Cellular Plasticity of Defa4Cre-Expressing Paneth Cells in Response to Notch Activation and Intestinal Injury
Cellular Plasticity of Defa4Cre-Expressing Paneth Cells in Response to Notch Activation and Intestinal Injury
复制标题
DOI：
10.1016/j.jcmgh.2018.11.004
复制
发表时间：
2019-01-01
影响因子：
7.2
通讯作者：
Dempsey, Peter J.
中科院分区：
文献类型：
--
作者：
Jones, Jennifer C.;Brindley, Constance D.;Dempsey, Peter J.
背景与目的：富含亮氨酸重复序列的G蛋白偶联受体5阳性隐窝基底柱状细胞的丢失为不同的兼性干细胞群体去分化和重新填充干细胞区室提供了允许的条件。在这项研究中，我们使用了防御素α 4-Cre重组酶，（Defa 4Cre）系来确定Paneth细胞去分化的潜力，并有助于在正常稳态和肠损伤后维持肠干细胞（ISC）。Rosa 26串联二聚体番茄（tdTomato），一种红色荧光蛋白，（或Rosa 26增强型黄色荧光蛋白（EYFP））报告基因，Notch功能获得性（Defa 4（Cre）; Rosa 26 Notch胞内结构域（NICD）-虹膜-核绿色荧光蛋白（nGFP）和Defa 4（Cre）; Rosa 26（反向四环素反式激活子）增强型绿色荧光蛋白（EGFP）; TetO（NICD）），一种含有解整合素和金属蛋白酶结构域的蛋白10（ADAM 10）功能丧失分析了小鼠（Defa 4（Cre）; ADAM 10（flox/flox））和腺瘤性结肠息肉病（APC）失活（Defa 4（Cre）; APC（flox/flox））。阿霉素治疗用作急性肠损伤模型。谱系追踪，增殖和分化进行了评估，在体外和invivo.RESULTS：Defa 4（Cre）-表达细胞注定成为成熟的潘氏细胞，并不有助于ISC维持在体内正常的稳态。然而，在类肠细胞中观察到自发谱系追踪，荧光激活细胞分选仪分选的Defa 4（Cre）标记的细胞显示克隆形成的类肠细胞生长。表达Defa 4（Cre）的细胞中的Notch激活导致体内多能ISC的去分化，并且是腺瘤形成所需的。ADAM 10基因缺失对隐窝内环境稳定性无明显影响。然而，急性阿霉素诱导的损伤后，Defa 4（Cre）-表达细胞有助于再生的ADAM 10-Notch依赖的mathematics.CONCLUSIONS：我们的研究表明，Defa 4（Cre）-表达潘氏细胞具有细胞可塑性，可以去分化成多能干细胞后，Notch激活，并可以促进肠再生在急性损伤模型。
BACKGROUND & AIMS: Loss of leucine-rich repeat-containing G-protein-coupled receptor 5-positive crypt base columnar cells provides permissive conditions for different facultative stem cell populations to dedifferentiate and repopulate the stem cell compartment. In this study, we used a defensin alpha 4-Cre recombinase (Defa4Cre) line to define the potential of Paneth cells to dedifferentiate and contribute to intestinal stem cell (ISC) maintenance during normal homeostasis and after intestinal injury.METHODS: Small intestine and enteroids from Defa4(Cre); Rosa26 tandem dimer Tomato (tdTomato), a red fluoresent protein, (or Rosa26 Enhanced Yellow Fluorescent Protein (EYFP)) reporter, Notch gain-of-function (Defa4(Cre); Rosa26 Notch Intracellular Domain (NICD)-ires-nuclear Green Fluorescent Protein (nGFP) and Defa4(Cre);Rosa26(reverse tetracycline transactivator-ires) Enhanced Green Fluorescent Protein (EGFP); TetO(NICD)), A Disintegrin and Metalloproteinase domain-containing protein 10 (ADAM10) loss-of-function (Defa4(Cre); ADAM10(flox/flox)), and Adenomatous polyposis coli (APC) inactivation (Defa4(Cre); APC(flox/flox)) mice were analyzed. Doxorubicin treatment was used as an acute intestinal injury model. Lineage tracing, proliferation, and differentiation were assessed in vitro and in vivo.RESULTS: Defa4(Cre)-expressing cells are fated to become mature Paneth cells and do not contribute to ISC maintenance during normal homeostasis in vivo. However, spontaneous lineage tracing was observed in enteroids, and fluorescent-activated cell sorter-sorted Defa4(Cre)-marked cells showed clonogenic enteroid growth. Notch activation in Defa4(Cre)-expressing cells caused dedifferentiation to multipotent ISCs in vivo and was required for adenoma formation. ADAM10 deletion had no significant effect on crypt homeostasis. However, after acute doxorubicin-induced injury, Defa4(Cre)-expressing cells contributed to regeneration in an ADAM10-Notch-dependent manner.CONCLUSIONS: Our studies have shown that Defa4(Cre)-expressing Paneth cells possess cellular plasticity, can dedifferentiate into multipotent stem cells upon Notch activation, and can contribute to intestinal regeneration in an acute injury model.