Engineering Improved Retroviral Stability

CBET-0828244M。这项拟议研究的总体目标是开发有效的策略，以提高逆转录病毒载体的稳定性和滴度，用于基因治疗应用。最近，相当大的努力致力于在一些临床试验中利用这些和其他病毒载体。尽管一些出版物描述了不同程度的成功，但逆转录病毒载体的临床成功受到病毒灭活速度快和病毒滴度有限的限制。因此，具有增强的内在稳定性和高滴度的逆转录病毒将缓解限制逆转录病毒载体在临床上成功的一些实际障碍。这些原理很可能也可以扩展到其他病毒载体。这项研究旨在完成以下工作：(1)确定逆转录病毒生物活性丧失的机制和未成熟/成熟病毒颗粒的相对稳定性；(2)优化培养条件和方法，以便生产高滴度病毒载体；以及(3)修饰病毒逆转录酶，以实现更高效率的病毒复制。为了实现这些目标，将使用物理、化学和分子操纵技术，以减缓逆转录病毒的衰变速度和增加逆转录病毒的滴度。拟议的工作将为参与该项目的研究生研究员以及通过调查人员举办的各种课程和培训方案接触到该项目的学生提供极好的培训。此外，PI还将为各种高中和大学本科研究项目提供良好的教育环境。科学方法的设计不仅是为了建立一个优秀的科学计划，而且还通过课堂教学和直接的实验室互动让学生接触到最先进的技术和方法。这项研究本身将提供有关逆转录病毒衰变机制和解决方法的有价值的新信息。实验结果可能会对利用或正在开发病毒载体传递方案的科学和临床场所产生广泛影响。总之，拟议的工作旨在提供基础研究和人员基础，以解决与实施提高病毒载体适用于基因治疗应用的技术有关的问题。

CBET-0828244M. Yarmush, Rutgers University New BrunswickThe overall goal of the proposed research is to develop effective strategies to increase the stability and titer of retroviral vectors for gene therapy applications. Recently, considerable effort has been dedicated toward utilizing these, and other viral, vectors in a number of clinical trials. Although several publications have described various degrees of success, the clinical success of retroviral vectors is limited by the rapid rate of viral inactivation and limited viral titers. Thus, a retrovirus with enhanced intrinsic stability and high titer would alleviate some of the practical obstacles limiting the success of retroviral vectors in the clinic. It is likely that these principles could also be extended to other viral vectors. The proposed research seeks to accomplish the following: (1) To determine the mechanisms underlying the loss of retroviral bioactivity and the relative stability of the immature / mature viral particles; (2) To optimize culture conditions and methods, in order to allow the production of high-titer viral vectors; and (3) To modify the viral reverse transcriptase to achieve higher efficiency viral replication. To achieve these goals, physical, chemical, and molecular manipulation techniques will be used with the objective of slowing the rate of retroviral decay and increasing retroviral titer. The proposed work will provide excellent training for the graduate fellows involved in the project, as well as to students exposed to it through various courses and training programs conducted by the investigators. In addition, the PI will provide an excellent educational environment for various high school and university-wide undergraduate research programs. The scientific approach was designed not only to establish an excellent scientific program, but also to expose students to state of the art technology and methodology through both classroom instruction and direct laboratory interaction. The research itself will provide valuable new information regarding the mechanisms of retroviral decay and methods to address them. The experimental results are likely to have a broad impact on the scientific and clinical venues which utilize or are developing viral vector delivery protocols. In summary, the proposed work seeks to provide fundamental research and a human base of personnel equipped to solve problems related to the implementation of techniques to improve suitability of viral vectors for gene therapy applications.