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A probiotic-derived protein regulates epigenetic programming in intestinal epithelial cells for long-term prevention of colitis

益生菌衍生的蛋白质调节肠上皮细胞的表观遗传编程，以长期预防结肠炎

基本信息

批准号：
10598497
负责人：
FANG YAN
金额：
$ 44.15万
依托单位：
VANDERBILT UNIVERSITY MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-04-01 至 2025-03-31
项目状态：
未结题

项目摘要

PROJECT SUMMARY Dysbiosis in infants and children is associated with increased susceptibility to inflammatory bowel disease (IBD) in adults. However, the mechanisms whereby the gut microbiota colonization in early life confers health outcomes throughout the lifespan remain unclear. Studies from the previous funding period isolated and cloned a Lactobacillus rhamnosus GG (LGG)-derived secretory protein, p40, and demonstrated that neonatal p40 supplementation prevents colitis in adult mice. Our preliminary studies discovered that p40 interacts with two transcriptional factors, Mga and Max, to regulate expression of Setd1β gene, which encodes a methyltransferase for catalyzing mono and trimethylation of histone 3-lysine 4 (H3K4me1/3). We identified TGFβ as a potential target of p40-up-regulated Setd1β. IECs are rapidly renewed and continuously regenerated from intestinal stem cells (ISCs). We found that p40 modulates H3K4m31/3 in ISCs in early stage, and p40 treatment in neonates, but not adult mice, stimulates sustained increase in TGFβ gene expression in IECs. TGFβ has been shown to have multiple functions against inflammation. Thus, we hypothesize that p40 up-regulates Setd1β gene expression in IECs through increasing Mga:Max dimerization. Supplementation with p40 in early life stimulates Setd1β-dependent H3K4me1/3 deposition at the TGFβ locus in ISCs, which is inherited by IECs to enable the sustained increase in TGFβ production, and subsequently prevention of intestinal inflammation in adulthood. In Aim 1, we will determine whether p40-stimulated Setd1β gene expression and H3K4me1/3 are required to drive the increase in expressing TGFβ gene in IECs, and elucidate whether the interaction between p40 and the Mga:Max dimers mediates up-regulation of Setd1β production. In Aim 2, we will identify the temporal window of p40 exposure in early life that causes the sustained increase in TGFβ production in IECs, and define whether p40-regulated sustained increase in TGFβ gene expression in IECs requires the increase of Setd1β gene expression inISCs in early life. Human and mouse enteroids and colonoids and 2D cultures and cell lines with silencing Setd1β or Mga genes, and mouse models of constitutive and inducible Setd1β gene knock down in IECs or ISCs will be utilized for these two aims. In Aim 3, we will determine whether the sustained increase in TGFβ production after p40 supplementation in early life is required to prevent colitis in adult mice. We will use neutralizing antibodies and inhibitors to block TGFβ function in mice with induced and spontaneously developed colitis. We will also determine whether sustained increase in TGFβ production enhances Tregs induction in the intestine and protective epithelial responses for the prevention of colitis in adulthood. This aim will be tested in mice with TGFβ receptor II deletion in CD4+ T cells and Smad4 deletion in IECs. Together, our proposed research will elucidate a novel mechanism underlying the consequences of long-lasting effects of p40 on prevention of colitis, and lay the foundation for developing early intervention with p40 as a novel therapy for prevention of IBD in adults.

项目概要婴儿和儿童的菌群失调与炎症性肠易感性增加有关成人疾病（IBD）。然而，生命早期肠道微生物群定植的机制赋予整个生命周期的健康结果仍不清楚。上一资助期的研究被隔离并克隆了鼠李糖乳杆菌 GG (LGG) 衍生的分泌蛋白 p40，并证明新生儿补充 p40 可预防成年小鼠的结肠炎。我们的初步研究发现 p40 与两个转录因子 Mga 和 Max，调节 Setd1β 基因的表达，该基因编码甲基转移酶，用于催化组蛋白 3-赖氨酸 4 (H3K4me1/3) 的单甲基化和三甲基化。我们确定了 TGFβ 作为 p40 上调 Setd1β 的潜在靶标。 IEC 快速更新且持续不断由肠干细胞（ISC）再生。我们发现p40在早期调节ISC中的H3K4m31/3，新生小鼠（而非成年小鼠）的 p40 治疗可刺激小鼠中 TGFβ 基因表达的持续增加 IEC。 TGFβ已被证明具有多种抗炎症功能。因此，我们假设 p40 通过增加 Mga:Max 二聚化上调 IEC 中的 Setd1β 基因表达。补充生命早期的 p40 会刺激 ISC 中 TGFβ 位点上依赖 Setd1β 的 H3K4me1/3 沉积，这是由 IEC 继承，以实现 TGFβ 产量的持续增加，并随后预防成年后肠道发炎。在目标 1 中，我们将确定 p40 刺激的 Setd1β 基因是否表达和 H3K4me1/3 是驱动 IEC 中 TGFβ 基因表达增加所必需的，并阐明 p40 和 Mga:Max 二聚体之间的相互作用是否介导 Setd1β 产生的上调。在目标 2，我们将确定生命早期 p40 暴露的时间窗口，该窗口导致 IEC 中 TGFβ 的产生，并确定 p40 调节的 TGFβ 基因表达是否持续增加 IECs 需要生命早期 ISCs 中 Setd1β 基因表达的增加。人类和小鼠肠类和具有沉默 Setd1β 或 Mga 基因的结肠和 2D 培养物和细胞系，以及组成型小鼠模型 IEC 或 ISC 中的诱导型 Setd1β 基因敲除将用于这两个目标。在目标 3 中，我们将确定生命早期补充 p40 后 TGFβ 产量的持续增加是否是预防成年小鼠结肠炎所必需的。我们将使用中和抗体和抑制剂来阻断 TGFβ 在患有诱导性和自发性结肠炎的小鼠中发挥作用。我们还将确定是否持续 TGFβ 产生的增加增强了肠道中 Tregs 的诱导和保护性上皮反应预防成年期结肠炎。该目标将在 CD4+ T 中 TGFβ 受体 II 缺失的小鼠中进行测试 IEC 中的细胞和 Smad4 缺失。我们提出的研究将共同阐明一种新机制揭示 p40 对预防结肠炎的长期影响的后果，并为开发 p40 早期干预作为预防成人 IBD 的新疗法。

项目成果

期刊论文数量（18）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Noncanonical NF-κB activation mediates STAT3-stimulated IDO upregulation in myeloid-derived suppressor cells in breast cancer.

非规范的NF-κB激活介导了髓样衍生的抑制细胞中STAT3刺激的IDO上调。

DOI：
10.4049/jimmunol.1400833
发表时间：
2014-09-01
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
0
作者：
Yu J;Wang Y;Yan F;Zhang P;Li H;Zhao H;Yan C;Yan F;Ren X
通讯作者：
Ren X

Disruption of NF-kappaB signalling by ancient microbial molecules: novel therapies of the future?

古代微生物分子对 NF-kappaB 信号传导的破坏：未来的新疗法？

DOI：
10.1136/gut.2009.179614
发表时间：
2010
期刊：
Gut
影响因子：
24.5
作者：
Yan,Fang;Polk,DBrent
通讯作者：
Polk,DBrent

Berberine induces caspase-independent cell death in colon tumor cells through activation of apoptosis-inducing factor.

DOI：
10.1371/journal.pone.0036418
发表时间：
2012
期刊：
PloS one
影响因子：
3.7
作者：
Wang L;Liu L;Shi Y;Cao H;Chaturvedi R;Calcutt MW;Hu T;Ren X;Wilson KT;Polk DB;Yan F
通讯作者：
Yan F

Interactions between the gut microbiota-derived functional factors and intestinal epithelial cells - implication in the microbiota-host mutualism.