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：
8878038
负责人：
Michael J. Passineau
金额：
$ 43.51万
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 2017-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8878038
关键词：
AQP1 gene Acinar Cell Acupuncture Therapy 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 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

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）：我们最近报道了唾液腺超声辅助基因转移（UAGT）方法的发展。该技术使用 FDA 批准的“现成的”微泡和临床超声设备，能够将瞬时、非侵入性、非病毒基因转移到唾液腺。我们认为，考虑到病毒载体系统的局限性，包括唾液腺炎和随后的唾液分泌不足，这项技术可能被证明能够应用于口干症的基因治疗。考虑到之前在小型猪中作为唾液腺功能的接近人类模型进行的大量工作，我们的首要目标是在小型猪中升级我们的 UAGT 技术，并将由此产生的基因表达转入病毒载体。第一个目标的总体目标是显示出与病毒载体的等效性，从而为它们的替代提供了理由。我们还将在我们的系统中测试新一代（小环）质粒，该质粒具有在非病毒基因转移后具有更持久基因表达的假定能力。我们的下一步将是利用 Aquaporin-1 (AQP1) 对我们的猪受试者进行恢复性基因治疗，这种治疗性转基因在正在进行的人类基因治疗临床试验中已显示出唾液恢复的希望。 UAGT 和病毒方法将并行使用，主要实验指标是功能性的，即唾液流量。将根据之前公布的方法对动物进行照射。使用病毒载体（腺病毒和腺相关病毒）进行 AQP1 治疗将如前所述进行，并将作为衡量我们基于 UAGT 的 AQP1 基因治疗功效的指标。与早期的研究相比，当唾液流量下降至对侧（对照）腺体唾液流量的 20% 以下时，我们将努力进行再治疗，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 将通过再治疗实现恢复性基因治疗的长期维持，而病毒载体则不会。在我们的最终目标中，我们将探索一个重要但迄今为止尚未解决的问题，即 AQP1 基因治疗产生的唾液的蛋白质组质量。为此，我们将对接受 AQP1 基因治疗的动物受试者产生的唾液进行全蛋白质组分析，并与未受辐射的猪和唾液腺部分幸存的受辐射猪的唾液进行比较。将使用二维差异凝胶电泳进行扫描，并辅以多维色谱和质谱方法。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Artificial Induction of Native Aquaporin-1 Expression in Human Salivary Cells.

人唾液细胞中天然 Aquaporin-1 表达的人工诱导。

DOI：
发表时间：
2017-04
期刊：
影响因子：
7.6
作者：
Wang, Z;Pradhan;Farach;Passineau, M J
通讯作者：
Passineau, M J

Ultrasound-assisted nonviral gene transfer of AQP1 to the irradiated minipig parotid gland restores fluid secretion.

超声辅助将 AQP1 非病毒基因转移至受辐射的小型猪腮腺可恢复液体分泌。

DOI：
发表时间：
2015-09
期刊：
影响因子：
5.1
作者：
Wang, Z;Zourelias, L;Wu, C;Edwards, P C;Trombetta, M;Passineau, M J
通讯作者：
Passineau, M J

Proteomic profiling of salivary gland after nonviral gene transfer mediated by conventional plasmids and minicircles.

由常规质粒和小环介导的非病毒基因转移后唾液腺的蛋白质组学分析。

DOI：
发表时间：
2014
期刊：
影响因子：
0
作者：
Geguchadze, Ramaz;Wang, Zhimin;Zourelias, Lee;Perez;Edwards, Paul C;Machen, Laurie;Passineau, Michael J
通讯作者：
Passineau, Michael J

CRISPR-Cas9 HDR system enhances AQP1 gene expression.

CRISPR-Cas9 HDR 系统增强 AQP1 基因表达。

DOI：
发表时间：
2017-12-19
期刊：
影响因子：
0
作者：
Wang, Zhimin;Wang, Yaohe;Wang, Songling;Zhang, Li;Zhang, Na;Cheng, Zhenguo;Liu, Qingshi;Shields, Kelly J;Hu, Baoli;Passineau, Michael J
通讯作者：
Passineau, Michael J

CERE-120 Prevents Irradiation-Induced Hypofunction and Restores Immune Homeostasis in Porcine Salivary Glands.

CERE-120 可预防辐射引起的功能减退并恢复猪唾液腺的免疫稳态。

DOI：
发表时间：
2020-09-11
期刊：
影响因子：
0
作者：
Lombaert, Isabelle M A;Patel, Vaishali N;Jones, Christina E;Villier, Derrick C;Canada, Ashley E;Moore, Matthew R;Berenstein, Elsa;Zheng, Changyu;Goldsmith, Corinne M;Chorini, John A;Martin, Daniel;Zourelias, Lee;Trombetta, Mark G;Edwards, Pau
通讯作者：
Edwards, Pau