Development of an Inducible Conditional Gene Deletion Mouse Model to Study Plasma Cell Development and Longevity

开发可诱导条件基因缺失小鼠模型来研究浆细胞发育和寿命

• 批准号: 10452247
• 负责人: Peter Dion Pioli
• 金额: $ 5.4万
• 依托单位: UNIVERSITY OF SASKATCHEWAN
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-15 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10452247
• 关键词: Acute Myelocytic Leukemia; Adopted; Age; Aging; Animals; Antibodies; Antibody Formation; Antibody Repertoire; B cell differentiation; B-Lymphocytes; Behavior; Biological Models; Biological Process; Blood Cells; Bone Marrow; Cell Count; Cell surface; Cells; Cellular Assay; Cellularity; DNA Binding; DNA Damage; Deoxyuridine; Derivation procedure; Detection; Development; Disease; Dose; Elderly; Enterobacteria phage P1 Cre recombinase; Event; Female; Flow Cytometry; Fluorescence; Gene Deletion; Generations; Genes; Genetic Recombination; Glucose; Health; Hematopoietic; Hematopoietic stem cells; Human; Humoral Immunities; Immune response; Incidence; Individual; Inflammation; Inflammatory; Label; Lead; Link; Longevity; Maintenance; Mature B-Lymphocyte; Methods; Modeling; Mouse Protein; Mus; Myelogenous; Myeloid Leukemia; Myelopoiesis; Phenotype; Physiologic pulse; Physiological Processes; Plasma Cells; Plasmablast; Play; Population; Population Dynamics; Process; Production; Proteins; Protocols documentation; Reporter; Research; Role; Sex Bias; Sex Differences; Site; Spleen; System; T-Lymphocyte; Tamoxifen; Techniques; Technology; Terminator Codon; Testing; Thymus Gland; Time; Work; Youth; adaptive immune response; age related; aged; aging population; base; cell age; cell behavior; experimental study; glucose analog; glucose uptake; healthspan; male; mouse Cre recombinase; mouse model; plasma cell development; progenitor; prospective; response; sex; sex disparity; therapy development; transcription factor; vaccine response

Project Summary/Abstract Aging is associated with an overall decline in the adaptive immune response with a bulk of the research focusing on the deficiencies surrounding not only the function, but also the generation, of naive B and T lymphocytes. In contrast, not much is known about how aging alters the functionality of plasmacytes. Plasmacytes represent the terminal differentiation step of mature B cells and consist of both short-lived, proliferative plasmablasts (PBs) and more mature, non-proliferative plasma cells (PCs). These cells are key facilitators of humoral immunity through their robust production of antibodies (Abs) as well as their potential to be long-lived. In humans, this longevity can span decades. Historically, these cells have been considered nothing more than Ab factories; however, recent work has demonstrated the ability of plasmacytes to regulate a variety of physiological processes. In the context of aging, PCs have been shown to evolve a pro-inflammatory phenotype and be key drivers of the increased bone marrow (BM) myelopoiesis normally observed in the elderly. This is in part through the PC-dependent expansion in the numbers of hematopoietic stem cells and myeloid progenitors which has been linked to the elevated incidence of myeloid leukemias in the aged population. More recently, we have identified a sex disparity that exists in regard to PB/PC populations within the young thymus (THY) in which 3 months old female mice have significantly higher numbers of these cells compared to males. Interestingly enough, these THY PBs/PCs express key proteins required for THY T cell selection and their numbers significantly correlate with overall THY cellularity. Similar to BM, we have observed changes in THY PB/PC populations in the context of aging. In particular, male mice accumulate THY PBs/PCs with age while numbers remain constant in females. This is in contrast to BM and spleen (SPL) where PBs/PCs increase with age in both sexes. Thus, understanding how PBs/PCs age will allow us to better evaluate their role(s) in adaptive immune responses in the elderly as well as their evolving repertoire of Ab-independent functions. Ultimately, this may lead to the development of therapies that alter PB/PC behavior and promote increased healthspan in aged individuals. It is currently difficult to identify bona fide long-lived, or aged, plasmacytes. While a variety of cell surface markers have been shown to identify plasmacytes, these determinants do not necessarily correlate with longevity but rather maturity. Alternatively, cell labeling techniques rely on efficient labeling of upstream B cells and label retention in the subsequently generated PBs/PCs. In some instances, detection of these labels requires cell permeabilization thus eliminating the ability to further study living, aged PBs/PCs. Aim 1 focuses on the development of an inducible system to indelibly mark, or timestamp, plasmacytes using a fluorescence reporter thus facilitating the ability to assess population dynamics and potentially isolate cells of various ages. Aim 2 will demonstrate the feasibility of this reporter system to specifically determine differential population turnover and/or maintenance in female versus male THY plasmacytes.

项目摘要/摘要 衰老与适应性免疫反应的全面下降有关，大部分研究集中在 关于幼稚B和T淋巴细胞的功能和生成方面的缺陷。在……里面 相比之下，人们对衰老如何改变浆细胞的功能知之甚少。浆细胞代表 成熟B细胞的终末分化步骤，由短暂的、增殖的浆母细胞(PBS)组成 和更成熟的、非增殖性浆细胞(PC)。这些细胞是体液免疫的关键促进者。 通过它们产生强大的抗体(Abs)以及它们长期存活的潜力。在人类身上，这是 寿命可以跨越几十年。从历史上看，这些细胞被认为只不过是抗体工厂； 然而，最近的研究表明，浆细胞能够调节多种生理功能。 流程。在衰老的背景下，PC已被证明进化为促炎表型，并成为关键 通常在老年人中可以观察到骨髓(BM)造血增加的驱动因素。这在一定程度上是通过 PC依赖的造血干细胞和髓系祖细胞数量的扩增 与老年人群中髓系白血病发病率的升高有关。最近，我们已经 确认了幼年胸腺(THY)内存在的PB/PC种群的性别差异，其中3 与雄性相比，几个月大的雌性小鼠的这些细胞数量要多得多。有趣的是 足够了，这些Thy PBS/PC表达了Thy T细胞选择及其数量所需的关键蛋白质 与你全身的细胞密度有显著的相关性。与黑石相似，我们观察到您的PB/PC发生了变化 老龄化背景下的人口。特别是，雄性小鼠随着年龄的增长而积累你的PBS/PC 在雌性中保持不变。这与骨髓和脾(SPL)形成对比，两者的PBS/PC都随着年龄的增长而增加 性别。因此，了解PBS/PC是如何老化的将使我们能够更好地评估它们在适应性免疫中的作用(S 老年人的反应以及他们进化的抗体非依赖性功能谱。最终，这可能会 导致改变PB/PC行为和促进老年人健康寿命延长的治疗方法的发展 个人。目前很难识别真正的长寿或老化的浆细胞。而各种细胞 表面标记已经被证明可以识别浆细胞，这些决定因素不一定与 长寿而不是成熟。或者，细胞标记技术依赖于上游B细胞的有效标记 以及在随后产生的PBS/PC中的标签保持。在某些情况下，检测这些标签需要 细胞通透性，从而消除了进一步研究活的、老化的PBS/PC的能力。目标1侧重于 开发一种使用荧光报告器不可磨灭地标记或时间标记血浆细胞的诱导系统 从而促进了评估种群动态和潜在地分离不同年龄的细胞的能力。目标2将 论证该报告系统具体确定不同人口流动率和/或 女性对男性的维持你的浆细胞。