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

• 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
• 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

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 先前临床试验失败的策略，再生疗法已增加了对大脑的关注 中风后修复和功能恢复。最近，突破性发现表明转导 非神经元细胞，例如带有panneuronal转录因子神经1（ND1）的反应性星形胶质细胞和 其他一些可以将这些细胞直接重新编程为神经祖细胞，甚至是成熟和功能性神经元 通过称为直接重编程/转换的过程，该过程绕过干细胞阶段。慢病毒矢量传递 反应性星形胶质细胞的ND1导致永久重编程的功能神经元无需 维持基因的异位表达。因此，内部直接重新编程意味着 内源性神经发生的前所未有的资源通过利用现有的增殖星形胶质细胞。这 提案是缺血性中风后直接重编程的新颖应用，并探讨了其在 老鼠。这种方法具有伤害引起的星形胶质细胞激活和积累的优势 侵入周围区域。在损伤部位重新编程的新神经元，称为诱导神经元（无神经元或INS） 是自体和有丝分裂后的，可以消除移植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 体外和小鼠局灶性缺血性卒中模型。甚至在大脑中发现了许多转换的INS 中风后一个月和转换。基于我们的体外和体内数据和新兴证据 从其他小组中，我们建议在老年的局灶性缺血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INS下的体外和体内重编程 低氧/缺血性条件。使用与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的功效，效率和时间窗口作为内源性 用于脑修复的神经元供应。特定目标2将测试直接重新编程的假设 时间可以减少神经发生的物理和化学障碍。利益的机制和 将测试具有神经保护性和允许再生的平衡微环境。 特定3将研究直接转换以及增加周围的康复策略 老年小鼠的活性，旨在克服老年人的神经创造和神经可塑性受损 脑。我们假设组合方法促进了活动依赖性神经可塑性，电路 维修和中风后的功能恢复。这三个目标是协调但不同的再生的目标 机制，以令人信服的证据和最先进的技术认可。我们预测每个目标 单独和/或一起将提供新的策略来进行再生疗法。

项目成果

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
• 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