Mutations in the Apc gene occur in many cancers but are especially frequent in colorectal cancer (CRC). The ApcMin/+ mouse is a highly studied model of intestinal tumorigenesis since the Apc gene mutation results in dysregulation of the Wnt signaling pathway. Our recent work using IL-17A deficient (KO) ApcMin/+ mice has identified IL-17A as a proinflammatory cytokine affecting tumorigenesis. Introduction of wildtype (WT) Tregs into ApcMin/+ mice regressed tumors markedly while Tregs from ApcMin/+ mice could not, suggesting an important factor from Tregs for tumor regression lacking in ApcMin/+ Tregs. In ApcMin/+ mice, the expression of Gata-3 was decreased in Tregs as well as effector T cells. Our analysis of Tregs and other immune cell types has identified the Wnt antagonist Dikkopf-1(Dkk1) as the most abundant Wnt family gene produced by Tregs. Importantly, IL-17A effectively inhibited Dkk1 expression in ApcMin/+ Tregs but not in WT Tregs. Dkk1 could induce Gata-3 and also IL-10, which was downregulated in ApcMin/+ mice. Dkk1 also stimulated Treg proliferation but did not impair Treg function. The lack of Treg-derived Dkk1 blocked the ability of Treg to suppress inflammatory bowel disease (IBD) in a standard murine IBD model. Based on these results, first we hypothesize that Treg-derived Dkk1 suppresses intestinal inflammation that is fueled by IL-17A under homeostatic conditions. Second, we argue that the loss of Dkk1 expression in ApcMin/+ Treg leads to the loss of IL-10, and also fails to regulate the Wnt pathway controlling ApcMin/+ intestinal stem cell proliferation. Third, we hypothesize that Treg-derived Dkk1 and IL-17A from Th17 cells are crucial regulators of the tumor microenvironment, directly targeting ISCs carrying the ApcMin/+ mutation.
Four aims will focus on testing these three hypotheses. First, Dkk-1 deficient Treg will be used in in vivo tumor regression experiments and in in vitro biologic studies to define the contribution of Dkk1 to Treg function. The mechanism that Dkk1 induces Treg proliferation will be studied in three different signaling pathways. Second, the role of IL-17A to downregulate Dkk1 in ApcMin/+ Tregs will be studied by utilizing IL-17 receptor-deficient Foxp3+ ApcMin/+ Tregs in vivo and in vitro. The mechanism which IL-17A and Dkk1 regulates IL-10 will be studied as well. Third, the effects of IL-17A and Dkk1 on intestinal stem cells (ISC) carrying the mutation in the Apc gene will be characterized. Conditional deletion of the IL-17 receptor in ISCs in the ApcMin/+ background will be studied in vivo. We will deliver Dkk1 into the intestine in ApcMin/+ mice, testing the potential for Dkk1 as a therapeutic. Finally, we will extend our findings to human FAP Treg utilizing a Treg/T cell expansion technique from colon biopsies, and compare them with cells from sporadic colon cancer patients and healthy donors. These studies should give important mechanistic insight into the specific inflammatory molecules that can compromise cells of the immune system. This mechanistic insight should provide a rational approach to translation of these results to the clinic.
This project will assess the proinflammatory mechanisms leading to induction of intestinal polyps via IL-17 and Wnt signaling. The Wnt antagonist Dkk1 is hypothesized to be a central regulator of intestinal homeostasis via regulatory T cells. There are distinct cell type specific signaling mechanisms that will be characterized.
|Chae, Wook-Jin; Bothwell, Alfred L M (2018) Therapeutic Potential of Gene-Modified Regulatory T Cells: From Bench to Bedside. Front Immunol 9:303|
|Jayakumar, Asha; Bothwell, Alfred L M (2017) Stat6 Promotes Intestinal Tumorigenesis in a Mouse Model of Adenomatous Polyposis by Expansion of MDSCs and Inhibition of Cytotoxic CD8 Response. Neoplasia 19:595-605|
|Chae, Wook-Jin; Park, Jong-Hyun; Henegariu, Octavian et al. (2017) Membrane-bound Dickkopf-1 in Foxp3+ regulatory T cells suppresses T-cell-mediated autoimmune colitis. Immunology 152:265-275|
|Chae, Wook-Jin; Bothwell, Alfred L M (2016) Snapshots of CD4(+) T cell plasticity in the pathogenesis of allergic asthma. J Thorac Dis 8:E1010-E1012|
|Chae, Wook-Jin; Ehrlich, Allison K; Chan, Pamela Y et al. (2016) The Wnt Antagonist Dickkopf-1 Promotes Pathological Type 2 Cell-Mediated Inflammation. Immunity 44:246-58|
|Chae, Wook-Jin; Bothwell, Alfred L M (2015) Spontaneous Intestinal Tumorigenesis in Apc (/Min+) Mice Requires Altered T Cell Development with IL-17A. J Immunol Res 2015:860106|
|Kidane, Dawit; Chae, Wook Jin; Czochor, Jennifer et al. (2014) Interplay between DNA repair and inflammation, and the link to cancer. Crit Rev Biochem Mol Biol 49:116-39|
|Tobiasova, Zuzana; Zhang, Lufeng; Yi, Tai et al. (2011) Peroxisome proliferator-activated receptor-? agonists prevent in vivo remodeling of human artery induced by alloreactive T cells. Circulation 124:196-205|