Tolerance Induction by Neonatal Gene Delivery

通过新生儿基因传递诱导耐受

• 批准号: 7224956
• 负责人: KARIN L GAENSLER
• 金额: $ 30.63万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-15 至 2011-04-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7224956
• 关键词: Adoptive Transfer; Adult; Affect; Animals; Antigen Presentation; Antigen-Presenting Cells; Antigens; Biodistribution; Bioluminescence; Cell Maturation; Cells; Development; Disease; Disease susceptibility; Exhibits; Factor IX; Funding; Gene Delivery; Gene Expression; Gene Transfer; Genes; Goals; Growth; Hemophilia A; Hemophilia B; Hemorrhage; Hereditary Disease; Heterophile Antigens; Histocompatibility Antigens Class II; Human; Human Development; Immune; Immune response; Immune system; Immunity; Immunologic Monitoring; Injection of therapeutic agent; Intervention; Invasive; Life; Liver; Luc Gene; Luciferases; Mediating; Modeling; Morbidity - disease rate; Mus; Muscle; Nature; Neonatal; Pattern; Peptides; Perinatal; Polymerase Chain Reaction; Population; Pre-Clinical Model; Pregnancy; Proteins; Reporter; Reporter Genes; Research Personnel; Route; Serotyping; T-Cell Development; T-Lymphocyte; Testing; Time; Tissues; Transgenes; Week; adeno-associated viral vector; base; critical developmental period; gene therapy; gene transfer vector; in utero; in vivo; mortality; neonate; nonhuman primate; postnatal; prevent; programs; promoter; research study; response; therapeutic gene; therapeutic protein; tissue tropism; transduction efficiency; transgene expression; uptake; vector; viral gene delivery

DESCRIPTION (provided by applicant): Development of inhibitory immune responses to vector and transgene products continues to be a major impediment to long-term correction of genetic disorders. To circumvent this problem, we propose to deliver and express genes early in the maturation of the immune system, with the goal of achieving tolerance to therapeutic gene products and gene delivery vectors. This would also enable pre-emptive treatment of genetic Disorders with perinatal morbidity and mortality. In this context, one of the important differences in ontogeny between mouse and human immune systems is that T cell maturation primarily occurs early during the postnatal period in the mouse, but during mid-gestation in human development. Thus, the'immunological maturity of postnatal mice within the first 3 weeks of life represents a more relevant preclinical model to test strategies for perinatal gene delivery and induction of immunotolerance in humans. Therefore, the overall goal of this revised proposal is to develop strategies for induction of immunotolerance to AAV vector- mediated delivery and expression of transgenes in neonatal mice. Throughout, we will (1) compare bio- distribution, level, and duration of gene expression after primary neonatal delivery and secondary adult re- challenge with highly efficient AAV 1 and 8 serotype vectors, (2) monitor immune responses to the vectors and an intracellular reporter, luciferase, or a secreted therapeutic protein, Factor IX, and (3) define the critical period in the first 3 weeks of life when induction of tolerance is feasible. We will also examine whether differences in AAV 1 vs. 8 tissue tropism affect immune response, and whether the route of administration (IV or IP delivery) affects immune responses to human Factor IX in normal and hemophilic animals. Finally, through adoptive transfer and immunodepletion experiments, we will identify specific immune mechanisms that may influence the persistence or abrogation of gene expression following neonatal gene delivery and adult re-challenge with AAV-murine F.IX vectors in hemophilic animals. RELEVANCE. These studies will define a period in immune ontogeny when viral gene delivery vectors can be repeatedly injected and expressed without eliciting inhibitory immune responses that limit therapeutic gene expression. Furthermore, for many genetic disorders, intervention in the perinatal period to correct the disorder would limit or abrogate the development of irrevocable, often lethal consequences of the disease.

描述（由申请人提供）：对载体和转基因产品产生抑制性免疫应答仍然是长期纠正遗传疾病的主要障碍。为了解决这个问题，我们建议在免疫系统成熟的早期递送和表达基因，目的是实现对治疗性基因产物和基因递送载体的耐受性。这也将使预防性治疗遗传疾病与围产期发病率和死亡率。在这种情况下，小鼠和人类免疫系统之间个体发育的重要差异之一是T细胞成熟主要发生在小鼠出生后早期，但在人类发育中在妊娠中期。因此，出生后小鼠在生命的前3周内的免疫成熟度代表了用于测试人类围产期基因递送和免疫耐受诱导的策略的更相关的临床前模型。因此，该修订提案的总体目标是开发用于在新生小鼠中诱导对AAV载体介导的转基因递送和表达的免疫耐受的策略。在整个过程中，我们将（1）比较初次新生儿分娩和二次成人用高效AAV 1和8血清型载体再攻击后基因表达的生物分布、水平和持续时间，（2）监测对载体和细胞内报告基因荧光素酶或分泌的治疗性蛋白因子IX的免疫应答，以及（3）确定在出生后的前3周内诱导耐受可行的关键时期。我们还将检查AAV 1与AAV 8组织嗜性的差异是否影响免疫应答，以及施用途径（IV或IP递送）是否影响正常和血友病动物中对人因子IX的免疫应答。最后，通过过继转移和免疫耗竭实验，我们将确定特定的免疫机制，可能会影响持久性或废除的基因表达后，新生儿基因传递和成人再挑战与AAV-鼠F.IX载体在血友病动物。本案无关这些研究将定义免疫个体发育的一个时期，此时病毒基因递送载体可以重复注射和表达，而不会引发限制治疗基因表达的抑制性免疫反应。此外，对许多遗传性疾病而言，在围产期进行干预以纠正疾病，将限制或消除疾病造成的不可挽回的、往往是致命的后果。