Mechanisms of axon guidance in laryngeal reinnervation following injury of the recurrent laryngeal nerve

喉返神经损伤后喉部神经支配的轴突引导机制

• 批准号: 10663855
• 负责人: Michael Pitman
• 金额: $ 62.78万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10663855
• 关键词: Adult; Axon; Businesses; Chronic Disease; Chronic Obstructive Pulmonary Disease; Chronology; Communication; Congestive Heart Failure; Controlled Environment; Cues; Data; Development; Disease; Dysphonia; Embryology; Exhibits; Fright; Frustration; Functional disorder; Future; Goals; Growth Cones; Impairment; In Vitro; Injury; Knowledge; Laboratories; Laryngeal Injury; Laryngeal muscle structure; Larynx; Mental Depression; Modeling; Modification; Molecular; Morbidity - disease rate; Motion; Motor Neurons; Muscle; NTN1 gene; Nerve Regeneration; Operative Surgical Procedures; Paralysed; Pathway interactions; Patients; Pattern; Persons; Play; Process; Productivity; Quality of life; Rattus; Receptor Signaling; Recurrent Laryngeal Nerve; Research; Role; Signal Transduction; Synkinesis; System; Testing; United States; Up-Regulation; Voice; Work; axon growth; axon guidance; axonal pathfinding; disability payment; experimental study; glial cell-line derived neurotrophic factor; in vivo Model; loved ones; muscle reinnervation; nerve injury; nerve supply; neuronal survival; nucleus ambiguus; optimal treatments; receptor; receptor expression; regenerative growth; reinnervation; restoration; virtual; vocal cord

PROJECT SUMMARY: Recurrent laryngeal nerve (RLN) injury occurs in tens of thousands of patients per year in the United States. It results in synkinetic reinnervation inducing vocal fold paralysis and severe dysphonia. This produces significant patient morbidity as the voice is a unique and integral part of each person's identity, impacting virtually every aspect of our daily lives, from speaking to loved ones, to engaging in business or simply completing daily tasks. Voice dysfunction leads to fear and isolation, as well as impaired quality of life, disability claims and lost worker productivity similar to severe chronic diseases, resulting in a significant societal burden. Current therapies are suboptimal and there are no treatments that can restore vocal fold motion and normal voice. There are critical gaps in our knowledge about the molecular mechanisms underlying axonal guidance during nerve regeneration. Our long-term goals are to identify the expression and actions of guidance cues and to manipulate them to guide selective, non-synkinetic reinnervation of the laryngeal muscles after RLN injury, with restoration of normal vocal fold function and normal voice. Our previous work has revealed that in both embryologic development and post-RLN injury, laryngeal muscles are innervated in similar sequences. Yet, post-RLN injury reinnervation is disordered, resulting in synkinesis with vocal fold dysfunction. Our laboratory has identified Netrin-1 and GDNF as the factors most likely to play a central role in axon guidance during RLN regeneration. We have shown that their expression in laryngeal muscles and the expression of their receptors in the nucleus ambiguus (NA) are chronologically coordinated with axonal reinnervation. We have demonstrated that manipulation of these guidance cues can alter the pattern of reinnervation. These findings suggest a controlled but dysfunctional expression of guidance cues resulting in synkinetic reinnervation and vocal fold paralysis after RLN injury. In comparison, developmental innervation, which is also tightly controlled chronologically, results in normal vocal fold function. Our central hypothesis is that Netrin-1 and GDNF are critical guidance cues in laryngeal reinnervation. Further, manipulation of their pathways post-RLN injury to mimic those seen during normal development will in result in selective, non- synkinetic reinnervation with restoration of normal vocal fold function. Guided by strong preliminary data, we seek to pursue the following three Specific Aims: 1) to describe the patterns of expression of GDNF, Netrin-1, and their receptors during innervation of the larynx in development 2) to describe the pathways that modulate the function of GDNF, Netrin-1 and their receptors during innervation in development and during reinnervation post-RLN injury and 3) to evaluate in vitro the effects of modulating pathways in the NA and muscle. This research will broadly impact the field of nerve regeneration, advancing our knowledge of the mechanisms of axonal pathfinding with the potential to transform the treatment of vocal fold paralysis.

项目摘要：每年有数万名患者发生喉返神经 (RLN) 损伤 年在美国。它会导致联动神经支配，引起声带麻痹和严重的声带麻痹。 发声困难。这会导致患者显着发病，因为声音是每个人独特且不可或缺的一部分。 人的身份，几乎影响着我们日常生活的方方面面，从与亲人交谈到参与 业务或只是完成日常任务。声音功能障碍会导致恐惧和孤立，以及受损 与严重慢性病类似的生活质量、残疾索赔和工人生产力损失，导致 重大的社会负担。目前的治疗方法并不理想，没有任何治疗方法可以恢复声音 折叠动作和正常声音。我们对分子机制的了解存在严重差距 神经再生过程中潜在的轴突引导。我们的长期目标是识别表达和 引导线索的行动并操纵它们来指导选择性的、非联动的神经再支配 RLN 损伤后喉部肌肉恢复正常声带功能和正常声音。 我们之前的工作表明，在胚胎发育和 RLN 损伤后，喉肌 以相似的序列受到神经支配。然而，RLN 损伤后神经支配紊乱，导致联带运动 伴有声带功能障碍。我们的实验室已确定 Netrin-1 和 GDNF 是最有可能发挥作用的因素 RLN 再生过程中轴突引导的核心作用。我们已经证明它们在喉部的表达 肌肉及其在疑核（NA）中受体的表达是按时间顺序协调的 与轴突神经支配。我们已经证明，操纵这些指导线索可以改变 神经再生的模式。 这些发现表明引导线索的表达受到控制但功能失调，导致联动 RLN 损伤后神经再支配和声带麻痹。相比之下，发育神经支配，即 也按时间顺序严格控制，导致正常的声带功能。我们的中心假设是 Netrin-1 和 GDNF 是喉神经支配的关键指导信号。此外，操纵他们的 RLN 损伤后的通路模仿正常发育过程中的通路将导致选择性的、非 联动神经支配恢复正常声带功能。在强有力的初步数据的指导下，我们 力求实现以下三个具体目标：1) 描述 GDNF、Netrin-1、 及其受体在发育过程中喉部神经支配过程中的作用 2) 描述调节途径 GDNF、Netrin-1 及其受体在发育过程中的神经支配和神经再支配过程中的功能 RLN 损伤后；3) 体外评估 NA 和肌肉中调节通路的影响。这 研究将广泛影响神经再生领域，增进我们对神经再生机制的了解 轴突寻路有可能改变声带麻痹的治疗方法。

项目成果

Expression of Glial Cell-Derived Neurotrophic Factor Receptors Within Nucleus Ambiguus During Rat Development.

• DOI: 10.1002/lary.30440
• 发表时间: 2023-09
• 期刊: LARYNGOSCOPE
• 影响因子: 2.6
• 作者: Blount, Quinton;Hernandez-Morato, Ignacio;Moayedi, Yalda;Pitman, Michael J.
• 通讯作者: Pitman, Michael J.

A review of the peripheral proprioceptive apparatus in the larynx.

• DOI: 10.3389/fnana.2023.1114817
• 发表时间: 2023
• 期刊: Frontiers in neuroanatomy
• 影响因子: 2.9

An optimized method for high-quality RNA extraction from distinctive intrinsic laryngeal muscles in the rat model.

从大鼠模型中，从独特的内在喉部肌肉中提取高质量RNA的优化方法。

• DOI: 10.1038/s41598-022-25643-y
• 发表时间: 2022-12-15
• 期刊: SCIENTIFIC REPORTS
• 影响因子: 4.6
• 作者: Kemfack, Angela M.;Hernandez-Morato, Ignacio;Moayedi, Yalda;Pitman, Michael J.
• 通讯作者: Pitman, Michael J.

Temporal expression of Laminin-111 in the developing rat larynx.

• DOI: 10.1016/j.neulet.2022.136658
• 发表时间: 2022-06-11
• 期刊: NEUROSCIENCE LETTERS
• 影响因子: 2.5
• 作者: Caplan, Ian F.;Hernandez-Morato, Ignacio;Pitman, Michael J.
• 通讯作者: Pitman, Michael J.