Ye Xiang, Hu Mengqi, Hu Yunli, Qiu Haibo, Li Ning
Regeneron Pharmaceuticals Inc., Tarrytown, NY, 10591-6707, USA.
Gene Ther. 2025 May 9. doi: 10.1038/s41434-025-00539-4.
Recombinant adeno-associated viruses (AAVs) have become increasingly popular as gene therapy vectors in recent years. Like all viruses, AAVs undergo dynamic structural changes in response to varying temperature and pH conditions. However, the specific capsid regions involved in these processes remain unknown. In this study, we employed Hydrogen-Deuterium Exchange Mass Spectrometry (HDX-MS) to investigate the impact of pH and temperature on the structure and conformational dynamics of AAV capsids. Our analysis identified specific regions of the capsid that are sensitive to these environmental changes. Additionally, our data elucidated the structural basis for DNA uncoating or leakage triggered by low pH or high temperature. Detailed structural characterization of AAVs by HDX-MS in this study deepens our understanding of viral capsid conformational dynamics and stability in AAV transduction and manufacturing and storage conditions, paving the way for formulation development and next-generation capsid engineering.
近年来，重组腺相关病毒（AAV）作为基因治疗载体越来越受欢迎。与所有病毒一样，AAV会根据不同的温度和pH条件发生动态结构变化。然而，参与这些过程的特定衣壳区域仍然未知。在本研究中，我们采用氢-氘交换质谱（HDX-MS）来研究pH和温度对AAV衣壳结构和构象动力学的影响。我们的分析确定了衣壳中对这些环境变化敏感的特定区域。此外，我们的数据阐明了低pH或高温引发的DNA脱壳或泄漏的结构基础。本研究通过HDX-MS对AAV进行详细的结构表征，加深了我们对AAV转导、制造和储存条件下病毒衣壳构象动力学和稳定性的理解，为制剂开发和下一代衣壳工程铺平了道路。
