权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Compacted DNA Nanoparticles for Ocular Therapy

用于眼部治疗的压缩 DNA 纳米颗粒

基本信息

批准号：
10246311
负责人：
MUAYYAD R AL-UBAIDI
金额：
$ 54.81万
依托单位：
UNIVERSITY OF HOUSTON
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-01-01 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10246311
关键词：
3-Dimensional Active Biological Transport Animals Binding Biochemical Biology Biomedical Engineering Caliber Cell Differentiation process Cell Nucleus Cell surface Cells Chemistry Clinical Clinical Trials Coupled DNA Defect Degenerative Disorder Development Diabetic Retinopathy Disease Disease model Dose Elements Engineering Enhancers Equipment Evaluation Eye Eye diseases Foundations Future Gene Expression Generations Genes Genomics Goals Hour Hyaluronic Acid Hydrogels Immune response Injections Inner Limiting Membrane Knowledge Lead Leber&apos s amaurosis Link Lung Matrix Attachment Regions Mediating Messenger RNA Methodology Mitotic Modeling Molecular Mus Mutation Nanotechnology Nuclear Oligonucleotides Onset of illness Papio Peptides Phenotype Photoreceptors Physiology Polyethylene Glycols Polylysine Proteins Resources Retina Retinal Degeneration Retinal Diseases Retinitis Pigmentosa Shapes Structure Structure of retinal pigment epithelium Surface Technology Testing Therapeutic Time Toxic effect Transfection Transgenes Treatment Efficacy Viral Vector Wild Type Mouse adeno-associated viral vector aptamer bioprinting capsule clinically significant design diabetic effective therapy efficacy testing epigenetic silencing experience gene delivery system gene therapy improved in vivo evaluation materials science mouse model nanoformulation nanoparticle nanoparticle delivery nanoscale non-viral gene delivery nucleolin plasmid DNA polycation prevent programs ranpirnase residence rho scaffold single molecule targeted treatment therapeutic evaluation therapeutic nanoparticles therapeutically effective transduction efficiency transgene expression tyrosine O-sulfate uptake vector

项目摘要

PROJECT SUMMARY We aim to advance our current DNA nanoparticle (NP) delivery platform and gene expression to develop safe and effective therapies targeting important photoreceptor-associated ocular disorders caused by defects in large genes. We are merging knowledge in molecular bioengineering, nanoformulation, material science, eye biology/physiology and chemistry to accelerate essential steps for the generation of effective non-viral gene delivery platform for eye diseases. The NPs consist of single molecules of DNA compacted with poly-lysine- PEG polycation and have a diameter of 8-11 nm. Their small size coupled with cellular uptake via cell-surface nucleolin, which efficiently traffics the NPs to the nucleus, accounts for their ability to transfect post-mitotic photoreceptors. Using NPs has led to efficient expression of large genes, an essential prerequisite for targeting hard-to-rescue diseases of the retina. Although upper gene size limitation has not been established, the largest size tested in the lung was 20 kb and in the eye was ~14 kb making NPs an ideal alternative to AAVs for delivery of large genes. We have showed that NP treatment led to efficient transfection of ocular cells including photoreceptors and retinal pigment epithelium, exerted no toxic effects on the eye even after multiple injections, distributed throughout the subretinal space, and mediated appreciable structural/functional rescue in mouse models of retinitis pigmentosa, Leber’s congenital amaurosis, Stargardt’s and diabetic retinopathy. Effective gene expression without toxicity has also been achieved in baboon eyes. These proof- of-principle studies confirms the potential clinical significance of this technology and highlights the value of a large capacity, but emphasized the need for prolonged high levels of gene expression. Our main goal is to enhance photoreceptor uptake of NPs to achieve long-term therapeutic levels of expression of large genes for full phenotypic rescue. We propose to accomplish this by targeted vector engineering to boost expression levels along with NP-delivery platform to enhance their delivery from the vitreous to photoreceptors, achieve pan retinal distribution, promote episomal stability in the nucleus and prevent epigenetic silencing. Subsequently, we will test these optimized NPs/delivery platform(s) for their ability to mediate full phenotypic rescue in a large gene disease model; specifically the Abca4-/- model of Stargardt’s associated with the lack of ABCA4 gene. ABCA4 is a large gene which has not been fully rescued by traditional viral vectors and as a result development of targeted therapeutics for Stargardt’s has lagged. We plan in aim1 to engineer vectors that can achieve therapeutic levels of expression in photoreceptors and in aim 2 to develop effective NP- delivery platforms to enhance photoreceptor uptake of NPs from the vitreous. Aim 3 will test the efficacy of the best delivery platform of the most effective NPs for therapeutically-effective levels of expression in photoreceptors of Abca4-/- mice before and after the onset of the disease. In summary, results from this application will facilitate the advancement of DNA NP use for ocular diseases associated with large genes.

项目概要我们的目标是推进我们当前的 DNA 纳米颗粒 (NP) 递送平台和基因表达，以开发安全的针对由光感受器缺陷引起的重要的光感受器相关眼部疾病的有效疗法大基因。我们正在融合分子生物工程、纳米配方、材料科学、眼科等方面的知识生物学/生理学和化学加速产生有效非病毒基因的重要步骤眼科疾病配送平台。纳米颗粒由单分子 DNA 与聚赖氨酸压实组成。 PEG聚阳离子，直径为8-11 nm。它们的体积小，并且通过细胞表面进行细胞摄取核仁素可有效地将 NP 转运至细胞核，这解释了它们转染有丝分裂后的能力光感受器。使用 NP 可以实现大基因的高效表达，这是针对难以挽救的视网膜疾病。尽管基因大小上限尚未确定，在肺中测试的最大尺寸为 20 kb，在眼睛中测试的最大尺寸约为 14 kb，这使得 NP 成为理想的替代品用于传递大基因的 AAV。我们已经证明 NP 治疗可有效转染眼部包括光感受器和视网膜色素上皮在内的细胞，即使在多次注射，分布在整个视网膜下腔，并介导明显的结构/功能拯救色素性视网膜炎、莱伯氏先天性黑蒙、斯塔加特氏病和糖尿病小鼠模型视网膜病变。在狒狒眼睛中也实现了无毒性的有效基因表达。这些证明—— 原理研究证实了该技术的潜在临床意义，并强调了该技术的价值大容量，但强调需要长时间高水平的基因表达。我们的主要目标是增强光感受器对纳米粒子的摄取，以达到大基因表达的长期治疗水平全表型拯救。我们建议通过靶向载体工程来增强表达来实现这一目标水平与 NP 传递平台一起增强它们从玻璃体到光感受器的传递，实现泛视网膜分布，促进细胞核中游离体的稳定性并防止表观遗传沉默。随后，我们将测试这些优化的 NP/递送平台介导完整表型的能力大型基因疾病模型中的拯救；特别是 Stargardt 的 Abca4-/- 模型与缺乏 ABCA4基因。 ABCA4是一个大基因，传统病毒载体尚未完全拯救，作为 Stargardt 靶向治疗的结果开发已经滞后。我们计划在 goal1 中设计向量可以在光感受器中达到治疗水平的表达，并在目标 2 中开发有效的 NP- 增强光感受器从玻璃体摄取纳米粒子的递送平台。目标 3 将测试效果最有效的 NP 的最佳递送平台，可实现治疗有效的表达水平 Abca4-/-小鼠发病前后的光感受器。综上所述，由此得出的结果该应用将促进DNA NP在与大基因相关的眼部疾病中的应用。

项目成果

期刊论文数量（12）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Co-Injection of Sulfotyrosine Facilitates Retinal Uptake of Hyaluronic Acid Nanospheres Following Intravitreal Injection.

DOI：
10.3390/pharmaceutics13091510
发表时间：
2021-09-18
期刊：
Pharmaceutics
影响因子：
5.4
作者：
Eblimit A;Makia MS;Strayve D;Crane R;Conley SM;Sinha T;Acharya G;Al-Ubaidi MR;Naash MI
通讯作者：
Naash MI

Yttrium oxide nanoparticles prevent photoreceptor death in a light-damage model of retinal degeneration.

DOI：
10.1016/j.freeradbiomed.2014.07.013
发表时间：
2014-10
期刊：
FREE RADICAL BIOLOGY AND MEDICINE
影响因子：
7.4
作者：
Mitra, Rajendra N.;Merwin, Miles J.;Han, Zongchao;Conley, Shannon M.;Al-Ubaidi, Muayyad R.;Naash, Muna I.
通讯作者：
Naash, Muna I.

Ocular delivery of compacted DNA-nanoparticles does not elicit toxicity in the mouse retina.

DOI：
10.1371/journal.pone.0007410
发表时间：
2009-10-12
期刊：
PloS one
影响因子：
3.7
作者：
Ding XQ;Quiambao AB;Fitzgerald JB;Cooper MJ;Conley SM;Naash MI
通讯作者：
Naash MI

Synthesis and characterization of glycol chitosan DNA nanoparticles for retinal gene delivery.

DOI：
10.1002/cmdc.201300371
发表时间：
2014-01
期刊：
CHEMMEDCHEM
影响因子：
3.4
作者：
Mitra, Rajendra N.;Han, Zongchao;Merwin, Miles;Al Taai, Muhammed;Conley, Shannon M.;Naash, Muna I.
通讯作者：
Naash, Muna I.

A review of therapeutic prospects of non-viral gene therapy in the retinal pigment epithelium.