The Role of surfactant protein genetic variants in cyst*

表面活性蛋白遗传变异在囊肿中的作用*

• 批准号: 6528004
• 负责人: JOANNA FLOROS
• 金额: $ 31.61万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-30 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6528004
• 关键词: CHO cells; Pseudomonas aeruginosa; chloride channels; chronic disease /disorder; clinical research; cystic fibrosis; family genetics; gene expression; gene mutation; genetic polymorphism; genetic susceptibility; genotype; host organism interaction; human genetic material tag; human subject; linkage disequilibriums; medical complication; molecular pathology; pathologic process; polymerase chain reaction; protein structure function; pulmonary surfactants; restriction fragment length polymorphism; statistics /biometry

DESCRIPTION (provided by applicant): Our long-term objectives are to study mechanisms by which modifier genes affect pulmonary disease severity in patients with cystic fibrosis (CF), with emphasis on the surfactant proteins (SP-), which are produced by lung epithelial cells. SP-A and SP-D play a role in the innate host defense and/or the regulation of inflammatory processes in the lung, and SP-B is essential for normal lung function. Preliminary findings indicate that SP-A genetic variants are associated with disease severity in CF. The overall rationale for the proposed studies is that significant heterogeneity exists in the severity of pulmonary disease in CF patients, even in those that are homozygous for the F508 mutation of the CF transmembrane conductance regulator gene (CFTR). This phenotypic diversity may be related to the genetic heterogeneity of the surfactant protein genes. Genetic variation in the SP-B gene may relate to differences in pulmonary function and genetic variation in SP-A and SP-D may relate to differences in innate host defense function in CF. The overall hypothesis states that SP-A, SP-B, and SP-D are modifiers of pulmonary disease severity in CF and that differences exist among SP-A genetic variants in their ability to modulate phagocytosis of Pseudomonas aeruginosa. In this proposal, we will carry out two major groups of experiments. First, we will study family-based associations by carrying out extended transmission disequilibrium test (ETDT) and/or TDT analyses, of the SP-A, SP-B, and SP-D marker loci, and CF, to determine whether these loci are linked to CF and identify susceptibility SP alleles for CF severity. The entire CF study group as well as a number of subgroups based on CFTR genotype, severity, or other criteria, will be studied. Logistic regression analysis will be used to identify factors that may be of particular significance in CF along with specific genotypes (Aim 1). In the second group of experiments, we will focus on SP-A genetic variants for which preliminary evidence of an association between SP-A alleles and CF severity exists. We will study differences in the ability of SP-A alleles to enhance phagocytosis, by a macrophage-like cell line, of laboratory strains of mucoid and non-mucoid P. aeruginosa grown under different environmental conditions that have been shown to reproduce certain biochemical and functional characteristics found in clinical isolates of P. aeruginosa (Aim 2) and of mucoid and non-mucoid isolates of P. aeruginosa from CF patients (Aim 3). The findings may help identify specific host defense mechanisms involving alleles associated with pulmonary disease severity in CF and develop a useful in vitro model to study the in vivo modifications of P. aeruginosa and its clearance, and provide the basis for further consideration of novel therapeutic strategies to treat CF-related lung disease.

描述（由申请人提供）：我们的长期目标是研究修饰基因的机制 影响囊性纤维化 (CF) 患者肺部疾病的严重程度 重点关注肺表面活性蛋白（SP-） 上皮细胞。 SP-A 和 SP-D 在宿主先天防御和/或 肺部炎症过程的调节，SP-B 至关重要 为了正常的肺功能。 初步研究结果表明 SP-A 遗传 变异与 CF 疾病的严重程度相关。 总体理由 对于拟议的研究来说，存在显着的异质性 CF 患者肺部疾病的严重程度，即使是那些患有 CF 的患者 CF 跨膜电导的 F508 突变纯合 调节基因（CFTR）。 这种表型多样性可能与 表面活性蛋白基因的遗传异质性。 遗传变异 SP-B基因可能与肺功能和遗传差异有关 SP-A 和 SP-D 的变异可能与先天宿主防御的差异有关 CF 中的功能。 总体假设表明 SP-A、SP-B 和 SP-D 是 CF 肺部疾病严重程度的修饰因素，并且之间存在差异 SP-A 基因变异调节假单胞菌吞噬作用的能力 铜绿假单胞菌。 在本提案中，我们将开展两大组 实验。 首先，我们将通过开展以下活动来研究以家庭为基础的协会： 扩展传递不平衡测试 (ETDT) 和/或 TDT 分析， SP-A、SP-B 和 SP-D 标记基因座以及 CF，以确定这些基因座是否是 与 CF 相关并确定 SP 等位基因对 CF 严重程度的易感性。 这 整个 CF 研究组以及基于 CFTR 基因型的多个亚组， 将研究严重程度或其他标准。 逻辑回归分析 将用于识别在 CF 中可能特别重要的因素 以及特定的基因型（目标 1）。 在第二组实验中，我们 将重点关注 SP-A 遗传变异，初步证据表明 SP-A 等位基因与 CF 严重程度之间存在关联。 我们将学习 SP-A 等位基因增强吞噬作用的能力差异 粘液型和非粘液型 P. 实验室菌株的巨噬细胞样细胞系。 已证明在不同环境条件下生长的铜绿假单胞菌 重现某些生物化学和功能特征 铜绿假单胞菌（目标 2）以及粘液和非粘液的临床分离株 从 CF 患者中分离出铜绿假单胞菌（目标 3）。 这些发现可能有助于确定涉及的特定宿主防御机制 与 CF 肺部疾病严重程度相关的等位基因，并开发一个有用的 体外模型研究铜绿假单胞菌及其体内修饰 清除，并为进一步考虑新颖性提供依据 治疗 CF 相关肺部疾病的治疗策略。