和细胞因子进行线性回归，以减少重要特征的数量，并使用偏最小二乘回归（PLSR）根据变量重要性投影得分获得 T 细胞抑制的预测标志物。

结果

三条 MSC 系之间观察到明显的功能异质性（在 T 细胞抑制和 IDO 活性方面），并且根据代次还存在供体依赖性差异。组学特征分析通过主成分分析显示细胞系之间存在明显差异。细胞系根据组织来源（骨髓源性与诱导多能干细胞源性）在主成分一上分离，用于 NMR、MS 和细胞因子图谱。使用 NMR（R=0.86）、MS（R=0.83）、细胞因子（R=0.70）和所有特征的组合（R=0.88）对重要特征进行 PLSR 建模，可预测 MSC 功能能力。