Characterizing proviral populations in brain tissues

 产品说明：本研究旨在满足RFA-DA-15-018的目标：开发监测HIV脑储库的技术和工具以及暴露于滥用物质如何改变它们。为了在R21阶段应对这一挑战，我们建议开发和验证下一代测序（Pacific Biosciences单分子真实的时间（SMRT）测序）和生物信息学平台，以准确测量和表征脑和淋巴结组织中的HIV DNA群体。特别地，该平台将能够测量和鉴定（i）完整且有复制能力的，（ii）完整但无复制能力的，和（iii）不完整的前病毒群体。我们将开发和验证基于诊断分子方法（即CAP和OIE）的国际标准程序的拟议平台，使用来自HIV感染者的脑和淋巴结组织，这些人在死亡前接受了至少2年的优化抗逆转录病毒治疗。一旦开发和验证，我们将在R33阶段使用这项技术来确定HIV DNA如何在兴奋剂使用者和非使用者的大脑中繁殖。

 DESCRIPTION: This study is designed to meet the goals of the RFA-DA-15-018: Developing Technologies and Tools to Monitor HIV Brain Reservoirs and How They May be Altered by Exposure to Substances of Abuse. To meet this challenge in the R21 phase, we propose to develop and validate a combined next generation sequencing (Pacific Biosciences single-molecule, real time (SMRT) sequencing) and bioinformatics platform to accurately measure and characterize HIV DNA populations in brain and lymph node tissues. In particular, this platform will be able to measure and identify proviral populations that are (i) intact and replication competent, (ii) intact but replication incompetent, and (iii) not intact. We will develop and validate the proposed platform based on international standard procedures for diagnostic molecular methods (i.e. CAP and OIE) using brain and lymph node tissues from HIV-infected individuals on optimized antiretroviral therapy for at least 2 years before death. Once developed and validated, we will use this technology in the R33 phase to determine how HIV DNA populates the brains of stimulant users and non-users.