Innovative Genetic Approaches to Enhance Liver Repopulation and Reduce Cancer Risk and Progression

增强肝脏再生并降低癌症风险和进展的创新遗传学方法

• 批准号: 10434813
• 负责人: KLAUS H KAESTNER
• 金额: $ 53.34万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-15 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10434813
• 关键词: Acute; Acute Liver Failure; Attenuated; BAY 54-9085; Biological Models; Clinical Treatment; Complementary DNA; DNA; Data; Development; Disease; Early Diagnosis; Evaluation; FDA approved; Genes; Genetic; Genetic Screening; Genotype; Grant; Growth; Health; Hepatic; Hepatocyte; High-Throughput Nucleotide Sequencing; Human; Individual; Injury; Investigation; Knockout Mice; Length; Libraries; Liver; Liver Extract; Liver Regeneration; Liver diseases; Malignant Neoplasms; Malignant neoplasm of liver; Maps; MicroRNAs; Modeling; Mus; Mutation; Natural regeneration; Neoplasms; Oncogenes; Oncogenic; Partial Hepatectomy; Pathway interactions; Patients; Pharmacotherapy; Phenotype; Plasmids; Porifera; Prevention; Primary carcinoma of the liver cells; Process; Prognosis; Recovery; Resistance; Risk; Role; Survival Rate; TP53 gene; Technology; Testing; The Cancer Genome Atlas; Toxic Hepatitis; Transforming Growth Factor alpha; Tumor Burden; Tyrosine Kinase Inhibitor; United States; Xenograft Model; Xenograft procedure; acetaminophen overdose; acute liver injury; base; cancer prevention; cancer risk; chronic liver disease; chronic liver injury; effective therapy; genetic approach; genetic makeup; hepatocellular carcinoma cell line; improved; in vivo; inhibitor; innovation; liver cancer model; liver cell proliferation; liver injury; mimetics; mouse model; nanoparticle; new combination therapies; novel; novel therapeutics; overexpression; prevent; response; screening; small hairpin RNA; therapeutic target; tumor; tumor growth; tumor initiation; tumor progression; tumorigenesis

A better understanding of the liver’s response to toxic injury, which includes hepatocyte proliferation, and – unfortunately – an increased risk for hepatocellular carcinoma (HCC), is a prerequisite for the development of novel clinical treatments for chronic liver disease and improved cancer prevention. Existing drug therapies for HCC such as sorafenib extend patient survival by only three months. We recently developed a massively parallel in vivo screening platform to test the impact of genetic factors such as full-length cDNAs or miRNAs on liver repopulation and tumorigenesis. We have used this screening technology to build a map of all miRNAs active in liver regeneration. Here, we propose to exploit this innovative paradigm to conduct a comprehensive evaluation of the effects of the 135 most abundant but evolutionarily conserved hepatic miRNAs on the processes of recovery from toxic liver injury and HCC tumorigenesis. In Specific Aim 1, we will determine the combined benefits of three miRNAs identified in our prior screen on liver repopulation following toxic injuries, as a step toward using miRNA-mimetic drug therapies for liver diseases. This will be accomplished through delivery of miRNA-encoding plasmids or nanoparticles singly and in all combination. In Specific Aim 2, we will determine the impact of hepatic miRNAs and miRNA combinations on HCC tumor development in vivo. To this end, we have developed two models of rapid HCC development in mice, in which we will screen our library of 135 ‘tough decoys’ (“TuD’s”), or inhibitors of miRNA action, on tumor formation. We will quantify the abundance of all TuD’s using high throughput sequencing in the tumor-loaded liver compared to the input library. TuD’s enriched in after tumor formation target miRNAs that normally limit tumor growth, and those found less abundant target miRNAs that promote tumorigenesis. We will then test the combinations of the most potent miRNA effectors on tumor formation following systemic delivery. In Specific Aim 3 we will perform a conditional screen of miRNAs that impact Sorafenib resistance to identify novel combination treatments for the prevention or treatment of HCC. Together, our powerful genetic screens promise to identify miRNA effectors that can be employed for the treatment of acute liver injury and, in combination with Sorafenib, as a more effective treatment to prevent HCC initiation and progression.

更好地了解肝脏对毒性损伤的反应，包括肝细胞增殖， 不幸的是，肝细胞癌（HCC）的风险增加，是治疗的先决条件。 开发慢性肝病的新型临床治疗方法和改善癌症预防。现有 用于HCC的药物疗法如索拉非尼仅延长患者存活3个月。我们最近开发 一个大规模平行的体内筛选平台，以测试遗传因子如全长cDNA或 miRNAs对肝脏再增殖和肿瘤发生的影响。我们用这种筛选技术建立了一个地图 miRNAs在肝再生中的活性。在这里，我们建议利用这种创新的范式来进行一个 对135个最丰富但进化上保守的肝细胞的影响进行了综合评价， miRNAs对中毒性肝损伤和HCC肿瘤发生恢复过程的影响具体目标1： 确定我们先前筛选中鉴定的三种miRNAs对肝脏再生的联合益处， 毒性损伤，作为使用miRNA模拟药物治疗肝脏疾病的一步。这将是 通过单独和全部组合递送编码miRNA的质粒或纳米颗粒来实现。在 具体目标2，我们将确定肝脏miRNA和miRNA组合对HCC肿瘤的影响 体内发育为此，我们开发了两种小鼠肝癌快速发展模型，其中 我们将筛选我们的135个“坚韧诱饵”（“TuD”）库，或抑制肿瘤形成的miRNA作用。 我们将使用高通量测序在肿瘤负载的肝脏中定量所有TuD的丰度 与输入库相比。在肿瘤形成后富集的TuD靶向通常限制肿瘤形成的miRNA 生长，以及那些发现不太丰富的促进肿瘤发生的靶miRNA。然后我们将测试 在全身性递送后对肿瘤形成最有效的miRNA效应物的组合。在特定 目标3我们将对影响索拉非尼耐药性的miRNA进行条件性筛选，以识别新的 用于预防或治疗HCC的组合治疗。我们强大的基因筛选系统 有望鉴定出可用于治疗急性肝损伤的miRNA效应物， 与索拉非尼联合，作为预防HCC发生和进展的更有效治疗。