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Published on: July 25, 2020
Molecular cloning of canine Wilms' tumor 1 for immunohistochemical analysis in canine tissues
Osamu Sakai1, Masashi Sakurai2, Hiroki Sakai3
1Laboratory of Molecular Diagnostics and Therapeutics, The United Graduate School of Veterinary Medicine, Yamaguchi University, Yamaguchi 753-8515, Japan.
Abstract:
Wilms' tumor 1 (WT1) expression has been investigated in various human cancers as a target molecule for cancer immunotherapy. However, few studies have focused on WT1 expression in dogs. Firstly, cDNA of canine WT1 (cWT1) was molecularly cloned from normal canine kidney. The cross-reactivity of the anti-human WT1 monoclonal antibody (6F-H2) with cWT1 was confirmed via Western blotting using cells overexpressing cWT1. Immunohistochemical staining revealed that cWT1 expression was detected in all canine lymphoma tissues and in some normal canine tissues, including the kidney and lymph node. cWT1 is a potential immunotherapy target against canine cancers.
Insights
Wilms
Area of Science:
- Veterinary Oncology
- Molecular Biology
- Immunotherapy
Background:
- The Wilms' tumor 1 (WT1) gene is a validated target for cancer immunotherapy in humans.
- WT1 expression in canine cancers, particularly lymphoma, remains understudied.
- Understanding WT1 expression is crucial for developing novel canine cancer treatments.
Purpose of the Study:
- To clone and characterize canine WT1 (cWT1).
- To investigate cWT1 expression in canine lymphoma and normal tissues.
- To evaluate cWT1 as a potential target for canine cancer immunotherapy.
Main Methods:
- Molecular cloning of canine WT1 cDNA from normal canine kidney.
- Western blotting to confirm antibody cross-reactivity with cWT1.
- Immunohistochemical staining of canine lymphoma and normal tissues for cWT1 expression.
Main Results:
- Canine WT1 cDNA was successfully cloned.
- The anti-human WT1 antibody (6F-H2) demonstrated cross-reactivity with cWT1.
- cWT1 was expressed in all analyzed canine lymphoma samples.
- cWT1 expression was also detected in normal canine kidney and lymph node tissues.
Conclusions:
- Canine WT1 is successfully cloned and characterized.
- WT1 is expressed in canine lymphoma, indicating its potential as a therapeutic target.
- WT1-targeted immunotherapy could be a viable strategy for treating canine cancers.
