Directed differentiation of embryonic stem cells for urologic applications

用于泌尿外科应用的胚胎干细胞定向分化

• 批准号: 7953503
• 负责人: JAMES A BASSUK
• 金额: $ 19.52万
• 依托单位: SEATTLE CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-13 至 2010-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7953503
• 关键词: Address; Autologous; Automobile Driving; Award; Beta Cell; Biliary; Biological Assay; Bladder; Bladder Tissue; Cardiac; Cardiac Myocytes; Cell Differentiation process; Cells; Clinical; Congenital Abnormality; Development; Disease; Duct (organ) structure; Electron Microscopy; Embryo; Endoderm; Epithelial; Epithelium; Experimental Designs; Fibrous capsule of kidney; Foundations; Goals; Human; Hypospadias; Implant; In Vitro; Infant; Institutes; Insulin; Interstitial Cystitis; K-18 conjugate; Keratin; Lead; Learning; Life; Malignant Neoplasms; Measurement; Measures; Membrane; Mentors; Mesenchymal; Mesenchyme; Multiple Sclerosis; Mus; Neurogenic Bladder; Nude Mice; Operative Surgical Procedures; Organ; Outcome; Pancreas; Patients; Pediatric Hospitals; Pluripotent Stem Cells; Publishing; Rattus; Recombinants; Regenerative Medicine; Reporting; Research; Resistance; Services; Solutions; Source; Spinal Dysraphism; Stem cells; Structure; Surgeon; Technology; Teratoma; Testing; Tight Junctions; Time; Tissue Engineering; Tissue Grafts; Tissues; Toxin; Training; Transitional Epithelium; Trauma; Tube; Undifferentiated; Universities; Ureter; Urethra; Urine; Urology; Urothelial Cell; Washington; Xenograft procedure; base; boys; career; diabetic; embryonic stem cell; experience; human embryonic stem cell; improved; in vitro Model; induced pluripotent stem cell; pathogen; professor; programs; public health relevance; reconstruction; repaired; self-renewal; stem cell technology; urologic

DESCRIPTION (provided by applicant): In this K18 Career Enhancement Award in Stem Cell Reseach application, the Candidate (Dr. James Bassuk) proposes to take time from his regular professional responsibilities in order to broaden his scientific background by acquiring new research capabilities. The specifiic new research capability to be learned is the use of embryonic stem cells (ESCs) in regenerative medicine applications such as tissue engineered repair of the urinary bladder and its associated tract. There is a significant need to develop (i) replacement bladders for patients suffering from cancer, interstitial cystitis or neurogenic bladder (spina bifida or multiple sclerosis) and (ii) epithelial tubes for reconstruction of the urethra due to trauma, stricture disease or hypospadias (a common birth defect in boys). The project represents a true career enhancement for him because it brings to his research program, for the first time, an experimental design that focuses on how developmental steps of bladder formation can be studied with a stem cell-based approach. The strategy involves finding the minimum number of ESCs needed to form a teratoma with the goal of redirecting these ESCs to form only one organ type. Embryonic mesenchyme is well established as an inductive tissue which dictates organ-specific programming of epithelial tissues. In Aim 1, the activity of rat embryonic bladder mesenchyme in driving mouse ESCs into functional bladder tissue will be measured by a stereological quantification of urothelial cell terminal differentiation. Under the guidance of Drs. Carol Ware (Mentor), C. Anthony Blau (Co-Mentor), Simon Hayward (Contributor) and Andrew Farr (Contributor), the Candidate will be trained in the proper handling of embryonic stem cells and in setting up xenograft assays where recombinant grafts are implanted under the renal capsule of athymic nude mice. In Aim 2a, human ESCs will be substituted for mouse ESCs. In Aim 2b, mesenchyme from human infant bladder will be substituted for rat embryonic mesenchyme. The source of infant mesenchyme will be the urology service at Seattle Children's Hospital, as procured by Drs. Richard Grady and Thomas Lendvay (Contributors). Thus, the question asked is will human ESCs respond appropriately to bladder mesenchymal developmental control? Over a 2-year project period, the Candidate will devote 75% of his time to train at the Institute for Stem Cell and Regenerative Medicine at the University of Washington. Once the project has been completed, then the Candidate's newly developed expertise will be applied to (i) the further study of mesenchymal factors necessary for developmental control of bladder tissue formation from inducible pluripotent stem cells, (ii) generating new in vitro models of urothelial cell differentiation and (iii) tissue engineered solutions for urologic use. PUBLIC HEALTH RELEVANCE: This project will provide a training experience in stem cell technology for Dr. James Bassuk, an Associate Professor of Urology at the University of Washington. Once the project is over, Dr. Bassuk will apply his newly acquired expertise to developing new solutions to assist the surgeon in repairing the urinary bladder and its associated tract. These efforts are expected to lead to improved outcomes for patients suffering with a non-functional bladder due to cancer, interstitial cystitis, multiple sclerosis or spina bifida.

描述（由申请人提供）：在STEM细胞Reseach申请中的K18职业增强奖中，候选人（James Bassuk博士）建议从他的常规专业责任中抽出时间，以通过获得新的研究能力来扩大其科学背景。要学习的特殊新研究能力是在再生医学应用中使用胚胎干细胞（ESC），例如膀胱的组织工程修复及其相关区域。对于患有癌症，间质性膀胱炎或神经源性膀胱（脊柱裂或多发性硬化症）的患者（ii）（ii）上皮管重建因创伤，狭窄疾病或炎症性疾病或男生常见的出生缺陷而引起的尿素的上皮管（ii）上皮管（男孩）的上皮管（ii），需要开发（i）（i）（i）替换膀胱替代膀胱。该项目对他来说是一个真正的职业增强，因为它首次为他的研究计划带来了一个实验设计，该设计重点介绍了如何通过基于干细胞的方法研究膀胱形成的发展步骤。该策略涉及发现形成畸胎瘤所需的最小ESC数量，目的是将这些ESC重定向到仅形成一种器官。胚胎间充质被很好地确定为一种诱导组织，它决定了上皮组织的器官特异性编程。在AIM 1中，将通过对尿路上皮细胞末端分化的立体定量来测量大鼠胚胎膀胱间充质在驱动小鼠ESC中的活性。在博士的指导下。 Carol Ware（导师），C。AnthonyBlau（Co-Mentor），Simon Hayward（贡献者）和Andrew Farr（贡献者），将在适当处理胚胎干细胞的适当处理以及建立重组移植物的适当处理中培训，其中重组移植物在肾囊中植入了Athymic Nude nude小鼠的肾囊中。在AIM 2A中，人类ESC将被代替老鼠ESC。在AIM 2B中，来自人类婴儿膀胱的间质将被取代用于大鼠胚胎间充质。婴儿间充质的来源将是西雅图儿童医院的泌尿外科服务，如Drs所述。理查德·格雷迪（Richard Grady）和托马斯·伦德维（Thomas Lendvay）（贡献者）。因此，问的问题是，人类ESC会对膀胱间充质发育控制做出适当反应吗？在2年的项目期间，候选人将花75％的时间在华盛顿大学的干细胞和再生医学研究所培训。该项目完成后，候选人的新开发的专业知识将应用于（i）进一步研究从诱导多能多能的干细胞中开发膀胱组织形成所必需的间充质因子，（ii）为尿路细胞分化和（III）组织工程溶液的新体外模型用于尿路尿路。 公共卫生相关性：该项目将为华盛顿大学泌尿外科副教授James Bassuk博士提供干细胞技术的培训经验。该项目结束后，Bassuk博士将应用其新获得的专业知识来开发新的解决方案，以帮助外科医生修复膀胱及其相关区域。预计这些努力将导致因癌症，间质性膀胱炎，多发性硬化症或脊柱裂伤引起的非功能性膀胱的患者改善结果。