Genetically engineered K cells provide sufficient insulin to correct hyperglycemia in a nude murine model.
A gene therapy-based treatment of type 1 diabetes mellitus requires the development of a surrogate beta cell that can synthesize and secrete functionally active insulin in response to physiologically relevant changes in ambient glucose levels. In this study, the murine enteroendocrine cell line STC-1 was genetically modified by stable transfection. Two clone cells were selected (STC-1-2 and STC-1-14) that secreted the highest levels of insulin among the 22 clones expressing insulin from 0 to 157.2 microIU/ml/10(6) cells/d. After glucose concentration in the culture medium was increased from 1 mM to 10 mM, secreted insulin rose from 40.3+/-0.8 to 56.3+/-3.2 microIU/ml (STC-1-2), and from 10.8+/-0.8 to 23.6+/-2.3 microIU/ml (STC-1-14). After STC-1-14 cells were implanted into diabetic nude mice, their blood glucose levels were reduced to normal. Body weight loss was also ameliorated. Our data suggested that genetically engineered K cells secrete active insulin in a glucose-regulated manner, and in vivo study showed that hyperglycemia could be reversed by implantation of the cells, suggesting that the use of genetically engineered K cells to express human insulin might provide a glucose-regulated approach to treat diabetic hyperglycemia.
Zhang
Yiqun
Y
Department of General Surgery, Zhongshan Hospital of Fudan University, Shanghai 200032, China.
Yao
Liqing
L
Shen
Kuntang
K
Xu
Meidong
M
Zhou
Pinghong
P
Yang
Weige
W
Liu
Xinyuan
X
Qin
Xinyu
X
D000818:Animals; D001483:Base Sequence; D001786:Blood Glucose / Q000378:metabolism; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D003921:Diabetes Mellitus, Experimental / Q000473:pathology / Q000503:physiopathology / Q000628:therapy; D003922:Diabetes Mellitus, Type 1 / Q000097:blood / Q000503:physiopathology / Q000628:therapy; D004195:Disease Models, Animal; D019858:Enteroendocrine Cells / Q000378:metabolism* / Q000637:transplantation*; D015870:Gene Expression; D005818:Genetic Engineering; D015316:Genetic Therapy; D005951:Glucose Tolerance Test; D006801:Humans; D006943:Hyperglycemia / Q000097:blood / Q000503:physiopathology / Q000628:therapy*; D007328:Insulin / Q000235:genetics* / Q000378:metabolism*; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000473:pathology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D001786:Blood Glucose; D017931:DNA Primers; D007328:Insulin; D011994:Recombinant Proteins