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Updated: Aug 5, 2026
Virus Delivery of CRISPR Guides to the Murine Prostate for Gene Alteration
Published on: April 27, 2018
The establishment of prostate-specific, SKP2 humanized mice by CRISPR knock-in method reveals neoplastic initiation
Liankun Song1, Yurong Song2, Vyvyan Nguyen1
1Department of Urology, University of California, Irvine, Orange, CA, USA.
Abstract:
Genetic inactivation of SKP2 has been shown to effectively prevent cancer initiation and block tumorigenesis. However, direct in vivo evidence for SKP2 on cancer initiation and prostatic microenvironment is still lacking and a SKP2 humanized mouse model is critical for developing prostate cancer immunoprevention approaches through targeting SKP2. We therefore have established a prostate-specific human SKP2 knock-in mouse model driven by an endogenous mouse probasin promoter. Overexpression of hSKP2 induces PIN and low-grade carcinoma. RNA-sequencing analysis revealed significant gene expression alterations in EMT, extracellular matrix, and interferon signaling. Single-cell deconvolution showed an increase of fibroblast population and a decrease of CD8+ T cell and B cell populations. Consistent with these results from the SKP2 humanized mouse, SKP2 protein is overexpressed in human prostatic hyperplasia, PIN and prostate adenocarcinoma compared to normal prostate tissues. Overexpression of SKP2 markedly increased cell migration and invasion and induced the gene expression of EMT and interferon pathways. Inhibition of SKP2 signaling by Flavokawain A and C1 reverses EMT and affects EMT and interferon-related gene expression. In addition, paired prostate organoids were derived from SKP2 humanized and wild-type mice for drug screening and validated by known SKP2 inhibitors, Flavokawain A and C1. Both of which selectively decreased viability and altered the morphologies of organoids of hSKP2 knock-in rather than wild-type mice. Our studies provide a well-characterized prostate-specific hSKP2 knock-in mouse model and offer new mechanistic insights for understanding the oncogenic role of SKP2 in shaping the prostatic microenvironment during early carcinogenesis.
Insights
A new human SKP2 mouse model shows SKP2 drives prostate cancer initiation and alters the tumor microenvironment. Targeting SKP2 may offer new prostate cancer immunoprevention strategies.
Area of Science:
- Oncology
- Molecular Biology
- Immunology
Background:
- Genetic inactivation of SKP2 prevents cancer, but its role in prostate cancer initiation and the microenvironment needs in vivo validation.
- A human SKP2 (hSKP2) mouse model is crucial for developing prostate cancer immunoprevention targeting SKP2.
Purpose of the Study:
- Establish a prostate-specific hSKP2 knock-in mouse model to investigate SKP2's role in prostate cancer initiation and microenvironment.
- Explore therapeutic strategies targeting SKP2 in prostate cancer.
Main Methods:
- Created a prostate-specific hSKP2 knock-in mouse model using an endogenous mouse probasin promoter.
- Utilized RNA-sequencing and single-cell deconvolution to analyze gene expression and cellular changes.
- Established prostate organoids for drug screening and validated SKP2 inhibitors (Flavokawain A and C1).
Main Results:
- Overexpression of hSKP2 induced prostatic intraepithelial neoplasia (PIN) and low-grade carcinoma.
- Observed significant alterations in EMT, extracellular matrix, and interferon signaling, with increased fibroblasts and decreased CD8+ T cells and B cells.
- SKP2 overexpression increased cell migration/invasion and EMT/interferon gene expression; SKP2 inhibitors reversed these effects.
Conclusions:
- The established hSKP2 knock-in mouse model provides insights into SKP2's oncogenic role in the prostate microenvironment during early carcinogenesis.
- SKP2 targeting with agents like Flavokawain A and C1 shows potential for selective inhibition of hSKP2-driven prostate cancer organoids.
