Intrasurgical tissue engineering of autologous grafts using irreversible electroporation for bladder reconstruction

使用不可逆电穿孔进行膀胱重建的自体移植物的术中组织工程

• 批准号: 10447720
• 负责人: Govindarajan Srimathveeravalli
• 金额: $ 34.21万
• 依托单位: UNIVERSITY OF MASSACHUSETTS AMHERST
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-12 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10447720
• 关键词: Agonist; Autologous Transplantation; Basement membrane; Biochemical; Biocompatible Materials; Bioreactors; Bladder; Blood Vessels; Cell Death; Cell Differentiation process; Cells; Cicatrix; Clinic; Clinical; Congenital Disorders; Data; Development; Electroporation; Engineering; Esophagus; Extracellular Matrix; Family suidae; Frequencies; Functional Regeneration; Gastrointestinal tract structure; Heterogeneity; Histocompatibility; Immunohistochemistry; Impairment; In Vitro; Inflammation; Intestines; Knowledge; Malignant Neoplasms; Malignant neoplasm of urinary bladder; Mechanics; Metabolic; Metabolic acidosis; Modeling; Molecular Biology Techniques; Morbidity - disease rate; Mucous Membrane; Natural regeneration; Neurogenic Bladder; Non-Malignant; Operative Surgical Procedures; Organ; Pathway interactions; Patients; Pelvis; Perfusion; Phenotype; Physiologic pulse; Physiological; Piezo 1 ion channel; Production; Rattus; Reconstructive Surgical Procedures; Regenerative Medicine; Research; Research Design; Risk; Role; Safety; Second Primary Cancers; Small Intestinal Submucosa; Smooth Muscle Myocytes; Source; Stretching; Techniques; Technology; Tissue Engineering; Tissues; Trachea; Translating; Translations; Tubular formation; Urinary Diversion; Urothelium; Work; bladder surgery; cell killing; electric field; improved; improved outcome; in silico; in vivo; innovation; knock-down; mechanotransduction; new technology; novel; porcine model; preservation; reconstruction; response; standard of care; tissue support frame; tool; tumor ablation; urinary; urogenital tract; voltage

PROJECT SUMMARY Bladder reconstruction (BR) is essential to restore urinary function in patients with neurogenic bladder, congenital disorders, and as sequalae to the surgical treatment of bladder or pelvic malignancies. The current standard of care is to use the patient's own intestinal tissue (ileal segment; IS) as graft material during BR to create a neobladder or urinary diversion. While IS grafts are non-immunogenic and readily available, post- surgical persistence of intestinal cells in the graft impedes regeneration into bladder wall, resulting in stone formation, metabolic acidosis and risk of secondary cancers. Our objective is to develop new technology for intrasurgical tissue engineering of the IS by knockdown of cellular components using irreversible electroporation (IRE) and identify factors fundamental for regeneration of functional bladder wall. IRE is used in patients for tumor ablation by inducing cell death with ultrashort electric pulses. Our proposed strategy builds upon our preliminary data showing (i) IRE can knockdown intestinal cells in an IS graft, aiding repopulation with urothelium in a rat model of BR, (ii) feasibility of new pulse application strategies for the focal knockdown of mucosa, or decellularization while preserving vasculature and ECM in the IS, and (iii) phenotypic changes in IRE treated IS following urothelialization, that were not observed in sham controls under physiologic conditions of bladder filling and voiding. In specific aim 1, we will Define the impact of graft perfusion on bladder wall regeneration by performing vasculature sparing IRE of the IS. In specific aim 2, Elucidate the role of IS mucosa in post-BR complications by knockdown with IRE. In specific aim 3, Investigate the role of mechanotransduction in bladder function development in IRE treated IS. Intrasurgical creation of a perfused, histocompatible graft (Aim 1) and focal decellularization of the mucosa while sparing the underlying layers in the IS (Aim 2) are the first examples of in vivo knockdown tissue engineering using IRE. The study and application of mechanotransduction principles to augment urinary barrier function development in IS grafts (Aim 3) is previously undescribed. Knowledge gained from proposed research will yield a simple intrasurgical technique (mt-IRE or vs-IRE) that combines technology (IRE) and grafting technique (with IS) that are already in the clinic, enabling rapid translation for the immediate benefit of patients undergoing BR. Eventually, we anticipate our work to advance the concept of in vivo production of functionalized grafts using the patient as the source of biomaterial and the bioreactor, with application to reconstructive surgery involving other tubular organs such as the esophagus, trachea or large blood vessels.

项目摘要 膀胱重建术（BR）是恢复神经源性膀胱患者排尿功能的关键， 先天性疾病，以及作为膀胱或盆腔恶性肿瘤手术治疗的后遗症。当前 标准治疗是在BR期间使用患者自身的肠组织（回肠段; IS）作为移植材料， 形成新膀胱或尿流改道虽然IS移植物是非免疫原性的并且容易获得，但术后 移植物中肠细胞的手术残留阻碍了膀胱壁的再生，导致结石 形成，代谢性酸中毒和继发性癌症的风险。我们的目标是开发新技术， IS的外科内组织工程，通过使用不可逆的 电穿孔（IRE）并鉴定功能性膀胱壁再生的基本因素。IRE用于 患者通过用超短电脉冲诱导细胞死亡进行肿瘤消融。我们提出的战略建立在 根据我们的初步数据显示：（i）IRE可以敲低IS移植物中的肠细胞， 在BR的大鼠模型中使用尿路刺激，（ii）新的局部敲除脉冲应用策略的可行性 在IS中保留脉管系统和ECM的同时进行粘膜或脱细胞化，以及（iii） IRE治疗的IS后，尿路上皮化，这是没有观察到在假对照组在生理条件下 膀胱充盈和排泄的过程在具体目标1中，我们将定义移植物灌注对膀胱壁的影响 通过进行保留IS的IRE的脉管系统再生。在具体目标2中，阐明信息系统的作用 通过用IRE敲低BR后并发症中的粘膜。在具体目标3中， 在IRE治疗的IS中膀胱功能发育中的机械转导。在手术中创造了一个灌注的， 组织相容性移植物（Aim 1）和粘膜局灶性脱细胞化，同时保留下层， IS（Aim 2）是使用IRE的体内敲低组织工程的第一个实例。学习 应用机械传导原理增强IS移植物中的尿屏障功能发育 (Aim（3）以前没有描述过。从拟议研究中获得的知识将产生一个简单的外科手术 技术（mt-IRE或vs-IRE），它结合了技术（IRE）和嫁接技术（与IS），这些技术已经 在临床上，能够快速翻译，使接受BR的患者立即受益。最终我们 预期我们的工作，以推进概念，在体内生产的功能化移植物使用的病人作为 生物材料源和生物反应器，应用于涉及其他管状结构的重建手术 器官如食道、气管或大血管。