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）：在这个K18干细胞研究职业提升奖申请中，候选人（James Bassuk博士）建议从他的常规专业职责中抽出时间，通过获得新的研究能力来扩大他的科学背景。需要学习的具体新研究能力是胚胎干细胞（ESCs）在再生医学应用中的应用，例如膀胱及其相关尿道的组织工程修复。目前迫切需要开发(i)用于癌症、间质性膀胱炎或神经性膀胱（脊柱裂或多发性硬化症）患者的替代膀胱，以及（ii）用于因创伤、狭窄疾病或尿道下裂（男孩常见的出生缺陷）而重建尿道的上皮管。这个项目代表了他真正的职业生涯提升，因为它第一次为他的研究项目带来了一个实验设计，专注于如何用基于干细胞的方法研究膀胱形成的发育步骤。该策略包括找到形成畸胎瘤所需的ESCs的最小数量，目标是将这些ESCs重定向到仅形成一种器官类型。胚胎间质是一种诱导组织，它决定了上皮组织的器官特异性编程。在Aim 1中，将通过尿路上皮细胞末梢分化的立体学定量来测量大鼠胚胎膀胱间充质在驱动小鼠ESCs进入功能性膀胱组织中的活性。在博士的指导下。Carol Ware（导师），C. Anthony Blau（联合导师），Simon Hayward（投注者）和Andrew Farr（投注者），候选人将接受关于正确处理胚胎干细胞和建立异种移植试验的培训，将重组移植物植入胸腺裸鼠的肾包膜下。在Aim 2a中，人类ESCs将取代小鼠ESCs。在Aim 2b中，人婴儿膀胱间质将取代大鼠胚胎间质。婴儿间质的来源将是西雅图儿童医院的泌尿科，由dr。Richard Grady和Thomas Lendvay（贡献者）。因此，问题是人类ESCs会对膀胱间充质发育控制做出适当的反应吗？在2年的项目期间，候选人将投入75%的时间在华盛顿大学干细胞和再生医学研究所进行培训。一旦项目完成，候选人新开发的专业知识将应用于(i)进一步研究从诱导多能干细胞发育控制膀胱组织形成所需的间充质因子，（ii）生成新的体外尿路上皮细胞分化模型，（iii）用于泌尿系统的组织工程解决方案。