TISSUE CULTURE MODEL FOR RETINITIS PIGMENTOSA--A MOLECULAR APPROACH

每年数以千计的美国人失明，超过一半的人 被诊断为有遗传缺陷。眼科疾病 视网膜色素上皮(PE)与视网膜色素变性有关 (RP)、回旋萎缩、脉络膜血症、脉络膜硬化症和Stargedt‘s 黄斑变性。该项目的长期目标是确定 以及描述眼部疾病中的遗传缺陷和/或缺陷 广泛归类为视网膜色素变性(RP)。视网膜色素变性是一种 视网膜退行性疾病的特征是最终丧失 感光细胞。正常和营养不良动物模型的研究 提示RP的视觉功能障碍可能是继发性的 视网膜和色素上皮(PE)。虽然RP是最常见的 发生视网膜退行性疾病的原因和信息 疾病的发病机制既缺乏又令人困惑。 现有信息的不足可能与以下方面有特殊关系 生物组织供应不足。初代外植体培养 在PE中，视网膜的寿命是有限的，一些永久性细胞系是 去分化。因此，它们不是研究的最佳模型 视网膜变性的症状。这项提议的目标是追求一种 交替入路建立正常和反相眼球细胞系 供体眼(常染色体隐性、常染色体显性和性别连锁) 使用各种原癌基因和病毒DNA(H-ras-(Val 12)v- MYC、SV-40病毒DNA和SV-40T抗原基因。我们已经这么做了 利用原癌基因建立多种正常PE细胞系 (见附录)，并已能够获得更多差异化 利用细胞外基质自发建立细胞系 (见附录)。在本研究期间，我们建议建立视网膜 来自RP供体的细胞系并与正常永生细胞系进行比较 用RP细胞系进行生长、形态和PE-1表达的研究 视杆外节和视网膜吞噬功能标志物 新陈代谢。我们还将探讨增长因素和 化学诱导剂在感光细胞分化中的作用 最终是光感受器细胞退化的疾病。长期的 目标是通过使用以下方法识别和纠正RP中的病变 组织培养法。

More than half of the thousands of Americans who become blind each year are diagnosed as having a genetic defect. Ocular diseases in which retinal pigment epithelium (PE) is implicated are retinitis pigmentosa (RP), gyrate atrophy, choroidemia, choroidal sclerosis, and Stargedt's macular degeneration. The long-term goal of this project is to identify and characterize the genetic defect and/or defects in ocular diseases broadly grouped as retinitis pigmentosa (RP). Retinitis pigmentosa is a degenerative retinal disease characterized by eventual loss of photoreceptor cells. Studies on normal and dystrophic animal models suggest that visual failure in RP may be secondary to dysfunction in the retina and pigment epithelium (PE). Although RP is the commonest occurring retinal degenerative disease, the information on etiology and mechanism of pathogenesis in disease is both scant and confusing. Deficiencies in available information could be singularly related to inadequate availability of biological tissues. Primary explant cultures of PE, retina have a finite lifespan and a few permanent cell lines are dedifferentiated. As such they are not an optimal model for the study of retinal degeneration. The goal of this proposal is to pursue an alternate approach and establish ocular cell lines from normal and RP donor eyes (autosomal recessive, autosomal dominant and sex linked) by use of a variety of protooncogenes and viral DNAs (H-ras - (Val 12) v- myc, SV-40 viral DNA and SV-40T antigen gene. We have already established a variety of normal PE cell lines by use of protooncogenes (see Appendix) and have also been able to obtain more differentiated spontaneously established cell lines by use of extracellular matrices (see Appendix). In this study period, we propose to establish retinal cell lines from RP donors and compare the normal immortal cell lines with RP cell lines for growth, morphology and expression of PE - functional markers like phagocytosis of rod outer segments and retinal metabolism. We will also explore the role of growth factors and chemical inducers in photoreceptor cell differentiation as RP is ultimately a disease of photoreceptor cell degeneration. The long-term goal will be to identify and correct the lesion in RP by using this tissue culture approach.