Re-engineered Mitochondrially Targeted p53 Gene Therapy in Liver Cancer

重新设计的线粒体靶向 p53 基因疗法治疗肝癌

基本信息

批准号：
10317129
负责人：
Carol S. Lim
金额：
$ 20.96万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-01-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10317129
关键词：
Adenoviruses Aflatoxins Antibodies Apoptosis Apoptotic BAY 54-9085 Biological Models Bypass CCL15 gene Cancer Etiology Caspase Cell Aging Cell Nucleus Cells Cessation of life Chimeric Proteins China Cirrhosis Clinical DNA Repair Development Distant Dominant-Negative Mutation Drug resistance Engineering Excision Failure Genetic Genetic Status Goals HERC1 gene Hep3B Hepatic artery Hepatitis B Virus Hepatitis C virus Histologic Homo Human Immunotherapy In Vitro Incidence Liver Liver neoplasms Malignant Epithelial Cell Malignant Neoplasms Malignant neoplasm of liver Malignant neoplasm of ovary Mitochondria Modeling Modification Mus Mutate Neoplasm Metastasis Nivolumab Nuclear Operative Surgical Procedures Organ Other Genetics Outer Mitochondrial Membrane Ovarian Pathway interactions Patients Pharmaceutical Preparations Phenotype Prevention Primary carcinoma of the liver cells Proteins Publishing Signal Transduction Signaling Protein Specificity Survival Rate TP53 gene Testing Translations Transplantation Work Xenograft Model Xenograft procedure Zebrafish alpha-Fetoproteins anti-PD-1/PD-L1 cancer cell cell killing chemotherapeutic agent cytochrome c design effective therapy gain of function gene therapy hepatocellular carcinoma cell line hybrid gene hybrid protein improved in vivo kinase inhibitor liver cancer model liver transplantation mouse model mutant novel novel therapeutics offspring pre-clinical promoter senescence standard of care success targeted agent therapy design transcription factor tumor

项目摘要

Abstract: Hepatocellular carcinoma (HCC), the most common cancer of the liver, is the third leading cause of cancer death worldwide. In the U.S., the overall 5-year survival rate for HCC is only 17% and only 3% for distantly metastasized HCC. Surgical resection and liver transplantation are the current standard of care for HCC, but only ~5% qualify for resection due to cirrhosis. Limited organ availability also severely restricts the number of transplants. New targeted agents for HCC including kinase inhibitors and immunotherapies show promise, but patients using targeted agents will inevitably develop drug resistance. Effective new therapies for HCC are urgently needed. We propose a new, re-engineered p53 tumor suppressor-apoptotic protein hybrid for gene therapy of HCC called p53-Bad*. Wild-type p53 was approved for gene therapy as Gendicine® for cancers in China with limited success. Limited efficacy may be related to the dominant negative effect that mutant p53 exerts over wt p53. As nuclear p53 activity is dependent on homo-tetramerization, mutant p53 in cancer cells can also tetramerize with wt p53 to cause its inactivation. Mutant p53 can also have gain-of-function and other activities that can counter the effects of wt p53. To overcome this limitation, a potent p53 fusion protein with novel mechanisms of action is proposed here that overcomes dominant negative inhibition and causes apoptosis regardless of the p53 status of cancer cells. By fusing p53 to an apoptotic BH3 protein with an inherent mitochondrial targeting signal, such as Bad, this increases apoptosis further by combining the apoptotic effects of mitochondrial p53 with those of pro-apoptotic BH3 proteins. p53-Bad* (a mutant version of Bad with increased mitochondrial localization) is our top cancer cell-killing construct. We hypothesize that the combination of p53 and BH3 proteins at the mitochondria, with liver cancer specificity driven by a modified alpha fetoprotein (AFP) promoter, will potently induce apoptosis in liver cancer both in vitro (regardless of p53 status), and in vivo in zebrafish and mouse models. Zebrafish are an ideal model system for studying human cancer, with “histologic and genetic similarities between zebrafish and human tumors” while an orthotopic xenograft mouse model is an important step toward translation. The following aims are proposed: Aim 1: a. p53-BH3 fusion (p53-Bad*) will localize to the mitochondria and cause potent apoptosis in relevant hepatocellular carcinoma (HCC) cell lines regardless of cellular p53 status; b. the use of modified/combination AFP promoters will confer liver cancer cell specificity and kill HCC cells and not normal human cells. Aim 2: Expression of p53-Bad* under the liver specific promoter fabp10a in zebrafish, crossed with a liver cancer zebrafish model will produce offspring with reduced expression of liver cancer specific phenotypes. Aim 3: p53-Bad* (expressed in adenovirus with the optimized HCC specific promoter) will shrink liver tumors in an orthotopic xenograft mouse model of HCC (using AFP- expressing human HCC liver cells, Hep3B). The ultimate goal of this study is the development of a new, effective therapy for treatment of HCC patients worldwide.

【摘要】：肝细胞癌(HCC)是最常见的肝癌,是导致肝癌的第三大原因。在美国，HCC 的总体 5 年生存率仅为 17%，远处的癌症死亡率仅为 3%。手术切除和肝移植是目前治疗 HCC 的标准方法。由于肝硬化，只有约 5% 的患者有资格进行切除，这也严重限制了器官的数量。包括激酶抑制剂和免疫疗法在内的 HCC 新靶向药物显示出前景，但使用靶向药物的患者将不可避免地产生耐药性，有效的 HCC 新疗法正在出现。我们提出了一种新的、重新设计的 p53 肿瘤抑制-凋亡蛋白杂交基因。 HCC 疗法称为 p53-Bad* 野生型 p53 被批准用于癌症基因疗法 Gendicine®。在中国取得的成功有限，可能与突变体 p53 的显性负面影响有关。由于核 p53 活性依赖于癌细胞中的同源四聚化，因此突变型 p53 发挥作用。还可以与wt p53四聚化，导致其失活，突变体p53还可以具有功能获得性等。为了克服这一限制，我们开发了一种有效的 p53 融合蛋白。这里提出了新的作用机制，克服了显性负性抑制和细胞凋亡的原因无论癌细胞的 p53 状态如何，通过将 p53 与具有固有的凋亡 BH3 蛋白融合。线粒体靶向信号，例如 Bad，这通过结合细胞凋亡效应进一步增加细胞凋亡线粒体 p53 与促凋亡 BH3 蛋白的 p53-Bad*（Bad 的突变版本，增加）。线粒体定位）是我们最重要的癌细胞杀伤结构，我们已经捕获了 p53 的组合。线粒体中的 BH3 蛋白，具有由修饰的甲胎蛋白 (AFP) 驱动的肝癌特异性启动子，将在体外（无论 p53 状态如何）和体内在肝癌中有效诱导细胞凋亡斑马鱼和小鼠模型是研究人类癌症的理想模型系统，具有“组织学特征”。以及斑马鱼和人类肿瘤之间的遗传相似性”，而原位异种移植小鼠模型是一种提出以下目标：目标 1：p53-BH3 融合（p53-Bad*）定位于线粒体并引起相关肝细胞癌 (HCC) 细胞系的有效凋亡无论细胞 p53 状态如何，使用修饰/组合 AFP 启动子都会产生肝癌细胞。特异性并杀死 HCC 细胞而不是正常人类细胞目标 2：p53-Bad* 在肝脏特异性下的表达。斑马鱼中的启动子 fabp10a 与肝癌斑马鱼模型杂交将产生具有减少的后代肝癌特异性表型的表达目标 3：p53-Bad*（在腺病毒中表达，并经过优化） HCC 特异性启动子）将缩小 HCC 原位异种移植小鼠模型中的肝脏肿瘤（使用 AFP- 这项研究的最终目标是开发一种新的、有效的表达人类 HCC 肝细胞（Hep3B）的方法。治疗全球 HCC 患者的疗法。