Abstract
BackgroundLipoprotein 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.MethodsThe 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.ResultsAfter 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.ConclusionThe 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.
Highlights
Lipoprotein lipase (LPL) deficiency is an autosomal recessive genetic disorder characterized by extreme hypertriglyceridemia, with no cure presently available
This paper reports the screening of gene vector constructs and ideal transplanting tissues, resulting in the successful construction of the retroviral vector MSCV-human LPL (hLPL), which led to the preliminary confirmation that induced adipocytes can carry foreign LPL genes and can stably secrete LPL to the surrounding tissue after transplantation
The muscle cell line C2C12, Preadipocyte cell line 3 T3-L1 and kidney cell line HEK293T efficiently produced LPL in vitro Using the calcium chloride transfection method, the MSCV-hLPL virion suspension was obtained using the retro-viral packaging system Phoenix-ECO developed by Stanford University [19]
Summary
Lipoprotein lipase (LPL) deficiency is an autosomal recessive genetic disorder characterized by extreme hypertriglyceridemia, with no cure presently available. This study set out to develop a strategy for the treatment of LPL deficiency using induced pluripotent stem cells. The feasibility of this idea was verified in a mouse model. The resulting stem cells were differentiated to yield highly efficient actively LPL-producing tissue cells, and were transplanted back into the mice in vivo, with the expectation that they could cure the LPL deficiency. This method avoids the problems of allograft immune rejection and has good prospects for clinical application [14]
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