Mitochondrially Targeted p53 DBD for Treatment of Ovarian Cancer

线粒体靶向 p53 DBD 治疗卵巢癌

基本信息

批准号：
8957167
负责人：
Carol S. Lim
金额：
$ 16.2万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-07 至 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8957167
关键词：
Address Adenovirus Vector Adenoviruses Apoptosis Apoptotic Area BCL2 gene Biological Bypass Cancer Model Cancer cell line Carboplatin Carcinoma Caring Caspase Cell Nucleus Cells Cessation of life Clinical Trials Coupled Cytotoxic agent DNA Binding Domain Diagnosis Dimerization Disease Dominant-Negative Mutation Engineering Epigenetic Process Failure Family member Genes Genetic Goals Gynecologic Heterogeneity Integrins Intraperitoneal Injections Malignant Neoplasms Malignant neoplasm of ovary Membrane Methods Mitochondria Mus Mutation Nuclear Oncogenes Outer Mitochondrial Membrane Ovarian Paclitaxel Pathway interactions Patients Protein p53 Proteins Recurrence Relapse Reporting Serous Signal Pathway Signal Transduction Staging System TP53 gene Testing Therapeutic Transcriptional Activation Tropism Tumor Suppressor Genes Woman Work base bcl-xlong protein cancer cell cancer surgery cancer therapy chemotherapy design gene therapy killings loss of function mutation monomer mutant neoplastic cell neutralizing antibody novel ovarian neoplasm poly(ethylenimine)-co-(N-(2-aminoethyl) ethyleneimin)-co-N-(N-cholesteryloxycarbonyl-(2-aminoethyl)ethylenimine)preclinical study promoter protein expression public health relevance receptor response standard of care transcription factor

项目摘要

DESCRIPTION (provided by applicant): Ovarian cancer continues to be the most lethal gynecological malignancy with 69% of patients succumbing to their disease. Heterogeneity of ovarian cancer is a formidable challenge, suggesting that targeting multiple biological pathways will almost certainly be required for disease treatment. However, mutations/loss of function in p53 tumor suppressor gene occurs in >96% of all cases of high-grade serous carcinomas (HGSC), the most common form of ovarian cancer. While much work using p53 has focused on its effects as a transcription factor in the nucleus, this proposal will exploit the direct, cell-klling effect that p53 has at the mitochondria. p53's rapid effects at the mitochondria represent the shortest pathway for executing p53 death signaling, which triggers apoptosis. The goal of this proposal is to exploit the p53 intrinsic, direct apoptotic pathway at the mitochondria for use in ovarian cancer therapy. Mitochondrially targeted domains of p53 have never been attempted for ovarian cancer therapy, unlike wild-type p53. We propose to: A) re-engineer a small, monomeric domain of p53 with a mitochondrial targeting signal (MTS) that is highly potent, that kills any cancer cell regardless of p53 status or genetics, and bypasses the dominant negative effect in cancer cells, and B) use a polymeric WSLP delivery system (currently in clinical trials) for effective delivery to cells. Our central hypothesis is that re-engineered, mitochondrially targeted p53 with a proven delivery method administered intraperitoneally will be effective against ovarian cancer. The advantage of using a mitochondrial targeted protein encoded by a gene, rather than a cytotoxic agent [21] is the ability to incorporate a promoter for cancer specific expression of that protein. Ultimately, correction of the p53 pathway and activation of apoptosis may be a universal approach: functional, mitochondrially targeted monomeric p53 re-introduced into cancer cells would act as a "sledgehammer," effective under any circumstances (regardless of genetics or the pathway upon which the cancer develops). Three aims are proposed: Aim 1: Design a novel apoptotic gene therapy construct (called DBD-MTS) based on the DBD of p53, coupled to an optimal mitochondrial targeting signal (MTS), driven by a cancer-specific promoter, capable of activating intrinsic apoptosis. Aim 2: Determine the apoptotic potential of DBD-MTS in ovarian cancer cell lines with varying p53 status, individually, and in combo with standard of care, carboplatin and paclitaxel (CPTX). Aim 3: Deliver DBD-MTS using water soluble lipopolymer (WSLP) currently being used in clinical trials by intraperitoneal injection, ino our new syngeneic orthotopic metastatic mouse ovarian cancer model (developed by our collaborator), alone and in combination with carboplatin and paclitaxel (CPTX). If successful, we anticipate this work to advance to a new clinical trial initiated for women diagnosed with late stage (>III) or recurrent high grade serous ovarian cancer and advance the largely unexplored area of mitochondrially targeted p53 gene therapy.

描述（由适用提供）：卵巢癌仍然是最致命的妇科恶性肿瘤，有69％的患者屈服于他们的疾病。卵巢癌的异质性是一个巨大的挑战，表明靶向多种生物学途径几乎肯定是疾病治疗所必需的。然而，p53肿瘤抑制基因中的突变/功能丧失发生在所有高级浆液性癌（HGSC）的情况中，> 96％是卵巢癌的最常见形式。尽管使用p53的大量工作集中在其作为核中转录因子的效果上，但该建议将利用p53在线粒体上具有的直接，细胞锁定作用。 p53在线粒体上的快速影响是执行p53死亡信号传导的最短途径，这会触发凋亡。该提案的目的是利用线粒体的p53固有的直接凋亡途径用于卵巢癌治疗。与野生型p53不同，从未尝试过p53的线粒体靶向结构域进行卵巢癌治疗。 We propose to: A) re-engineer a small, monomeric domain of p53 with a mitochondrial targeting signal (MTS) that is highly potential, that kills any cancer cell regardless of p53 status or genetics, and bypasses the dominant negative effect in cancer cells, and B) use a polymeric WSLP delivery system (currently in clinical trials) for effective delivery to cells.我们的中心假设是重新设计的线粒体靶向 p53采用腹膜内施用的经过验证的递送方法将有效地抵抗卵巢癌。使用由基因编码的线粒体靶向蛋白而不是细胞毒性剂[21]的优点是，能够将启动子纳入该蛋白的癌症特异性表达。最终，对p53途径的校正和凋亡的激活可能是一种普遍的方法：功能性的，线粒体靶向的单体p53重新引入癌细胞的p53在任何情况下都可以作为“大锤”，在任何情况下有效（无论遗传学或癌症发展的途径如何）。提出了三个目的：目标1：基于p53的DBD设计一种新型的凋亡基因治疗构建体（称为DBD-MTS），与能够激活固有凋亡的癌症特异性启动子驱动的最佳线粒体靶向信号（MTS）耦合。 AIM 2：确定DBD-MT在具有不同状态的卵巢癌细胞系中的凋亡潜力，单独，并与标准的护理，卡泊肽和紫杉醇（CPTX）组合。 AIM 3：使用腹膜内注射中当前在临床试验中使用的水固体脂聚合物（WSLP）传递DBD-MT，INO INO我们的新型Syngeneic Orthopic转移性小鼠卵巢癌模型（由我们的合作者开发），单独开发，与Carboplatin和carboplatin和pacplitaxel（CPTX）相结合。如果成功的话，我们预计这项工作将促进针对诊断为晚期（> III）或经常性高级浆液卵巢癌的妇女发起的新临床试验，并推进了线粒体靶向p53基因疗法的意外领域。