Human Umbilical Cord as a Regenerative Meningeal Patch to Prevent Tethering and Improve Neurological Function Following In-Utero Spina Bifida Repair

总结 超过一半接受子宫内脊柱裂修复术的儿童无法行走，超过四分之一的儿童无法行走。 他们需要在学龄前进行脊髓栓系手术。改善长期脊髓功能的方法 以及这些患者的合并症仍然难以捉摸。长期目标是找到子宫内脊柱裂 治疗方法，解决方法的完善，将促进神经和 在以后的生活中，神经发育的收益，从而在功能上调整患者的生活质量更好。的 本发明的目的是使用一种新的方法在绵羊模型系统中修复子宫内脊柱裂， 随着时间的推移，可能会比目前的修复方法对羔羊产生更有益的结果。冷冻保存的 人脐带（HUC）是一种来自健康足月妊娠分娩后的同种异体移植物， 炎症和抗瘢痕形成的特性，同时重要的过程，直接影响是否脊柱 会发生脊髓栓系，并因此产生长期结果。HUC目前已被FDA批准用于 眼科疾病在我们的临床前研究中，我们已经表明，使用冷冻保存的人 脐带作为脑膜补片有助于再生蛛网膜层，这是脊髓浸润的关键屏障 瘢痕形成细胞，最终减少组织学束缚并改善脊髓功能。我们的中央 假设HUC在子宫内脊柱裂期间用作双层闭合的脑膜补片时， 修复，将改善长期临床结果相比，传统的修复或预测的生物材料。这 是根据我们的初步数据制定的，代表了脊柱裂治疗的重大进展 鉴于临床后遗症的改善，有可能垂直推进子宫内治疗。的理由 对于这项研究来说，迫切需要一种抗瘢痕形成和再生基质， 作为脊髓和皮肤层之间的中间层以减少束缚。核心假设将 通过在具有脊髓切开术的外科脊柱裂绵羊模型系统中追求三个特定目标来进行测试：1） 比较使用HUC的子宫内SB修复方法与 实质等同（DurepairTM）生物材料作为脑膜补片。2)量化和比较长期的空间和 利用3T技术对脊髓损伤修复部位的时间变化进行研究 MRI和扩散张量成像。3)定量比较长期的潜在机制， SB缺陷修复方法的治疗效果。我们将采用创新的 由于使用了冷冻保存的人脐带， 脐带通过将焦点转移到抗炎， 脊柱裂脑膜覆盖物的抗瘢痕形成和再生修复。其意义在于 这项工作的预期结果是，人脐带补片作为脑膜补片应该是可行的， 脊柱裂修复的纵向治疗选择并支持人体临床试验。

SUMMARY Over half the children who underwent in-utero spina bifida repair are unable to walk, and over one-fourth of them require surgery for tethered spinal cord by school age. Methods to improve long-term spinal cord function and its co-morbidities for these patients, remains elusive. The long-term goal is to find in-utero spina bifida treatment approaches that address refinement of methodologies that will promote neurological and neurodevelopmental gains later in life, thereby functionally adjusting patients’ quality of life for the better. The objective of this proposal is to use a new approach to repair spina bifida in-utero in a sheep model system, which may render more beneficial outcomes for lambs over time than current methods of repair. A cryopreserved human umbilical cord (HUC), an allograft from a healthy term pregnancy following delivery, exerts anti- inflammatory and anti-scarring properties, synonymously crucial processes that directly impact whether spinal cord tethering will occur, and hence long-term outcomes. The HUC is currently FDA approved for ophthalmological diseases. In our preclinical studies, we have shown that use of the cryopreserved human umbilical cord as a meningeal patch helps regenerate the arachnoid layer, a key barrier to spinal cord infiltrating scar forming cells, ultimately reducing histological tethering and improving upon spinal cord function. Our central hypothesis is that the HUC, when used as a meningeal patch with two-layer closure during in-utero spina bifida repair, will improve long-term clinical outcomes compared to conventional repair or predicated biomaterials. This was formulated based on our preliminary data, representing a significant advancement in spina bifida treatment given clinical sequelae improvements, with the potential to vertically advance in-utero treatment. The rationale for this study is that there is an urgent critical need for an anti-scarring and regenerative matrix that can be placed as an intermediary layer between the spinal cord and skin layer to reduce tethering. The central hypothesis will be tested by pursuing three specific aims in a surgical spina bifida sheep model system with a myelotomy: 1) To compare long-term functional outcome differences between in-utero SB repair methodologies using a HUC vs. predicate (DurepairTM) biomaterial as a meningeal patch. 2) To quantify and compare long-term spatial and temporal changes of the spinal cord at defect repair sites for tethering, syringomyelia and inflammation using 3T MRI and diffusion tensor imaging. 3) To quantitatively compare the underlying mechanisms for long-term therapeutic efficacies of SB defect repair methodologies. We will pursue these aims using an innovative combination of surgical and biological manipulative techniques, since use of the cryopreserved human umbilical cord represents a new and substantive departure from the status quo by shifting focus to the anti-inflammatory, anti-scarring and regenerative repair of the meningeal coverings of spina bifida. The significance lies in the expected outcome from this work, that the human umbilical cord patch as a meningeal patch should be a viable longitudinal therapeutic option for spina bifida repair and support a human clinical trial.