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Updated: Jun 11, 2025
Comparing Metastatic Clear Cell Renal Cell Carcinoma Model Established in Mouse Kidney and on Chicken Chorioallantoic Membrane
Published on: February 8, 2020
Toward a CRISPR-based mouse model of Vhl-deficient clear cell kidney cancer: Initial experience and lessons learned
Laura A Stransky1, Wenhua Gao1, Laura S Schmidt2,3
1Division of Molecular and Cellular Oncology, Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02215.
Abstract:
CRISPR is revolutionizing the ability to do somatic gene editing in mice for the purpose of creating new cancer models. Inactivation of the VHL tumor suppressor gene is the signature initiating event in the most common form of kidney cancer, clear cell renal cell carcinoma (ccRCC). Such tumors are usually driven by the excessive HIF2 activity that arises when the VHL gene product, pVHL, is defective. Given the pressing need for a robust immunocompetent mouse model of human ccRCC, we directly injected adenovirus-associated viruses (AAVs) encoding sgRNAs against VHL and other known/suspected ccRCC tumor suppressor genes into the kidneys of C57BL/6 mice under conditions where Cas9 was under the control of one of two different kidney-specific promoters (Cdh16 or Pax8) to induce kidney tumors. An AAV targeting Vhl, Pbrm1, Keap1, and Tsc1 reproducibly caused macroscopic ccRCCs that partially resembled human ccRCC tumors with respect to transcriptome and cell of origin and responded to a ccRCC standard-of-care agent, axitinib. Unfortunately, these tumors, like those produced by earlier genetically engineered mouse ccRCCs, are HIF2 independent.
Insights
CRISPR technology enables new mouse cancer models. Researchers created a clear cell renal cell carcinoma (ccRCC) model using CRISPR gene editing, which partially mimics human tumors and responds to axitinib treatment.
Area of Science:
- Oncology
- Genetics
- Molecular Biology
Background:
- Clear cell renal cell carcinoma (ccRCC) is the most common kidney cancer.
- VHL gene inactivation drives ccRCC, leading to excessive HIF2 activity.
- There is a need for immunocompetent mouse models of human ccRCC.
Purpose of the Study:
- To develop a robust immunocompetent mouse model of human ccRCC using CRISPR-Cas9 somatic gene editing.
- To investigate the efficacy of targeting multiple tumor suppressor genes in ccRCC development.
- To assess the response of the developed ccRCC model to standard-of-care treatment.
Main Methods:
- Adenovirus-associated viruses (AAVs) encoding sgRNAs against VHL and other ccRCC-associated genes were injected into mouse kidneys.
- Cas9 expression was controlled by kidney-specific promoters (Cdh16 or Pax8).
- Tumor development, transcriptome, cell of origin, and response to axitinib were analyzed.
Main Results:
- Targeting VHL, Pbrm1, Keap1, and Tsc1 reproducibly induced macroscopic ccRCCs in mice.
- The generated ccRCC tumors partially resembled human ccRCC in transcriptome and cell of origin.
- The mouse ccRCC model responded to the standard-of-care agent axitinib.
- The developed ccRCC tumors were HIF2 independent.
Conclusions:
- CRISPR-mediated somatic gene editing can create a relevant immunocompetent mouse model for ccRCC.
- This model partially recapitulates human ccRCC characteristics and treatment response.
- The model's HIF2 independence highlights a difference from typical human ccRCC pathogenesis.
