Targeting the microtubule cytoskeleton to promote cavernous nerve regeneration and erectile function after injury

靶向微管细胞骨架促进损伤后海绵体神经再生和勃起功能

基本信息

批准号：
10719124
负责人：
KELVIN P DAVIES
金额：
$ 66.27万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-01 至 2028-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10719124
关键词：
Acceleration Affect Age Aging Animal Model Animals Antigen-Presenting Cells Apoptosis Architecture Axon Bilateral Blood flow Cells Chronic Complex Cytoskeleton Electric Stimulation Electron Microscopy Endothelial Cells Endothelium Enzymes Erectile dysfunction Fibrosis Functional Regeneration Immune Immunohistochemistry In Vitro Injury Maintenance Measurement Microscopic Microtubules Modeling Morphology NOS2A gene Nerve Nerve Regeneration Neurons Nitroxidergic Nerves Operative Surgical Procedures Outcome Peripheral Nerves Process Proteins Publishing Quantitative Reverse Transcriptase PCR Radical Prostatectomy Rattus Recovery Research Rodent Model Schwann Cells Site Small Interfering RNA Systemic blood pressure TdT-Mediated dUTP Nick End Labeling Assay Testing Time Tissues Trichrome stain Visual Western Blotting age effect aged axon growth axon regeneration behavioral response cell motility cell type density improved in vivo in vivo Model men migration molecular marker nerve injury nerve repair nerve supply neural neuron loss neuronal cell body novel novel therapeutic intervention older men penis preservation pressure prevent recruit repair function response response to injury

项目摘要

Abstract We recently published in vitro studies identifying Fidgetin-like 2 (FL2) as a novel regulator of microtubule dynamics, with an inhibitory effect on axonal growth and nerve regeneration [1]. Since FL2-depletion had axonal growth-promoting effects in vitro, we tested whether FL2 could be targeted to promote nerve regeneration in a rodent model of cavernous nerve (CN) injury. The CNs are parasympathetic nerves that regulate blood flow to the penis and are commonly damaged during the surgical procedures of radical prostatectomy (RP), resulting in erectile dysfunction (ED). Remarkably, in these rodent models, FL2-depletion at the site and time of CN injury led to accelerated visible nerve regeneration, accompanied by improved erectile function outcomes. However, axonal outgrowth is only one component in the complex process of regenerating functional peripheral nerves, which also involves recruitment and activity of multiple accessory cell-types, such as Schwann cells, immune cells, and endothelial cells [2, 3]. In other published studies, we have demonstrated that FL2 can regulate motility and migration of several cell types [4]. Therefore, we hypothesize that the mechanism by which FL2- depletion promotes CN regeneration and improves erectile function outcomes following injury is multi- faceted, involving accelerated migration of key cell types to the site of injury, in addition to enhancing axonal growth. We will test this hypothesis using both in vitro and in vivo models and confirm that the mechanisms of CN repair function in both young and older animals (more appropriate to the age at which men undergo RP). We will test our hypothesis through two Specific Aims. In Aim 1, we will determine if accelerated nerve regeneration by FL2-depletion is associated with increased migration of Schwann cells, immune cells, and endothelial cells to the site of CN injury and if aging affects the recruitment of these accessory cell-types. In Aim 2 we will determine the ability of targeted FL2-depletion to promote CN-regeneration, preserve penile architecture and recover erectile function in rat models of RP. Our research will lead to greater understanding of the underlying mechanisms by which FL2-depletion promotes CN repair and recovery of erectile function after RP, and if aging affects these mechanisms. By understanding these mechanisms, we will potentially identify novel therapeutic strategies not only to treat, but also to prevent, ED following RP.

抽象的我们最近发表了体外研究，鉴定出类似于fidgetin的2（FL2）是微管的新调节剂动力学，对轴突生长和神经再生有抑制作用[1]。由于fl2止动具有轴突在体外，我们测试了FL2是否可以针对促进神经再生海绵神经（CN）损伤的啮齿动物模型。中枢神经系统是副交感神经，调节血液流向阴茎通常在根治性前列腺切除术（RP）的手术过程中受损，导致勃起功能障碍（ED）。值得注意的是，在这些啮齿动物模型中，CN损伤的位点和时间处的FL2消耗导致可见的神经再生，并伴随着改善的勃起功能结果。然而，轴突产物在复杂的功能周围神经的复杂过程中只是一个成分，这也涉及多种辅助细胞类型的募集和活动，例如雪旺细胞，免疫细胞和内皮细胞[2，3]。在其他已发表的研究中，我们证明了FL2可以调节运动能力和几种细胞类型的迁移[4]。因此，我们假设FL2-的机制耗竭促进CN再生并改善受伤后的勃起功能结果是多数刻画，涉及关键细胞类型的加速迁移到伤害部位，此外还可以增强轴突生长。我们将使用体外和体内模型测试这一假设，并确认年轻动物和年龄较大的动物中CN修复功能的机制（更适合男性的年龄进行RP）。我们将通过两个具体目标检验我们的假设。在AIM 1中，我们将确定是否加速神经通过FL2止血的再生与雪旺细胞，免疫细胞和内皮细胞到CN损伤部位，如果衰老会影响这些辅助细胞类型的募集。目标 2我们将确定靶向FL2止动物促进CN恢复，保留阴茎的能力 RP大鼠模型中的架构和恢复勃起功能。我们的研究将导致对FL2消耗促进的潜在机制有更深入的了解 RP后的CN修复和恢复勃起功能，如果衰老会影响这些机制。通过理解这些机制，我们将有潜在地确定新颖的治疗策略，不仅可以治疗，还可以预防，预防， ED遵循RP。