Vector engineering for non-viral delivery of large genomic DNA to the RPE
用于将大基因组 DNA 非病毒传递至 RPE 的载体工程
基本信息
- 批准号:10667049
- 负责人:
- 金额:$ 23.25万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2023
- 资助国家:美国
- 起止时间:2023-06-01 至 2025-05-31
- 项目状态:未结题
- 来源:
- 关键词:AffectBiochemicalBlindnessCellsClinicalClinical TrialsCloningConeCytomegalovirus InfectionsDNADependovirusDiseaseDistalEarly treatmentElementsEngineeringEnhancersEnsureEquilibriumEquipmentExonsEyeGene DeliveryGene ExpressionGene Expression RegulationGene Transduction AgentGenesGeneticGenomic DNAGenomicsGoalsHereditary DiseaseHigh PrevalenceHousingHumanHuman CloningInheritedInjectionsIntronsLeber&aposs amaurosisLongevityMatrix Attachment RegionsMediatingMethodologyMethodsModelingMolecular BiologyMusOnset of illnessOutcomeOutcome MeasurePapioParentsPeptidesPhenotypePhysiologicalPolyethylene GlycolsPolylysinePositioning AttributeQuantitative Reverse Transcriptase PCRRPE65 proteinRegulationRegulatory ElementResearchResourcesRetinaRetinal DiseasesRetinal gene therapyRetinitis PigmentosaRodStructureStructure of retinal pigment epitheliumTestingTherapeuticToxic effectTranscriptTranslationsTreatment EfficacyUntranslated RNAUntranslated RegionsViralVisionVisualWestern Blottingadeno-associated viral vectorcis acting elementclinical applicationcytotoxicdelivery vehicledesigndisease phenotypeearly childhoodefficacious treatmentefficacy evaluationexperienceexperimental studyexpression vectorextracellulargene therapyimprovedin vivoinnovationmouse modelnanoparticlenon-viral gene therapynovelpreservationpromoterscaffoldtargeted treatmenttherapeutic genetherapeutically effectivetherapy developmenttooltransduction efficiencytransgene expressiontranslational impactuptakevector
项目摘要
Summary
The vast number of inherited blinding disorders has made ocular gene therapy an active research field in recent
years, culminating in well-publicized clinical trials. However, efficacious therapies are still elusive for two
reasons: gene expression often remains inadequate in duration and levels, and the limited packaging capacity
of standard vectors prohibits the inclusion of disease genes with their cis-regulatory elements. Overcoming
these barriers is critical for the advancement of the field and widespread clinical application. Current gene
delivery strategies use cDNA-based vectors that lack the non-coding and cis-elements found in genomic DNA
that regulate gene expression. These genomic sequences can preserve the stability of the transcript, improve
translation and produce physiologically relevant levels of expression. In this application, we test the hypothesis
that providing the entire corrective gene including its authentic promoter, enhancer, introns and untranslated
regions (UTRs) improves the levels and duration of transgene expression in mouse model of RPE65-associated
Leber Congenital Amaurosis (LCA), a disease that affects the retinal pigment epithelium (RPE). This disease
manifests in early childhood leading to a gradual vision loss often resulting in blindness. Due to the high
prevalence of RPE65-associated LCA, several clinical trials have been conducted using adeno-associated
viruses (AAV) to deliver human RPE65 cDNA. However, almost all attempts failed to halt the ongoing visual
loss. We thus aim to establish an effective therapeutic approach by delivering the whole human RPE65 gene
with its 5’/3’ regulatory elements, exons and introns to treat a mouse model of LCA (RPE65-/-). We aim to achieve
full regulation and long-term of expression in a cell-specific manner to ensure rescue of the LCA disease
phenotype. We have cloned the human RPE65 genomic fragment into an expression vector and will develop
an effective delivery platform utilizing naked DNA or DNA formulated as nanoparticles (NPs) with polylysine
peptides conjugated to polyethylene glycol (CK30PEG). We will test the longevity and levels of gene expression
after RPE65 delivery using these platforms, and evaluate their ability to mediate full phenotypic rescue in the
RPE65-/- mice. In aim1, we will engineer vectors that can achieve therapeutic levels of expression in the RPE
and in aim 2 we will evaluate long-term rescue and transduction efficiency in the RPE of the LCA model before
and after the onset of the disease phenotype. In summary, results from this application will facilitate the
advancement of non-viral gene therapy for RPE-associated diseases.
摘要
近年来,大量的遗传性致盲疾病使眼部基因治疗成为一个活跃的研究领域
几年,最终在广为人知的临床试验中达到顶峰。然而,对于这两个人来说,有效的治疗方法仍然难以捉摸。
原因:基因表达在持续时间和水平上往往仍然不足,而且包装能力有限
标准载体的使用禁止将疾病基因及其顺式调控元件包括在内。克服困难
这些障碍对该领域的发展和临床的广泛应用至关重要。现行基因
递送策略使用基于cdna的载体,这种载体缺乏基因组DNA中的非编码元件和顺式元件。
来调节基因的表达。这些基因组序列可以保持转录本的稳定性,提高
翻译并产生生理上相关的表达水平。在此应用程序中,我们检验假设
提供完整的纠正基因,包括其真正的启动子、增强子、内含子和未翻译的
区域(UTRs)改善RPE65相关小鼠模型中转基因表达的水平和持续时间
Leber先天性黑色素沉着症(LCA),一种影响视网膜色素上皮(RPE)的疾病。这种病
表现在儿童早期,导致逐渐的视力丧失,常常导致失明。由于高企
RPE65相关LCA的患病率,已经进行了几项临床试验,使用腺相关
携带人RPE65基因的病毒(AAV)。然而,几乎所有的尝试都未能阻止正在进行的视觉
损失。因此,我们的目标是通过传递完整的人类RPE65基因来建立一种有效的治疗方法
利用其5‘/3’调控元件、外显子和内含子治疗LCA小鼠模型(RPE65-/-)。我们的目标是实现
以细胞特异性的方式全面调控和长期表达,以确保拯救LCA疾病
表型。我们已经将人RPE65基因组片段克隆到表达载体中,并将开发出
利用裸露的DNA或以纳米粒子(NPs)形式与多赖氨酸复合的DNA的有效传递平台
聚乙二醇偶联肽(CK30PEG)。我们将测试寿命和基因表达水平
在使用这些平台交付RPE65之后,并评估它们在
RPE65-/-小鼠。在AIM 1中,我们将设计能够在RPE中达到治疗性表达水平的载体
在目标2中,我们将评估LCA模型在RPE中的长期救援和转导效率
和发病后的疾病表型。总之,这项申请的结果将有助于
RPE相关疾病的非病毒基因治疗进展。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Muna I. Naash其他文献
Characterization of glutathione peroxidase in frog retina.
青蛙视网膜谷胱甘肽过氧化物酶的表征。
- DOI:
- 发表时间:
1984 - 期刊:
- 影响因子:2
- 作者:
Muna I. Naash;Robert E. Anderson - 通讯作者:
Robert E. Anderson
882. Non-Viral Ocular Gene Transfer for Hereditary Retinal Degeneration
- DOI:
10.1016/j.ymthe.2006.08.971 - 发表时间:
2006-01-01 - 期刊:
- 影响因子:
- 作者:
Muna I. Naash;Ron M. Ballard;Jeff Skaggs;Zack Nash;Alexander B. Quiambao;Mark J. Cooper;Rafal Farjo - 通讯作者:
Rafal Farjo
The regional distribution of vitamins E and C in mature and premature human retinas.
维生素 E 和 C 在成熟和早产人类视网膜中的区域分布。
- DOI:
- 发表时间:
1988 - 期刊:
- 影响因子:4.4
- 作者:
Jorl C. Nielsen;Muna I. Naash;Robert E. Anderson - 通讯作者:
Robert E. Anderson
Correction: Comparative study of PRPH2 D2 loop mutants reveals divergent disease mechanism in rods and cones
- DOI:
10.1007/s00018-023-04929-y - 发表时间:
2023-09-12 - 期刊:
- 影响因子:6.200
- 作者:
Larissa Ikelle;Mustafa Makia;Tylor Lewis;Ryan Crane;Mashal Kakakhel;Shannon M. Conley;James R. Birtley;Vadim Y. Arshavsky;Muayyad R. Al-Ubaidi;Muna I. Naash - 通讯作者:
Muna I. Naash
Muna I. Naash的其他文献
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{{ truncateString('Muna I. Naash', 18)}}的其他基金
Non-viral gene delivery platforms for the treatment of Usher Syndrome Type 2A.
用于治疗 2A 型亚瑟综合症的非病毒基因递送平台。
- 批准号:
10578428 - 财政年份:2023
- 资助金额:
$ 23.25万 - 项目类别:
Compacted DNA Nanoparticles for Ocular Therapy
用于眼部治疗的压缩 DNA 纳米颗粒
- 批准号:
8504140 - 财政年份:2013
- 资助金额:
$ 23.25万 - 项目类别:
Compacted DNA Nanoparticles for Ocular Therapy
用于眼部治疗的压缩 DNA 纳米颗粒
- 批准号:
8677899 - 财政年份:2013
- 资助金额:
$ 23.25万 - 项目类别:
DNA nanoparticle formulations for optimal ocular gene delivery
用于最佳眼部基因传递的 DNA 纳米颗粒配方
- 批准号:
8545860 - 财政年份:2012
- 资助金额:
$ 23.25万 - 项目类别:
DNA nanoparticle formulations for optimal ocular gene delivery
用于最佳眼部基因传递的 DNA 纳米颗粒配方
- 批准号:
8734431 - 财政年份:2012
- 资助金额:
$ 23.25万 - 项目类别:
DNA nanoparticle formulations for optimal ocular gene delivery
用于最佳眼部基因传递的 DNA 纳米颗粒配方
- 批准号:
8365445 - 财政年份:2012
- 资助金额:
$ 23.25万 - 项目类别:
Compacted DNA Nanoparticles for Ocular Therapy
用于眼部治疗的压缩 DNA 纳米颗粒
- 批准号:
7353945 - 财政年份:2008
- 资助金额:
$ 23.25万 - 项目类别:
Compacted DNA Nanoparticles for Ocular Therapy
用于眼部治疗的压缩 DNA 纳米颗粒
- 批准号:
8007344 - 财政年份:2008
- 资助金额:
$ 23.25万 - 项目类别:
Compacted DNA Nanoparticles for Ocular Therapy
用于眼部治疗的压缩 DNA 纳米颗粒
- 批准号:
8204931 - 财政年份:2008
- 资助金额:
$ 23.25万 - 项目类别:
Compacted DNA Nanoparticles for Ocular Therapy
用于眼部治疗的压缩 DNA 纳米颗粒
- 批准号:
8134621 - 财政年份:2008
- 资助金额:
$ 23.25万 - 项目类别:
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