Cell-based Platform for Gene Delivery to the Brain

基于细胞的基因传递至大脑的平台

• 批准号: 10333329
• 负责人: ELENA BATRAKOVA
• 金额: $ 33.86万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-03-01 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10333329
• 关键词: Acute; Adverse effects; Affect; Anti-Inflammatory Agents; Astrocytes; Autologous; Behavioral; Biodistribution; Biological Availability; Blood - brain barrier anatomy; Blood Cells; Brain; Brain Injuries; Cell Communication; Cell Line; Cell Survival; Cells; Central Nervous System Diseases; Chimeric Proteins; Corpus striatum structure; Development; Diagnosis; Diagnostic; Disease; Dopamine; Dose; Drug Delivery Systems; Electrophoretic Mobility Shift Assay; Embryo; Emission-Computed Tomography; Encephalitis; Engineering; Evaluation; Fluorescence-Activated Cell Sorting; Formulation; Freezing; Gene Delivery; Gene Expression; Gene Targeting; Gene Transfer; Genes; Genetic Materials; Glial Fibrillary Acidic Protein; Goals; Histologic; Home; Homing; Horizontal Gene Transfer; Human; Image; Immune system; In Vitro; Individual; Inflammation; Inflammatory; Infusion procedures; Injections; Integrins; Intoxication; Knock-out; Lead; Ligands; Link; Lymphocyte Function-Associated Antigen-1; Measures; Mediating; Messenger RNA; Microdialysis; Microglia; Modality; Modeling; Mus; NF-kappa B; Nerve Degeneration; Nerve Regeneration; Neurodegenerative Disorders; Neuroglia; Neurons; Neurotransmitters; Nuclear Extract; Organ; Outcome; Palliative Care; Parkin; Parkinson Disease; Patients; Photons; Procedures; Production; Protein Biosynthesis; Proteins; Proteomics; Reporter Genes; Reporting; Research; Reverse Transcriptase Polymerase Chain Reaction; Role; Route; Schedule; Site; Substantia nigra structure; Synapsin I; T-Lymphocyte; Testing; Therapeutic; Therapeutic Agents; Therapeutic Index; Toxicology; Transfection; Transgenic Mice; Treatment Efficacy; Treatment Factor; Viral Vector; Western Blotting; Work; base; behavioral study; bioimaging; brain cell; brain endothelial cell; brain tissue; dopaminergic neuron; exosome; experimental study; extracellular vesicles; gene delivery system; glial cell-line derived neurotrophic factor; high risk; immunogenicity; improved; in vivo; induced pluripotent stem cell; infrared spectroscopy; insight; intercellular cell adhesion molecule; macromolecule; macrophage; magnetic resonance spectroscopic imaging; monocyte; mouse model; neuroimaging; neuronal survival; neuroprotection; neurotrophic factor; novel; novel therapeutic intervention; novel therapeutics; peripheral blood; plasmid DNA; promoter; real-time images; receptor; regeneration function; restoration; safety study; single photon emission computed tomography; therapeutic gene; therapeutic protein; therapy development; transcription factor; vector

ABSTRACT Parkinson’s disease (PD) affects more than a million individuals in the U.S. with up to 60,000 new cases diagnosed each year. Currently, there are no treatments that can halt or reverse the course of PD; only palliative therapies, such as replacement strategies for missing neurotransmitters, exist. The pathobiology of PD is associated with the loss of dopaminergic (DA) neurons. Thus, the successful delivery of neurotrophic factors, in particular, glial cell line-derived neurotrophic factor (GDNF), that promote neuronal survival and reverse the progression of PD is of great importance. Regrettably, the blood brain barrier (BBB) remains a seemingly insurmountable obstacle to the routine use of systemically administered macromolecules - including GDNF 1. To circumvent this problem, we propose using genetically modified peripheral blood monocytes (PBM) for systemic gene delivery to the brain. It is well established that specialized cells of the immune system, including monocytes, macrophages, and T cells, can easily penetrate the BBB and migrate rapidly to sites of brain inflammation and degeneration 2,3. Our research has previously demonstrated that macrophages transfected ex vivo with therapeutic protein-encoding DNA plasmid (pDNA), can deliver therapeutic gene in intoxicated mice with acute brain inflammation, and in the transgenic mice, Parkin-Q311X(A). Mechanistic studies revealed that genetically modified macrophages release extracellular vesicles, exosomes, packed with protein-encoding genetic material, pDNA and mRNA, as well as a transcription factor involved in the encoded gene expression. Importantly, multiple lines of evidence for therapeutic efficacy were observed in PD mouse models, including decreased brain inflammation, significant neuroprotection, and improved locomotor functions. Planned studies include the evaluation of brain bioavailability for engineered PBM and GDNF gene transfer in Parkin-Q311X(A) mouse model (SA1). To enforce outcomes of the new formulation, PBM will be differentiated to a specific subset of “alternatively activated” (M2) macrophages with regenerative functions. The mechanism of macrophage-mediated gene transfer including involvement cell-cell interactions and/or exosomes secreted by GDNF-transfected PBM will be elucidated (SA2). We will then assess the therapeutic potential of this novel product by measuring its anti-inflammatory and neuroprotective effects and, lastly, by extensive behavioral analysis in Parkin-Q311X(A) mice (SA3). To provide translational link, human induced pluripotent stem cells (iPSCs) with almost unlimited expandability will be tested. Furthermore, to obtain a universal cell-carrier, iPSCs with knockout MHC class II (MHC-II) receptor will be utilized. Our studies will provide fundamental insights into how PBM interact with brain cells and facilitate horizontal gene transfer upon neurodegeneration, potentially opening up other cell-based gene delivery systems to the CNS and beyond.

摘要 帕金森病（PD）在美国影响着100多万人，新发病例多达60，000例 每年诊断。目前，还没有治疗方法可以阻止或逆转PD的进程;只有姑息治疗。 存在诸如用于缺失的神经递质的替代策略的疗法。PD的病理学是 与多巴胺能（DA）神经元的损失有关。因此，神经营养因子的成功递送， 特别是胶质细胞源性神经营养因子（GDNF），它促进神经元存活并逆转神经元凋亡。 PD的进展非常重要。遗憾的是，血脑屏障（BBB）仍然是一个表面上， 这是常规使用全身施用的大分子（包括GDNF 1）的不可逾越的障碍。 为了避免这个问题，我们建议使用遗传修饰的外周血单核细胞（PBM）， 系统性基因传递到大脑。众所周知，免疫系统的特化细胞，包括 单核细胞、巨噬细胞和T细胞可以容易地穿透血脑屏障并迅速迁移到脑组织部位。 炎症和变性2，3.我们以前的研究已经证明， 体内携带治疗性蛋白编码DNA质粒（pDNA），可将治疗性基因导入中毒小鼠体内 急性脑炎症，在转基因小鼠中，Parkin-Q311 X（A）。机制研究表明， 基因修饰的巨噬细胞释放细胞外囊泡，外泌体， 遗传物质、pDNA和mRNA，以及参与编码基因表达的转录因子。 重要的是，在PD小鼠模型中观察到了治疗功效的多条证据，包括 减少脑炎症、显著的神经保护和改善的运动功能。 计划中的研究包括评估工程PBM和GDNF基因转移在脑中的生物利用度。 Parkin-Q311 X（A）小鼠模型（SA 1）。为了执行新公式的结果，将区分PBM 具有再生功能的“交替激活”（M2）巨噬细胞的特定子集。机制 巨噬细胞介导的基因转移，包括参与细胞-细胞相互作用和/或 将阐明GDNF转染的PBM（SA 2）。然后我们将评估这部小说的治疗潜力 产品通过测量其抗炎和神经保护作用，最后，通过广泛的行为， 在Parkin-Q311 X（A）小鼠中的分析（SA 3）。为了提供翻译连接，人诱导多能干细胞 具有几乎无限可扩展性的iPSC将被测试。此外，为了获得通用细胞载体，iPSC 将使用具有敲除的MHC II类（MHC-II）受体的细胞。 我们的研究将为PBM如何与脑细胞相互作用提供基本见解， 神经退行性变时的水平基因转移，可能打开其他基于细胞的基因递送 系统到CNS和更远。

项目成果

Extracellular Vesicles Released by Genetically Modified Macrophages Activate Autophagy and Produce Potent Neuroprotection in Mouse Model of Lysosomal Storage Disorder, Batten Disease.

• DOI: 10.3390/cells12111497
• 发表时间: 2023-05-29
• 期刊: CELLS
• 影响因子: 6
• 作者: El-Hage, Nazira;Haney, Matthew J.;Zhao, Yuling;Rodriguez, Myosotys;Wu, Zhanhong;Liu, Mori;Swain, Carson J.;Yuan, Hong;Batrakova, Elena V.
• 通讯作者: Batrakova, Elena V.

Using Extracellular Vesicles Released by GDNF-Transfected Macrophages for Therapy of Parkinson Disease.

• DOI: 10.3390/cells11121933
• 发表时间: 2022-06-15
• 期刊: Cells
• 影响因子: 6

Targeting Beclin1 as an Adjunctive Therapy against HIV Using Mannosylated Polyethylenimine Nanoparticles.

• DOI: 10.3390/pharmaceutics13020223
• 发表时间: 2021-02-06
• 期刊: Pharmaceutics
• 影响因子: 5.4
• 作者: Rodriguez M;Soler Y;Muthu Karuppan MK;Zhao Y;Batrakova EV;El-Hage N
• 通讯作者: El-Hage N

Biodistribution of Biomimetic Drug Carriers, Mononuclear Cells, and Extracellular Vesicles, in Nonhuman Primates.

• DOI: 10.1002/adbi.202101293
• 发表时间: 2022-03
• 期刊: Advanced biology
• 影响因子: 3.7
• 作者: Haney MJ;Yuan H;Shipley ST;Wu Z;Zhao Y;Pate K;Frank JE;Massoud N;Stewart PW;Perlmutter JS;Batrakova EV
• 通讯作者: Batrakova EV

Mannosylated Cationic Copolymers for Gene Delivery to Macrophages.

• DOI: 10.1002/mabi.202000371
• 发表时间: 2021-04
• 期刊: Macromolecular bioscience
• 影响因子: 4.6
• 作者: Lopukhov AV;Yang Z;Haney MJ;Bronich TK;Sokolsky-Papkov M;Batrakova EV;Klyachko NL;Kabanov AV
• 通讯作者: Kabanov AV