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：肝癌（HCC）是肝癌最常见的癌症，是第三个主要原因全球癌症死亡。在美国，HCC的总5年生存率仅为17％，距离仅为3％转移的HCC。手术切除和肝移植是HCC的当前护理标准，但由于肝硬化，只有约5％的人有资格切除。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的新靶向剂显示出希望，但使用靶向剂的患者不可避免地会发展出耐药性。 HCC有效的新疗法是迫切需要。我们提出了一种新的，重新设计的p53肿瘤 - 抑制型凋亡蛋白杂交基因 HCC的治疗称为p53-bad*。野生型p53被批准用于基因治疗，作为癌症的Gendicine® 中国成功有限。有限的有效性可能与突变体p53的主要负面影响有关在WT P53上发挥作用。由于核p53活性取决于同型四聚体，因此癌细胞中的突变体p53 还可以用WT p53四焦点引起其失活。突变体p53也可以具有功能奖励和其他可以应对WT p53影响的活动。为了克服这一限制，潜在的p53融合蛋白与这里提出了新的作用机制，以克服主导的负面抑制并导致凋亡不管癌细胞的p53状态如何。通过将p53融合到凋亡的BH3蛋白与继承线粒体靶向信号（例如不良）通过结合凋亡作用而进一步增加凋亡线粒体p53和促凋亡BH3蛋白的p53。 p53-bad*（不良的突变版，增加线粒体定位）是我们最大的癌细胞杀伤构建体。我们假设p53的组合线粒体处的BH3蛋白，其特异性由改性α胎儿蛋白（AFP）驱动启动子，可能会在体外诱导肝癌凋亡（无论p53状态如何）和体内斑马鱼和鼠标模型。斑马鱼是研究人类癌症的理想模型系统，“组织学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。提出了以下目的：目标1： p53-BH3融合（p53-bad*）将位于线粒体并引起相关肝细胞癌（HCC）细胞系的潜在凋亡不论细胞p53状态如何； b。修改/组合AFP启动子的使用将召集肝癌细胞特异性并杀死HCC细胞，而不是正常的人类细胞。 AIM 2：肝脏特异性下的p53-bad*的表达斑马鱼中的启动子Fabp10a与肝癌斑马模型交叉，将产生后代，并减少肝癌特异性表型的表达。 AIM 3：P53-BAD*（用优化的腺病毒在腺病毒中表达 HCC特异性启动子）将在HCC的原位型小鼠模型中收缩肝肿瘤（使用AFP- 表达人HCC肝细胞，Hep3b）。这项研究的最终目标是开发新的有效的全球HCC患者治疗的治疗。