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. 2020 Nov;38(11):1479-1491.
doi: 10.1002/stem.3253. Epub 2020 Aug 8.

The Sca-1+ and Sca-1- mouse prostatic luminal cell lineages are independently sustained

Oh-Joon Kwon  1 , Jong Min Choi  2 , Li Zhang  1 , Deyong Jia  1 , Zhouyihan Li  3 , Yiqun Zhang  4 , Sung Yun Jung  2 , Chad J Creighton  4 , Li Xin  1   5
Affiliations

The Sca-1+ and Sca-1- mouse prostatic luminal cell lineages are independently sustained

Oh-Joon Kwon et al. Stem Cells. 2020 Nov.

Abstract

The phenotypic and functional heterogeneity of the mouse prostate epithelial cell lineages remains incompletely characterized. We show that the Sca-1+ luminal cells at the mouse proximal prostate express Sox2. These cells are replicative quiescent, castration resistant, and do not possess secretory function. We use the Probasin-CreERT2 and Sox2-CreERT2 models in concert with a fluorescent reporter line to label the Sca-1- and Sca-1+ luminal cells, respectively. By a lineage tracing approach, we show that the two luminal cell populations are independently sustained. Sox2 is dispensable for the maintenance of the Sca-1+ luminal cells but is essential for their facultative bipotent differentiation capacity. The Sca-1+ luminal cells share molecular features with the human TACSTD2+ luminal cells. This study corroborates the heterogeneity of the mouse prostate luminal cell lineage and shows that the adult mouse prostate luminal cell lineage is maintained by distinct cellular entities rather than a single progenitor population.

Keywords: Sca-1; Sox2; castration resistance; prostate stem cells.

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Conflict of interest statement

CONFLICT OF INTEREST

The authors declared no potential conflicts of interest.

Figures

FIGURE 1
FIGURE 1
Molecular characterization of mouse Sca-1+ luminal cells. A, Heatmap of RNA-seq analysis shows expression of representative lineage markers in fluorescence activated cell sorting-isolated mouse prostate basal (Ba), Sca-1− luminal (S−Lu), Sca-1+ luminal (S+Lu), and stromal cells (Str). B, Quantitative reverse transcription polymerase chain reaction analysis shows mean ± SD of expression of nine genes differentially expressed between S−Lu and S+Lu cells. N = 3. C, Gene ontology analysis of RNA-Seq of S−Lu and S+Lu cells based on genes differentially expressed with P < .01, fold > 1.4. D, Heatmap of RNA-seq analysis shows expression of prostatic secretion-related genes in S−Lu and S+Lu cells. E, Volcano plot of proteomic analysis in S−Lu and S+Lu cells. F, Bar graph shows means ± SD of intensity-based fraction of total level of prostatic secretory proteins in S−Lu and S+Lu cells. N = 3. G, Gene ontology analysis of proteomic analysis of S−Lu and S+Lu cells
FIGURE 2
FIGURE 2
Mouse Sca-1− luminal cells are self-sustained. A, Bar graph shows means ± SD of expression of Pbsn in fluorescence activated cell sorting-sorted mouse basal (Ba), stromal (Str), Sca-1+ (S+Lu), and Sca-1− (S−Lu) luminal cells by quantitative reverse transcription polymerase chain reaction N = 3. B, Coimmunostaining of cytokeratin 5 (K5)/eYFP (upper) and Nkx3.1/eYFP (lower) in proximal and distal anterior lobe of tamoxifen-treated Pbsn-eYFP mice. Scale bars = 25 μm. C, Schematic illustration of lineage tracing experiment during ageing of Pbsn-eYFP mice. Tmx, tamoxifen. D, Dot plot shows means ± SD of percentage of eYFP+ luminal cells during ageing of Pbsn-eYFP mice (N = 7). E, Schematic illustration of lineage tracing experiment with three cycles of androgen deprivation and replacement of Pbsn-eYFP mice. F, Dot plot shows means ± SD of percentage of eYFP+ cells in prostate basal, luminal, and stromal cells of Pbsn-eYFP mice after mock surgery (control) or three cycles of castration-regeneration(C-R) (N = 7). G,H, Immunostaining of Nkx3.1/eYFP in proximal and distal regions of control (G, upper panels) or castration-regeneration (C-R) (G, lower panels, and H) group. White bars = 25 μm; yellow bars = 200 μm. I, Coimmunostaining of Nkx3.1/eYFP/BrdU in anterior lobes of Pbsn-eYFP mice after androgen replacement. Yellow bar = 75 μm; white bar = 25 μm. J, Dot plot shows means ± SD of percentage of BrdU+ cells in eYFP−Nkx3.1+ and eYFP+Nkx3.1+ cells of Pbsn-eYFP mice (N = 6)
FIGURE 3
FIGURE 3
Sox2+ luminal cells in mouse proximal prostate are quiescent and self-sustained. A, Coimmunostaining of Sox2/K8 (left) and Sox2/Trp63 (right) in proximal and distal anterior prostate of C57/Bl6 mice. Scale bars = 25 μm. B, Fluorescent images (left) and coimmunostaining of Nkx3.1/eYFP in proximal (middle) and distal (right) anterior prostate of tamoxifen-treated Sox2-eYFP mice. Yellow bar = 2 mm; white bars = 25 μm. C, Dot plot shows means ± SD of percentage of eYFP+Nkx3.1+ cells in tamoxifen-treated Sox2-eYFP mice during ageing (N = 6). D, Coimmunostaining of Nkx3.1/eYFP of anterior prostate of Sox2-eYFP mice at 6 months after tamoxifen treatment. Scale bar = 200 μm. E, Schematic illustration of lineage tracing experiment with two cycles of androgen deprivation and replacement using Sox2-eYFP mice. Dot plot shows means ± SD of percentage of eYFP+Nkx3.1+ cells in control and castration-regeneration (C-R) group (N = 6). F, Coimmunostaining of cleaved caspase3 (CC3)/eYFP and eYFP/Nkx3.1 in proximal and distal anterior prostate of tamoxifen-treated Sox2-eYFP mice after castration. White arrows indicate CC3+ cells. White bars = 50 μm. G, Dot plot shows means ± SD of percentage of eYFP+CC3+ and eYFP−CC3+ cells in tamoxifen-treated Sox2-eYFP mice after castration (N = 4). H, Coimmunostaining of BrdU/eYFP/Nkx3.1 in proximal and distal prostate of tamoxifen-treated Sox2-eYFP mice after androgen replacement. Scale bars = 25 μm. I, Dot plot shows means ± SD of percentage of BrdU+ cells in eYFP+Nkx3.1− and eYFP−Nkx3.1+ cells in tamoxifen-treated Sox2-eYFP mice after androgen replacement (N = 4)
FIGURE 4
FIGURE 4
Sox2 is necessary for facultative bipotent differentiation capacity of Sca-1+ luminal cells. A, H&E staining (upper) and coimmunostaining of Sox2/K8 (lower) in proximal prostates of tamoxifen-treated control (Sox2fl/fl) and K8-Sox2 mice. Black bars = 100 μm; white bars = 50 μm. B,C, Dot plots show means ± SD of organoid forming unit (B) and organoid size (C) from Sca-1− (S−Lu) and Sca-1+ (S+Lu) luminal cells of control and K8-Sox2 mice. N.S., no significance. D, Pie charts show percentage of four types of organoids in cultures of S+Lu cells of control and K8-Sox2 mice. P < .0001. E, Dot plot shows means ± SD of percentage of gland-forming unit from S+Lu cells of tamoxifen-treated control and K8-Sox2 mice in prostate regeneration assay (N = 3). F, H&E staining and coimmunostaining of Sox2/K14, K5/K8, and Ar/Trp63 of regenerated tissues from S+Lu cells of tamoxifen-treated control and K8-Sox2 mice. Black bars = 100 μm; white bars = 50 μm
FIGURE 5
FIGURE 5
Mouse Sca-1+ luminal cells share features with human TROP2+ luminal cells. A, Heatmap shows expression of genes differentially expressed between TROP2+ and TROP2− luminal cells in Sca-1+ and Sca-1− luminal cells. Bar graph shows scores for degree of manifestation by t score metric. B, Box plots show signature scores of Sca-1+ luminal cell signature with increasing Gleason grade in two human prostate cancer data sets
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References

    1. Xin L Cells of origin for prostate cancer. Adv Exp Med Biol. 2019; 1210:67–86. - PubMed
    1. Abate-Shen C, Shen MM. Molecular genetics of prostate cancer. Genes Dev. 2000;14(19):2410–2434. - PubMed
    1. Hepburn AC, Curry EL, Moad M, et al. Propagation of human prostate tissue from induced pluripotent stem cells. Stem Cells Translational Medicine. 2020;9:734–745. - PMC - PubMed
    1. Choi N, Zhang B, Zhang L, Ittmann M, Xin L. Adult murine prostate basal and luminal cells are self-sustained lineages that can both serve as targets for prostate cancer initiation. Cancer Cell. 2012;21(2): 253–265. - PMC - PubMed
    1. Wang ZA, Mitrofanova A, Bergren SK, et al. Lineage analysis of basal epithelial cells reveals their unexpected plasticity and supports a cell-of-origin model for prostate cancer heterogeneity. Nat Cell Biol. 2013; 15(3):274–283. - PMC - PubMed

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