The role of cytomegalovirus antiapoptotic proteins in replication in macrophages.

描述(申请人提供)：人巨细胞病毒(HCMV)是一种高度流行的疱疹病毒，通过输血或移植有效地传播。通过减少白细胞和病原体灭活技术(例如补骨脂素/长波紫外线)进行绝育是可行的，但开发用于移植的HCMV安全的细胞疗法需要更复杂的策略。作为制定依赖于减少细胞死亡的病毒控制的策略的长期目标的一部分，该提案探索病毒细胞死亡抑制基因UL36在巨噬细胞中HCMV生长中的作用，目的是验证UL36作为选择性抑制细胞治疗成分中HCMV的靶点。单核细胞及其衍生的巨噬细胞很可能是这种操作的靶细胞，基于以下原因：1)粒细胞、巨噬细胞和树突状细胞(GM-Ps)的前体细胞是巨细胞病毒传播的重要贡献者，因为它们携带作为感染源的潜伏病毒；2)这些前体细胞向巨噬细胞的分化允许病毒的裂解生长；以及3)巨噬细胞的生长被认为是巨细胞病毒传播和随后疾病的关键步骤。小鼠巨细胞病毒(MCMV)UL36同源物M36对巨噬细胞的复制是必不可少的，预计HCMV中的UL36对巨噬细胞的生长也是必不可少的。据预测，UL36的性质，结合原天冬氨酸蛋白酶-8和防止激活，也使它在分化过程中成为一个有吸引力的靶点，产生允许细胞，因为这种蛋白酶在单核细胞分化过程中以及在凋亡过程中都是重要的。此外，由UL36基因编码的新的细胞死亡抑制功能已经被发现，这扩大了该基因在这一重要细胞类型中的预期作用。长期目标和近期目标将在两个特定目标中启动和解决，这两个目标旨在1)确定差异剪接对巨噬细胞UL36基因功能的贡献，以及2)确定microRNA miR 36-1对巨噬细胞UL36基因功能的贡献。重组病毒和强大的、基于转基因的策略将被纳入一项重点工作，以确定UL36基因产物，这些基因产物抑制巨噬细胞部署的多条细胞死亡途径，以抑制HCMV复制。公共卫生相关性：人类巨细胞病毒(CMV)是一种高度流行的疱疹病毒，可导致危及生命的疾病，特别是在输血、移植或将其传播给未出生的婴儿后。用于移植的巨细胞病毒安全细胞疗法的发展需要新的方法；这些策略的发展很可能表明在其他情况下预防传播的有效手段。这项提议将确定病毒机制，以防止杀死对疾病传播至关重要的受感染细胞的内在防御。

DESCRIPTION (provided by applicant): Human cytomegalovirus (HCMV) is a highly prevalent herpesvirus efficiently transmitted by transfusion or transplantation. Sterilization techniques through leukoreduction and pathogen inactivation techniques (e.g., psoralen/UVA) are available but the development of HCMV-safe cellular therapies for transplantation requires more sophisticated strategies. As part of a long-term goal to devise strategies that rely on reducing viral control of cell death, this proposal explores the role of the viral cell death suppression gene, UL36, in HCMV growth in macrophages, with the objective of validating UL36 as a target for selectively suppressing HCMV in cellular therapy components. Monocytes and the macrophages derived from them are highly likely target cells for this manipulation based on the following: 1) progenitors of granulocytes, macrophages, and dendritic cells (GM-Ps) are important contributors to HCMV transmission because they harbor latent virus that serves as the source for infection, 2) differentiation of these progenitors to macrophages allows for lytic growth of the virus, and 3) growth in macrophages is considered to be a critical step for HCMV transmission and ensuing disease. The murine CMV (MCMV) UL36 homolog, M36, is essential for replication in macrophages, and it is anticipated that UL36 in HCMV will also be essential for growth in macrophages. It is anticipated that properties of UL36, binding procaspase-8 and preventing activation, also makes it an attractive target during the differentiation process that yields permissive cells because this protease is important in monocytes undergoing macrophage differentiation as well as during apoptosis. Further, novel cell death suppression functions encoded by the UL36 gene have been uncovered that expand the anticipated role of this gene in this important cell type. Long-term goals will be initiated and immediate goals addressed in two specific aims designed to 1) determine the contribution of differential splicing to UL36 gene function in macrophages, and 2) determine the contribution of the microRNA miR 36-1 to UL36 gene function in macrophages. Recombinant viruses and robust, transfection-based strategies will be incorporated in a focused effort to identify the UL36 gene products that inhibit multiple cell death pathways deployed by macrophages to inhibit HCMV replication. PUBLIC HEALTH RELEVANCE: Human cytomegalovirus (CMV) is a highly prevalent herpes virus that causes life- threatening disease, especially following blood transfusions, transplantations, or if it is transmitted to an unborn child. The development of CMV-safe cellular therapies for transplantation requires novel approaches; the development of these strategies will likely indicate effective means of preventing transmission in other situations. This proposal will determine viral mechanisms that prevent intrinsic defenses that kill infected cells important to disease transmission.