骨髓间充质干细胞上的CD44岩藻糖基化增强归巢能力并促进糖尿病小鼠的肠神经系统重塑。

CD44 fucosylation on bone marrow-derived mesenchymal stem cells enhances homing and promotes enteric nervous system remodeling in diabetic mice.

作者信息

Shi Huiying, Jiang Chen, Yao Hailing, Zhang Yurui, Zhang Qin, Hou Xiaohua, Lin Rong

机构信息

Department of Gastroenterology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.

Department of Pathology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.

出版信息

Cell Biosci. 2021 Jun 30;11(1):118. doi: 10.1186/s13578-021-00632-2.

DOI:10.1186/s13578-021-00632-2

PMID:34193268

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8243650/

Abstract

BACKGROUND

Diabetes can cause extensive enteric nervous system (ENS) injuries and gastrointestinal motility disorder. In developing possible treatments, researchers have engaged in tissue regeneration engineering with the very promising bone marrow-derived mesenchymal stem cells (BMSCs). However, BMSCs have poor homing ability to the targeted tissues after intravenous injection. Thus, we aimed to investigate whether enhancing the expression of E-selectin ligand on BMSCs could improve their homing ability and subsequently influence their role in ENS remodeling in diabetic mice.

METHODS

First, we constructed the fucosylation modification of CD44 on BMSCs through a fucosyltransferase VII (FTVII) system to generate a Hematopoietic Cell E-/L-selectin Ligand (HCELL) property, a fucosylated sialyllactosaminyl glycovariant of CD44 that potently binds E-selectin. Next, FTVII-modified and unmodified BMSCs labeled with green fluorescent protein (GFP) were injected into diabetic mice through the tail vein to compare their homing ability to the gastrointestinal tract and their effect on ENS remodeling, respectively. A bioluminescent imaging system was used to evaluate the homing ability of GFP-labeled BMSCs with and without FTVII modification, to the gastrointestinal tract. Gastrointestinal motility was assessed by gastrointestinal transient time, defecation frequency, stool water content and colon strips contractility. Immunofluorescence staining and western blotting were used to assess the expression levels of protein gene product 9.5 (PGP9.5), glial fibrillary acidic protein (GFAP) and glial cell line-derived neurotrophic factor (GDNF).

RESULTS

The FTVII-mediated α(1,3)-fucosylation modification of CD44 on BMSCs generated a HCELL property. Bioluminescent imaging assays showed that FTVII-modified BMSCs had enhanced homing ability to gastrointestinal tract, mainly to the colon, 24 h after injection through the tail vein. Compared with diabetic mice, FTVII-modified BMSCs significantly promoted the gastrointestinal motility and the ENS remodeling, including intestinal peristalsis (P < 0.05), increased feces excretion (P < 0.05) and the water content of the feces (P < 0.05), restored the spontaneous contraction of the colon (P < 0.05), and upregulated the protein expression levels of PGP9.5 (P < 0.01), GFAP (P < 0.001), and GDNF (P < 0.05), while unmodified BMSCs did not (P > 0.05).

CONCLUSIONS

CD44 fucosylation modification on murine BMSCs promotes homing ability to the gastrointestinal tract and ENS remodeling in diabetic mice.

摘要

背景

糖尿病可导致广泛的肠神经系统（ENS）损伤和胃肠动力障碍。在研发可能的治疗方法时，研究人员致力于利用非常有前景的骨髓间充质干细胞（BMSCs）进行组织再生工程。然而，静脉注射后BMSCs向靶组织的归巢能力较差。因此，我们旨在研究增强BMSCs上E-选择素配体的表达是否能提高其归巢能力，进而影响其在糖尿病小鼠ENS重塑中的作用。

方法

首先，我们通过岩藻糖基转移酶VII（FTVII）系统构建了BMSCs上CD44的岩藻糖基化修饰，以产生造血细胞E-/L-选择素配体（HCELL）特性，即CD44的一种岩藻糖基化唾液酸化乳糖胺聚糖变体，它能有效结合E-选择素。接下来，将用绿色荧光蛋白（GFP）标记的FTVII修饰和未修饰的BMSCs通过尾静脉注射到糖尿病小鼠体内，分别比较它们对胃肠道的归巢能力及其对ENS重塑的影响。使用生物发光成像系统评估有无FTVII修饰的GFP标记BMSCs对胃肠道的归巢能力。通过胃肠转运时间、排便频率、粪便含水量和结肠条收缩性评估胃肠动力。采用免疫荧光染色和蛋白质印迹法评估蛋白基因产物9.5（PGP9.5）、胶质纤维酸性蛋白（GFAP）和胶质细胞源性神经营养因子（GDNF）的表达水平。

结果

FTVII介导的BMSCs上CD44的α(α(1,3)-岩藻糖基化修饰产生了HCELL特性。生物发光成像分析显示，通过尾静脉注射24小时后，FTVII修饰的BMSCs对胃肠道，主要是对结肠的归巢能力增强。与糖尿病小鼠相比，FTVII修饰的BMSCs显著促进了胃肠动力和ENS重塑，包括肠道蠕动（P<0.05）、粪便排泄增加（P<0.05）和粪便含水量增加（P<0.05），恢复了结肠的自发收缩（P<0.05），并上调了PGP9.5（P<0.01）、GFAP（P<0.001）和GDNF（P<0.05）的蛋白表达水平，而未修饰的BMSCs则没有（P>0.05）。