Nanoparticles for Selective Theranostic DNA Delivery to Liver Cancer

用于选择性治疗诊断 DNA 递送至肝癌的纳米颗粒

• 批准号: 10160637
• 负责人: Hannah Vaughan
• 金额: $ 2.41万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-16 至 2021-06-11
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10160637
• 关键词: AFP gene; Acyclovir; Address; Biocompatible Materials; Biodistribution; Cancer Etiology; Cell Death; Cells; Cessation of life; Chemotherapy and/or radiation; Cirrhosis; Clinical; DNA; DNA delivery; Dangerousness; Data; Development; Diagnosis; Diagnostic; Engineering; Enhancers; Esters; Excision; Formulation; Ganciclovir; Gene Delivery; Gene Expression; Genes; Genetic; Genetic Transcription; Goals; Guanine; Hepatocyte; Imaging Techniques; In Vitro; Incidence; Intravenous; Liver; Liver Failure; Liver diseases; Liver neoplasms; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of liver; Mediating; Methods; Modeling; Molecular Genetics; Mus; Nanotechnology; Operative Surgical Procedures; Outcome; PET/CT scan; Pathology; Patients; Plasmids; Positron-Emission Tomography; Pre-Clinical Model; Primary carcinoma of the liver cells; Prodrugs; Radiolabeled; Reporter Genes; Research; Research Training; Risk; Role; Solid Neoplasm; Specificity; System; TK Gene; Testing; Therapeutic; Thymidine Kinase; Tissues; Toxic effect; Transfection; United States; Work; X-Ray Computed Tomography; Xenograft procedure; alpha-Fetoproteins; anti-cancer; anticancer research; base; biodegradable polymer; cancer cell; career; cell killing; cell type; clinical development; clinical translation; cytotoxic; cytotoxicity; design; gene delivery system; gene product; gene therapy; genotoxicity; human model; imaging genetics; immunogenicity; in vitro activity; in vivo; innovation; insight; intravenous injection; liver transplantation; liver xenograft; mortality; nanocarrier; nanoparticle; nanoparticle delivery; neoplastic cell; next generation; novel; nucleic acid delivery; nucleoside analog; palliative; personalized approach; promoter; radiotracer; skills; suicide gene; targeted cancer therapy; targeted delivery; targeted treatment; theranostics; therapeutic DNA; therapeutic gene; therapeutically effective; tool; transgene expression; tumor; tumor ablation; tumor xenograft; vector; viral gene delivery

Our overall goal of this project is to develop a next-generation nanoparticle for targeted treatment of hepatocellular carcinoma (HCC). Liver cancer, including HCC, has the sixth highest cancer incidence and is the second leading cause of cancer mortality worldwide. While surgery or liver transplantation can be curative, most patients present with invasive HCC and are only eligible for palliative and locoregional treatments. Broadly cytotoxic treatments, including chemotherapy and radiation, have off-target toxicities to healthy hepatocytes, which is particularly dangerous to HCC patients due to a high incidence of underlying liver disease. Therefore, there is a critical need for a potent and highly targeted treatment option for liver cancer. To this end, we propose to develop a highly targeted nanoparticle gene delivery strategy for HCC tumors. Our preliminary data suggests that the biodegradable polymeric nanocarrier PBAE 536 can be used to selectively transfect HCC cells over healthy hepatocytes in vitro. Tumor-specific delivery of a therapeutic gene would enable targeted cancer cell killing with reduced off-target cytotoxicity to the surrounding healthy liver tissue. However, the therapeutic potential of this targeted nanotechnology has yet to be realized. The progress of clinical development is dependent on 1) optimization of nanoparticles for in vivo use and 2) the development of a safe, effective therapeutic DNA which is compatible with this delivery system. In this proposal, we will address both hurdles and test the final therapeutic in a preclinical model of human HCC. In Specific Aim 1, we will formulate and evaluate PBAE nanoparticles for gene delivery to orthotopic murine liver tumors. Nanoparticles will be systemically administered by intravenous injection, and off-target delivery and transfection will also be assessed. In Specific Aim 2, we will develop of a theranostic DNA vector targeted to HCC. We will use a thymidine kinase suicide gene (SR39), a dual-functional gene product that (1) converts the prodrug ganciclovir into a cancer-killing compound and (2) phosphorylates radiolabeled nucleoside analogs to enable PET imaging of gene expression. To enhance tumor targeting, we will employ α-fetoprotein (AFP) transcriptional targeting, which we expect will restrict SR39 expression to HCC cells. Engineered AFP- SR39 vectors will be assessed in vitro for therapeutic gene expression in HCC cells and off-target healthy cell types. In Specific Aim 3, we will combine PBAE nanoparticles with our novel transcriptionally targeted theranostic plasmid for testing in a murine xenograft liver tumor model. We hypothesize that systemically administered PBAE nanoparticles will enable SR39 delivery to HCC tumors for safe and effective cancer killing and molecular genetic imaging. If successful, this interdisciplinary project will result in a novel precision approach to HCC treatment which may be adapted for clinical use.

我们该项目的总体目标是开发下一代纳米颗粒，用于靶向治疗 肝细胞癌（HCC）。肝癌（包括 HCC）是第六位癌症发病率最高的癌症 全球癌症死亡的第二大原因。虽然手术或肝移植可以治愈，但大多数 患有侵袭性 HCC 的患者仅适合接受姑息治疗和局部治疗。广义上 细胞毒性治疗，包括化疗和放疗，对健康肝细胞具有脱靶毒性， 由于潜在肝病的发生率很高，这对 HCC 患者尤其危险。所以， 迫切需要一种有效且高度针对性的肝癌治疗方案。为此，我们建议 开发针对 HCC 肿瘤的高度靶向的纳米颗粒基因递送策略。我们的初步数据表明 生物可降解聚合物纳米载体PBAE 536可用于选择性转染HCC细胞 体外健康肝细胞。治疗基因的肿瘤特异性递送将使靶向癌细胞成为可能 杀伤并减少对周围健康肝组织的脱靶细胞毒性。然而，治疗 这种有针对性的纳米技术的潜力尚未被认识到。临床开发进展情况 取决于 1) 体内使用纳米颗粒的优化，以及 2) 安全、有效的纳米颗粒的开发 与该递送系统兼容的治疗性DNA。 在本提案中，我们将解决这两个障碍并在人类临床前模型中测试最终的治疗方法 肝癌。在具体目标 1 中，我们将制定和评估 PBAE 纳米粒子，用于将基因传递至原位 小鼠肝脏肿瘤。纳米颗粒将通过静脉注射进行全身给药，并且脱靶 还将评估递送和转染。在具体目标 2 中，我们将开发一种治疗诊断 DNA 载体 针对 HCC。我们将使用胸苷激酶自杀基因 (SR39)，这是一种双功能基因产物，(1) 将前药更昔洛韦转化为抗癌化合物，并且 (2) 磷酸化放射性标记的核苷 类似物可实现基因表达的 PET 成像。为了增强肿瘤靶向性，我们将采用甲胎蛋白 （法新社）转录靶向，我们预计这将限制 SR39 在 HCC 细胞中的表达。工程 AFP- 将在体外评估 SR39 载体在 HCC 细胞和脱靶健康细胞中的治疗基因表达 类型。在特定目标 3 中，我们将 PBAE 纳米粒子与我们的新型转录靶向结合起来 用于在小鼠异种移植肝肿瘤模型中进行测试的治疗诊断质粒。我们假设系统地 施用的 PBAE 纳米粒子将使 SR39 能够递送至 HCC 肿瘤，从而安全有效地杀死癌症 和分子遗传成像。如果成功，这个跨学科项目将带来一种新颖的精确方法 HCC 治疗可能适合临床使用。