Novel Strategies in Bladder Tissue Engineering: Stem Cells and Silk Biomaterials

描述（由申请人提供）：我在这个提案中的总体目标是解决目前膀胱重建手术选择的局限性，同时发展我的专业能力，以过渡到一个独立的研究者。我的职业目标是在一家一流的学术机构担任实验室主任，这将使我能够继续拓展再生医学的前沿，通过创造创新的基于干细胞的泌尿组织重建疗法。我的主要研究训练目标将是获得泌尿系统组织工程系统和人类多能干细胞生物学的新技术技能。这将通过额外的基础课程和实验室协议和技术的实践指导来完成，如提案的目标所述。职业发展机制将包括指导性的研讨会和在基金写作、学术工作面试、实验室管理和论文准备方面的指导。一个咨询委员会将评估我的科学和职业发展里程碑的完成情况，并帮助我过渡到一个独立的研究者。膀胱的组织工程方法最近得到了发展，利用3D支架植入自体平滑肌细胞（SMCs）。在短期临床试验中，这些策略已经导致器官功能的部分恢复。然而，这些结构对完全缺陷修复的功效可能受到传统生物材料载体的次优特性和功能性细胞来源的可用性的限制。丝基生物材料提供了一种特殊的物理特性组合，非常适合支持膀胱功能，因此可以解决当前支架材料的局限性。多能干细胞群体，如胚胎干细胞（ESCs）和/或诱导多能干细胞（iPS）细胞可以满足对能够产生器官和患者特异性SMCs的替代细胞来源的需求。为了解决膀胱功能组织工程（FTE）目前的局限性，我们将挑战这样一个总体假设：使用丝基生物材料植入原代或干细胞衍生的SMCs的FTE模式将促进平滑肌缺陷的巩固和膀胱功能的恢复。该研究的具体目的是：目的1：确定以原代SMCs为种子的丝基生物材料对膀胱平滑肌组织再生的功效。目的2：评估ESC和ips衍生的SMCs作为功能性膀胱组织工程细胞来源的效用。

DESCRIPTION (provided by applicant): My overall goal in this proposal is to address the current limitations in surgical options for bladder reconstruction while developing my professional abilities to transition into an independent investigator. My professional objective is to have a position as a lab director in a superb academic institution that will allow me to continue to expand the frontiers of regenerative medicine by creating innovative stem cell-based therapies for urological tissue reconstruction. My primary research training objectives will be to acquire new technical skills in urological tissue engineered systems and human pluripotent stem cell biology. This will be accomplished through additional fundamental coursework and hands on instruction in laboratory protocols and techniques as detailed in the aims of the proposal. Career development mechanisms will include instructive seminars and mentored guidance in grant writing, academic job interviewing, laboratory management, and manuscript preparation. An advisory committee will evaluate the completion of both my scientific and career development milestones and facilitate my transition into an independent investigator. Tissue engineering approaches for the urinary bladder have recently been developed utilizing 3D scaffolds seeded with autologous smooth muscle cells (SMCs). These strategies have led to partial restoration of organ functionality in short-term clinical trials. However, the efficacy of these constructs for complete defect repair may be limited by the suboptimal properties of conventional biomaterial carriers and availability of functionally competent cell sources. Silk-based biomaterials provide an exceptional combination of physical characteristics that are well suited to support bladder function and therefore may address limitations associated with current scaffold materials. Pluripotent stem cell populations such as embryonic stem cells (ESCs) and/or induced pluripotent stem (iPS) cells may fulfill the need for alternative cell sources capable of generating organ and patient-specific SMCs. In order to address the current limitations in bladder functional tissue engineering (FTE), we will challenge the overall hypothesis that: FTE modalities using silk-based biomaterials seeded with primary or stem cell-derived SMCs will promote smooth muscle defect consolidation and restoration of bladder function. The specific aims of the study are: Aim 1: Determine the efficacy of silk-based biomaterials seeded with primary SMCs for bladder smooth muscle tissue regeneration. Aim 2: Evaluate the utility of ESC and iPS-derived SMCs as cell sources for functional bladder tissue engineering. PUBLIC HEALTH RELEVANCE: The results of this proposal are anticipated to produce new tissue regeneration strategies that will overcome current biomaterial and cell source limitations for urinary bladder reconstruction. This study will provide new information that is wider in scope than our specific goals and which may be applicable for repair of other urogenital tissues (e.g. urethra) and possibly other hollow organs.