Ultrasound-Assisted AQP1 Gene Therapy for Functional Restoration of Salivary Glan

超声辅助 AQP1 基因治疗唾液腺功能恢复

基本信息

批准号：
8514570
负责人：
Michael J. Passineau
金额：
$ 43.19万
依托单位：
ALLEGHENY-SINGER RESEARCH INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-20 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8514570
关键词：
AQP1 gene Acinar Cell Acupuncture procedure Adenovirus Vector Adenoviruses Affect Animals Cancer Survivor Capsid Cell Therapy Chromatography Chronic Clinical Clinical Trials Complex Complication Contralateral Deglutition Dependovirus Development Devices Disease Eating Equipment FDA approved Family suidae Gene Expression Gene Transfer Gene Transfer Techniques Generations Genes Gland Goals Head and neck structure Hereditary Disease Human Immune response Injury Ionizing radiation Laboratories Life Maintenance Mass Spectrum Analysis Measures Mediating Methodology Methods Metric Microbubbles Miniature Swine Modeling Mus Non-Viral Vector Opportunistic Infections Oral Oral cavity Outpatients Patients Pharmaceutical Preparations Plasmids Procedures Proteome Proteomics Publishing Quality of life Radiation Rattus Rehydrations Relative (related person)Reporting Research Design Retreatment Saliva Salivary Salivary Glands Scanning Secondary to Sialadenitis Sjogren&apos s Syndrome Structure System Techniques Technology Testing Tooth Loss Toxic effect Ulcer Ultrasonography Validation Viral Viral Vector Work Xerostomia advanced system base clinical application conventional therapy design functional improvement functional restoration gel electrophoresis gene therapy gene therapy clinical trial improved indexing insight man next generation novel public health relevance restoration saliva proteome sonoporation success therapeutic transgene two-dimensional water channel

AQP1 gene Acinar Cell Acupuncture procedure Adenovirus Vector Adenoviruses Affect Animals Cancer Survivor Capsid Cell Therapy Chromatography Chronic Clinical Clinical Trials Complex Complication Contralateral Deglutition Dependovirus Development Devices Disease Eating Equipment FDA approved Family suidae Gene Expression Gene Transfer Gene Transfer Techniques Generations Genes Gland Goals Head and neck structure Hereditary Disease Human Immune response Injury Ionizing radiation Laboratories Life Maintenance Mass Spectrum Analysis Measures Mediating Methodology Methods Metric Microbubbles Miniature Swine Modeling Mus Non-Viral Vector Opportunistic Infections Oral Oral cavity Outpatients Patients Pharmaceutical Preparations Plasmids Procedures Proteome Proteomics Publishing Quality of life Radiation Rattus Rehydrations Relative (related person)Reporting Research Design Retreatment Saliva Salivary Salivary Glands Scanning Secondary to Sialadenitis Sjogren&apos s Syndrome Structure System Techniques Technology Testing Tooth Loss Toxic effect Ulcer Ultrasonography Validation Viral Viral Vector Work Xerostomia advanced system base clinical application conventional therapy design functional improvement functional restoration gel electrophoresis gene therapy gene therapy clinical trial improved indexing insight man next generation novel public health relevance restoration saliva proteome sonoporation success therapeutic transgene two-dimensional water channel

展开

项目摘要

DESCRIPTION (provided by applicant): We have recently reported the development of an ultrasound-assisted gene transfer (UAGT) method in the salivary gland. This technique enables transient, non-invasive, non-viral gene transfer to the salivary gland using "off the shelf" FDA-approved microbubbles and clinical ultrasonography equipment. We submit that this technology may prove enabling for applications in gene therapy for Xerostomia given the limitations of viral vector systems, which include sialoadenitis and consequent hyposalivation. Considering the substantial prior work that has been performed in miniature swine as a close-to- man model of salivary gland function, our first aim will be to upscale our UAGT technique in miniature swine and index resultant gene expression to viral vectors. The overall goal of this first aim will be to show equivalence to viral vectors, thereby providing a rationale for their replacement. We will also test in our system an advanced-generation (minicircle) plasmid with the putative capability of longer-lasting gene expression following non-viral gene transfer. Our next step will be to undertake a restorative gene therapy treatment in our swine subjects utilizing Aquaporin-1 (AQP1), the therapeutic transgene that has shown salivary restorative promise in an ongoing human gene therapy clinical trial. UAGT and viral methods will be utilized in parallel, with primary experimental metrics being functional, namely salivary flow. Animals will be irradiated according to previously published methodologies. AQP1 treatment with viral vectors (Adenovirus and Adeno- associated virus) will be undertaken as previously described and will serve as an index by which to measure the efficacy of our UAGT-based AQP1 gene therapy. In contrast to earlier studies, we will endeavor retreatment when salivary flow declines to <20% of that in the contralateral (control) gland, with our goal being maintenance of functional improvement for 12 months. We will thereby test our hypothesis that UAGT will enable chronic maintenance of restorative gene therapy through retreatment, whereas viral vectors will not. In our final aim, we will explore an important but heretofore unaddressed issue, that of the proteomic quality of saliva produced by AQP1 gene therapy. To do this, we will undertake full- proteome profiling of the saliva produced in our animal subjects with AQP1 gene therapy, compared with saliva from non-irradiated swine and irradiated swine wherein partial sparing of the salivary gland has occurred. Scans will be performed using 2-dimensional difference gel electrophoresis, supplemented by multi- dimensional chromatography and mass spectrometry methods.

描述（由申请人提供）：我们最近报告了唾液腺中超声辅助基因转移（UAGT）方法的发展。该技术可以使用“ Off the Benfl” FDA批准的微泡和临床超声检查设备，使瞬时，非侵入性，非病毒基因转移至唾液腺。我们认为，鉴于病毒载体系统的局限性，该技术可能证明可以在基因治疗中应用，包括病毒载体系统，其中包括唾液腺炎和随之而来的缺乏症。考虑到在微型猪中所做的大量先前工作是唾液腺功能的接近模型，我们的第一个目的是在微型猪中提高我们的UAGT技术，并将基因表达产生为病毒载体。第一个目标的总体目标是表现出与病毒载体的等效性，从而为其替换提供了理由。我们还将在我们的系统中测试一个晚期（微量圆）质粒，其推定的能力具有持久基因转移后持久基因表达。我们的下一步将是利用Aquaporin-1（AQP1）（AQP1）的猪受试者进行恢复性基因治疗，这是一种治疗转基因，在持续的人类基因治疗临床试验中表现出唾液恢复有望。 UAGT和病毒方法将并行使用，主要实验指标是功能性的，即唾液流动。动物将根据先前发表的方法进行照射。将按照先前描述的病毒载体（腺病毒和腺相关病毒）处理AQP1处理，并将作为衡量基于UAGT的AQP1基因治疗功效的指数。与较早的研究相反，当唾液流量下降到对侧（对照）腺体中的20％时，我们将努力退缩，而我们的目标是维持功能改善12个月。因此，我们将检验我们的假设，即UAGT将通过恢复长期维持恢复性基因治疗，而病毒载体则不能。在我们的最终目标中，我们将探讨一个重要但迄今未解决的问题，即AQP1基因疗法产生的唾液的蛋白质组学质量。为此，与非辐照猪和辐照猪的唾液相比，我们将对我们动物受试者中产生的唾液的全蛋白质组分析，其中发生了唾液腺的部分保留。扫描将使用二维差异凝胶电泳进行，并通过多维色谱和质谱法补充。