College of Biological and Chemical Science and Engineering, Jiaxing University, Lianglin Campus,118 Jiahang Road, Jiaxing, 314001, China.
College of life science and biotechnology, Hebei Normal University of Science and Technology, Qinhuangdao, 066004, China.
Lipids Health Dis. 2017 Oct 2;16(1):189. doi: 10.1186/s12944-017-0577-4.
Lipoprotein lipase (LPL) deficiency is an autosomal recessive genetic disorder characterized by extreme hypertriglyceridemia, with no cure presently available. The purpose of this study was to test the possibility of using cell therapy to alleviate LPL deficiency.
The LPL coding sequence was cloned into the MSCV retrovirus vector, after which MSCV-hLPL and MSCV (empty construct without LPL coding sequence) virion suspensions were made using the calcium chloride method. A muscle cell line (C2C12), kidney cell line (HEK293T) and pre-adipocyte cell line (3 T3-L1) were transfected with the virus in order to express recombinant LPL in vitro. Finally, each transfected cell line was injected subcutaneously into nude mice to identify the cell type which could secret recombinant LPL in vivo. Control cells were transfected with the MSCV empty vector. LPL activity was analyzed using a radioimmunoassay.
After virus infection, the LPL activity at the cell surface of each cell type was significantly higher than in the control cells, which indicates that all three cell types can be used to generate functional LPL. The transfected cells were injected subcutaneously into nude mice, and the LPL activity of the nearby muscle tissue at the injection site in mice injected with 3 T3-L1 cells was more than 5 times higher at the injection sites than at non-injected control sites. The other two types of cells did not show this trend.
The subcutaneous injection of adipocytes overexpressing LPL can improve the LPL activity of the adjacent tissue of nude mice. This is a ground-breaking preliminary study for the treatment of LPL deficiency, and lays a good foundation for using cell therapy to correct LPL deficiency.
脂蛋白脂肪酶（LPL）缺乏症是一种常染色体隐性遗传疾病，其特征为极度高甘油三酯血症，目前尚无治愈方法。本研究旨在探讨细胞治疗缓解 LPL 缺乏症的可能性。
将 LPL 编码序列克隆到 MSCV 逆转录病毒载体中，然后使用氯化钙法制备 MSCV-hLPL 和 MSCV（不含 LPL 编码序列的空载体）病毒悬液。将病毒转染肌肉细胞系（C2C12）、肾细胞系（HEK293T）和前脂肪细胞系（3T3-L1），以体外表达重组 LPL。最后，将每种转染的细胞系皮下注射到裸鼠中，以鉴定体内可分泌重组 LPL 的细胞类型。对照细胞用 MSCV 空载体转染。用放射免疫法分析 LPL 活性。
病毒感染后，各细胞类型细胞表面的 LPL 活性明显高于对照细胞，表明这三种细胞类型均可产生功能性 LPL。将转染的细胞皮下注射到裸鼠中，注射 3T3-L1 细胞的裸鼠注射部位附近肌肉组织的 LPL 活性比未注射对照部位高 5 倍以上。其他两种类型的细胞没有表现出这种趋势。
皮下注射过表达 LPL 的脂肪细胞可提高裸鼠邻近组织的 LPL 活性。这是治疗 LPL 缺乏症的开创性初步研究，为使用细胞治疗纠正 LPL 缺乏症奠定了良好基础。
