BBB Protection in HIV Infection: Barrier-shielding effects of PARP inhibition

HIV 感染中的 BBB 保护：PARP 抑制的屏障屏蔽作用

基本信息

批准号：
8415634
负责人：
Yuri Persidsky
金额：
$ 49.58万
依托单位：
TEMPLE UNIV OF THE COMMONWEALTH
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-04-15 至 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8415634
关键词：
Address Adhesions Animal Model Anti-Inflammatory Agents Anti-inflammatory Attention Attenuated Blood Blood - brain barrier anatomy Brain CD40 Ligand Cell Adhesion Cell Adhesion Molecules Cells Chronic Cytoskeleton DNA Sequence Rearrangement Data Disease Encephalitis Endothelial Cells Endothelium Event Functional disorder Funding Genome Glycogen Synthase Kinase 3 Glycogen Synthase Kinases HIV HIV Infections HIV-1 Human Immune Immune response Immunotherapy Impaired cognition Impairment In Vitro Individual Infection Infiltration Inflammatory Inflammatory Response Injury Integrins Leukocytes Light Link Lymphocyte Maintenance Mediating Microglia Modality Modeling Molecular Monomeric GTP-Binding Proteins Mononuclear Multiple Sclerosis NOD/SCID mouse Neurocognitive Neurons Neurotoxins PARP inhibition Pathogenesis Pathway interactions Permeability Phosphorylation Physiology Poly(ADP-ribose) Polymerases Prevalence Production Property Proteins Publishing Reaction Role Signal Pathway Signal Transduction Staging Stroke Surface System TNFRSF5 gene Testing Therapeutic Tight Junctions Translations Viral Virus Diseases Virus Replication antiretroviral therapy base cancer therapy cytokine improved in vivo inhibitor/antagonist innovation intravital microscopy macrophage migration monocyte neuroinflammation novel strategies prevent protective effect reconstitution research clinical testing response

项目摘要

DESCRIPTION (provided by applicant): Prevalence of HIV-associated neurocognitive disorders (HAND) remains high despite the introduction of combined antiretroviral therapy. HAND is associated with elevation of pro-inflammatory factors in blood and subsets of activated infected monocytes, both shown to cause blood brain barrier (BBB) impairment that contributes to HAND. Therapeutic strategies that disrupt monocyte migration can slow progression of HIV infection and BBB injury, thereby ameliorating HAND. During the previous period of funding, we focused on studies of (1) molecular mechanisms of BBB injury and (2) anti-inflammatory and barrier protective properties of glycogen synthase kinase (GSK) 3? inhibition in neuroinflammation driven by HIV infection. We uncovered molecular mechanisms of BBB dysfunction [tight junction (TJ) phosphorylation, CD40/CD40 ligand interactions at BBB and signaling events behind the direct effects of HIV on brain endothelium]. We demonstrated barrier tightening following GSK3? suppression in brain endothelium due to TJ stabilization. We uncovered potent anti-inflammatory effects of GSK3? inhibition in brain endothelium (suppression of monocyte migration, diminution of inflammatory factor production, and BBB protection) in vitro and in vivo. GSK? inhibition in monocytes attenuated their adhesion/migration across the BBB, down regulated active integrin expression via suppression of the small GTPase, Rac1, and protected the BBB. Yet, BBB shielding properties or inhibition of monocyte migration/adhesion were not fully attained in vitro or in vivo, suggesting that additional pathways complimentary to GSK3? are necessary for the restitution of BBB function. In search of such molecules, we turned our attention to poly(ADP-ribose) polymerase-1 (PARP-1) and its inhibitors, recently recognized as anti-inflammatory/immune modulating agents with significant neuroprotective properties. Based upon preliminary data, we propose that PARP inhibition will attenuate BBB injury caused by HIV-1 via effects on monocytes, brain endothelium, activated microglia and HIV-1 infected macrophages. Indeed, preliminary data indicate that PARP suppression in primary human brain microvascular endothelial cells (BMVEC) improved BBB integrity and augmented expression of TJ proteins. PARP inhibitors prevented barrier disruption caused by inflammatory factors, diminished monocyte adhesion/migration across a BBB model, down regulated adhesion molecules/pro-inflammatory molecules and decreased activity of RhoA/Rac1. In monocytes, PARP inhibitors down regulated the active ?-integrin that paralleled RhoA/Rac1 suppression. PARP inhibitors decreased expression of pro-inflammatory molecules and diminished HIV replication in human macrophages. Although the modulatory effects of PARP inhibitors on immune cells have been studied to some extent, nothing is known about their effects in the setting of HIV CNS infection. PARP inhibitors have now reached the stage of clinical testing for cancer treatment, assuring quick translation to therapy of immune/inflammatory disorders. PUBLIC HEALTH RELEVANCE: Prevalence of HIV-associated neurocognitive disorders (HAND) remains high despite the introduction of combined antiretroviral therapy, and adjunctive treatment modalities are necessary. We propose novel approach to ameliorate HAND via poly (ADP-ribose) polymerase-1 (PARP-1) inhibition. We hypothesized PARP-1 suppression will diminished injury of blood brain barrier caused by HIV-1 due its effects in infected leukocytes (monocytes and macrophages) and brain endothelium. Although the modulatory effects of PARP inhibitors on immune cells have been studied to some extent, nothing is known about their effects in the setting of HIV brain infection. PARP inhibitors have now reached the stage of clinical testing for cancer treatment, assuring quick translation to therapy of immune/inflammatory disorders.

描述（由申请人提供）：尽管引入了抗逆转录病毒疗法，但与HIV相关的神经认知障碍（手）的患病率仍然很高。手与激活感染的单核细胞的血液和亚群中促炎因子的升高有关，这两者都证明会导致血液脑屏障（BBB）损害有助于手。破坏单核细胞迁移的治疗策略可以减缓HIV感染和BBB损伤的进展，从而改善手。在上一期的资金期间，我们重点研究了（1）BBB损伤的分子机制以及（2）糖原合酶激酶（GSK）3的抗炎和屏障保护特性？受HIV感染驱动的神经炎症的抑制作用。我们发现了BBB功能障碍[紧密连接（TJ）磷酸化，CD40/CD40配体相互作用的分子机制以及HIV对HIV对脑内皮的直接影响背后的信号事件]。我们证明了GSK3之后的障碍拧紧？由于TJ稳定而导致脑内皮抑制。我们发现了GSK3的有效抗炎作用？体外和体内抑制脑内皮（抑制单核细胞迁移，炎症因子产生减少和BBB保护）。 gsk？单核细胞中的抑制作用减弱了其在BBB跨BBB的粘附/迁移，通过抑制小GTPase，Rac1抑制活性整联蛋白表达，并保护BBB。然而，BBB屏蔽特性或单核细胞迁移/粘附的抑制尚未在体外或体内完全获得，这表明gsk3的其他途径是补充的？对于恢复BBB功能是必需的。为了寻找这种分子，我们将注意力转移到了多（ADP-核糖）聚合酶-1（PARP-1）及其抑制剂上，最近被认为是具有重要神经保护特性的抗炎/免疫调节剂。基于初步数据，我们建议PARP抑制作用会通过对单核细胞，脑内皮，活化的小胶质细胞和HIV-1感染的巨噬细胞的影响来减轻HIV-1引起的BBB损伤。实际上，初步数据表明，原发性人脑微血管内皮细胞（BMVEC）的PARP抑制改善了BBB完整性和TJ蛋白的增强表达。 PARP抑制剂可预防由炎症因子引起的屏障破坏，单核细胞粘附/跨BBB模型的迁移减少，降低了调节的粘附分子/促炎分子的粘附分子和RHOA/RAC1的活性降低。在单核细胞中，PARP抑制剂下调了与RhoA/Rac1抑制平行的活性？积聚蛋白。 PARP抑制剂降低了促炎性分子的表达，并减少了人类巨噬细胞中的HIV复制。尽管已经研究了PARP抑制剂对免疫细胞的调节作用，但在HIV CNS感染的情况下，对它们的影响一无所知。现在，PARP抑制剂已经达到了癌症治疗的临床测试阶段，可快速转化为免疫/炎症性疾病的治疗。公共卫生相关性：尽管引入了抗逆转录病毒疗法，并且需要辅助治疗方式，但与HIV相关的神经认知障碍（手）的患病率仍然很高。我们提出了通过聚（ADP-核糖）聚合酶1（PARP-1）抑制的新方法来改善手。我们假设PARP-1抑制作用将减少由HIV-1引起的血液屏障的损伤，因为它在感染的白细胞（单核细胞和巨噬细胞）和脑内皮中的影响。尽管在某种程度上研究了PARP抑制剂对免疫细胞的调节作用，但在HIV脑感染的情况下，对它们的影响一无所知。现在，PARP抑制剂已经达到了癌症治疗的临床测试阶段，可快速转化为免疫/炎症性疾病的治疗。