Unité de Thérapie Cellulaire et Centre d'Investigation Clinique en Biothérapies CIC-BT501, Assistance Publique-Hôpitaux de Paris, Hôpital Saint Louis, Paris, France.
BMJ Open. 2013 Jan 7;3(1):e001890. doi: 10.1136/bmjopen-2012-001890.
The present work aimed to evaluate the expression of transforming growth factor-β (TGF-β) receptors on bone marrow-derived multipotent mesenchymal stromal cells (MSCs) in patients with systemic sclerosis (SSc) and the consequences of TGF-β activation in these cells, since MSC have potential therapeutic interest for SSc patients and knowing that TGF-β plays a critical role during the development of fibrosis in SSc.
This is a prospective research study using MSC samples obtained from SSc patients and compared with MSC from healthy donors.
One medical hospital involving collaboration between an internal medicine department for initial patient recruitment, a clinical biotherapeutic unit for MSC preparation and an academic laboratory for research.
9 patients with diffuse SSc for which bone marrow (BM) aspiration was prescribed by sternum aspiration before haematopoietic stem cell transplantation, versus nine healthy donors for normal BM.
TGF-β, TGF-β receptor types I (TBRI) and II (TBRII) mRNA and protein expression were assessed by quantitative PCR and flow cytometry, respectively, in MSC from both SSc patients and healthy donors. MSC were exposed to TGF-β and assessed for collagen 1α2 synthesis and Smad expression. As positive controls, primary cultures of dermal fibroblasts were also analysed.
Compared with nine controls, MSC from nine SSc patients showed significant increase in mRNA levels (p<0.002) and in membrane expression (p<0.0001) of TBRII. In response to TGF-β activation, a significant increase in collagen 1α synthesis (p<0.05) and Smad-3 phosphorylation was upregulated in SSc MSC. Similar results were obtained on eight SSc-derived dermal fibroblasts compared to six healthy controls.
TBRII gene and protein expression defect in MSC derived from SSc patients may have pathological significance. These findings should be taken into account when considering the use of MSC-based therapies in an autologous setting.
本研究旨在评估系统性硬化症（SSc）患者骨髓来源的多能间充质基质细胞（MSC）中转化生长因子-β（TGF-β）受体的表达及其在这些细胞中 TGF-β激活的后果，因为 MSC 对 SSc 患者具有潜在的治疗意义，且 TGF-β 在 SSc 纤维化的发展中起着关键作用。
这是一项使用 SSc 患者和健康供体的 MSC 样本进行的前瞻性研究。
一家医疗医院，涉及内科部门进行初步患者招募、临床生物治疗单位进行 MSC 准备以及学术实验室进行研究的协作。
9 例弥漫性 SSc 患者，在造血干细胞移植前经胸骨抽吸进行骨髓（BM）抽吸，与 9 例健康供体的正常 BM 进行比较。
通过定量 PCR 和流式细胞术分别评估 TGF-β、TGF-β 受体 I（TBRI）和 II（TBRII）在 SSc 患者和健康供体的 MSC 中的 mRNA 和蛋白表达。将 MSC 暴露于 TGF-β 中，并评估胶原 1α2 合成和 Smad 表达。真皮成纤维细胞的原代培养也作为阳性对照进行分析。
与 9 例对照相比，9 例 SSc 患者的 MSC 显示出明显的 mRNA 水平升高（p<0.002）和膜表达升高（p<0.0001）。在 TGF-β 激活后，SSc MSC 中胶原 1α 合成（p<0.05）和 Smad-3 磷酸化显著增加。与 6 例健康对照相比，8 例 SSc 衍生的真皮成纤维细胞也得到了类似的结果。
SSc 患者来源的 MSC 中 TBRII 基因和蛋白表达缺陷可能具有病理意义。在考虑在自体环境中使用基于 MSC 的治疗方法时，应考虑这些发现。
