Mitochondrial-Targeted Antioxidant-Encapsulating Nanoparticles as a Promising Therapeutic Strategy in Regulating Outflow Resistance

线粒体靶向抗氧化剂封装纳米颗粒作为调节流出阻力的有前景的治疗策略

• 批准号: 10046288
• 负责人: Evan B. Stubbs
• 金额: --
• 依托单位: EDWARD HINES JR VA HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-10-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10046288
• 关键词: Affect; Age-Years; Anterior; Antioxidants; Aqueous Humor; Attenuated; Blindness; Cells; Clinical; Deposition; Development; Dose; Encapsulated; Extracellular Matrix; Eye; Family suidae; Functional disorder; Gene Expression; Glaucoma; Goals; Histology; Human; Immunohistochemistry; In Vitro; Individual; Intervention; Investigation; Mediating; Medication Management; Methodology; Mitochondria; Molecular; Neurodegenerative Disorders; Neurons; Oxidative Stress; Patients; Physiologic Intraocular Pressure; Prevalence; Primary Open Angle Glaucoma; Production; Quality of life; Quantitative Reverse Transcriptase PCR; Reactive Oxygen Species; Reporting; Resistance; Role; Signal Pathway; Signal Transduction; Source; Surgical Management; Technology; Testing; Therapeutic; Time; Tissues; Trabecular meshwork structure; Transforming Growth Factor Beta 2; Transforming Growth Factor beta; Translations; Veterans; cytokine; immunocytochemistry; inhibitor/antagonist; laser capture microdissection; nanoparticle; novel strategies; optic nerve disorder; oxidative damage; palliative; socioeconomics; targeted treatment

Neurodegenerative diseases, including glaucoma, substantially alter quality of life of the affected Veteran. Glaucoma remains a leading cause of irreversible blindness. Currently affecting over 60 million individuals, this insidious optic neuropathy is characterized by a gradual loss of RGC neurons and is projected to impact nearly 80 million people by the year 2020. The prevalence of primary open angle glaucoma (POAG), the most common form of glaucoma, is approximately 1.86%, or nearly 2 million individuals, 45 years of age and older in the US. Despite being an extraordinarily significant socioeconomic burden to the DVA, the treatment of US Veterans with POAG remains limited and palliative. Current treatment options are restricted to non-specific interventions aimed at lowering intraocular pressure (IOP), a poorly-understood hallmark of POAG. For many glaucomatous Veterans, however, pharmacological and surgical management of IOP remains clinically refractive. The development of targeted therapeutic strategies directed at the cause of elevated IOP is critical for the advanced management of glaucomatous Veterans. In healthy eyes, IOP is maintained through balanced production and outflow of aqueous humor (AH). Increased resistance to AH outflow through the trabecular meshwork/juxtacanalicular tissue (TM/JCT) is a major contributor of aberrant elevation of IOP in POAG. The molecular mechanisms responsible for elevated IOP remain elusive, but most likely involve aberrant expression and signaling of transforming growth factor-β2 (TGF-β2). Numerous studies demonstrate that TGF-β2, a multifunctional cytokine that promotes TM cell contractility and increased extracellular matrix (ECM) synthesis and deposition within the TM, is markedly elevated in the AH of patients with POAG. While the cellular origin of TGF-β2 is unclear, we reported that human TM cells constitutively express and secrete active TGF-β2, highlighting the TM as a viable targetable source of active TGF-β2. Whereas the development of selective TGF-β2 downstream signaling pathway inhibitors continues to be the focus of intense investigations, targeted disruption of constitutive TGF-β2 expression and release from the TM represents an underexploited therapeutic strategy for the management of IOP in POAG. Numerous studies also report elevated levels of oxidative stress markers in AH of POAG patients, along with altered expression of antioxidant defenses in the TM. Selective oxidative damage to TM mitochondria (Mt) elicits TM cell dysfunction. Mt-generated reactive oxygen species (ROS) are required for TGF-β induced gene expression. Here, we present unpublished findings demonstrating Mt-targeted antioxidants significantly attenuate expression and release of TGF-β2 from cultured human TM cells. Collectively, a penultimate role of Mt-generated ROS in TGF-β2 mediated decreases in outflow facility and increased IOP in POAG begins to emerge. Hypothesis: Targeted disruption of constitutive TGF-β2 expression or signaling within the TM/JCT with mitochondrial-targeted antioxidant-encapsulating nanoparticles will increase outflow facility and lower IOP. The hypothesis of this study will be tested with the following three Specific Aims using a combination of ex vivo and in vitro established experimental approaches: Aim 1 will determine, ex vivo, whether perfusing human or porcine anterior segments with mitochondrial-targeted antioxidant-encapsulating nanoparticles will protect against experimentally induced decreases in outflow facility. Aim 2 will determine, ex vivo, whether perfusing human or porcine anterior segments with mitochondrial-targeted antioxidant-encapsulating nanoparticles will alter TM tissue integrity, endogenous TGF-β2 expression, or TGF-β2 mediated changes in ECM composition. Aim 3 will elucidate the mechanism(s) by which mitochondrial-targeted antioxidant-encapsulating nanoparticles alter TGF-β2 expression or TGF-β2 mediated changes in ECM composition in cultured human TM cells. Successful completion of the proposed study will serve as a critical first step toward developing this technology for the management of Veterans with POAG.

包括青光眼在内的神经退行性疾病极大地改变了受影响退伍军人的生活质量。 青光眼仍然是导致不可逆失明的主要原因。目前影响超过 6000 万人， 隐匿性视神经病变的特点是 RGC 神经元逐渐丧失，预计会影响近 到 2020 年，这一数字将达到 8000 万人。原发性开角型青光眼 (POAG) 的患病率是最常见的 青光眼的常见形式，约占 1.86%，即 45 岁及以上的近 200 万人 美国。尽管对 DVA 来说是极其重大的社会经济负担，但美国的待遇 患有 POAG 的退伍军人仍然有限且姑息治疗。目前的治疗方案仅限于非特异性 旨在降低眼内压（IOP）的干预措施，这是人们对 POAG 的一个鲜为人知的标志。对于很多人来说 然而，对于患有青光眼的退伍军人来说，眼压的药物和手术治疗仍处于临床阶段 折射。针对眼压升高原因制定针对性治疗策略至关重要 用于青光眼退伍军人的高级管理。 在健康的眼睛中，眼压是通过房水（AH）的平衡产生和流出来维持的。 通过小梁网/近小管组织 (TM/JCT) 对 AH 流出的阻力增加是 POAG 中 IOP 异常升高的主要原因。导致升高的分子机制 IOP 仍然难以捉摸，但很可能涉及转化生长因子-β2 的异常表达和信号传导 （TGF-β2）。大量研究表明TGF-β2是一种多功能细胞因子，可促进TM细胞 收缩性和增加细胞外基质 (ECM) 合成和 TM 内沉积，显着 POAG 患者的 AH 升高。虽然 TGF-β2 的细胞起源尚不清楚，但我们报道称 人类 TM 细胞组成型表达并分泌活性 TGF-β2，凸显 TM 是一种可行的靶向药物 活性TGF-β2的来源。鉴于选择性TGF-β2下游信号通路的发展 抑制剂仍然是深入研究的焦点，有针对性地破坏 TGF-β2 的组成型表达 TM 的释放代表了 POAG 中 IOP 管理的尚未充分利用的治疗策略。 许多研究还报告了 POAG 患者 AH 中氧化应激标志物水平升高，以及 TM 中抗氧化防御表达的改变。 TM 线粒体 (Mt) 选择性氧化损伤 TM细胞功能障碍。 Mt 产生的活性氧 (ROS) 是 TGF-β 诱导基因所必需的 表达。在这里，我们提出了未发表的研究结果，证明 Mt 靶向抗氧化剂显着减弱 培养的人 TM 细胞中 TGF-β2 的表达和释放。总的来说，Mt 产生的倒数第二个角色 TGF-β2 中的 ROS 介导的流出设施减少和 POAG 中的 IOP 增加开始出现。 假设：有针对性地破坏 TM/JCT 内的组成型 TGF-β2 表达或信号传导 靶向线粒体的抗氧化剂封装纳米粒子将增加流出能力并降低眼压。这 本研究的假设将通过以下三个具体目标结合离体和体内进行检验 体外建立的实验方法：目标 1 将确定体外灌注是人还是猪 具有靶向线粒体的抗氧化剂封装纳米颗粒的眼前节将防止 实验导致流出设施减少。目标 2 将确定离体是否灌注人类或 猪前段带有靶向线粒体的抗氧化剂封装纳米粒子将改变 TM 组织 完整性、内源性 TGF-β2 表达或 TGF-β2 介导的 ECM 成分变化。目标 3 将阐明 线粒体靶向抗氧化剂封装纳米颗粒改变 TGF-β2 的机制 表达或 TGF-β2 介导培养的人 TM 细胞中 ECM 成分的变化。 成功完成拟议的研究将成为开发这一项目的关键的第一步 使用 POAG 管理退伍军人的技术。

项目成果

Poly(lactic-co-glycolic acid) Nanoparticles Encapsulating the Prenylated Flavonoid, Xanthohumol, Protect Corneal Epithelial Cells from Dry Eye Disease-Associated Oxidative Stress.

• DOI: 10.3390/pharmaceutics13091362
• 发表时间: 2021-08-30
• 期刊: Pharmaceutics
• 影响因子: 5.4
• 作者: Ghosh AK;Thapa R;Hariani HN;Volyanyuk M;Ogle SD;Orloff KA;Ankireddy S;Lai K;Žiniauskaitė A;Stubbs EB Jr;Kalesnykas G;Hakkarainen JJ;Langert KA;Kaja S
• 通讯作者: Kaja S

TGF-β2 Promotes Oxidative Stress in Human Trabecular Meshwork Cells by Selectively Enhancing NADPH Oxidase 4 Expression.

• DOI: 10.1167/iovs.62.4.4
• 发表时间: 2021-04-01
• 期刊: Investigative ophthalmology & visual science
• 影响因子: 4.4
• 作者: Rao VR;Stubbs EB Jr
• 通讯作者: Stubbs EB Jr

Isoprenylation of Monomeric GTPases in Human Trabecular Meshwork Cells.

人小梁网细胞中单体 GTP 酶的异戊二烯化。

• DOI: 10.1007/978-1-4939-6996-8_18
• 发表时间: 2017
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: StubbsJr,EvanB