Salivary Gland-Based Gene Therapy for Lysosomal Storage Diseases

基于唾液腺的溶酶体贮积病基因治疗

基本信息

批准号：
7848134
负责人：
Michael J. Passineau
金额：
$ 24.65万
依托单位：
ALLEGHENY-SINGER RESEARCH INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-06-01 至 2012-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7848134
关键词：
Address Alpha-galactosidase Biochemical Biodistribution Bioreactors Blood Blood Vessels Catabolism Clinical Trials Code Defect Disease Dose Emerging Technologies Endocrine Engineering Enzymes Evaluation Fabry Disease Face Failure Family member Funding Galactosidase Gene Transduction Agent Human Immune Immune Response Genes Immune response Intervention Knockout Mice Lead Light Longevity Lysosomal Storage Diseases Measures Modeling Mus Occupations Organ Pathology Pathway interactions Patients Pharmaceutical Preparations Physiological Physiology Plasma Proteins Recombinants Saliva Salivary Salivary Glands Site Symptoms System Techniques Technology Testing Therapeutic Therapeutic Studies Tissues Transgenes Translating Translational Research Validation Viral Vector Virus adeno-associated viral vector base enzyme activity enzyme replacement therapy gene therapy human disease in vivo insight interest macromolecule meetings pre-clinical research study response therapeutic protein therapeutic transgene transgene expression uptake vector

项目摘要

Recent studies have demonstrated that an intrinsic endocrine pathway is present in salivary glands, distinct from the classical exocrine pathway whereby digestive enzymes are produced and secreted into the saliva by the salivary glands under normal physiological conditions. As such, this distinct endocrine pathway has recently been shown to be capable of robust synthesis and secretion of non-regulated (i.e. constitutive) gene products into the vascular space as a result of retroductal administration of a gene therapy vector. Fabry disease is a member of the family of lysosomal storage diseases, a group of monogenic disorders characterized by deficiency or lack of a class of enzymes (e.g. alpha-Galactosidase A in Fabry disease) involved in intra-lysosomal catabolism of macromolecules. An interesting feature of these diseases is the observation that tissues lacking the specific lysosomal enzyme implicated in the disease are capable of uptake of the specific lysosomal enzyme from the vascular space. This physiology has led to the proposal and subsequent validation of "Enzyme Replacement Therapy" wherein the missing enzyme is produced syntheically and infused intravenously to Fabry patients, resulting in a mitigation of disease symptoms. We seek to combine these emergent technologies into a salivary gland-based gene therapy strategy for Fabry disease in highly relevant mouse knockout model, namely B6;129-Gla(tm1/Kul/J), an alpha- Galactosidase A (-/O) model. We seek to test the hypothesis that an AAV vector containing the sequence for human alpha-Gal A delivered to the salivary glands in alpha-Gal A deficient "Fabry" mice will result in systemic replacement of the enzyme and reversal of the specific systemic biochemical defect underlying Fabry disease. Our specific aims involve: 1) delivery of an alpha-Gal A containing vector to the salivary glands of Fabry mice and biochemical testing of systemic delivery of the enzyme, 2) careful examination of the organ biodistribution of the enzyme, and 3) optimization of dosing and immune physiology to achieve long-term expression of the enzyme. These studies will produce important insights into the technical paramaters of this system, including: longevity of transgene expression, host immune response, and mitigation of Fabry-associated pathology, and form the basis whereby other systemic disorders could be addressed using salivary glands as the biosynthetic site for systemic therapeutics.

最近的研究表明，唾液腺中存在固有的内分泌途径，不同从经典的外分泌途径中产生并分泌到唾液中在正常的生理条件下，唾液腺。因此，这种独特的内分泌途径有最近被证明能够鲁棒的合成和分泌非调节（即本构）基因由于基因疗法载体的卷轴施用，产物进入血管空间。法布里病是溶酶体储存疾病家族的成员，这是一组单基因疾病以缺乏或缺乏一类酶的特征（例如，在Fabry疾病中α-半乳糖苷酶A）参与大分子内溶质体内分解代谢。这些疾病的一个有趣特征是观察到缺乏与疾病有关的特定溶酶体酶的组织能够从血管空间吸收特定的溶酶体酶。这种生理学导致了该建议并随后验证“酶替代疗法”，其中丢失的酶被产生合成性并静脉注入Fabry患者，导致疾病症状的缓解。我们试图将这些新兴技术结合到基于唾液腺的基因治疗策略中高度相关小鼠敲除模型的Fabry病，即B6； 129-GLA（TM1/KUL/J），α- 半乳糖苷酶A（ - /O）模型。我们试图检验以下假设：AAV向量包含序列的序列人α-gal a将α-gal中的唾液腺传递给A Alpha-gal的唾液腺不足的“ Fabry”小鼠将导致全身替代酶和特定系统性生化缺陷的逆转法布里病。我们的具体目的涉及：1）将含有alpha-gal a的载体交付给唾液 Fabry小鼠的腺体和酶全身递送的生化测试，2）仔细检查酶的器官生物分布和3）优化剂量和免疫生理学以实现酶的长期表达。这些研究将对技术产生重要的见解该系统的参数体，包括：转基因表达的寿命，宿主免疫反应和缓解Fabry相关的病理学，并构成其他系统疾病的基础使用唾液腺作为全身治疗剂的生物合成部位。