Role of the Macrophage in Developmentally Programmed NAFLD

具体目标1。检验妊娠期和哺乳期母体WD通过不同的代谢和/或转录机制调节后代骨髓细胞功能的假设，这些机制在后代寿命期间加速NAFLD风险。 1a）我们将测试妊娠或哺乳期间的WD是否诱导胎肝单核细胞（Mo）/Mφ和造血干细胞和祖细胞（HSPC）或成年后代肝Mφ的数量、活化状态或功能反应的变化，以及这是否可以通过与食物（CH）喂养的母鼠交叉培养来挽救。肝脏组织学和代谢表型将用于评价16周龄时NAFLD的严重程度和代谢适应。 1b）我们将分析后代肝脏Mφ和BMDM中的代谢物和糖酵解/TCA循环呼吸活性，以测试免疫代谢的重编程。 1c）我们将确定母体WD暴露对分离的肝脏Mo/Mφ和BMDM在生命早期和成年期的转录和表观遗传适应的影响。 具体目标2。检验AHR信号传导减少提供了一种机制联系的假设 母体WD生态失调、Mφ代谢重编程和NAFLD风险之间的关系。 2a）我们将用基于吲哚的AHR激动剂补充WD喂养的母鼠，以确定恢复AHR信号传导是否保护后代免受母体WD诱导的骨髓功能障碍。 2b）我们将在具有AHR的髓样缺失（KO）的WD暴露后代中描述肝脏Mo/Mφ中的转录和表观遗传适应，并评估Mφ功能变化和NAFLD进展。 具体目标3。测试膳食PQQ促进肠道微生物代谢的假设， 保护暴露于母体WD后代中Mφ功能的吲哚。 3a）我们将测试在妊娠或哺乳期间转移到CH喂养的GF母鼠的暴露于PQQ的微生物是否增加后代中吲哚的细菌产生。

Specific Aim 1. Test the hypothesis that maternal WD during gestation and lactation regulates offspring myeloid cell function through distinct metabolic and/or transcriptional mechanisms that accelerate NAFLD risk across the offspring lifespan. 1a) We will test if WD during gestation or lactation induces changes in number, activation state, or functional responses of fetal liver monocytes (Mo)/Mφ and hematopoietic stem and progenitor cells (HSPCs) or adult offspring liver Mφ and if this can be rescued by cross-fostering to a chow (CH)- fed dam. Liver histology and metabolic phenotyping will be used to evaluate severity of NAFLD and metabolic adaptations at 16 wks of age. 1b) We will profile metabolites and glycolytic/TCA cycle respiratory activity in offspring liver Mφ and BMDM to test for re-programming of immunometabolism. 1c) We will determine the impact of maternal WD exposure on transcriptional and epigenetic adaptations in isolated liver Mo/Mφ and BMDMs in early life and during adulthood. Specific Aim 2. Test the hypothesis that decreased AHR signaling provides a mechanistic link between maternal WD dysbiosis, Mφ metabolic reprogramming, and NAFLD risk. 2a) We will supplement WD-fed dams with an indole-based AHR agonist to determine if restoring AHR signaling protects against maternal WD-induced myeloid dysfunction in offspring. 2b) We will profile transcriptional and epigenetic adaptations in liver Mo/Mφ in WD-exposed offspring with myeloid deletion (KO) of AHR and assess for Mφ functional changes and NAFLD progression. Specific Aim 3. Test the hypothesis that dietary PQQ promotes gut microbial metabolism of indoles that preserve Mφ function in offspring exposed to maternal WD. 3a) We will test if PQQ-exposed microbes transferred to CH-fed GF dams during gestation or lactation increases bacterial production of indoles in offspring.