Glucose and Amino Acid Catabolism in Plasma Cell Biology

摘要： 代谢途径的使用是定制的，以满足特定细胞类型的特定功能和需求。 特别感兴趣的是代谢如何支持浆细胞的存活和抗体分泌， 负责体液免疫的主要细胞类型。这些细胞的寿命决定了 感染或疫苗后抗体介导的免疫-这是一个特别相关的主题， 流行病在上一个资助期间，我们的工作表明， 在控制浆细胞寿命的转录途径。相反，代谢途径在功能上区分 不同寿命的浆细胞利用新发明的遗传学工具，我们将快速定义和剖析 体内必需的浆细胞代谢途径。我们将使用新产生的等离子体细胞Cre-drivers， 慢病毒骨髓嵌合体和CRISPR/Cas9方法来功能性地定义线粒体动力学， 必需的代谢途径和促进浆细胞寿命的囊泡成熟途径， 抗体分泌这些方法将与灵敏的成像质谱法相结合， 同位素示踪实验，以提供机制的见解。具体来说，我们的实验将定义 线粒体分裂和融合在浆细胞能量代谢、抗体产生和 生存使用病毒感染和免疫的生理实验将确定促进免疫的关键因素。 随着这些细胞的寿命逐渐延长，浆细胞代谢重新编程。第二，基于 根据完成的全基因组CRISPR/Cas9筛选的结果，我们将继续研究V型ATP酶的重要性。 在体内氨基酸摄取、浆细胞寿命和抗体分泌方面。

Abstract: The usage of metabolic pathways is tailored to meet the specific functions and demands of a given cell type. Of particular interest is how metabolism supports the survival and antibody secretion of plasma cells, the primary cell type that is responsible for humoral immunity. The lifespan of these cells dictates the duration of antibody-mediated immunity after infections or vaccines—a particularly relevant topic in the midst of this pandemic. During the previous funding period, our work suggested a surprisingly minimal role for transcriptional pathways in controlling plasma cell lifespan. Instead, metabolic pathways functionally distinguish plasma cells of differing lifespans. Using newly created genetic tools, we will rapidly define and dissect essential plasma cell metabolic pathways in vivo. We will use newly generated plasma cell Cre-drivers, lentiviral bone marrow chimeras, and CRISPR/Cas9 approaches to functionally define mitochondrial dynamics, essential metabolic pathways, and vesicular maturation pathways that promote plasma cell lifespan and antibody secretion. These approaches will be coupled with sensitive imaging mass spectrometry and stable isotope-tracing experiments to provide mechanistic insight. Specifically, our experiments will define the importance of mitochondrial fission and fusion in plasma cell energy metabolism, antibody production, and survival. Physiological experiments using viral infections and immunizations will define key factors that promote plasma cell metabolic re-programming as these cells become progressively longer lived. Second, based upon results of a completed genome-wide CRISPR/Cas9 screen, we will pursue the importance of V-type ATPases in amino acid uptake, plasma cell lifespan, and antibody secretion in vivo.