PUBLICATION
p53-Bad* Fusion Gene Therapy Induces Apoptosis In Vitro and Reduces Zebrafish Tumor Burden in Hepatocellular Carcinoma
- Authors
- Bowman, K.E.R., Ahne, L., O'Brien, L., Vander Mause, E.R., Lu, P., Wallis, B., Evason, K.J., Lim, C.S.
- ID
- ZDB-PUB-221210-7
- Date
- 2022
- Source
- Molecular pharmaceutics 20(1): 331-340 (Journal)
- Registered Authors
- Evason, Kimberley
- Keywords
- Bcl-2 antagonist of cell death, liver cancer, mitochondrial apoptosis, p53-Bad* fusion, tumor suppressor p53
- MeSH Terms
-
- Animals
- Apoptosis/genetics
- Apoptosis Regulatory Proteins/genetics
- Carcinoma, Hepatocellular*/genetics
- Carcinoma, Hepatocellular*/metabolism
- Carcinoma, Hepatocellular*/therapy
- Cell Line, Tumor
- Genetic Therapy
- Liver Neoplasms*/genetics
- Liver Neoplasms*/metabolism
- Liver Neoplasms*/therapy
- Tumor Burden
- Tumor Suppressor Protein p53/genetics
- Tumor Suppressor Protein p53/metabolism
- Zebrafish/genetics
- PubMed
- 36490361 Full text @ Mol. Pharm.
Citation
Bowman, K.E.R., Ahne, L., O'Brien, L., Vander Mause, E.R., Lu, P., Wallis, B., Evason, K.J., Lim, C.S. (2022) p53-Bad* Fusion Gene Therapy Induces Apoptosis In Vitro and Reduces Zebrafish Tumor Burden in Hepatocellular Carcinoma. Molecular pharmaceutics. 20(1):331-340.
Abstract
With few curative treatments and a global yearly death rate of over 800,000, hepatocellular carcinoma (HCC) desperately needs new therapies. Although wild-type p53 gene therapy has been shown to be safe in HCC patients, it has not shown enough efficacy to merit approval. This work aims to show how p53 can be re-engineered through fusion to the pro-apoptotic BH3 protein Bcl-2 antagonist of cell death (Bad) to improve anti-HCC activity and potentially lead to a novel HCC therapeutic, p53-Bad*. p53-Bad* is a fusion of p53 and Bad, with two mutations, S112A and S136A. We determined mitochondrial localization of p53-Bad* in liver cancer cell lines with varying p53 mutation statuses via fluorescence microscopy. We defined the apoptotic activity of p53-Bad* in four liver cancer cell lines using flow cytometry. To determine the effects of p53-Bad* in vivo, we generated and analyzed transgenic zebrafish expressing hepatocyte-specific p53-Bad*. p53-Bad* localized to the mitochondria regardless of the p53 mutation status and demonstrated superior apoptotic activity over WT p53 in early, middle, and late apoptosis assays. Tumor burden in zebrafish HCC was reduced by p53-Bad* as measured by the liver-to-body mass ratio and histopathology. p53-Bad* induced significant apoptosis in zebrafish HCC as measured by TUNEL staining but did not induce apoptosis in non-HCC fish. p53-Bad* can induce apoptosis in a panel of liver cancer cell lines with varying p53 mutation statuses and induce apoptosis/reduce HCC tumor burden in vivo in zebrafish. p53-Bad* warrants further investigation as a potential new HCC therapeutic.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping