eukaryotic expression vectors resistant to transgene silencing

抗转基因沉默的真核表达载体

• 批准号: 8057175
• 负责人: James Williams
• 金额: $ 58.02万
• 依托单位: NATURE TECHNOLOGY CORPORATION
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-10 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8057175
• 关键词: American; Amputation; Antibiotics; Area; Biology; Clinical; Clinical Trials; Collaborations; Cosmetics; Cutaneous; Dermatology; Development; Diabetes Mellitus; Diabetic Foot Ulcer; Electroporation; Enhancers; FGF7 gene; Foot Ulcer; Future; Gene Delivery; Gene Expression; Gene Transduction Agent; Genes; Human; Hypoxia Inducible Factor; Intramuscular; Licensing; Licensure; Lower Extremity; Mediating; Medicine; Muscle; National Institute of General Medical Sciences; Neuropathy; Pharmaceutical Preparations; Phase; Plasmid Cloning Vector; Plasmids; Public Health; Resistance; Rhytidoplasty; Skeletal Muscle; Skin; Skin Aging; Small Business Innovation Research Grant; Solutions; Technology; Transcriptional Silencer Elements; Transfection; Transgenes; Universities; Validation; Vertebral column; Wound Healing; base; commercialization; design; diabetic; expression vector; gene therapy; gene transfer vector; health application; hypoxia inducible factor 1; improved; in vivo; innovation; interest; keratinocyte growth factor; non-viral gene therapy; novel; prevent; therapeutic development; transgene expression; vector

DESCRIPTION (provided by applicant): Plasmid-directed gene expression is now near the efficacy barrier that to date has prevented commercialization of plasmid-based therapies for human health applications. Previous innovations such as electroporation (EP) increased transgene expression through more effective gene delivery. In Phase I we developed potent minimalized antibiotic-free expression vectors, and demonstrated dramatic improvements in transgene expression may be obtained through vector design innovations. Two platform technologies were developed. " Novel compositions that prevent vector-backbone mediated transgene silencing from plasmid vectors (Anti-Silencing Elements: ASE platform) " Novel vector backbone functionalities that improve transgene expression from plasmid vectors after transient transfection (transient expression enhancers: TEE platform) In Phase II we hypothesize that combining ASE and TEE vectors with state of the art plasmid delivery will create enabling vector-delivery platforms for gene therapy. In Specific Aims 1 and 2 optimal ASE/TEE antibiotic-free vector - EP delivery platforms for skeletal muscle and cutaneous gene therapy, respectively, are identified. In Specific Aim 3 the cutaneous gene therapy platform is applied to create a hypoxia-inducible factor 11 (HIF-11) based gene medicine for diabetic foot ulcer treatment. In Specific Aim 4 a dermatological gene therapy to treat skin aging is developed using a keratinocyte growth factor (KGF) vector- microdermabrasion delivery combination. Specific Aims 3 and 4 are performed in collaboration with wound healing gene therapy expert Dr. John Harmon at Johns Hopkins University and dermatology gene therapy expert Dr Aaron Tabor at Gene Facelift, LLC. The vector-delivery platforms developed herein will further improve gene expression to levels that will enable gene medicine licensure for multiple applications for unmet public health needs. In Phase III the gene therapies for diabetic foot ulcers and skin cosmetics will undergo clinical development. PUBLIC HEALTH RELEVANCE: The objective of this proposal is to validate a novel antibiotic-free non-viral gene therapy platform, and as such is responsive to NIGMS SBIR high-priority area of interest in development of improved vectors for gene transfer. The vectors contain transient expression enhancers that improve transgene expression level and duration after gene delivery to skin or muscle. The platform will be applied to create gene therapy products to treat diabetic neuropathic foot ulcers and skin aging.

描述（由申请人提供）：质粒定向基因表达现已接近功效障碍，迄今为止，该障碍阻碍了基于质粒的疗法在人类健康应用中的商业化。电穿孔 (EP) 等先前的创新通过更有效的基因传递增加了转基因表达。在第一阶段，我们开发了有效的最小化无抗生素表达载体，并证明通过载体设计创新可以显着改善转基因表达。开发了两种平台技术。 “ 防止质粒载体介导的载体主链介导的转基因沉默的新型组合物（反沉默元件：ASE 平台） “ 提高瞬时转染后质粒载体转基因表达的新型载体主链功能（瞬时表达增强剂：TEE 平台） 在第二阶段，我们假设将 ASE 和 TEE 载体与最先进的质粒递送相结合将创造出可能的效果。 用于基因治疗的载体递送平台。在具体目标 1 和 2 中，确定了分别用于骨骼肌和皮肤基因治疗的最佳 ASE/TEE 无抗生素载体 - EP 递送平台。在具体目标 3 中，应用皮肤基因治疗平台创建基于缺​​氧诱导因子 11 (HIF-11) 的基因药物，用于治疗糖尿病足溃疡。在具体目标 4 中，使用角质形成细胞生长因子 (KGF) 载体-微晶换肤术递送组合开发了一种治疗皮肤衰老的皮肤病基因疗法。具体目标 3 和 4 是与约翰霍普金斯大学的伤口愈合基因治疗专家 John Harmon 博士和 Gene Facelift, LLC 的皮肤病基因治疗专家 Aaron Tabor 博士合作执行的。本文开发的载体递送平台将进一步提高基因表达水平，从而使基因医学许可能够用于多种应用，以满足未满足的公共卫生需求。在第三阶段，糖尿病足溃疡和皮肤化妆品的基因疗法将进行临床开发。 公共健康相关性：该提案的目的是验证一种新型的无抗生素非病毒基因治疗平台，因此响应 NIGMS SBIR 在开发改进的基因转移载体方面的高优先级兴趣领域。该载体含有瞬时表达增强剂，可提高转基因表达水平和基因递送至皮肤或肌肉后的持续时间。该平台将用于创建基因治疗产品，以治疗糖尿病神经性足部溃疡和皮肤老化。