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: 10217062
• 负责人: KLAUS H KAESTNER
• 金额: $ 54.43万
• 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/10217062
• 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; 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/antagonist; innovation; 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，其中包括肝细胞增殖， 而且 - 不幸的是 - 肝细胞癌（HCC）的风险增加，是​​该癌的先决条件 开发用于慢性肝病和改善癌症预防的新型临床治疗方法。现存的 HCC（例如索拉非尼）的药物疗法仅将患者的生存率延长了三个月。我们最近开发了 一个大规模平行的体内筛查平台，用于测试遗传因素的影响，例如全长cDNA或 miRNA在肝脏再生和肿瘤发生上。我们已经使用了这种筛选技术来构建所有人的地图 活跃于肝脏再生的miRNA。在这里，我们建议利用这种创新的范式进行 对135个最丰富但进化上构成肝的影响的全面评估 miRNA在毒性肝损伤和HCC肿瘤发生中恢复的过程中。在特定目标1中，我们将 确定我们先前屏幕上有关肝回生量的三个miRNA的综合益处 有毒损伤是迈向使用miRNA模拟药物治疗肝脏疾病的一步。这将是 通过单独和所有组合的miRNA编码质粒或纳米颗粒的递送完成。在 具体目标2，我们将确定肝miRNA和miRNA组合对HCC肿瘤的影响 体内发展。为此，我们开发了两种在小鼠中快速HCC开发的模型，其中 我们将筛选我们的135个“强硬诱饵”（“ Tud's”）或miRNA作用抑制剂的图书馆。 我们将使用肿瘤的肝脏中使用高吞吐量测序来量化所有TUD的抽象 与输入库相比。 TUD在肿瘤形成靶标miRNA后富集，通常限制肿瘤 生长，发现促进肿瘤发生的靶标miRNA较少。然后我们将测试 全身传递后，最潜在的miRNA对肿瘤形成的影响。具体 AIM 3我们将执行有条件的miRNA屏幕，该屏幕会影响索拉非尼的抵抗以识别新颖 预防或治疗HCC的组合处理。在一起，我们强大的遗传屏幕 承诺确定可用于治疗急性肝损伤的miRNA效应，并在 结合索拉非尼，作为防止HCC计划和进展的更有效的治疗方法。