TY - JOUR
T1 - A novel model of urinary tract differentiation, tissue regeneration, and disease
T2 - reprogramming human prostate and bladder cells into induced pluripotent stem cells
AU - Moad, Mohammad
AU - Pal, Deepali
AU - Hepburn, Anastasia C
AU - Williamson, Stuart C
AU - Wilson, Laura
AU - Lako, Majlinda
AU - Armstrong, Lyle
AU - Hayward, Simon W
AU - Franco, Omar E
AU - Cates, Justin M
AU - Fordham, Sarah E
AU - Przyborski, Stefan
AU - Carr-Wilkinson, Jane
AU - Robson, Craig N
AU - Heer, Rakesh
N1 - Publisher webpage states OA under CC-BY 4.0
PY - 2013/11
Y1 - 2013/11
N2 - BACKGROUND: Primary culture and animal and cell-line models of prostate and bladder development have limitations in describing human biology, and novel strategies that describe the full spectrum of differentiation from foetal through to ageing tissue are required. Recent advances in biology demonstrate that direct reprogramming of somatic cells into pluripotent embryonic stem cell (ESC)-like cells is possible. These cells, termed induced pluripotent stem cells (iPSCs), could theoretically generate adult prostate and bladder tissue, providing an alternative strategy to study differentiation.OBJECTIVE: To generate human iPSCs derived from normal, ageing, human prostate (Pro-iPSC), and urinary tract (UT-iPSC) tissue and to assess their capacity for lineage-directed differentiation.DESIGN, SETTING, AND PARTICIPANTS: Prostate and urinary tract stroma were transduced with POU class 5 homeobox 1 (POU5F1; formerly OCT4), SRY (sex determining region Y)-box 2 (SOX2), Kruppel-like factor 4 (gut) (KLF4), and v-myc myelocytomatosis viral oncogene homolog (avian) (MYC, formerly C-MYC) genes to generate iPSCs.OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: The potential for differentiation into prostate and bladder lineages was compared with classical skin-derived iPSCs. The student t test was used.RESULTS AND LIMITATIONS: Successful reprogramming of prostate tissue into Pro-iPSCs and bladder and ureter into UT-iPSCs was demonstrated by characteristic ESC morphology, marker expression, and functional pluripotency in generating all three germ-layer lineages. In contrast to conventional skin-derived iPSCs, Pro-iPSCs showed a vastly increased ability to generate prostate epithelial-specific differentiation, as characterised by androgen receptor and prostate-specific antigen induction. Similarly, UT-iPSCs were shown to be more efficient than skin-derived iPSCs in undergoing bladder differentiation as demonstrated by expression of urothelial-specific markers: uroplakins, claudins, and cytokeratin; and stromal smooth muscle markers: α-smooth-muscle actin, calponin, and desmin. These disparities are likely to represent epigenetic differences between individual iPSC lines and highlight the importance of organ-specific iPSCs for tissue-specific studies.CONCLUSIONS: IPSCs provide an exciting new model to characterise mechanisms regulating prostate and bladder differentiation and to develop novel approaches to disease modelling. Regeneration of bladder cells also provides an exceptional opportunity for translational tissue engineering.
AB - BACKGROUND: Primary culture and animal and cell-line models of prostate and bladder development have limitations in describing human biology, and novel strategies that describe the full spectrum of differentiation from foetal through to ageing tissue are required. Recent advances in biology demonstrate that direct reprogramming of somatic cells into pluripotent embryonic stem cell (ESC)-like cells is possible. These cells, termed induced pluripotent stem cells (iPSCs), could theoretically generate adult prostate and bladder tissue, providing an alternative strategy to study differentiation.OBJECTIVE: To generate human iPSCs derived from normal, ageing, human prostate (Pro-iPSC), and urinary tract (UT-iPSC) tissue and to assess their capacity for lineage-directed differentiation.DESIGN, SETTING, AND PARTICIPANTS: Prostate and urinary tract stroma were transduced with POU class 5 homeobox 1 (POU5F1; formerly OCT4), SRY (sex determining region Y)-box 2 (SOX2), Kruppel-like factor 4 (gut) (KLF4), and v-myc myelocytomatosis viral oncogene homolog (avian) (MYC, formerly C-MYC) genes to generate iPSCs.OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: The potential for differentiation into prostate and bladder lineages was compared with classical skin-derived iPSCs. The student t test was used.RESULTS AND LIMITATIONS: Successful reprogramming of prostate tissue into Pro-iPSCs and bladder and ureter into UT-iPSCs was demonstrated by characteristic ESC morphology, marker expression, and functional pluripotency in generating all three germ-layer lineages. In contrast to conventional skin-derived iPSCs, Pro-iPSCs showed a vastly increased ability to generate prostate epithelial-specific differentiation, as characterised by androgen receptor and prostate-specific antigen induction. Similarly, UT-iPSCs were shown to be more efficient than skin-derived iPSCs in undergoing bladder differentiation as demonstrated by expression of urothelial-specific markers: uroplakins, claudins, and cytokeratin; and stromal smooth muscle markers: α-smooth-muscle actin, calponin, and desmin. These disparities are likely to represent epigenetic differences between individual iPSC lines and highlight the importance of organ-specific iPSCs for tissue-specific studies.CONCLUSIONS: IPSCs provide an exciting new model to characterise mechanisms regulating prostate and bladder differentiation and to develop novel approaches to disease modelling. Regeneration of bladder cells also provides an exceptional opportunity for translational tissue engineering.
KW - Aged
KW - Biomarkers/metabolism
KW - Cell Differentiation/genetics
KW - Cell Lineage/genetics
KW - Cell Separation
KW - Cells, Cultured
KW - Cellular Reprogramming
KW - Female
KW - Gene Expression Regulation, Developmental
KW - Humans
KW - Induced Pluripotent Stem Cells/metabolism
KW - Kallikreins/metabolism
KW - Kruppel-Like Transcription Factors/genetics
KW - Male
KW - Middle Aged
KW - Octamer Transcription Factor-3/genetics
KW - Prostate/cytology
KW - Prostate-Specific Antigen/metabolism
KW - Proto-Oncogene Proteins c-myc/genetics
KW - Receptors, Androgen/metabolism
KW - Regeneration/genetics
KW - SOXB1 Transcription Factors/genetics
KW - Time Factors
KW - Tissue Engineering/methods
KW - Transfection
KW - Ureter/cytology
KW - Urinary Bladder/cytology
KW - Uroplakins/metabolism
U2 - 10.1016/j.eururo.2013.03.054
DO - 10.1016/j.eururo.2013.03.054
M3 - Article
C2 - 23582880
SN - 0302-2838
VL - 64
SP - 753
EP - 761
JO - European Urology
JF - European Urology
IS - 5
ER -