ORAL GENE THERAPY WITH NiMOS FOR INFLAMMATORY BOWEL DISEASE

NiMOS 口服基因疗法治疗炎症性肠病

• 批准号: 7779418
• 负责人: Mansoor M Amiji
• 金额: $ 33.03万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2012-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7779418
• 关键词: Acute; Affect; Age; American; Anti-Inflammatory Agents; Anti-inflammatory; Biodistribution; Biology; Caliber; Cells; Charge; Chronic; Clinical; Clinical Investigator; Clinical Research; Colitis; Crohn' s disease; DNA; DNA Maintenance; Development; Disease; Disease Management; Disorder by Site; Drug Formulations; Emulsions; Encapsulated; Endocytosis; Enzymes; Epithelium; Equilibrium; Etiology; Evaluation; Exposure to; Fasting; Gelatin; Gene Delivery; Gene Expression; Immune; In Vitro; Inflammatory; Inflammatory Bowel Diseases; Interdisciplinary Study; Interleukin-10; Intestines; Laboratories; Large Intestine; Lead; Lipase; Literature; Medicine; Methods; Microspheres; Morphology; Mus; Names; Nanotechnology; Non-Viral Vector; Oral; Oral Administration; Particle Size; Pattern; Peptide Hydrolases; Pharmaceutical Preparations; Physiological; Plasmids; Principal Investigator; Proteins; Rattus; Rectal Administration; Reporter; Request for Applications; Research; Research Personnel; Resources; Role; Solvents; Surface; System; TNFR-Fc fusion protein; Therapeutic; Time; Transfection; Treatment Efficacy; Tumor Necrosis Factor Receptor; Tumor Necrosis Factor-alpha; Ulcerative Colitis; Vaccination; Viral; alternative treatment; base; beta-Galactosidase; crosslink; cytokine; effective therapy; enhanced green fluorescent protein; gene therapy; human TNFRSF1A protein; innovation; interleukin-23; nanoparticle; nanoscience; non-viral gene delivery; novel; particle; plasmid DNA; poly(epsilon-caprolactone); pre-clinical; programs; receptor; residence; response; therapeutic effectiveness; therapeutic gene; therapeutic protein; transgene expression; treatment strategy; tumor necrosis factor alpha receptor; uptake; vector

DESCRIPTION (provided by applicant): Inflammatory bowel disease (IBD) affects as many as 100 per 100,000 Americans under the age of 30. Recent studies suggest that IBD may be caused by alterations in the delicate balance between pro- and anti- inflammatory cytokines in the gastro-intestinal (GI) tract. The role of tumor necrosis factor-alpha (TNF) and interleukin-10 (IL-10) as pro- and anti-inflammatory cytokines, respectively, has been examined in preclinical and clinical studies. In the proposed study, an interdisciplinary team of investigators will evaluate oral gene therapy using soluble TNF receptor-1 (sTNF-R1) and IL-10 expressing plasmid DNA in acute colitis-induced Balb/c mice. The plasmid DNA will be encapsulated in gelatin nanoparticles, which are further protected by a poly(epsilon- caprolactone (PCL) matrix to form Nanoparticles-in-Microsphere Oral System (NiMOS) of 1-5 mm in diameter. Our innovative strategy using non-viral gene delivery vector relies on the fact that gelatin nanoparticles can efficiently encapsulate plasmid DNA, are internalized in cells by endocytosis, protects against DNA degradation in cells, and allows for efficient transfection. The PCL matrix is expected to protect the DNA- containing nanoparticles upon oral administration and allow the release to occur in the small and large intestine due to enzymatic degradation with lipases. Our preliminary studies show that optimized NiMOS formulation have longer residence time in the small and large intestine upon administration in fasted rats and can afford transfection of reporter plasmids in the small and large intestine. The specific aims of this R01 application are: (1) to prepare, characterize, and optimize NiMOS formulations containing plasmid DNA encoding for sTNF-R1 and IL-10, (2) evaluate the biodistribution pattern and residence profile of NiMOS upon oral administration in naove and acute colitis-induced Balb/c mice, (3) determine qualitative and quantitative transfection efficiency upon and oral and rectal administration of DNA- containing NiMOS in naove and colitis-induced mice, and (4) evaluate the therapeutic efficacy of expressed sTNF-R1 and IL-10 in acute colitis-induced mice upon oral and rectal administration. This study will provide important understanding of the barriers to oral therapeutic gene delivery and the potential of NiMOS as safe and effective non-viral vector system. The potential of oral gene therapy for the treatment of IBD using this clinically-translatable strategy has tremendous promise in the effective management of this disease. Project Narrative: Inflammatory bowel disease (IBD) is a significant clinical problem affecting as many as 100 per 100,000 Americans under the age of 30. Recent studies suggest that IBD may be caused by the alternations in the delicate balance between pro- and anti-inflammatory protein molecules, called cytokines, in the gastro- intestinal (GI) tract. Due to the presence of proteolytic degrading enzymes, oral administration of protein drugs does not provide an effective therapy. In the proposed study, an interdisciplinary group of investigators will evaluate if oral gene therapy with plasmid DNA that encodes for soluble receptor (soluble tumor necrosis factor receptor-1, sTNF-R1) and anti-inflammatory cytokine interleukin-10 (IL-10) can be used as a clinically translatable alternative for treatment of IBD. We have formulated Nanoparticles-in-Microsphere Oral System (NiMOS) to provide a safe and effective gene delivery mechanism to the small and large intestine upon oral administration. The sTNF- R1- and IL-10-encoding plasmid DNA will be encapsulated in gelatin nanoparticles, which are further protected with a poly(epsilon-caprolactone) (PCL) matrix to form 1-5 micron particles for effective delivery. Once the NiMOS reach small and large intestine, the PCL shell is degraded and the DNA-containing gelatin nanoparticles are released for cellular uptake and efficient gene expression. Our preliminary studies show that NiMOS, optimized for oral gene delivery, can localize the encapsulated nanoparticles in the small and large intestine in fasted rats and lead to transfection of the encapsulated reporter plasmid. In this study we will develop NiMOS with plasmid DNA encoding for sTNF-R1 and IL-10, characterize, and optimize them for oral delivery. Following the optimization of DNA-containing NiMOS, we will examine distribution and residence of NiMOS upon oral administration in naove and colitis-induced Balb/c mice. The qualitative and quantitative transfection efficiency of sTNF-R1 and IL-10 will be examined next. Lastly, we will carefully evaluate the therapeutic effectiveness of orally-administered NiMOS containing plasmid DNA that encodes for sTNF-R1 and IL-10 in colitis-induced mice. The results of this study will be extremely beneficial, since the expressed therapeutic protein can be produced locally at the disease site for a long period of time. In addition, NiMOS would be useful for other oral gene therapy strategies for treatment of diseases or oral DNA vaccination.

描述（由申请人提供）：炎症性肠病（IBD）影响30岁以下每10万美国人中多达100人。最近的研究表明，IBD可能是由胃肠道（GI）中促炎细胞因子和抗炎细胞因子之间微妙平衡的改变引起的。肿瘤坏死因子-α（TNF）和白细胞介素-10（IL-10）分别作为促炎细胞因子和抗炎细胞因子的作用已在临床前和临床研究中进行了研究。在拟议的研究中，一个跨学科的研究小组将评估口服基因治疗使用可溶性TNF受体-1（sTNF-R1）和IL-10表达质粒DNA在急性结肠炎诱导的Balb/c小鼠。质粒DNA将被封装在明胶纳米颗粒中，其进一步被聚（ε-己内酯（PCL）基质保护以形成直径为1-5 mm的微球口腔系统中的纳米颗粒（NiMOS）。我们使用非病毒基因递送载体的创新策略依赖于明胶纳米颗粒可以有效地封装质粒DNA，通过内吞作用内化在细胞中，防止细胞中的DNA降解，并允许有效转染。预期PCL基质在口服给药时保护含DNA的纳米颗粒，并且由于脂肪酶的酶促降解而允许在小肠和大肠中发生释放。我们的初步研究表明，优化的NiMOS制剂在禁食大鼠中给药后在小肠和大肠中具有更长的停留时间，并且可以在小肠和大肠中转染报告质粒。本R 01应用程序的具体目标是：（1）制备、表征和优化含有编码sTNF-R1和IL-10的质粒DNA的NiMOS制剂，（2）评估在未患结肠炎和急性结肠炎诱导的Balb/c小鼠中口服给药后NiMOS的生物分布模式和驻留特征，（3）在未感染和结肠炎诱导的小鼠中测定口服和直肠施用含DNA的NiMOS后的定性和定量转染效率，和（4）评价表达的sTNF-R1和IL-10经口和直肠给药对急性结肠炎诱导小鼠的治疗效果。这项研究将提供重要的理解的障碍，口服治疗基因传递和潜力的NiMOS作为安全和有效的非病毒载体系统。使用这种临床上可翻译的策略治疗IBD的口服基因疗法的潜力在有效管理这种疾病方面具有巨大的前景。 项目叙述：炎症性肠病（IBD）是一种重要的临床问题，影响30岁以下每10万美国人中多达100人。最近的研究表明，IBD可能是由胃肠道（GI）中促炎和抗炎蛋白分子（称为细胞因子）之间微妙平衡的变化引起的。由于蛋白水解降解酶的存在，口服蛋白质药物不能提供有效的治疗。在拟议的研究中，一个跨学科的研究人员小组将评估编码可溶性受体（可溶性肿瘤坏死因子受体-1，sTNF-R1）和抗炎细胞因子白细胞介素-10（IL-10）的质粒DNA的口服基因治疗是否可用作IBD治疗的临床可翻译替代方案。我们已经制定了纳米微球口服系统（NiMOS），以提供一个安全有效的基因传递机制，以小肠和大肠口服给药。将编码sTNF-R1和IL-10的质粒DNA包封在明胶纳米颗粒中，其进一步用聚（ε-己内酯）（PCL）基质保护以形成1-5微米颗粒用于有效递送。一旦NiMOS到达小肠和大肠，PCL壳被降解，含有DNA的明胶纳米颗粒被释放用于细胞摄取和有效的基因表达。我们的初步研究表明，NiMOS，优化的口服基因传递，可以本地化的封装纳米粒子在小肠和大肠禁食大鼠，并导致转染的封装报告质粒。在这项研究中，我们将开发NiMOS与质粒DNA编码的sTNF-R1和IL-10，表征，并优化它们的口服给药。在优化含DNA的NiMOS之后，我们将检查NiMOS在未患和结肠炎诱导的Balb/c小鼠中口服给药后的分布和驻留。接下来将检查sTNF-R1和IL-10的定性和定量转染效率。最后，我们将仔细评估口服NiMOS的治疗效果，该NiMOS含有编码sTNF-R1和IL-10的质粒DNA。这项研究的结果将是非常有益的，因为表达的治疗性蛋白质可以在疾病部位长时间局部产生。此外，NiMOS将用于治疗疾病或口服DNA疫苗的其他口服基因治疗策略。