Single-cell RNA sequencing of equine mesenchymal stromal cells from primary donor-matched tissue sources reveals functional heterogeneity in immune modulation and cell motility.
Single-cell RNA sequencing of equine mesenchymal stromal cells from primary donor-matched tissue sources reveals functional heterogeneity in immune modulation and cell motility.
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DOI：
10.1186/s13287-020-02043-5
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发表时间：
2020-12-04
影响因子：
7.5
通讯作者：
Van de Walle GR
中科院分区：
文献类型：
--
作者：
Harman RM;Patel RS;Fan JC;Park JE;Rosenberg BR;Van de Walle GR
间充质基质细胞(MSC)治疗的有效性被认为取决于来自不同组织来源的MSC培养的内在异质性以及来自同一组织来源的单个MSCs的内在异质性，这两者都不是很好的理解。为了研究这一点，我们使用了从马身上分离的MSC培养物。马被认为是适合于翻译MSC研究的生理上相关的大型动物模型。此外，由于它的大小，马允许同时从同一动物的多个组织中收集足够的样本，因此，可以从不同来源的捐赠者匹配的MSC培养中进行独特的收集。由于体型和伦理的限制，后者在老鼠和人类身上的挑战要大得多。在本研究中，我们对来自三个供者匹配的组织来源的原代马间充质干细胞进行了单细胞RNA测序(scRNA-seq)：脂肪组织(AT)、骨髓(BM)和外周血(PB)。基于scRNA-seq检测到的转录差异，我们进行了功能实验，以检测不同MSC群体的运动性和免疫调节功能。我们观察到了三个来源的马间充质干细胞的来源间和来源内的异质性。功能实验表明，转录差异与细胞运动和免疫调节功能的表型差异相对应。明确地说，我们发现(I)来自三个供者匹配的组织来源的MSC培养物之间的连接黏附分子2(JAM2)的差异表达转化为当使用RNA干扰来敲除该基因时骨髓来源的MSCs的细胞运动能力的改变，以及(Ii)来自相同组织来源的克隆MSC系中C-X-C基序趋化因子配体6(CXCL6)的表达差异与来自PB来源的MSCs的趋化能力相关。最终，这些发现将加强我们对MSC异质性的理解，并将导致MSCs治疗潜力的改善，加速从长凳到床边的过渡。网上版载有补充材料，可在10.1186/s13287-020-02043-5查阅。
The efficacy of mesenchymal stromal cell (MSC) therapy is thought to depend on the intrinsic heterogeneity of MSC cultures isolated from different tissue sources as well as individual MSCs isolated from the same tissue source, neither of which is well understood. To study this, we used MSC cultures isolated from horses. The horse is recognized as a physiologically relevant large animal model appropriate for translational MSC studies. Moreover, due to its large size the horse allows for the simultaneous collection of adequate samples from multiple tissues of the same animal, and thus, for the unique collection of donor matched MSC cultures from different sources. The latter is much more challenging in mice and humans due to body size and ethical constraints, respectively. In the present study, we performed single-cell RNA sequencing (scRNA-seq) on primary equine MSCs that were collected from three donor-matched tissue sources; adipose tissue (AT), bone marrow (BM), and peripheral blood (PB). Based on transcriptional differences detected with scRNA-seq, we performed functional experiments to examine motility and immune regulatory function in distinct MSC populations. We observed both inter- and intra-source heterogeneity across the three sources of equine MSCs. Functional experiments demonstrated that transcriptional differences correspond with phenotypic variance in cellular motility and immune regulatory function. Specifically, we found that (i) differential expression of junctional adhesion molecule 2 (JAM2) between MSC cultures from the three donor-matched tissue sources translated into altered cell motility of BM-derived MSCs when RNA interference was used to knock down this gene, and (ii) differences in C-X-C motif chemokine ligand 6 (CXCL6) expression in clonal MSC lines derived from the same tissue source correlated with the chemoattractive capacity of PB-derived MSCs. Ultimately, these findings will enhance our understanding of MSC heterogeneity and will lead to improvements in the therapeutic potential of MSCs, accelerating the transition from bench to bedside. The online version contains supplementary material available at 10.1186/s13287-020-02043-5.
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影响因子：
11.1
作者：
Birnbaum KD
通讯作者：
Birnbaum KD
影响因子：
7.5
作者：
Bussche L;Harman RM;Syracuse BA;Plante EL;Lu YC;Curtis TM;Ma M;Van de Walle GR
通讯作者：
Van de Walle GR
影响因子：
7
作者：
Derr A;Yang C;Zilionis R;Sergushichev A;Blodgett DM;Redick S;Bortell R;Luban J;Harlan DM;Kadener S;Greiner DL;Klein A;Artyomov MN;Garber M
通讯作者：
Garber M
影响因子：
4
作者：
Barrett, Angela N.;Fong, Chui-Yee;Bongso, Ariff
通讯作者：
Bongso, Ariff
影响因子：
7.5
作者：
Geburek F;Mundle K;Conrad S;Hellige M;Walliser U;van Schie HT;van Weeren R;Skutella T;Stadler PM
通讯作者：
Stadler PM