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

基本信息

批准号：
10652429
负责人：
Govindarajan Srimathveeravalli
金额：
$ 33.79万
依托单位：
UNIVERSITY OF MASSACHUSETTS AMHERST
依托单位国家：
美国
项目类别：
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/10652429
关键词：
Acceleration Agonist Autologous Transplantation Basement membrane Biochemical Biocompatible Materials Bioreactors Bladder Blood Vessels Cell Death Induction 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 Population 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 efficacy evaluation electric field improved improved outcome in silico in vivo innovation knock-down manufacture mechanotransduction new technology novel porcine model preservation reconstruction response standard of care tissue support frame tool tumor ablation urinary urogenital tract voltage

项目摘要

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（BR）对于恢复神经源性膀胱患者的尿功能至关重要，先天性疾病，作为膀胱或骨盆恶性肿瘤手术治疗的序列。电流护理标准是将患者自己的肠组织（iLeal片段; is）用作BR期间的移植物材料创建新蓝色或尿液转移。虽然是移植物是非免疫原性的，而且很容易获得移植物中肠细胞的手术持久性阻碍了再生到膀胱壁，导致石头形成，代谢性酸中毒和继发性癌症的风险。我们的目标是为通过使用不可逆的细胞成分敲低细胞成分电穿孔（IRE）并确定功能膀胱壁再生的因素。 ire被用通过用超短脉冲诱导细胞死亡来消融肿瘤的患者。我们提出的策略建立在我们的初步数据上，显示（i）可以敲除肠道中的肠细胞，以帮助重生在BR大鼠模型中使用尿皮上皮，（ii）新的脉冲应用策略的可行性在IS中保留脉管系统和ECM的同时，（iii）表型变化经过治疗的尿路上皮化是在生理条件下在假对照中观察到的膀胱填充和排尿。在特定目标1中，我们将定义移植物灌注对膀胱壁的影响通过进行脉管系统的再生，使IS的更新。在特定目标2中，阐明了IS的作用通过与IRE敲除后BR并发症中的粘膜。在特定目标3中，调查 IRE处理的IRE的膀胱功能发育的机械转导IS。灌注，组织兼容的移植物（AIM 1）和粘膜的局灶性脱细胞，同时占据下层层 IS（AIM 2）是使用IRE的体内敲低组织工程的第一个例子。研究和机械转移原理在增加尿路屏障功能开发中的应用是移植物（AIM 3）以前未描述。从提议的研究中获得的知识将产生简单的外科内部结合了技术（IRE）和嫁接技术的技术（MT-IRE或VS-IRE）（使用IS）在诊所中，可以快速翻译为接受BR的患者的直接益处。最终，我们预期我们的工作以利用患者为生物材料和生物反应器的来源，并应用于涉及其他管状的重建手术器官，例如食道，气管或大血管。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Directional Cell Migration Guided by a Strain Gradient.

由应变梯度引导的定向细胞迁移。

DOI：
10.1002/smll.202302404
发表时间：
2024
期刊：
Small (Weinheim an der Bergstrasse, Germany)
影响因子：
0
作者：
Yang,Feiyu;Chen,Pengcheng;Jiang,Han;Xie,Tianfa;Shao,Yue;Kim,Deok-Ho;Li,Bo;Sun,Yubing
通讯作者：
Sun,Yubing

Acute ATP loss during irreversible electroporation mediates caspase independent cell death.

不可逆电穿孔过程中急性 ATP 损失介导不依赖半胱天冬酶的细胞死亡。

DOI：
10.1016/j.bioelechem.2022.108355
发表时间：
2023
期刊：
Bioelectrochemistry (Amsterdam, Netherlands)
影响因子：
0
作者：
Razakamanantsoa,Leo;Rajagopalan,NeerajR;Kimura,Yasushi;Sabbah,Michele;Thomassin-Naggara,Isabelle;Cornelis,FrançoisH;Srimathveeravalli,Govindarajan
通讯作者：
Srimathveeravalli,Govindarajan

Imaging and detecting intercellular tensile forces in spheroids and embryoid bodies using lipid-modified DNA probes.