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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
Xiaolin Zi1, Liankun Song2, Yurong Song1
1Department of Urology, University of California, Irvine, Orange, CA 92868, 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. Consistently 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. 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
Genetic inactivation of SKP2 prevents cancer. A new humanized mouse model shows SKP2 drives prostate cancer initiation and alters the tumor microenvironment, offering targets for immunoprevention.
Area of Science:
- Oncology
- Molecular Biology
- Immunology
Background:
- Genetic inactivation of SKP2 inhibits cancer initiation and tumorigenesis.
- Direct in vivo evidence for SKP2's role in prostate cancer initiation and the prostatic microenvironment is limited.
- A humanized mouse model targeting SKP2 is crucial for developing prostate cancer immunoprevention strategies.
Purpose of the Study:
- To establish a prostate-specific human SKP2 (hSKP2) knock-in mouse model.
- To investigate the in vivo effects of hSKP2 overexpression on prostate cancer initiation and the tumor microenvironment.
- To provide mechanistic insights into SKP2's oncogenic role in prostate carcinogenesis.
Main Methods:
- Established a prostate-specific hSKP2 knock-in mouse model using an endogenous mouse probasin promoter.
- Performed RNA-sequencing and single-cell deconvolution to analyze gene expression and cellular composition changes.
- Utilized prostate organoids derived from hSKP2 knock-in and wild-type mice for drug screening with SKP2 inhibitors.
Main Results:
- Overexpression of hSKP2 induced prostatic intraepithelial neoplasia (PIN) and low-grade carcinoma.
- Significant alterations in EMT, extracellular matrix, and interferon signaling pathways were observed.
- Increased fibroblast populations and decreased CD8+ T cell and B cell populations were identified.
- SKP2 overexpression in human prostate tissues correlated with hyperplasia, PIN, and adenocarcinoma.
- SKP2 inhibitors selectively reduced the viability of hSKP2-knockin organoids.
Conclusions:
- The developed hSKP2 knock-in mouse model is well-characterized for studying prostate cancer.
- SKP2 plays a critical oncogenic role in shaping the prostatic microenvironment during early carcinogenesis.
- Targeting SKP2 presents a potential therapeutic strategy for prostate cancer immunoprevention.
