Engineered neuroprotective stem-cell exosomes for in utero spina bifida therapy

用于子宫内脊柱裂治疗的工程神经保护干细胞外泌体

• 批准号: 10271311
• 负责人: Diana Lee Farmer
• 金额: $ 48.99万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-30 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10271311
• 关键词: Achievement; Animal Model; Animals; Binding; Biocompatible Materials; Biomedical Engineering; Birth; Bladder Dysfunction; Bone Diseases; Bone Regeneration; Caring; Cell Communication; Cells; Chemical Injury; Child; Childhood; Clinical; Clinical Trials; Collagen; Defect; Deformity; Disease; Engineering; Environment; Exposure to; Fetal Development; Fetal Sheep; Fetal Tissues; Glycolic-Lactic Acid Polyester; Goals; Gold; Growth; Health Care Costs; Human; Hydrogels; Hydroxyapatites; Immobilization; In Vitro; Intestines; Kyphosis deformity of spine; Life; Limb structure; Longevity; Mechanics; Meningomyelocele; Mesenchymal; Methods; Modeling; Molecular; Molecular Weight; Morbidity - disease rate; Motor; Muscle; Musculoskeletal; Neural Tube Closure; Neurodegenerative Disorders; Operative Surgical Procedures; Oryctolagus cuniculus; Osteogenesis; Outcome; Paralysed; Patients; Peptides; Physical therapy; Play; Preventive care; Quality of life; Recovery of Function; Regimen; Rodent; Role; Speed; Spinal Cord; Spinal Dysraphism; Spinal cord injury; Structure; Surface; System; Testing; Therapeutic; Therapeutic Uses; Transplantation; Trauma; United States; United States National Institutes of Health; Vertebral column; Walking; base; biodegradable polymer; bioscaffold; bone; cognitive disability; density; design; dosage; exosome; experimental study; extracellular vesicles; fetal; functional group; hindbrain; improved; in utero; in vivo; independent ambulation; mechanical properties; neuroprotection; novel; novel strategies; novel therapeutics; paracrine; postnatal; pre-clinical; prenatal therapy; prevent; regeneration function; regeneration potential; regenerative; regenerative therapy; repaired; scaffold; sheep model; spinal cord compression; standard of care; stem cell exosomes; stem cells

ABSTRACT Myelomeningocele (MMC) is the most severe form of spina bifida (SB) and the most common congenital cause of lifelong paralysis in the United States, where approximately four children are born daily with this devastating disease. MMC results from the incomplete closure of the neural tube and absent overlying spine leaving the spinal cord exposed to intrauterine mechanical and chemical trauma. This trauma results in lifelong paralysis, bowel and bladder dysfunction, musculoskeletal deformities, and cognitive disabilities due to hindbrain herniation. In utero surgical repair improves morbidity, but functional recovery is incomplete and the majority of children are still unable to walk independently. We developed a treatment for MMC that augments the standard of care, in utero MMC surgical repair, with placental mesenchymal stromal/stem cells (PMSCs). We found that treatment with PMSCs, during in utero repair, prevents hind limb paralysis in the well-established fetal ovine model of MMC, due to PMSC paracrine secretion of neuroprotective factors. However, we confirmed that PMSCs did not engraft long-term and treated lambs developed severe kyphosis causing spinal cord compression and tethering due to the lack of bone and adjacent paraspinal muscles, which is consistent with human MMC musculoskeletal deformities. To meet this need for a long-lasting therapy for MMC, we explored the use of bioengineered multifunctional combination scaffolds. Exosomes are extracellular vesicles that play significant roles in cell-to cell communication. We confirmed that exosomes secreted by PMSCs (PMSC-exosomes) exert significant neuroprotective functions, similar in magnitude to the live PMSCs from which they are derived. In this study, we propose to develop an engineered hydrogel system that will allow for both local and sustained release of PMSC-exosomes to the spinal cord, which in turn will provide sustained neuroprotection to treat MMC before birth. In addition, we aim to increase the longevity of the in utero treatment by covering the defect with a biomaterial-based bony scaffold to provide structural and functional support. We plan to optimize the neuroprotective and regenerative functions of the bioscaffold using our well-established fetal rodent and rabbit models of MMC. We will evaluate the final combination multifunctional bioscaffold product in our gold-standard fetal ovine model of MMC. This therapeutic will be cell-free, off-the-shelf, and easy-to-use. If successful, this novel approach will be used to treat MMC before birth, and due to its regenerative qualities, the treatment will improve the quality of life of these patients, as well as significantly lower the healthcare costs associated with the current treatment.

抽象的 脊髓脊膜膨出 (MMC) 是脊柱裂 (SB) 最严重的形式，也是最常见的先天性病因 在美国，每天大约有四个孩子出生时患有这种毁灭性的终身瘫痪 疾病。 MMC 是由神经管不完全闭合和上覆脊柱缺失导致的 脊髓暴露于宫内机械和化学创伤。这种创伤导致终身瘫痪 肠和膀胱功能障碍、肌肉骨骼畸形以及后脑引起的认知障碍 疝气。子宫内手术修复可改善发病率，但功能恢复不完全，并且大多数 孩子们仍然无法独立行走。我们开发了一种 MMC 治疗方法，可提高标准 护理，使用胎盘间充质基质/干细胞 (PMSC) 进行子宫内 MMC 手术修复。我们发现 在子宫内修复过程中使用 PMSC 进行治疗可预防已成熟的胎羊的后肢瘫痪 MMC模型，由于PMSC旁分泌神经保护因子。然而，我们证实 PMSC 没有长期移植，经过治疗的羔羊出现严重的后凸畸形，导致脊髓受压 由于缺乏骨骼和邻近的椎旁肌肉而导致束缚，这与人类 MMC 一致 肌肉骨骼畸形。为了满足 MMC 长期治疗的需求，我们探索了使用 生物工程多功能组合支架。外泌体是发挥重要作用的细胞外囊泡 在细胞间通讯中发挥作用。我们证实 PMSC 分泌的外泌体（PMSC-exosome）发挥 显着的神经保护功能，其强度与它们来源的活 PMSC 相似。在这个 研究中，我们建议开发一种工程水凝胶系统，该系统将允许局部和持续释放 的 PMSC-外泌体到脊髓，这反过来将为治疗 MMC 提供持续的神经保护 出生。此外，我们的目标是通过用覆盖物覆盖缺陷来延长子宫内治疗的寿命。 基于生物材料的骨支架提供结构和功能支持。我们计划优化 使用我们成熟的胎儿啮齿动物和兔子的生物支架的神经保护和再生功能 MMC 模型。我们将按照我们的黄金标准评估最终组合的多功能生物支架产品 MMC胎羊模型。这种疗法是无细胞的、现成的且易于使用。如果成功的话，这 新方法将用于在出生前治疗 MMC，并且由于其再生特性，该治疗将 改善这些患者的生活质量，并显着降低与相关的医疗费用 目前的治疗。