Direct Reprogramming of the brain after ischemic stroke in the aged mouse

老年小鼠缺血性中风后大脑的直接重编程

基本信息

批准号：
10612413
负责人：
LING WEI
金额：
$ 42.05万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-15 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10612413
关键词：
Affect Age Animals Area Astrocytes Attention Autologous Blood Vessels Brain Brain Hypoxia-Ischemia Brain Ischemia Bypass Cell Death Cell Proliferation Cell Survival Cell Transplantation Cells Cerebral Ischemia Cessation of life Chemicals Cicatrix Clinical Trials Data Deterioration Ectopic Expression Electrophysiology (science)Face Female Gene Expression Genes Glial Fibrillary Acidic Protein Graft Rejection Homing Human Impairment In Vitro Infarction Inflammatory Injury Investigation Ischemia Ischemic Stroke Lentivirus Lentivirus Vector Measures Microglia Molecular Morphology Mus Natural regeneration Nerve Regeneration Neuroglia Neuronal Plasticity Neurons Pathway interactions Patients Peripheral Process Proliferating Recovery of Function Regenerative research Rehabilitation therapy Reperfusion Therapy Resources Risk Signal Transduction Site Source Stroke Techniques Technology Testing Therapeutic Therapeutic Studies Time Tissues Transplantation Vibrissae aged aging brain astrogliosis axon growth behavior test brain repair cell regeneration chemokine combinatorial cytokine delivery vehicle design direct application disability efficacy validation enhancing factor experimental study human mortality human old age (65+)in vivo male migration mortality mouse model nerve stem cell neural network neurogenesis neuroprotection novel novel strategies novel therapeutic intervention optogenetics permissiveness post stroke postmitotic programs promoter regeneration function regenerative regenerative approach regenerative repair regenerative therapy release factor repaired restraint stem cells stroke model stroke patient stroke therapy timeline tissue repair transcription factor tumorigenesis

项目摘要

Ischemic stroke is a leading cause of human death and disability. In addition to neuroprotective strategies that have failed previous clinical trials, regenerative therapies have gain escalating attention for brain repair and functional recovery after stroke. Recently, a breakthrough discovery demonstrates that transduction of non-neuronal cells such as reactive astrocytes with the panneuronal transcription factor NeuroD1 (ND1) and a few others can reprogram these cells directly into neural progenitors or even mature and functional neurons via a process called direct reprogramming/conversion that bypasses stem cell stage. Lentiviral vector delivery of ND1 to reactive astrocytes results in permanently reprogrammed functional neurons without the need for maintained ectopic expression of the gene. Thus, intra-lineage direct reprogramming implicates an unprecedented resource of endogenous neurogenesis by leveraging existing proliferative astrocytes. The proposal is a novel application of the direct reprogramming after ischemic stroke and explores its application in aged mice. This approach takes the advantages of injury-induced astrocyte activation and accumulation in the peri-infarct region. Reprogrammed new neurons, termed induced neurons (iNeurons or iNs), at the injury site are autologous and post-mitotic, which eliminate the risk of rejection and tumorigenesis of transplanted exogenous cells. In our preliminary experiments, we successfully converted astrocytes into mature neurons in vitro and in focal ischemic stroke models of the mouse. Many converted iNs were identified in the brain even one months after stroke and the conversion. Based on our in vitro and in vivo data and emerging evidence from other groups, we propose to test this regenerative therapy in a focal ischemic stroke model of aged mouse. Specific Aim 1 will study the in vitro and in vivo reprogramming of astrocytes into iNs under hypoxic/ischemic conditions. Using ND1 lentivirus packaged with the GFAP promoter and mCherry marker, we will validate the efficacy, efficiency and time windows of reprogramming reactive astrocytes as an endogenous neuronal supply for brain repair. Specific Aim 2 will test the hypothesis that direct reprogramming at the right time can reduce the physical and chemical barriers for neurogenesis. The mechanism of the benefits and a balanced microenvironment that is neuroprotective as well as permissive for regeneration will be tested. Specific 3 will study the direct conversion combined with a rehabilitative strategy of increased peripheral activities in aged mice, designed to overcome impaired neuroregeneration and neural plasticity in the aged brain. We hypothesize that the combinatorial approach promotes activity-dependent neural plasticity, circuitry repair, and functional recovery after stroke. These three Aims target coordinated but distinct regenerative mechanisms, endorsed by compelling evidence and state-of-the-art technologies. We predict that each Aim alone and/or together will provide novel strategies for a regenerative therapy.

缺血性中风是人类死亡和残疾的主要原因。除了神经保护作用先前临床试验失败的策略，再生疗法越来越受到大脑的关注中风后的修复和功能恢复。最近，一项突破性发现表明，转导非神经元细胞，例如具有全神经元转录因子 NeuroD1 (ND1) 的反应性星形胶质细胞和其他一些细胞可以将这些细胞直接重新编程为神经祖细胞，甚至成熟且有功能的神经元通过绕过干细胞阶段的直接重编程/转换过程。慢病毒载体递送 ND1 与反应性星形胶质细胞的结合会导致功能性神经元永久重新编程，而不需要维持该基因的异位表达。因此，谱系内直接重编程意味着通过利用现有的增殖性星形胶质细胞获得前所未有的内源性神经发生资源。这该提案是缺血性中风后直接重编程的一种新颖应用，并探索了其在以下领域的应用：年老的老鼠。该方法利用了损伤诱导的星形胶质细胞激活和积累的优点梗塞周围区域。在损伤部位重新编程的新神经元，称为诱导神经元（iNeurons 或 iNs）是自体和有丝分裂后的，消除了移植物的排斥和肿瘤发生的风险外源细胞。在我们的初步实验中，我们成功地将星形胶质细胞转化为成熟的神经元小鼠的体外和局灶性缺血性中风模型。甚至在大脑中也发现了许多转化的 iN 中风和转换后一个月。基于我们的体外和体内数据以及新出现的证据与其他小组相比，我们建议在老年人的局灶性缺血性中风模型中测试这种再生疗法老鼠。具体目标 1 将研究星形胶质细胞在体外和体内重编程为 iN 的情况缺氧/缺血状况。使用带有 GFAP 启动子和 mCherry 标记包装的 ND1 慢病毒，我们将验证重编程反应性星形胶质细胞作为内源性星形胶质细胞的功效、效率和时间窗大脑修复的神经元供应。具体目标 2 将检验直接在右侧进行重编程的假设时间可以减少神经发生的物理和化学障碍。效益及机制将测试具有神经保护作用且允许再生的平衡微环境。具体3将研究直接转换结合增加外周血的康复策略老年小鼠的活动，旨在克服老年小鼠的神经再生和神经可塑性受损脑。我们假设组合方法促进活动依赖性神经可塑性、电路修复和中风后的功能恢复。这三个目标的目标是协调但又不同的再生机制，并得到令人信服的证据和最先进技术的支持。我们预测每个目标单独和/或一起将为再生治疗提供新策略。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Pathogenesis of sporadic Alzheimer's disease by deficiency of NMDA receptor subunit GluN3A.

DOI：
10.1002/alz.12398
发表时间：
2022-03
期刊：
Alzheimer's & dementia : the journal of the Alzheimer's Association
影响因子：
0
作者：
Zhong W;Wu A;Berglund K;Gu X;Jiang MQ;Talati J;Zhao J;Wei L;Yu SP
通讯作者：
Yu SP

Extrasynaptic NMDA receptors in acute and chronic excitotoxicity: implications for preventive treatments of ischemic stroke and late-onset Alzheimer's disease.

DOI：
10.1186/s13024-023-00636-1
发表时间：
2023-07-03
期刊：
MOLECULAR NEURODEGENERATION
影响因子：
15.1
作者：
Yu, Shan P.;Jiang, Michael Q.;Shim, Seong S.;Pourkhodadad, Soheila;Wei, Ling
通讯作者：
Wei, Ling