Maturation of Adult β-Cells Revealed Using a Pdx1/Insulin Dual-Reporter Lentivirus

被引:63
|
作者
Szabat, Marta [2 ,3 ]
Luciani, Dan S. [1 ,4 ]
Piret, James M. [2 ,5 ]
Johnson, James D. [1 ,4 ]
机构
[1] Univ British Columbia, Dept Cellular & Physiol Sci, Vancouver, BC V6T 1Z3, Canada
[2] Univ British Columbia, Michael Smith Labs, Vancouver, BC V6T 1Z3, Canada
[3] Univ British Columbia, Grad Program Genet, Vancouver, BC V6T 1Z3, Canada
[4] Univ British Columbia, Dept Surg, Vancouver, BC V6T 1Z3, Canada
[5] Univ British Columbia, Dept Chem & Biol Engn, Vancouver, BC V6T 1Z3, Canada
基金
加拿大健康研究院; 加拿大自然科学与工程研究理事会;
关键词
DIFFERENTIAL GENE-EXPRESSION; PANCREATIC B-CELLS; INSULIN-SECRETION; TRANSCRIPTION FACTOR; HOMEODOMAIN PROTEIN; MIN6; SUBLINES; ISLET CELLS; IN-VITRO; GLUCOSE; HETEROGENEITY;
D O I
10.1210/en.2008-1224
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
The enigmatic process of beta-cell maturation has significant implications for diabetes pathogenesis, and potential diabetes therapies. This study examined the dynamics and heterogeneity of insulin and pancreatic duodenal homeobox (Pdx)-1 gene expression in adult beta-cells. Insulin and Pdx1 expression were monitored in human and mouse islet cells and MIN6 cells using a Pdx1-monomeric red fluorescent protein/insulin-enhanced green fluorescent protein dual-reporter lentivirus. The majority of fluorescent cells were highly positive for both Pdx1 and insulin. Cells expressing Pdx1 but little or no insulin (Pdx1(+)/Ins(low)) comprised 15-25% of the total population. Time-lapse imaging demonstrated that Pdx1(+)/Ins(low) primary beta-cells and MIN6 cells could convert to Pdx1(+)/Ins(+) cells without cell division. Genes involved in the mature beta-cell phenotype (Glut2, MafA) were expressed at higher levels in Pdx1(+)/Ins(+) cells relative to Pdx1(+)/Ins(low) cells. Conversely, genes implicated in early beta-cell development (MafB, Nkx2.2) were enriched in Pdx1(+)/Ins(low) cells. Sorted Pdx1(+)/Ins(low) MIN6 cells had a higher replication rate and secreted less insulin relative to double-positive cells. Long-term phenotype tracking of Pdx1(+)/Ins(low) cells showed two groups, one that matured into Pdx1(+)/Ins(+) cells and one that remained immature. These results demonstrate that adult beta-cells pass through distinct maturation states, which is consistent with previously observed heterogeneity in insulin and Pdx1 expression in adult beta-cells. At a given time, a proportion of adult beta-cells share similar characteristics to functionally immature embryonic beta-cell progenitors. The maturation of adult beta-cells recapitulates development in that Pdx1 expression precedes the robust expression of insulin and other mature beta-cell genes. These results have implications for harnessing the maturation process for therapeutic purposes. (Endocrinology 150: 1627-1635, 2009)
引用
收藏
页码:1627 / 1635
页数:9
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