Identifying novel regenerative treatments for CNS injury in adult mammals

确定成年哺乳动物中枢神经系统损伤的新型再生疗法

• 批准号: 10735524
• 负责人: SHUXIN LI
• 金额: $ 49.9万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10735524
• 关键词: Actomyosin; Adult; Axon; Binding; Cervical; Chronic; Clinical; Clinical Trials; Combined Modality Therapy; Cytoskeletal Proteins; Cytoskeleton; Disabled Persons; Dorsal; Environment; Failure; Gene Targeting; Genes; Genetic; Growth; Growth Cones; Individual; Knockout Mice; Lesion; Locomotor Recovery; Mammals; Microfilaments; Microtubules; Modeling; Molecular Target; Motor; Mus; Myosin Type II; Natural regeneration; Neuronal Injury; Neurons; Nonmuscle Myosin Type IIA; Nonmuscle Myosin Type IIB; Paralysed; Pathway interactions; Patients; Peptides; Pilot Projects; Population; Proteins; Recovery; Recovery of Function; Rodent; Role; Sensory; Signal Pathway; Signal Transduction; Spinal Cord; Spinal Cord Contusions; Spinal cord injury; Synapsins; Systemic Therapy; Therapeutic; Thoracic spinal cord structure; Tracer; Translating; Work; Zinc Fingers; antagonist; axon growth; axon injury; axon regeneration; axonal degeneration; cell growth; central nervous system injury; combinatorial; conditional knockout; design; effective therapy; inhibitor; injury and repair; mutant; neuronal growth; neurotransmission; non-muscle myosin; novel; pharmacologic; regenerative approach; regenerative treatment; repaired; safety assessment; targeted agent; therapeutic gene; transcription factor; translational study

Abstract We propose to study whether targeting both the transcription factor ZNF362 and cytoskeletal non-muscle myosin IIA and IIB (NMIIA&B) will promote better axon regeneration and functional recovery after spinal cord injury (SCI) than either approach alone. After SCI, severed axons fail to regenerate largely because of the reduced intrinsic growth capacity of adult CNS neurons and the poor environment for axon extension. Treatments to recover paralysis and other lost functions are not available and patients with SCI are often permanently disabled. Many genes have been determined to control the regrowth failure of mature neurons, but none have been translated to clinical use. There is a persistent need to identify better gene targets and therapeutic strategies. ZNF362 is highly expressed in the CNS, but its major function in mammals remains largely unknown. The PI’s group generated ZNF362 conditional knockout (cKO) mice and designed novel small peptides to block ZNF362 function selectively. Our pilot studies suggest that ZNF362 strongly suppresses the growth capacity of CNS neurons in adult rodents. After CNS injury, various inhibitory molecules around the lesion activate neuronal RhoA, which, in turn, activates cytoskeletal NMIIA&B to condense actin filaments and restricts microtubule protrusion and axon elongation. Because neuronal cytoskeleton is the major machinery to drive axon growth and the converging targets of multiple signaling pathways that control axon growth, manipulating neuronal cytoskeleton is also very attractive to promote significant CNS axon regeneration. We hypothesize that inhibiting both ZNF362 and NMIIA&B represents a dual approach for enhancing neuronal growth capacity and reducing environmental inhibition around the lesion. We propose to dissect potential critical roles of ZNF362 and NMIIA&B for controlling regrowth of mature CNS neurons and to develop novel and effective strategies for promoting CNS axon regeneration. We aim to stimulate robust axon regrowth and functional recovery in SCI rodents by suppressing these genes using cKO mice and new selective antagonist peptides designed in our lab. In Aim 1, we will study whether transgenically deleting ZNF362 and NMIIA&B acts synergistically to promote axon regeneration and recovery in adult mice with SCI. Aim 2 proposes to determine whether blocking individual ZNF362 and NMIIA&B signals pharmacologically with novel selective antagonists promotes axon regeneration and recovery in adult rodents with SCI. In Aim 3, we plan to develop combination therapies that block both ZNF362 and NMIIA&B signals, aiming to yield robust axon regrowth and functional recovery in adult rodents with SCI. Based on the promising results of our pilot studies, we anticipate that our novel regenerative strategies will significantly advance our ability to treat SCI. If our peptides are successful with rodent SCI models, we plan to move this work to peptide safety assessments and further translational studies (e.g., cervical/chronic SCI and clinical trials). Therefore, this project may facilitate identifying novel molecular targets for CNS repair and highly effective strategies for treating CNS lesions.

摘要 我们建议研究是否同时靶向转录因子ZNF 362和细胞骨架非肌肉细胞， 肌球蛋白IIA和II B（NMIIA&B）可促进脊髓损伤后轴突再生和功能恢复 （一）单方面的伤害。脊髓损伤后，切断的轴突不能再生，主要是因为 成年中枢神经系统神经元内在生长能力下降，轴突延伸环境恶劣。治疗 恢复瘫痪和其他丧失的功能是不可用的，SCI患者往往是永久性的 禁用.许多基因已被确定控制成熟神经元的再生失败，但没有一个 已转化为临床应用。一直需要确定更好的基因靶点和治疗方法。 战略布局ZNF 362在中枢神经系统中高度表达，但其在哺乳动物中的主要功能仍不清楚。 PI的研究小组产生了ZNF 362条件性敲除（cKO）小鼠，并设计了新的小肽来阻断 ZNF 362选择性地发挥作用。我们的初步研究表明，ZNF 362强烈抑制生长能力， 成年啮齿类动物的中枢神经系统神经元。中枢神经系统损伤后，病变周围的各种抑制性分子激活神经元， RhoA，其反过来激活细胞骨架NMIIA和B以使肌动蛋白丝凝聚并限制微管 突起和轴突伸长。由于神经元细胞骨架是驱动轴突生长的主要机制， 控制轴突生长的多种信号通路的会聚目标，操纵神经元 细胞骨架也非常有吸引力，以促进显着的CNS轴突再生。我们假设抑制 ZNF 362和NMIIA&B都代表了增强神经元生长能力和降低神经元生长能力的双重途径。 病变周围的环境抑制。我们建议剖析ZNF 362和NMIIA&B的潜在关键作用 用于控制成熟CNS神经元的再生，并开发新的和有效的策略， 轴突再生我们的目标是刺激脊髓损伤啮齿动物的轴突再生和功能恢复， 使用cKO小鼠和我们实验室设计的新的选择性拮抗肽抑制这些基因。在目标1中， 我们将研究转基因缺失ZNF 362和NMIIA&B是否协同作用促进轴突生长， 脊髓损伤成年小鼠的再生和恢复。目标2建议确定是否阻止个人 ZNF 362和NMIIA&B信号转导与新型选择性拮抗剂联合促进轴突再生 以及成年脊髓损伤啮齿类动物的恢复。在目标3中，我们计划开发联合疗法， ZNF 362和NMIIA&B信号，目的是在成年啮齿动物中产生稳健的轴突再生和功能恢复， SCI.基于我们的试点研究的有希望的结果，我们预计我们的新再生策略将 大大提高了我们治疗SCI能力。如果我们的肽在啮齿动物SCI模型上取得成功，我们计划 将这项工作转移到肽安全性评估和进一步的翻译研究（例如，颈椎/慢性SCI和 临床试验）。因此，该项目可能有助于确定CNS修复的新分子靶点， 治疗CNS病变的高效策略。