BK Virus Receptor and polyomavirus nephropathy

BK病毒受体与多瘤病毒肾病

• 批准号: 8043794
• 负责人: ANDREY SOROKIN
• 金额: $ 22.95万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8043794
• 关键词: Affect; Agonist; Applications Grants; BK Virus; BK Virus Nephritis; Binding; Caveolae; Cells; DNA; Data; Disease Progression; Distal; Endocytosis; Endothelin; Endothelin B Receptor; Endothelin Receptor; Endothelin-1; Environment; Epidemiology; Epithelial Cells; Epithelium; G-Protein-Coupled Receptors; Genes; Glycoproteins; Graft Survival; Human; Immunofluorescence Immunologic; Immunosuppressive Agents; Infection; Intervention; JC Virus; Kidney; Kidney Diseases; Kidney Transplantation; Knowledge; Laboratories; Large T Antigen; Link; Mediating; Mediator of activation protein; Molecular; Monkeys; Nature; Nephritis; Outcome; Pathogenesis; Pathway interactions; Pharmacologic Substance; Polyomavirus; Proteins; Research; Research Personnel; Risk; Role; Sialic Acids; Small Interfering RNA; Staging; Techniques; Testing; Therapeutic; Time; Transplant Recipients; Tubular formation; Viral; Viral Load result; Virion; Virus Diseases; Virus Receptors; Western Blotting; Work; base; experience; improved; kidney epithelial cell; novel strategies; particle; prevent; receptor; research study; serotonin receptor; therapy development; trafficking; treatment strategy; uptake

DESCRIPTION (provided by applicant): In recent years, nephritis induced by BK virus (BKV), a non-enveloped double-stranded DNA polyomavirus, has become a severe problem after renal transplantation. There is a critical need to better define the molecular mechanisms of BKV entry into its target cells and to develop efficient treatment strategies for BKN. An objective of the proposed study is to identify the protein component of the BKV receptor and to develop pharmaceutical agents capable of mitigating BKV entry into human renal proximal tubular epithelial cells (HRPTEC), considered to be one of the main natural targets of BKV. The identity of the BKV receptor is unknown. At present, no specific pharmacological agent preventing BKV nephritis is available. Currently, the only efficient therapy against BKV nephritis appears to be a reduction/change of immunosuppressive agents, and this may increase the inherent risk of rejection. In our preliminary studies we have established that caveolar endocytosis is critical for BKV infection of HRPTEC and revealed several important steps of the BKV intracellular trafficking pathway in HRPTEC. Furthermore, our data suggest that the antagonist of Endothelin-1 binding to Endothelin B receptor (ETRB) prevents BKV infection of HRPTEC. Endothelins (ET), agonists of G- protein coupled receptors; act as important mediators of renal disease progression. Our working hypothesis is that binding of BKV to ETRB in HRPTEC is the critical step in the BKV entry into HRPTEC and that ETRB antagonists can be efficient as agents against BKV nephritis. Specific Aim 1 will test the hypothesis that N- linked glycoprotein ETBR serves as a receptor for BKV particles in HRPTEC. To establish that ETRB serves as the binding molecule for BKV particles in HRPTEC, siRNA mediated silencing of the ETRB gene will be employed. We will also carry out experiments to determine direct association between ETRB and BKV particles and evaluate necessity for ETRB presence for BKV infection. BKV infection will be detected using the following markers: the percentage of infected cells, as detected by immunofluorescence, the cellular levels of BKV large T antigen expression, as detected by western blot analysis and viral load, as detected by real-time PCR. Proposed study will provide the basis for a novel strategy for therapeutical intervention in BKN, ultimately improving long-term graft survival after renal transplantation. PUBLIC HEALTH RELEVANCE: In recent years nephritis induced by BK virus (BKV), non-enveloped double-strand deoxyribonucleic acid polyomavirus, has become a severe problem after renal transplantation. Polyomavirus nephropathy, also termed BK-virus nephropathy (BKN) affects 1% to 10% of all kidney transplant recipients and is emerging as an escalating threat. Since graft loss due to BKN ranged from 10% to more than 80% in kidney transplant recipients with BKN there is a critical need now to better define the molecular mechanisms of BKV entry into its target cells and to develop efficient treatment strategies of BKN. The identity of the BKV receptor is unknown and no specific pharmacological agent preventing BKV nephritis is available. This proposal will uncover the protein component of BKV receptor and provide the basis for a novel strategy for therapeutical intervention in BKN to ultimately improve long-term graft survival after renal transplantation.

描述（由申请人提供）：近年来，由非发育的双链DNA多瘤病毒BK病毒（BKV）诱导的肾炎已成为肾移植后的严重问题。迫切需要更好地定义BKV进入其靶细胞的分子机制并制定有效的BKN治疗策略。拟议研究的目的是鉴定BKV受体的蛋白质成分，并开发能够减轻BKV进入人肾近端肾小管上皮细胞（HRPTEC）的药物，被认为是BKV的主要自然靶标之一。 BKV受体的身份尚不清楚。目前，尚无预防BKV肾炎的特定药理剂。目前，针对BKV肾炎的唯一有效疗法似乎是免疫抑制剂的减少/变化，这可能会增加排斥的固有风险。在我们的初步研究中，我们确定Caveolar内吞作用对于HRPTEC的BKV感染至关重要，并揭示了HRPTEC的BKV细胞内贩运途径的几个重要步骤。此外，我们的数据表明内皮素-1与内皮素B受体（ETRB）的拮抗剂可防止HRPTEC的BKV感染。内皮蛋白（ET），G蛋白偶联受体的激动剂；充当肾脏疾病进展的重要介体。我们的工作假设是，在HRPTEC中，BKV与ETRB的结合是BKV进入HRPTEC的关键步骤，并且ETRB拮抗剂可以作为反对BKV肾炎的药物有效。具体目标1将检验以下假设：N连接的糖蛋白ETBR用作HRPTEC中BKV颗粒的受体。为了确定ETRB用作HRPTEC中BKV颗粒的结合分子，将采用siRNA介导的ETRB基因沉默。我们还将进行实验，以确定ETRB和BKV颗粒之间的直接关联，并评估ETRB存在BKV感染的必要性。将使用以下标记检测BKV感染：如通过蛋白质印迹分析和病毒载量检测到的BKV大T抗原表达的细胞水平，被感染细胞的百分比，如实时PCR所检测到的。拟议的研究将为BKN治疗干预的新策略提供基础，最终改善肾移植后的长期移植物存活率。 公共卫生相关性：近年来，由BK病毒（BKV）诱导的肾炎，非发育的双链脱氧核糖核酸多瘤病毒，在肾移植后已成为一个严重的问题。多瘤病毒肾病也称为BK病毒肾病（BKN）影响所有肾脏移植受者的1％至10％，并且正成为升级的威胁。由于BKN造成的移植物损失范围从10％到具有BKN的肾脏移植受者的80％以上，因此现在至关重要的需要更好地定义BKV进入其靶细胞的分子机制并制定BKN的有效治疗策略。 BKV受体的身份尚不清楚，没有可预防BKV肾炎的特定药理剂。该建议将揭示BKV受体的蛋白质成分，并为BKN治疗干预的新策略提供基础，以最终改善肾移植后的长期移植物存活率。