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

• 批准号: 8496874
• 负责人: Yuri Persidsky
• 金额: $ 44.05万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 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/8496874
• 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

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.

描述（由申请人提供）：尽管采用了联合抗逆转录病毒疗法，但 HIV 相关神经认知障碍 (HAND) 的患病率仍然很高。 HAND 与血液中促炎因子和激活的感染单核细胞亚群的升高有关，这两种因子均会导致血脑屏障 (BBB) 损伤，从而导致 HAND。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 HAND。在前期资助期间，我们重点研究（1）BBB损伤的分子机制和（2）糖原合酶激酶（GSK）3？的抗炎和屏障保护特性。抑制 HIV 感染引起的神经炎症。我们揭示了 BBB 功能障碍的分子机制 [紧密连接 (TJ) 磷酸化、BBB 上的 CD40/CD40 配体相互作用以及 HIV 对脑内皮直接影响背后的信号事件]。我们在 GSK3 之后展示了壁垒收紧？由于 TJ 稳定而抑制脑内皮细胞。我们发现了 GSK3 的强效抗炎作用？体外和体内脑内皮细胞抑制（抑制单核细胞迁移、减少炎症因子产生和 BBB 保护）。葛兰素史克？单核细胞的抑制作用减弱了它们在 BBB 上的粘附/迁移，通过抑制小 GTP 酶 Rac1 下调活性整合素表达，并保护 BBB。然而，BBB 屏蔽特性或单核细胞迁移/粘附的抑制在体外或体内并未完全实现，这表明有其他途径与 GSK3 互补吗？是恢复 BBB 功能所必需的。为了寻找此类分子，我们将注意力转向聚（ADP-核糖）聚合酶-1 (PARP-1) 及其抑制剂，它们最近被认为是具有显着神经保护特性的抗炎/免疫调节剂。根据初步数据，我们认为 PARP 抑制将通过对单核细胞、脑内皮、激活的小胶质细胞和 HIV-1 感染的巨噬细胞的影响来减轻 HIV-1 引起的 BBB 损伤。事实上，初步数据表明，原代人脑微血管内皮细胞 (BMVEC) 中的 PARP 抑制改善了 BBB 完整性并增强了 TJ 蛋白的表达。 PARP 抑制剂可防止炎症因子引起的屏障破坏、减少单核细胞在 BBB 模型中的粘附/迁移、下调粘附分子/促炎分子以及降低 RhoA/Rac1 的活性。在单核细胞中，PARP 抑制剂下调活性 β-整合素，与 RhoA/Rac1 抑制平行。 PARP 抑制剂降低促炎分子的表达并减少人类巨噬细胞中的 HIV 复制。尽管 PARP 抑制剂对免疫细胞的调节作用已在一定程度上进行了研究，但其在 HIV 中枢神经系统感染中的作用尚不清楚。 PARP 抑制剂现已进入癌症治疗的临床测试阶段，确保快速转化为免疫/炎症性疾病的治疗。