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.

项目摘要婴儿和儿童的营养不良与炎症性肠的易感性增加有关成人疾病（IBD）。但是，早期肠道菌群殖民化的机制赋予了整个生命周期的健康结果尚不清楚。前一个资金期的研究孤立，克隆了rhamnosus lamnosus gg（LGG）衍生的秘密蛋白P40的克隆，并证明了新生儿补充P40可防止成年小鼠的结肠炎。我们的初步研究发现P40与 MGA和MAX的两个转录因子，用于调节SetD1β基因的表达，该基因编码A 甲基转移酶用于催化单声道和组蛋白3-赖氨酸4（H3K4ME1/3）的三甲基化。我们确定了 TGFβ是P40-UP调节的SETD1β的潜在靶标。 IEC迅速更新和连续从肠道干细胞（ISC）再生。我们发现P40在早期ISC中调节H3K4M31/3，在新生儿（而不是成年小鼠）中的P40治疗刺激TGFβ基因表达持续增加 IEC。 TGFβ已显示具有针对炎症的多个功能。那我们假设p40 通过增加MGA：最大二聚化来上调IEC中的SETD1β基因表达。补充早期生命中的p40刺激了ISC中TGFβ基因座的SETD1β依赖性H3K4ME1/3沉积， IEC遗传以使TGFβ产生的持续增加以及随后的预防成年时肠炎。在AIM 1中，我们将确定P40刺激的SETD1β基因是否需要表达和H3K4ME1/3才能驱动IEC中表达TGFβ基因的增加并阐明 P40与MGA：最大二聚体之间的相互作用是否介导SETD1β产生的上调。在 AIM 2，我们将确定早期生命中P40暴露的临时窗口，这会导致持续增加 IEC中的TGFβ产生，并确定P40调节的TGFβ基因表达是否持续增加 IEC需要在早期生命中增加setD1β基因表达。人类和小鼠肠内结石和2D培养物和带有沉默的SETD1β或MGA基因的细胞系，以及一致性的小鼠模型这两个目标将使用诱导的setD1β基因敲低IEC或ISC。在AIM 3中，我们将确定早期补充p40后TGFβ产生的持续增加是否为需要预防成年小鼠结肠炎。我们将使用中和抗体和抑制剂来阻断TGFβ 在具有诱导和赞助的结肠炎的小鼠中功能。我们还将确定是否持续 TGFβ产生的增加增强了肠道的Treg诱导和受保护的上皮反应的诱导预防成年后结肠炎。该目标将在CD4+ T中的TGFβ受体II缺失的小鼠中测试 IEC中的细胞和SMAD4删除。我们提出的研究将共同阐明一种新的机制 p40对预防结肠炎的持久影响的后果是基础的，并为用P40进行早期干预，作为一种新型疗法，用于预防成人IBD。

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.

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.