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Targeting lung tumors expressing mutant p53 with oncolytic viruses and suicide genes

用溶瘤病毒和自杀基因靶向表达突变型 p53 的肺部肿瘤

基本信息

批准号：
10159871
负责人：
Sumitra Deb
金额：
$ 47.41万
依托单位：
VIRGINIA COMMONWEALTH UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10159871
关键词：
Adenovirus Vector Adenoviruses Affect Alleles Amino Acids Automobile Driving Base Sequence Biological Assay Biological Models Bystander Effect Cell Death Cells Cessation of life Cloning Cytolysis Cytomegalovirus Cytosine deaminase DNA Sequence Early Promoters Exposure to Ganciclovir Gene Activation Gene Expression Gene Proteins Genes Goals Growth Herpesvirus 1 Human Lung Lung Neoplasms Lytic Lytic Phase Malignant Neoplasms Malignant neoplasm of lung Mediating Modeling Mus Mutation Mutation Analysis Normal Cell Oncogenic Oncogenic Viruses Oncolytic Oncolytic viruses Phenotype Process Prodrugs Property Proteins Reporter Research Response Elements Specificity Suicide System TP53 gene Technology Testing Therapeutic Thymidine Kinase Time Transactivation Viral Virus Virus Replication Work Xenograft procedure base cancer cell cell type gain of function gene cloning improved individual response innovation lung cancer cell mouse model mutant novel null mutation oncolysis oncolytic adenovirus patient derived xenograft model promoter response suicide gene suicide virus targeted treatment therapeutic target thymidine kinase 1 tumor tumor xenograft vector weapons

项目摘要

Mutations of the p53 gene are found in the majority of lung cancers and most of these mutations are single amino acid changes instilling gain-of-function (GOF) oncogenic phenotypes, making the GOF p53 oncoprotein an excellent cancer therapeutic target. We propose a radically different approach to current GOF p53 targeting therapeutic concepts in which instead of inhibiting the protein, we weaponize GOF p53 in promoting lung cancer cell death, either by suicide, viral lysis, or both, while leaving normal cells unscathed. This innovative strategy is possible based on our discovery of a unique transactivation mechanism for GOF p53, and from that, our creation of a GOF p53 inducible promoter. Our GOF p53 inducible promoter directs expression of any gene cloned downstream only if the cell has a GOF p53 mutation, with wild-type (WT) p53 having no effect on the promoter and cells with WT p53 or p53 null mutations showing no expression. For our first major goal, we propose using an exciting new oncolytic virus that only replicates, propagates, and kills cancer cells with GOF p53 while having no effect on normal cells. We have placed two adenoviral early genes, E1A and E1B, the genes needed for adenoviral replication, under the control of the GOF p53 inducible promoter within an adenoviral vector. Initial studies show that this virus has remarkable oncolytic ability and specificity for lung cancer cells with GOF p53, with no effect or viral growth whatsoever in cells with WT p53. The killing effects in xenograft tumors with GOF p53 appear as though there is sustained accelerated tumor killing after a short delay of when the virus is injected. We propose to enhance the oncolytic virus by adding additional lysis abilities and by combining the suicide strategy with the oncolytic strategy. Preliminary results look very promising for this combination. For our second goal, we propose devising a means of specifically killing lung cancer cells with GOF p53 mutations by cloning a suicide gene downstream of our GOF p53 inducible promoter. This construct will be introduced into an adenoviral vector so that when the virus infects cells, only cells with a GOF p53 mutation (cancer cells) will die from prodrug treatment. We have created such a virus using the Herpes Thymidine Kinase suicide gene and show striking killing effects and specificity for lung cancer cells with GOF p53 both in culture and in xenograft tumors. We aim to further discover how this strategy works and ways to improve it. We propose the use of the bacterial Cytosine Deaminase suicide gene (bCD) to enhance the bystander effect of our GOF p53 specific suicide virus. In addition, we propose to improve the inducibility of our GOF p53 inducible promoter to further enhance the suicide and oncolytic viruses. The potential impact of this work is far-reaching since these strategies should be applicable for any cancer with GOF p53 mutations, which constitutes over half of all cancers.

p53基因的突变在大多数肺癌中都有发现，而且这些突变大多是单一的