Combinatorial Manipulation of Transcription Factors to Promote CNS Regeneration

项目总结 在成年人中，中枢神经系统(CNS)中的轴突在受伤或死亡后通常不能再生 疾病，导致永久性和无法治愈的残疾。轴突的生长受到不利的生长环境的阻碍， 以及随着中枢神经系统神经元的老化，轴突生长的内在能力的发育性丧失。转录 因子(TF)与DNA相互作用，协调产生广泛的细胞材料，并具有 成为提高受损神经元再生能力的重要治疗靶点。例如，强制 在成年神经元中重新表达一种名为KLF6的促再生因子可以改善其轴突生长能力 在脊椎受伤后。我们现在将测试三个相辅相成和相辅相成的战略，以加强 KLF6具有良好的促再生性能。首先，使用一种新的生物信息学管道，我们预测 与KLF6在功能上相互作用并验证了它们协同促进轴突生长的其他TF 当与KLF6联合应用于轴突生长的细胞培养模型时。因此，我们将在体内进行三项测试 选定的因子，Eome，NR5A2和RARB，能够增强KLF6的S促再生特性 脊髓损伤的动物模型。其次，我们将用生长移植来补充这些TF干预- 允许干细胞进入脊柱损伤部位。这些移植物将缓解脊髓中持续的生长抑制。 因此，Tf治疗的促再生作用被揭示出来。最后，我们将利用一个新的 开发了基因治疗载体，能够以前所未有的效率逆行传递基因。注射法 将该载体导入脊髓可导致损伤神经元中广泛的基因表达。 脑干、中脑和运动皮质。该交付系统涉及更多数量和更广泛的多样性 细胞类型比目前直接脑注射的做法更多，从而最大限度地实现功能 脊椎损伤后的收益。通过这些目标，组织清除和3D显微镜将揭示新的解剖学 唤起的再生的细节。将这些尖端改进整合到以TF为中心的战略中 将把这一领域推向治疗衰弱症患者的新的有效疗法 中枢神经系统损伤的后果。

PROJECT SUMMARY In adults, axons in the central nervous system (CNS) generally fail to regenerate after they are lost to injury or disease, leading to permanent and incurable disability. Axon growth is prevented by a hostile growth environment, as well as a developmental loss in the intrinsic capacity for axon growth as CNS neurons age. Transcription factors (TFs) interact with DNA and coordinate the production of broad sets of cellular materials, and have emerged as important therapeutic targets to boost regenerative ability within injured neurons. For example, forced re-expression of a pro-regenerative TF called KLF6 in adult neurons can improve their capacity for axon growth after spinal injury. We will now test three complementary and mutually supportive strategies to enhance the promising pro-regenerative properties of KLF6. First, using a novel bioinformatics pipeline, we have predicted additional TFs that functionally interact with KLF6 and verified their ability to synergistically enhance axon growth when combined with KLF6 in cell culture models of axon growth. We will therefore perform in vivo tests of three selected factors, EOMES, NR5A2, and RARB, for the ability to enhance KLF6’s pro-regenerative properties in animal models of spinal cord injury. Second, we will supplement these TF interventions with transplants of growth- permissive stem cells into sites of spinal injury. These grafts will alleviate persistent growth inhibition in the spinal cord environment, and thus unmask the pro-regenerative effects of TF treatments. Finally, we will harness a newly developed gene therapy vector that enables retrograde delivery of genes with unprecedented efficiency. Injection of this vector to the spinal cord results in widespread gene expression in injured neurons throughout the brainstem, midbrain, and motor cortex. This delivery system engages a larger number and a wider diversity of cell types than the current practice of direct brain injection, thus maximizing the chance of achieving functional gains after spinal injury. Throughout these aims, tissue clearing and 3D microscopy will reveal new anatomical details of the evoked regeneration. Bringing together these cutting-edge improvements to a TF-centered strategy will move the field toward novel and effective treatments for individuals suffering from the debilitating consequences of CNS injury.