Environmental Pollutants and AHR pathway in Uterine Leiomyoma

环境污染物与子宫平滑肌瘤的 AHR 通路

• 批准号: 10567192
• 负责人: Serdar E. Bulun
• 金额: $ 63.22万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-13 至 2027-10-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10567192
• 关键词: ARNT gene; ATAC-seq; Affect; Age; Amino Acid Transporter; Anemia; Apoptosis; Aryl Hydrocarbon Receptor; Bioinformatics; Biological Assay; Cell Proliferation; Cell Survival; Cells; ChIP-seq; Chemical Exposure; Chromatin; Development; Diet; Diethylhexyl Phthalate; Disease; Endocrine Disruptors; Environmental Pollutants; Environmental Risk Factor; Enzymes; Epidemiology; Exposure to; FDA approved; Fibroid Tumor; Future; Gene Expression; Gene Expression Profile; Gene Mutation; Genes; Genetic; Goals; Growth; Health; Human; Hysterectomy; In Vitro; Inhibition of Apoptosis; Kynurenine; Leiomyoma; Ligands; Link; Malignant Neoplasms; Maps; Masks; Mediating; Medical; Metabolism; Molecular; Mutation; Obstruction; Pathogenesis; Pathway interactions; Perimenopause; Pregnancy loss; Premenopause; Process; Production; Progesterone Receptors; Proliferating; Receptor Activation; Recurrence; Reporting; Risk Factors; Role; Sampling; Small Interfering RNA; Smooth Muscle Myocytes; Testing; Time; Tissues; Translational Research; Tryptophan; Tryptophan Metabolism Pathway; Tryptophanase; Up-Regulation; Urine; Uterine Fibroids; Uterine hemorrhage; Uterus; Woman; Xenograft procedure; amino acid metabolism; antagonist; aryl hydrocarbon receptor ligand; cell growth; consumer product; epidemiology study; genome-wide analysis; histone modification; in vivo; inhibitor; knock-down; mouse genome; mouse model; mutant; myometrium; novel; novel therapeutic intervention; novel therapeutics; pharmacologic; phthalates; reproductive; reproductive system disorder; research and development; response; small hairpin RNA; steroid hormone; therapeutic target; transcriptome; transcriptome sequencing; tumor; tumor growth; tumorigenesis; uptake; ARNT gene; ATAC-seq; Affect; Age; Amino Acid Transporter; Anemia; Apoptosis; Aryl Hydrocarbon Receptor; Bioinformatics; Biological Assay; Cell Proliferation; Cell Survival; Cells; ChIP-seq; Chemical Exposure; Chromatin; Development; Diet; Diethylhexyl Phthalate; Disease; Endocrine Disruptors; Environmental Pollutants; Environmental Risk Factor; Enzymes; Epidemiology; Exposure to; FDA approved; Fibroid Tumor; Future; Gene Expression; Gene Expression Profile; Gene Mutation; Genes; Genetic; Goals; Growth; Health; Human; Hysterectomy; In Vitro; Inhibition of Apoptosis; Kynurenine; Leiomyoma; Ligands; Link; Malignant Neoplasms; Maps; Masks; Mediating; Medical; Metabolism; Molecular; Mutation; Obstruction; Pathogenesis; Pathway interactions; Perimenopause; Pregnancy loss; Premenopause; Process; Production; Progesterone Receptors; Proliferating; Receptor Activation; Recurrence; Reporting; Risk Factors; Role; Sampling; Small Interfering RNA; Smooth Muscle Myocytes; Testing; Time; Tissues; Translational Research; Tryptophan; Tryptophan Metabolism Pathway; Tryptophanase; Up-Regulation; Urine; Uterine Fibroids; Uterine hemorrhage; Uterus; Woman; Xenograft procedure; amino acid metabolism; antagonist; aryl hydrocarbon receptor ligand; cell growth; consumer product; epidemiology study; genome-wide analysis; histone modification; in vivo; inhibitor; knock-down; mouse genome; mouse model; mutant; myometrium; novel; novel therapeutic intervention; novel therapeutics; pharmacologic; phthalates; reproductive; reproductive system disorder; research and development; response; small hairpin RNA; steroid hormone; therapeutic target; transcriptome; transcriptome sequencing; tumor; tumor growth; tumorigenesis; uptake

Uterine leiomyomas (LM, fibroids) disrupt uterine function and cause recurrent pregnancy loss, excessive uterine bleeding, and anemia in 15-30% of reproductive-age women. No long-term medical treatment is available. Understanding how a LM develops is essential for identifying new non-surgical treatments. MED12 mutations (mut-MED12) occur in 70% of all LM and drive LM growth in a steroid hormone-dependent manner. Pre- and peri-menopausal women are widely exposed to endocrine disrupting chemicals (EDCs), e.g., phthalates, which are found in many consumer products and associated with a number of reproductive diseases. How a mut- MED12 influences LM growth and whether this process is enhanced by exposure to EDCs remain unknown. We recently found that tryptophan (Trp) 2,3-dioxygenase (TDO2) gene expression is strikingly upregulated in mut- but not wild-type (wt)-MED12 LM. TDO2 catalyzes a critical step in Trp breakdown to kynurenine (Kyn), an endogenous ligand for the aryl hydrocarbon receptor (AHR) whose activation stimulates the expression of pro- growth genes to promote cell survival and proliferation in various tissues. Kyn levels are markedly higher in LM vs myometrium, particularly in mut-MED12 LM. Trp and Kyn treatments of LM cells activated AHR and increased cell survival; blocking the Trp-Kyn-AHR pathway by siRNA knockdown or inhibitors of TDO2 or AHR abolished these effects, with mutant LM cells showing higher sensitivity to the treatments. Epidemiological studies have shown positive associations between LM and exposure to di(2-ethylhexyl) phthalate (DEHP). In vitro, mono(2- ethyl-5-hydroxyhexyl) phthalate (MEHHP), a major metabolite of DEHP, stimulated the expression of Trp transporters (LAT1 and LAT2), increased Trp uptake and Kyn production, activated AHR, and promoted LM cell survival. In addition, progesterone receptor is crucial for the expression of AHR and its nuclear translocator ARNT. Thus, the Trp-Kyn-AHR pathway appears to be a hub at which mut-MED12, steroid hormone action, and EDC effects converge enabling LM growth. Our overall hypothesis is that increased Trp uptake and metabolism, Kyn production, and AHR pathway activation, as a result of mut-MED12 or high exposure to the environmental pollutant DEHP, promote cell survival and proliferation and lead to LM growth. Using a xenograft mouse model and genome-wide studies, we will test our hypothesis in the following Aims: (1) Define the functional role of the Trp-Kyn-AHR pathway in LM tumorigenesis. Hypothesis: elevated expression of the TDO2 enzyme in mut- MED12 LM causes Kyn overproduction that activates AHR and promotes proliferation and survival of smooth muscle cells and tumor growth. (2) Determine whether DEHP stimulates LM growth via activation of the Trp- Kyn-AHR pathway. Hypothesis: exposure to DEHP and its metabolite MEHHP upregulates the expression of Trp transporters to increase its uptake, resulting in increased Kyn production and AHR activation leading to LM cell survival and tumor growth. The study will link, for the first time, abnormal amino acid metabolism and LM growth, opening a new avenue for translational research and the development of novel therapeutics for LM.

子宫平滑肌瘤（LM，肌瘤）破坏子宫功能并导致复发性妊娠丧失，子宫过多 出血和贫血在15-30％的生殖年龄妇女中。没有长期的医疗。 了解LM如何发展对于识别新的非手术治疗至关重要。 Med12突变 （mut-med12）以类固醇激素依赖性方式出现在所有LM的70％中，并驱动LM生长。前和 绝经期妇女被广泛暴露于内分泌破坏化学物质（EDC），例如邻苯二甲酸盐，其中 在许多消费产品中发现，并与多种生殖疾病有关。如何mut- MED12影响LM增长，并且通过接触EDC来增强该过程仍然未知。我们 最近发现，色氨酸（TRP）2,3-二氧酶（TDO2）基因表达在mut-中显着上调 但不是野生型（WT）-Med12 lm。 TDO2催化了TRP分解至Kynurenine（Kyn）的关键步骤 芳基碳氢化合物受体（AHR）的内源配体，其活化刺激了促进的表达 生长基因以促进各种组织中的细胞存活和增殖。 LM的Kyn水平明显更高 与子宫肌，特别是在Mut-Med12 LM中。 LM细胞的TRP和KYN处理激活AHR并增加 细胞存活；通过siRNA敲低或TDO2或AHR的抑制剂阻止TRP-KYN-AHR途径 这些影响，突变的LM细胞对治疗的敏感性更高。流行病学研究已有 显示LM与暴露于DI（2-乙基己基）邻苯二甲酸酯（DEHP）之间的正相关。在体外，单声道（2- 乙二醇乙酸乙酯（MEHHP），一种主要的DEHP代谢物，刺激了TRP的表达 转运蛋白（LAT1和LAT2），提高TRP吸收和Kyn产生，激活AHR并促进LM细胞 生存。此外，孕酮受体对于AHR及其核转运剂的表达至关重要 arnt。因此，TRP-KYN-AHR途径似乎是一个枢纽，在该枢纽中，Mut-Med12，类固醇激素作用和 EDC效应收敛于LM生长。我们的总体假设是，TRP的摄取和代谢增加， 由于MUT-MED12或高暴露于环境，Kyn生产和AHR途径激活 污染物DEHP，促进细胞存活和增殖，并导致LM生长。使用异种移植小鼠模型 和全基因组研究，我们将在以下目的中检验我们的假设：（1）定义该假设 LM肿瘤发生中的TRP-KYN-AHR途径。假设：TDO2酶在mut-中的表达升高 MED12 LM导致Kyn生产过量激活AHR并促进平滑的增殖和存活 肌肉细胞和肿瘤生长。 （2）确定DEHP是否通过激活TRP刺激LM生长 Kyn-Ahr途径。假设：暴露于DEHP及其代谢物MEHP上调TRP的表达 转运蛋白增加其摄取，从而增加Kyn产生和AHR激活，从而导致LM细胞 生存和肿瘤生长。该研究将首次联系异常的氨基酸代谢和LM生长， 为LM开辟了新的转化研究和新型治疗剂的途径。