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Evaluating the Differentiation Capacity of Mouse Prostate Epithelial Cells Using Organoid Culture
Published on: November 22, 2019
Miz1, a Novel Target of ING4, Can Drive Prostate Luminal Epithelial Cell Differentiation
Penny L Berger1, Mary E Winn2, Cindy K Miranti1
1Laboratory of Integrin Signaling, Van Andel Research Institute, Grand Rapids, Michigan.
Background:
How prostate epithelial cells differentiate and how dysregulation of this process contributes to prostate tumorigenesis remain unclear. We recently identified a Myc target and chromatin reader protein, ING4, as a necessary component of human prostate luminal epithelial cell differentiation, which is often lost in primary prostate tumors. Furthermore, loss of ING4 in the context of oncogenic mutations is required for prostate tumorigenesis. Identifying the gene targets of ING4 can provide insight into how its loss disrupts differentiation and leads to prostate cancer.
Methods:
Using a combination of RNA-Seq, a best candidate approach, and chromatin immunoprecipitation (ChIP), we identified Miz1 as a new ING4 target. ING4 or Miz1 overexpression, shRNA knock-down, and a Myc-binding mutant were used in a human in vitro differentiation assay to assess the role of Miz1 in luminal cell differentiation.
Results:
ING4 directly binds the Miz1 promoter and is required to induce Miz1 mRNA and protein expression during luminal cell differentiation. Miz1 mRNA was not induced in shING4 expressing cells or tumorigenic cells in which ING4 is not expressed. Miz1 dependency on ING4 was unique to differentiating luminal cells; Miz1 mRNA expression was not induced in basal cells. Although Miz1 is a direct target of ING4, and its overexpression can drive luminal cell differentiation, Miz1 was not required for differentiation.
Conclusions:
Miz1 is a newly identified ING4-induced target gene which can drive prostate luminal epithelial cell differentiation although it is not absolutely required. Prostate 77:49-59, 2017. © 2016 Wiley Periodicals, Inc.
Insights
ING4 protein loss disrupts prostate luminal cell differentiation and drives cancer. Miz1 is a new target gene induced by ING4 that can promote differentiation but is not essential.
Area of Science:
- Molecular biology
- Cell differentiation
- Cancer research
Background:
- Prostate epithelial cell differentiation mechanisms and their dysregulation in prostate cancer are not fully understood.
- ING4 (Inhibitor of Growth family, member 4), a Myc target and chromatin reader, is crucial for human prostate luminal epithelial cell differentiation.
- Loss of ING4, particularly with oncogenic mutations, is implicated in prostate tumorigenesis.
Purpose of the Study:
- To identify gene targets of ING4 to understand how its loss impairs differentiation and contributes to prostate cancer.
- To investigate the role of Miz1 as a potential ING4 target in prostate luminal cell differentiation.
Main Methods:
- RNA sequencing (RNA-Seq) and chromatin immunoprecipitation (ChIP) were employed to identify ING4 targets.
- In vitro differentiation assays using human prostate cells were performed with manipulations of ING4 and Miz1 expression (overexpression, shRNA knockdown).
Main Results:
- Miz1 was identified as a direct transcriptional target of ING4, with ING4 required for Miz1 mRNA and protein induction during luminal cell differentiation.
- Miz1 induction by ING4 was specific to differentiating luminal cells, not observed in basal cells or in cells lacking ING4.
- While Miz1 overexpression could drive luminal cell differentiation, Miz1 itself was not essential for this process.
Conclusions:
- Miz1 is a novel ING4-induced gene that can promote prostate luminal epithelial cell differentiation.
- ING4's role in regulating Miz1 is critical for differentiation, and its loss contributes to prostate tumorigenesis.
