Inflamation-Gene Expression by Monocytes in Frailty

描述（由申请人提供）：与非虚弱老年人相比，虚弱老年人的功能下降、福尔斯、住院、疗养院安置和早期死亡风险增加。新出现的证据表明，以CRP和IL-6水平升高为标志的炎症与虚弱有关。然而，人们对虚弱的老年人炎症系统的激活范围和潜在机制知之甚少。单核细胞在局部和全身炎症的激活和调节中起关键作用，并且研究编码细胞因子介质和受体的基因的单核细胞表达可以提供对脆弱中炎症系统激活的分子机制的相当深入的了解。这项建议的短期目标是建立从虚弱的老年人分离的单核细胞的炎症特异性基因表达的特异性调节的初步初步数据。该项目的长期目标是利用这些发现进行更深入的虚弱病因学和干预研究。我们假设年老体弱的人在组成型和LPS诱导的单核细胞编码IL-6、TNF-α、IL-1 β和IL-1受体拮抗剂（IL-1 ra）的基因表达方面有显著改变。本提案的主要目的是检验这一假设，并进一步探索在脆弱综合征中编码80种其他细胞因子介质和细胞表面分子的基因的组成性和LPS诱导的单核细胞表达的调制的初步证据。为了实现这些目标，我们提出了一个探索性的调查（R21）组成和LPS诱导的单核细胞表达的基因编码84细胞因子介质和细胞表面分子在社区居住的体弱和非体弱的老年人。一个经过验证的脆弱筛选工具将被用来确定15个年龄，种族和性别匹配的对脆弱和非脆弱的老年人。将在存在和不存在LPS的情况下分离和培养单核细胞。将使用市售的炎症特异性基因阵列（含有IL-6、TNF-α、IL-1 β、IL-1 ra和80种其他细胞因子介质和细胞表面分子的cDNA），评价并比较虚弱和非虚弱受试者的组成型和LPS诱导的单核细胞基因表达。具有显著差异的候选基因将通过定量RT-PCR进一步评估。对编码IL-6、TNF-α、IL-1 β、IL-1 ra的基因的组成性和LPS诱导的单核细胞表达的初步评价将为对体弱老年人中这些重要细胞因子介质的单核细胞产生和调节进行更有洞察力的机制研究奠定基础。该项目还将探索在体弱老年人中编码80种其他细胞因子介质和细胞表面分子的基因的组成性和LPS诱导的单核细胞表达的调制的初步证据。因此，它将作为一个假设生成项目，并为假设细化的基础，这将为未来的机制和潜在的老年虚弱的干预性研究开辟新的途径。

DESCRIPTION(provided by applicant): Frail older adults are at increased risk for functional decline, falls, hospitalization, nursing home placement, and early mortality as compared to non-frail older adults. Emerging evidence suggests that inflammation, as marked by elevated levels of CRP and IL-6, is associated with frailty. However, little is known about the scope of, and the underlying mechanisms that contribute to, the activation of the inflammation system in frail older adults. Monocytes play pivotal roles in the activation and regulation of local and systemic inflammation, and investigating monocytic expression of genes encoding cytokine mediators and receptors can provide considerable insight into molecular mechanisms that underlie activation of the inflammation system in frailty. The short-term goal of this proposal is to establish initial preliminary data for specific modulations in inflammation-specific gene expression of isolated monocytes from frail older adults. The long-term goal of this project is to utilize these findings for the development of more in-depth etiologic and interventional studies of frailty. We hypothesize that frail older adults have significant alterations in constitutive and LPS-induced monocytic expression of genes encoding IL-6, TNF-alpha, IL-1beta, and IL-1 receptor antagonist (IL-1ra). The primary objectives of this proposal are to test this hypothesis and to further explore initial evidence for modulations in constitutive and LPS-induced monocytic expression of genes encoding 80 other cytokine mediators and cell surface molecules in the frailty syndrome. To accomplish these objectives, we propose an exploratory investigation (R21) into constitutive and LPS-induced monocytic expression of genes encoding 84 cytokine mediators and cell surface molecules in community-dwelling frail and non-frail older adults. A validated frailty-screening tool will be used to identify 15 age-, race-, and sex-matched pairs of frail and nonfrail older adults. Monocytes will be isolated and cultured in the presence and absence of LPS. Constitutive and LPS-induced monocytic gene expression will be evaluated and compared between frail and non-frail subjects, using a commercially available, inflammation-specific gene array containing cDNAs of IL-6, TNF-alpha, IL-1beta, IL-1ra, and 80 other cytokine mediators and cell surface molecules. Candidate genes with significant differences will be further evaluated by quantitative RT-PCR. This initial evaluation of the constitutive and LPS-induced monocytic expression of genes encoding IL-6, TNF-alpha, IL-1beta, IL-1ra will set the stage for more discerning mechanistic studies on monocytic production and regulation of these important cytokine mediators in frail older adults. This project will also explore initial evidence for modulations in constitutive and LPS-induced monocytic expression of genes encoding 80 other cytokine mediators and cell surface molecules in frail older adults. Therefore, It will serve as a hypothesis-generating project, and a basis for hypothesis refinement, that will open new pathways for future mechanistic and, potentially, interventional investigations of frailty in old age.