Muscle progenitor cell-based implants for dynamic laryngeal muscle reconstruction

 描述（由申请人提供）：毁灭性的声音丧失（发音困难或失声）影响美国每年数千人接受创伤或肿瘤部分喉切除术，或因声带麻痹而遭受肌肉体积损失。这些患者的声音恢复选择是次优的，因此，大多数患者留下永久性的声音丧失和沟通障碍。本申请介绍了一种用于在直接声带损伤和/或去神经支配后恢复声带肌肉体积和功能的新方法。结果可能会导致改善声带麻痹和/或经历了半喉切除术，声带切除术，或创伤性撕脱的患者的声音恢复的手术选择。本申请的第一个目标是开发一种肌肉祖细胞衍生的植入物（MI），其在植入动物模型后接受强神经支配。具体地，将确定肌纤维运动终板表达对MI的植入后神经支配的影响。为此，我们将在体外用诱导MI肌肉表达运动终板的因子制造和预处理MI。MI将用于替换同系大鼠模型中的部分喉切除术缺损，并将根据喉肌电图和神经接触运动终板的定量确定植入后神经支配状态。使用该动物植入模型，将在喉返神经完整性和不完整性的环境中比较研究MI与对照MI的结局。本申请的下一个目标是在定制的胶原蛋白基质中开发MI，并确定该方法在用于修复喉肌缺损时是否有助于改善肌纤维的对齐和增强肌肉体积。为了实现这些目标，将用胶原聚合物产生MI，所述胶原聚合物表现出胶原原纤维基质的自组装和分级工程设计以引导细胞命运，从而促进具有模拟天然声带肌肉的特征的大小、肌纤维密度和排列的肌纤维的形成。同样，使用动物模型，将基于胶原的MI的结果与具有和不具有喉返神经完整性的环境中的对照MI的结果进行比较。从拟议的研究结果应克服目前的主要障碍，开发一个功能性组织工程MCC的半喉重建，这些障碍是神经支配的肌肉，次优组织的肌纤维，和异步发射的肌肉与本地收肌。此外，这些初步实验的结果应该会导致具有里程碑意义的临床创新，这些创新将与全球的声音恢复应用和肌肉修复概念相关。

 DESCRIPTION (provided by applicant): Devastating voice loss (dysphonia or aphonia) impacts thousands of individuals in the United States each year undergoing traumatic or oncologic partial laryngectomies, or suffering muscle volume loss due to vocal fold paralysis. Voice restoration options for these patients are suboptimal, and, as a result, most patients are left with permanent voice loss and communication impairment. This application introduces a novel approach for restoring vocal fold muscle volume and function after direct vocal fold injury and/or denervation. Results may lead to improved surgical options for voice restoration in patients who have vocal paralysis and/or have undergone hemilaryngectomies, cordectomies, or traumatic avulsions. The first goal of this application is to develop a muscle progenitor cell-derived implant (MI) that, after implantation in an animal model, receives strong innervation. Specifically, the effect of myofiber motor endplate expression on post-implantation innervation of MIs will be determined. To do this, we will fabricate and pre-treat MIs with factors in vitro that induce the MI muscle to express motor endplates. The MIs will be used to replace a partial laryngectomy defect in a syngeneic rat model, and post-implantation innervation status, based on laryngeal electromyography and quantification of motor endplates with nerve contact, will be determined. Using this animal implant model, outcomes with the study MIs will be compared to those of control MIs in environments with and without recurrent laryngeal nerve integrity. The next goal of this application is to develop a MI within a customized collagen matrix, and determine if this approach will facilitate improved alignment of muscle fibers and enhanced muscle volume when used to repair laryngeal muscle defects. To reach these goals, MIs will be created with collagen polymers exhibiting self-assembly and hierarchical engineering design of collagen fibril matrices to guide cell fate, and thereby facilitating formation of muscle fibers wih size, myofiber density, and alignment that mimics the characteristics of the native vocal fold muscle. Again, using an animal model, outcomes with the collagen based MIs will be compared to those of control MIs in environments with and without recurrent laryngeal nerve integrity. Findings from the proposed studies should overcome current major hurdles to developing a functional tissue engineered MCC for hemilaryngeal reconstruction-those hurdles being inadequate innervation of the muscle, suboptimal organization of myofibers, and asynchronous firing of the muscle with the native adductor muscle. Furthermore, results from these initial experiments should lead to landmark clinical innovations that will be relevant to both voice restoration applications, and muscle repair concepts globally.