STROMAL/EPITHELIAL INTERACTIONS IN THE BLADDER

膀胱中的间质/上皮相互作用

• 批准号: 2414930
• 负责人: GERALD R CUNHA
• 金额: $ 17.75万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-05-01 至 2001-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2414930
• 关键词: RNase protection assay; biological signal transduction; cell cell interaction; cell differentiation; connective tissue stroma; epidermal growth factor; gene targeting; growth factor; hormone regulation /control mechanism; immunocytochemistry; in situ hybridization; laboratory mouse; laboratory rat; mesenchyme; muscle hypertrophy; paracrine; receptor expression; smooth muscle; stromal cells; transforming growth factors; urinary bladder epithelium; urinary tract obstruction; wound healing

The main goal of this proposal is to investigate the role of growth factors as mediators of stromal-epithelial interactions in bladder development and dysfunction. Our previous work has shown that bladder smooth muscle (SM) is induced from undifferentiated bladder mesenchyme by bladder epithelium. This proposal is based on the hypothesis that interactions between bladder epithelial and mesenchymal cells regulate and maintain SM cell differentiation and growth in developing and dysfunctional bladders. These cell-cell interactions are mediated by the local production and action of growth factors an other paracrine acting mediators. This hypothesis will be tested using an experimental model of urethral obstruction in which bladder SM responds by undergoing hypertrophy. We have also created an experimental model of bladder augmentation in which a grafted acellular bladder tissue matrix serves as a scaffold for the ingrowth of native bladder smooth muscle and urothelial cells. During normal bladder development and in two experimental systems, cellular signaling mechanisms regulating both normal and abnormal growth and differentiation of bladder SM will be examined through pursuit of the following specific aims. (Specific aim "1) Role of growth factors as mediators of cell-cell interactions in normal bladder development and during bladder round repair. Expression of growth factors (TGF-alpha, EGF, KGF, and TGFbeta1, beta2 and beta3) and growth factor receptors will be defined in normal bladder development and during bladder wound repair by RNase protection and localized by in-situ hybridization and immunohistochemistry. (Specific aim #2) Role of growth factors and cell- cell signaling in pathogenesis of bladder hypertrophy resulting from partial obstruction of the bladder. This specific aim will be pursued by defining growth factor and growth factor receptor expression following partial bladder obstruction in rats. The hypothesis of this specific aim is that SM cells are direct targets of the physical signal eliciting fibromuscular hypertrophy and that diffusible growth factors. (TGFalpha, EGF, KGF, TGFbeta1, beta2 and beta3) are involved. To accomplish this specific aim chimeric bladders will be surgically created by grafting whole embryonic bladders or strips of neonatal or adult bladders (epithelium intact or removed) into the sub-detrusor space of adult bladders. This unique experimental design will allow study of SM development and hypertrophy in bladders from transgenic mice in which growth factor genes have been deleted or impaired (TGFalpha knockout, EGF receptor knockout, TGFbeta1 knockout and FGFR2 dominant negative (Keratin 14 promoter) mice). (Specific aim #3) Use of an acellular bladder matrix patch to study normal and abnormal development of the bladder. The strategy of this specific aim is to characterize detrusor neo-formation and fibromuscular hypertrophy following grafting of acellular bladder matrix. Additional studies will assess regional and temporal differences in growth factor expression during bladder regeneration in bladders grafted with acellular bladder matrix. Finally, the performance of transgenic mouse bladder transplants during matrix-driven bladder regeneration will elucidate the role of growth factors in bladder regeneration. The long-term objective of this research is to develop effective therapeutic strategies for treating bladder dysfunction based upon stromal-epithelial interactions.

这项提案的主要目标是研究增长因素的作用。 作为基质-上皮相互作用的介质在膀胱发育和 功能障碍。我们以前的工作表明，膀胱平滑肌(SM) 是由膀胱上皮未分化的膀胱间充质诱导而来的。 这一建议是基于这样的假设，即膀胱之间的相互作用 上皮细胞和间充质细胞对SM细胞的调节和维持 发育和功能障碍的膀胱的分化和生长。这些 细胞间的相互作用是由局部产生和作用于 生长因子是旁分泌的另一种作用介体.这一假说将 使用尿路梗阻的实验模型进行测试 膀胱SM的反应是肥大。我们还创建了一个 脱细胞移植物膀胱扩大术的实验模型 膀胱组织基质作为天然组织工程支架的实验研究 膀胱平滑肌和尿路上皮细胞。 在正常的膀胱发育过程中，在两个实验系统中，细胞 调节正常和异常生长的信号机制和 膀胱SM的辨证分型将通过追求 遵循特定的目标。(特定目标“1)生长因素的作用 细胞-细胞相互作用的介体在正常膀胱发育和 在膀胱环修复过程中。生长因子(转化生长因子-α、表皮生长因子、 KGF、TGFbeta1、Beta2和Beta3)和生长因子受体 在正常的膀胱发育和膀胱伤口修复过程中定义为 核糖核酸酶保护和定位的原位杂交和 免疫组织化学。(具体目标2)生长因子和细胞的作用- 细胞信号转导在慢性前列腺炎致膀胱肥大发病机制中的作用 膀胱的部分梗阻。这一具体目标将通过以下方式实现 定义生长因子和生长因子受体的表达 大鼠部分性膀胱梗阻模型。这一特定目的的假设 SM细胞是物理信号引发的直接目标 纤维肌肉肥大和弥散性生长因子。(TGFAlpha， 涉及EGF、KGF、TGFbeta1、Beta2和Beta3)。要做到这一点 特定目标嵌合体膀胱将通过手术移植完整 胚胎膀胱或新生儿或成人膀胱条(上皮 完整或移除)进入成人膀胱的逼尿肌下间隙。这 独特的实验设计将允许研究SM的发展和 携带生长因子基因的转基因小鼠膀胱肥大 已被删除或受损(TGFAlpha基因敲除，EGF受体基因敲除， TGFbeta1基因敲除和FGFR2显性阴性(角蛋白14启动子)小鼠)。 (特定目标#3)使用无细胞膀胱基质贴片研究正常 以及膀胱的异常发育。这一特定目标的战略 是描述逼尿肌新生和纤维肌肉肥大的特征 无细胞膀胱基质移植后。更多的研究将 评估生长因子表达的地区和时间差异 无细胞膀胱基质移植后膀胱再生的实验研究 最后，转基因小鼠膀胱移植在 基质驱动的膀胱再生将阐明生长的作用 影响膀胱再生的因素。这项研究的长期目标是 是开发有效的治疗策略来治疗膀胱 基于间质-上皮相互作用的功能障碍。