Synthetic hydrogel for satellite cell delivery to the dystrophic diaphragm

摘要 杜氏肌营养不良症是一种X连锁遗传疾病，会导致衰弱的健康后果。最 受影响的患者主要由于呼吸系统并发症而不能活过其早期成年期（例如，降低 呼吸、肺炎和肺不张）。当前呼吸 护理治疗，如机械辅助通气，仍然是姑息性的，是一种有效的治疗策略 靶向膈肌以恢复呼吸能力是一个关键的需求。本项目的目标 是开发一种生物材料介导的策略，将肌肉卫星细胞输送并移植到膈肌， 通过恢复肌营养不良蛋白、兴奋-收缩偶联来恢复呼吸功能， 机械稳定性这一目标将通过以下方式实现：（1）工程化合成生物材料，其能够 维持和指导3D中的肌肉卫星细胞功能，以及（2）递送包裹在3D中的卫星细胞。 工程生物材料，以刺激供体细胞的植入，从而挽救肌营养不良蛋白的表达， 神经肌肉接头的形成，以及年轻人（2个月）营养不良膈肌的收缩功能， 老年（12个月）mdx/mTR小鼠。指导这个项目的核心假设是，工程利基- 含有设计用于增加卫星细胞粘附、存活、增殖和分化的生物材料， 能够将细胞精确地输送到隔膜的下表面，增强随后的局部细胞保留， 供体细胞的存活、扩增、迁移和植入，并进一步为供体细胞提供机械支持。 老年/萎缩营养不良的膈肌。这项工作的结果将是重要的，在推进呼吸 治疗策略的背景下杜氏肌营养不良症。挽救衰竭的横膈膜肌肉， 因此，呼吸功能，将积极影响患者的健康和生活质量， 有机会绕过机械辅助通气的需要。这项工作的顺利完成将 还提供了进一步协同该平台与诱导多能干细胞和紧急 基因编辑技术来治疗患有各种形式的疾病的患者的患病隔膜功能， 神经肌肉疾病

ABSTRACT Duchenne muscular dystrophy is an X-linked genetic disorder that begets debilitating health consequences. Most affected patients do not live past their early adulthood primarily due to respiratory complications (e.g., decreased breathing, pneumonia and atelectasis) caused by deterioration of the diaphragm muscle. Current respiratory care therapies, such as mechanically assisted ventilation, remain palliative, and an effective therapeutic strategy targeting the diaphragm muscle to restore the respiratory capacity is a critical need. The objective of this project is to develop a biomaterial-mediated strategy to deliver and engraft muscle satellite cells to the diaphragm to reinstate the respiratory function through restoration of dystrophin, excitation-contraction coupling, and mechanical stability. This objective will be achieved by (1) engineering a synthetic biomaterial that is capable of maintaining and directing muscle satellite cell function in 3D and (2) delivering satellite cells encapsulated in the engineered biomaterial to stimulate engraftment of donor cells, thereby rescuing dystrophin expression, neuromuscular junction formation, and contractile function of the dystrophic diaphragm of young (2 months) and aged (12 months) mdx/mTR mice. The central hypothesis guiding this project is that the engineered niche- containing biomaterial designed to increase satellite cell adhesion, survival, proliferation, and differentiation will enable precise delivery of cells to the inferior surface of the diaphragm, enhance subsequent local cell retention, survival, expansion, migration, and engraftment of the donor cells, and further provide mechanical support to the aged/atrophied dystrophic diaphragm. The outcomes of this work will be significant in advancing the respiratory treatment strategy in the context of Duchenne muscular dystrophy. Salvaging the failing diaphragm muscle, and thereby the respiratory function, will positively impact patients healthspan and quality of life by providing opportunities to bypass the need for mechanically assisted ventilation. Successful completion of this work will also provide an opportunity to further synergize this platform with induced pluripotent stem cells and emergent gene editing technologies to treat diseased diaphragm function of patients suffering from various forms of neuromuscular diseases.