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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。全球癌症死亡人数。在美国，HCC 的总体 5 年生存率仅为 17%，远处转移的生存率仅为 3% 转移性肝癌。手术切除和肝移植是目前 HCC 的标准治疗方法，但只有约 5% 的患者因肝硬化而有资格接受切除术。有限的器官可用性也严重限制了移植。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不可避免地会产生耐药性。 HCC 的有效新疗法是急需。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 被批准用于基因治疗，称为 Gendicine®，用于治疗癌症中国的成功有限。有限的功效可能与突变体 p53 的显性负效应有关作用于wt p53。由于核 p53 活性依赖于同源四聚化，癌细胞中的突变 p53 也可与 wt p53 四聚化，导致其失活。突变的 p53 还可以具有功能获得性和其他功能可以对抗 wt p53 影响的活动。为了克服这一限制，一种有效的 p53 融合蛋白本文提出了克服显性负性抑制并引起细胞凋亡的新作用机制无论癌细胞的 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 蛋白的关系。 p53-Bad*（Bad 的突变版本，增加线粒体定位）是我们顶级的癌细胞杀伤结构。我们假设 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。斑马鱼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迈向翻译的重要一步。建议实现以下目标：目标 1：a． p53-BH3 融合 (p53-Bad*) 将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； b.使用修饰/组合 AFP 启动子将赋予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：肝脏特异性下p53-Bad*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- 表达人 HCC 肝细胞，Hep3B）。本研究的最终目标是开发一种新的、有效的治疗全球 HCC 患者的疗法。