eukaryotic expression vectors resistant to transgene silencing

抗转基因沉默的真核表达载体

• 批准号: 8256740
• 负责人: James Williams
• 金额: $ 52.46万
• 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 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8256740
• 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; public health relevance; 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）：质粒指导的基因表达现在接近迄今为止阻止基于质粒的人类健康应用疗法商业化的疗效障碍。诸如电穿孔（EP）之类的创新通过更有效的基因递送增加了转基因表达。在第一阶段，我们开发了有效的最小化抗生素表达载体，并且可以通过向量设计创新获得转基因表达的显着改善。开发了两种平台技术。 " 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用于基因治疗的载体分娩平台。在特定目的中，分别确定了1和2最佳的ASE/TEE抗生素载体 - 分别确定骨骼肌和皮肤基因治疗的EP输送平台。在特定目标3中，使用皮肤基因治疗平台来创建基于缺​​氧的因子11（HIF-11）基因医学，用于糖尿病足溃疡治疗。在特定的目标4中，使用角质形成细胞生长因子（KGF）载体 - 微晶递送组合开发了皮肤衰老的皮肤病学疗法。特定目的3和4是与约翰·霍普金斯大学（Johns Hopkins University）的伤口愈合基因治疗专家John Harmon博士合作进行的，而Dermatology Gene Therapy专家Aaron Tabor博士在Gene Facelift，LLC上进行。此处开发的矢量交付平台将进一步将基因表达提高到水平，这将使基因医学许可能够针对未满足的公共卫生需求进行多种应用。在第三阶段，用于糖尿病足溃疡和皮肤化妆品的基因疗法将经历临床发育。 公共卫生相关性：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，对Nigms SBIR高优先级的兴趣敏感，以开发改进的基因转移载体。向量包含瞬时表达增强子，可改善基因递送至皮肤或肌肉后的转基因表达水平和持续时间。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。

项目成果

Strengthening the skin with topical delivery of keratinocyte growth factor-1 using a novel DNA plasmid.

• DOI: 10.1038/mt.2014.2
• 发表时间: 2014-04
• 期刊: MOLECULAR THERAPY
• 影响因子: 12.4
• 作者: Dou, Chunqing;Lay, Frank;Ansari, Amir Mehdi;Rees, Donald J.;Ahmed, Ali Karim;Kovbasnjuk, Olga;Matsangos, Aerielle E.;Du, Junkai;Hosseini, Sayed Mohammad;Steenbergen, Charles;Fox-Talbot, Karen;Tabor, Aaron T.;Williams, James A.;Liu, Lixin;Marti, Guy P.;Harmon, John W.
• 通讯作者: Harmon, John W.