Development of a Primary Human Intestinal Epithelium Enriched in L-Cells for Assay of GLP-1 Secretion.
Development of a Primary Human Intestinal Epithelium Enriched in L-Cells for Assay of GLP-1 Secretion.
复制标题
用于GLP-1分泌测定的富含L细胞的原代人肠上皮细胞的开发。
DOI：
10.1021/acs.analchem.2c00912
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发表时间：
2022-07-12
影响因子：
7.4
通讯作者：
Allbritton, Nancy L.
中科院分区：
文献类型：
--
作者：
Villegas-Novoa, Cecilia;Wang, Yuli;Sims, Christopher E.;Allbritton, Nancy L.
2型糖尿病是一种与肥胖和人类摄食行为失调有关的慢性疾病。胰升糖素样肽1(GLP-1)是一种调节体重、食物摄入量和血糖水平的重要激素，由肠内分泌L细胞分泌。在原代培养的肠道细胞中，L细胞(包括类器官细胞和单层细胞)的数量很少，这使得对原代人类细胞分泌GLP-1的分析具有挑战性。在本论文中，我们建立了一个由优化的L细胞中的人肠道组织结构和基于标准抗体的GLP-1检测组成的分析流水线，为开发调节L细胞信号转导的药物筛选化合物。加入5-羟色胺受体激动剂Bimu-8，优化R-响应蛋白和Noggin浓度，以及使用血管活性肠肽，可增加原代人结肠上皮单层L细胞的密度。此外，气液界面(ALI)培养模式的加入增加了L细胞的数量，使传统的酶联免疫分析方法的信噪比可用于监测复合筛选中GLP-1的分泌。为了验证优化分析方法的实用性，对21种饮料甜味剂进行了刺激GLP-1分泌的能力筛选。甜菊糖苷和甜蜜素是最有效的GLP-1分泌诱导剂。该平台实现了对人原代L细胞分泌GLP-1的量化，将在理解L细胞的形成和生理方面有广泛的应用，并将改进对人类摄食行为调节因子的识别。
Type 2 diabetes mellitus is a chronic disease associated with obesity and dysregulated human feeding behavior. The hormone glucagon-like peptide 1 (GLP-1), a critical regulator of body weight, food intake, and blood glucose levels, is secreted by enteroendocrine L-cells. The paucity of L cells in primary intestinal cell cultures including organoids and monolayers has made assays of GLP-1 secretion from primary human cells challenging. In the current paper, an analytical assay pipeline consisting of an optimized human intestinal tissue construct enriched in L cells paired with standard antibody-based GLP-1 assays was developed to screen compounds for the development of pharmaceuticals to modulate L-cell signaling. The addition of the serotonin receptor agonist Bimu 8, optimization of R- spondin and Noggin concentrations, and utilization of vasoactive intestinal peptide (VIP) increased the density of L-cells in a primary human colonic epithelial monolayer. Additionally, the incorporation of an air-liquid interface (ALI) culture format increased L-cell number so that the signal to noise of conventional enzyme-linked immunoassays could be used to monitor GLP-1 secretion in compound screens. To demonstrate the utility of the optimized analytical method, 21 types of beverage sweeteners were screened for their ability to stimulate GLP-1 secretion. Stevioside and cyclamate were found to be the most potent inducers of GLP-1 secretion. This platform enables quantification of GLP-1 secretion from human primary L cells and will have broad application in understanding L-cell formation and physiology and will improve the identification of modulators of human feeding behavior.
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影响因子：
14.8
作者：
Hinman SS;Wang Y;Kim R;Allbritton NL
通讯作者：
Allbritton NL
影响因子：
8.1
作者：
Kuhre RE;Wewer Albrechtsen NJ;Larsen O;Jepsen SL;Balk-Møller E;Andersen DB;Deacon CF;Schoonjans K;Reimann F;Gribble FM;Albrechtsen R;Hartmann B;Rosenkilde MM;Holst JJ
通讯作者：
Holst JJ
影响因子：
2.7
作者：
Shyangdan DS;Royle PL;Clar C;Sharma P;Waugh NR
通讯作者：
Waugh NR
影响因子：
3.7
作者：
Oren DA;Wei Y;Skrabanek L;Chow BK;Mommsen T;Mojsov S
通讯作者：
Mojsov S
影响因子：
3.5
作者：
Kuhre RE;Wewer Albrechtsen NJ;Deacon CF;Balk-Møller E;Rehfeld JF;Reimann F;Gribble FM;Holst JJ
通讯作者：
Holst JJ