A Self-Delivery siRNA for Promoting Regenerative Healing in the Eye

用于促进眼部再生愈合的自我递送 siRNA

基本信息

批准号：
10664929
负责人：
AUDREY M BERNSTEIN
金额：
--
依托单位：
SYRACUSE VA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10664929
关键词：
3-Dimensional Actins Acute Aging Animals Apoptosis Award Binding Biological Blindness Brain Injuries Bulla Burr hole procedure C-terminal Cell Adhesion Cell Proliferation Cells Cholesterol Chronic Cicatrix Clinical Clinical Trials Collagen Cornea Corneal Injury Cost Savings Cytoskeleton Data Development Disease Dose Endothelium Exhibits Extracellular Matrix Eye Eye Injuries Family suidae Fibroblasts Fibronectins Fibrosis Filtering Surgery Filtration Flow Cytometry Freedom G3BP1 gene Gel General Population Genes Glaucoma Goals Health Hour Human Immunohistochemistry Induction of Apoptosis Inflammation Injury Integral Membrane Protein Integrin alphaV Integrins Legal patent Length Link Mediating Methodology Microscopy Military Personnel Mitomycin C Modification Myofibroblast N-terminal Nerve Regeneration Operative Surgical Procedures Organ Culture Techniques Oryctolagus cuniculus Outcome Pathologic Pathway interactions Penetration Pharmaceutical Preparations Physiologic Intraocular Pressure Pilot Projects Play Population Proliferating Publishing Quality of life Recycling Role Sclera Services Speed Structure TP53 gene Techniques Testing Therapeutic Therapeutic Intervention Thick Time Tissues Toxin Translations Trauma Ubiquitin Ubiquitination Veterans Viral Vision Visual Visual impairment Work age related antifibrotic treatment corneal burn corneal regeneration corneal scar cost disability experimental study glaucoma surgery healing immune cell infiltrate improved in vivo inhibitor irritation knock-down neovascularization novel operation overexpression pressure prevent regenerative slit lamp imaging standard of care surgery outcome targeted treatment therapeutic target tonometry ubiquitin isopeptidase wound wound closure wound healing

项目摘要

Project Summary Corneal scarring and glaucoma accounts for ocular disability in millions of veterans, active military, and civilians. Injuries to the eye remain a battlefield and clinical challenge with the potential to severely reduce vision and quality of life. Ocular trauma and brain injury with resulting visual deficits are major causes of vision loss among our veterans and troops engaged in Operations Enduring Freedom and Iraqi Freedom. In addition to acute trauma, aging veterans have a significant increase in age-related glaucoma compared to the general population. The total cost, including treatment is estimated at 2.4 billion annually for the Armed Services. The link between cornea and glaucoma is that a) almost any severe eye injury because of the inflammation generated during wound healing and the drugs administered to treat the inflammation will lead to the onset of glaucoma, and b) the two scarring indications have similar biological underpinnings and thus it is a cost savings to work on both indications at the same time. We are proposing the use of a self-delivery siRNA to prevent and reverse scarring in the cornea and to prevent scarring in the sclera bleb made to reduce intraocular pressure in the eye. Our self-delivery (modified with cholesterol to gain entry into cells) siRNA modified for in vivo use is protected with a utility patent to prevent scarring in the eye and a second provisional patent for the modified self-delivery siRNA sequence and structure. Acute scarring, similar to chronic fibrosis, is characterized by immune cell infiltration and the persistence of cells termed myofibroblasts. Pathological myofibroblasts exhibit increased cell adhesion and tissue contraction through the force generated by binding to the extracellular matrix and the intracellular actin cytoskeleton via integrins (transmembrane proteins). We have discovered a key point in the healing pathway that can be therapeutically targeted to control cell apoptosis, immune infiltration, and integrin- mediated pathological myofibroblast development. Collectively, our studies in primary human corneal cells, pig corneal organ culture, and in rabbits demonstrate that wounding induces the expression of the deubiquitinase (DUB), USP10. USP10 removes ubiquitin from both p53 and αv-integrins. Knockdown of USP10 with one dose of self-delivery siRNA in vivo after corneal wounding significantly reduced apoptosis, immune infiltration, fibrotic markers, and corneal scarring, and a pilot study demonstrated regenerative healing in the glaucoma filtration bleb. Towards the goal of realizing the most effective and specific USP10-targeted therapeutic, in Aim 1, we propose to elucidate the USP10 domains that lead to scarring outcomes and the effect of USP10 binding partner, G3BP2 on USP10 DUB activity. In Aim 2, we will expand our studies on USP10 knockdown to determine if corneal inflammation and scarring can be totally prevented with a second dose of self-delivery siRNA and reversed by novel techniques to permit siRNA entry into a scar. In Aim 3, we will test if USP10 knockdown can prevent inflammation and scarring in the bleb after glaucoma surgery that is performed to relieve intraocular pressure elevation associated with glaucoma. Successful completion of these Aims will significantly improve our understanding of the central mechanisms that promote scarring, and will lead to the development of novel self-delivery siRNA approaches to preventing corneal and glaucoma-related blindness in the veteran and civilian populations.

项目概要角膜疤痕和青光眼导致数百万退伍军人、现役军人和军人眼部残疾眼睛受伤仍然是一个战场和临床挑战，有可能造成严重后果。导致视力缺陷的眼外伤和脑损伤是导致视力和生活质量下降的主要原因。参与持久自由行动的退伍军人和部队视力丧失的原因除了急性创伤之外，老年退伍军人与年龄相关的疾病也显着增加。与一般人群相比，青光眼的总费用（包括治疗）估计为 2.4。每年为武装部队提供 10 亿美元角膜和青光眼之间的联系是：a) 几乎任何疾病。由于伤口愈合过程中产生的炎症和药物造成的严重眼损伤治疗炎症会导致青光眼的发作，并且b）这两种疤痕适应症具有相似的生物学基础，因此研究这两种适应症可以节省成本同时，我们建议使用自传递 siRNA 来预防和逆转疤痕形成。角膜并防止巩膜形成疤痕，降低眼内压。我们的自我递送（用胆固醇修饰以进入细胞）经过修饰用于体内使用的 siRNA 是受一项实用专利保护，以防止眼睛留下疤痕，并获得第二项临时专利保护修饰的自递送siRNA序列和结构。急性疤痕与慢性纤维化类似，其特点是免疫细胞浸润和持续存在称为肌成纤维细胞的细胞表现出细胞粘附和组织增加。通过与细胞外基质和细胞内肌动蛋白结合产生的力进行收缩通过整合素（跨膜蛋白）的细胞骨架我们发现了愈合的关键点。可以治疗靶向控制细胞凋亡、免疫浸润和整合素的途径总的来说，我们对原代人角膜的研究。细胞、猪角膜器官培养物和兔子证明，受伤会诱导表达去泛素酶 (DUB)，USP10 可去除 p53 和 αv 整合素中的泛素。角膜受伤后体内一剂自递送 siRNA 显着敲低 USP10 减少细胞凋亡、免疫浸润、纤维化标记物和角膜疤痕，以及一项试点研究青光眼滤过泡的再生愈合正在朝着实现这一目标的方向发展。最有效和特异性的 USP10 靶向治疗，在目标 1 中，我们建议阐明 USP10 导致疤痕结果的结构域以及 USP10 结合伴侣 G3BP2 对 USP10 的影响在目标 2 中，我们将扩大对 USP10 敲低的研究，以确定角膜是否有效。通过第二剂自我递送 siRNA 可以完全预防炎症和疤痕形成通过新技术逆转，允许 siRNA 进入疤痕。在目标 3 中，我们将测试 USP10 是否有效。敲低可以预防青光眼手术后滤泡内的炎症和疤痕缓解与青光眼相关的眼压升高。目标将显着提高我们对促进疤痕形成的核心机制的理解，以及将导致新型自我递送 siRNA 方法的开发，以预防角膜和退伍军人和平民中与青光眼相关的失明。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

USP10 Promotes Fibronectin Recycling, Secretion, and Organization.

USP10 促进纤连蛋白的回收、分泌和组织。

DOI：
发表时间：
2021
期刊：
影响因子：
4.4
作者：
Phillips, Andrew T;Boumil, Edward F;Castro, Nileyma;Venkatesan, Arunkumar;Gallo, Eugenio;Adams, Jarrett J;Sidhu, Sachdev S;Bernstein, Audrey M
通讯作者：
Bernstein, Audrey M

TGFβ2 Regulates Human Trabecular Meshwork Cell Contractility via ERK and ROCK Pathways with Distinct Signaling Crosstalk Dependent on the Culture Substrate.

TGFβ2 通过 ERK 和 ROCK 途径调节人类小梁网细胞收缩性，并具有依赖于培养基质的不同信号串扰。

DOI：
发表时间：
2022-08
期刊：
影响因子：
2
作者：
Li, Haiyan;Henty;Bernstein, Audrey M;Ganapathy, Preethi S;Herberg, Samuel
通讯作者：
Herberg, Samuel

TGFβ1-driven SMAD2/3 phosphorylation and myofibroblast emergence are fully dependent on the TGFβ1 pre-activation of MAPKs and controlled by maternal leucine zipper kinase.

TGFβ1 驱动的SMAD2/3 磷酸化和肌成纤维细胞的出现完全依赖于MAPK 的TGFβ1 预激活并由母体亮氨酸拉链激酶控制。

DOI：
10.1016/j.cellsig.2023.110963
发表时间：
2023-11-04
期刊：
Cellular signalling
影响因子：
4.8
作者：
Zhang Wang;Nileyma Castro;Audrey M Bernstein;J. Wolosin
通讯作者：
J. Wolosin

The formin DAAM1 regulates the deubiquitinase activity of USP10 and integrin homeostasis.

福尔马林 DAAM1 调节 USP10 的去泛素酶活性和整合素稳态。

DOI：
发表时间：
2023-12
期刊：
影响因子：
6.6
作者：
Phillips, Andrew T;Boumil, Edward F;Venkatesan, Arunkumar;Tilstra;Castro, Nileyma;Knox, Barry E;Henty;Bernstein, Audrey M
通讯作者：
Bernstein, Audrey M