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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.

项目摘要角膜瘢痕形成和青光眼导致数百万退伍军人、现役军人和平民眼睛受伤仍然是战场和临床的挑战，有可能严重降低视力和生活质量。主要是眼外伤和脑损伤导致的视力障碍我们的退伍军人和参加持久自由行动的部队中视力丧失的原因，伊拉克自由除了急性创伤，老年退伍军人有一个显着增加与年龄有关的青光眼与普通人群相比。包括治疗在内的总费用估计为2.4 每年为军队提供10亿美元。角膜和青光眼之间的联系是a）几乎任何由于伤口愈合过程中产生的炎症和药物，施用以治疗炎症将导致青光眼的发作，和B）两种瘢痕形成适应症具有相似的生物学基础，因此在两种适应症上开展工作可以节省成本同时还研究与讨论我们建议使用一种自我递送的siRNA来预防和逆转瘢痕形成，角膜和防止疤痕的巩膜泡，以降低眼内压。我们的自我递送（用胆固醇修饰以进入细胞）siRNA被修饰用于体内使用，受实用专利保护，以防止眼睛疤痕和第二个临时专利，修饰的自递送siRNA序列和结构。急性瘢痕形成与慢性纤维化相似，其特征在于免疫细胞浸润和持续性免疫细胞浸润。称为肌成纤维细胞。病理性肌成纤维细胞表现出增加的细胞粘附和组织粘附。通过与细胞外基质和细胞内肌动蛋白结合产生的力收缩细胞骨架通过整合素（跨膜蛋白）。我们发现了治愈的关键点途径，可以治疗靶向控制细胞凋亡，免疫浸润，和整合素- 介导的病理性肌成纤维细胞发育。总的来说，我们对原发性人类角膜炎的研究细胞，猪角膜器官培养，并在兔子证明，创伤诱导表达去泛素化酶（DUB），USP 10. USP 10从p53和α v-整联蛋白中去除泛素。在角膜创伤后用一个剂量的自递送siRNA在体内显著地敲低USP 10 减少细胞凋亡、免疫浸润、纤维化标志物和角膜瘢痕形成，以及一项初步研究证明了青光眼滤过泡的再生愈合。朝着实现最有效和特异性的USP 10靶向治疗，在目标1中，我们建议阐明USP 10 导致瘢痕结果的结构域以及USP 10结合伴侣G3 BP 2对USP 10的影响 DUB活动在目标2中，我们将扩大我们对USP 10敲除的研究，以确定角膜是否炎症和瘢痕形成可以用第二剂量的自递送siRNA完全预防，通过新技术逆转，允许siRNA进入疤痕。在目标3中，我们将测试USP 10是否敲除可以防止青光眼手术后滤过泡的炎症和瘢痕形成，以缓解与青光眼相关的眼内压升高。成功完成这些目的将大大提高我们对促进瘢痕形成的中心机制的理解，将导致新的自我递送siRNA方法的发展，以防止角膜和退伍军人和平民人群中与脊髓灰质炎相关的失明。

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.

DOI：
10.1167/iovs.62.13.15
发表时间：
2021-10-04
期刊：
Investigative ophthalmology & visual science
影响因子：
4.4
作者：
Phillips AT;Boumil EF;Castro N;Venkatesan A;Gallo E;Adams JJ;Sidhu SS;Bernstein AM
通讯作者：
Bernstein AM

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