Papadakis Konstantinos A, Krempski James, Svingen Phyllis, Xiong Yuning, Sarmento Olga F, Lomberk Gwen A, Urrutia Raul A, Faubion William A
Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, Minnesota; and
Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, Minnesota; and.
Am J Physiol Gastrointest Liver Physiol. 2015 Dec 1;309(11):G900-9. doi: 10.1152/ajpgi.00309.2015. Epub 2015 Oct 15.
Krüppel-like factor (KLF)-10 is an important transcriptional regulator of TGF-β1 signaling in both CD8(+) and CD4(+) T lymphocytes. In the present study, we demonstrate a novel role for KLF10 in the regulation of TGFβRII expression with functional relevance in macrophage differentiation and activation. We first show that transfer of KLF10(-/-) bone marrow-derived macrophages into wild-type (WT) mice leads to exacerbation of experimental colitis. At the cell biological level, using two phenotypic strategies, we show that KLF10-deficient mice have an altered colonic macrophage phenotype with higher frequency of proinflammatory LyC6(+)MHCII(+) cells and a reciprocal decrease of the anti-inflammatory LyC6(-)MHCII(+) subset. Additionally, the anti-inflammatory CD11b(+)CX3CR1(hi) subset of colonic macrophages is significantly decreased in KLF10(-/-) compared with WT mice under inflammatory conditions. Molecularly, CD11b(+) colonic macrophages from KLF10(-/-) mice exhibit a proinflammatory cytokine profile with increased production of TNF-α and lower production of IL-10 in response to LPS stimulation. Because KLF10 is a transcription factor, we explored how this protein may regulate macrophage function. Consequently, we analyzed the expression of TGFβRII expression in colonic macrophages and found that, in the absence of KLF10, macrophages express lower levels of TGFβRII and display an attenuated Smad-2 phosphorylation following TGF-β1 stimulation. We further show that KLF10 directly binds to the TGFβRII promoter in macrophages, leading to enhanced gene expression through histone H3 acetylation. Collectively, our data reveal a critical role for KLF10 in the epigenetic regulation of TGFβRII expression in macrophages and the acquisition of a "regulatory" phenotype that contributes to intestinal mucosal homeostasis.
Krüppel样因子(KLF)-10是CD8(+)和CD4(+) T淋巴细胞中TGF-β1信号的重要转录调节因子。在本研究中,我们证明了KLF10在调节TGFβRII表达方面具有新的作用,这与巨噬细胞分化和激活具有功能相关性。我们首先表明,将KLF10(-/-)骨髓来源的巨噬细胞转移到野生型(WT)小鼠中会导致实验性结肠炎加剧。在细胞生物学水平上,我们使用两种表型策略表明,KLF10缺陷小鼠的结肠巨噬细胞表型发生改变,促炎LyC6(+)MHCII(+)细胞频率更高,而抗炎LyC6(-)MHCII(+)亚群则相应减少。此外,在炎症条件下,与WT小鼠相比,KLF10(-/-)小鼠结肠巨噬细胞的抗炎CD11b(+)CX3CR1(hi)亚群显著减少。在分子水平上,来自KLF10(-/-)小鼠的CD11b(+)结肠巨噬细胞表现出促炎细胞因子谱,对LPS刺激的反应是TNF-α产生增加而IL-10产生减少。由于KLF10是一种转录因子,我们探讨了这种蛋白质如何调节巨噬细胞功能。因此,我们分析了结肠巨噬细胞中TGFβRII的表达,发现缺乏KLF10时,巨噬细胞表达较低水平的TGFβRII,并且在TGF-β1刺激后Smad-2磷酸化减弱。我们进一步表明,KLF10直接与巨噬细胞中的TGFβRII启动子结合,通过组蛋白H3乙酰化导致基因表达增强。总的来说,我们的数据揭示了KLF10在巨噬细胞中TGFβRII表达的表观遗传调控以及获得有助于肠道黏膜稳态的“调节性”表型中起关键作用。