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; 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 衰老与自适应免疫反应的总体下降有关，大部分研究重点 在围绕功能的不足，而且还产生了幼稚的B和T淋巴细胞的缺陷。在 对比，对于衰老如何改变浆细胞的功能并不了解。浆细胞代表 成熟B细胞的末端分化步骤，并由两个短寿命的增殖性质质体（PBS）组成 以及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因子 通过强大的抗体生产（ABS）以及长寿的潜力。在人类中，这个 寿命可以跨越数十年。从历史上看，这些细胞被认为只不过是工厂。 但是，最近的工作表明了浆细胞调节多种生理的能力 过程。在衰老的背景下，PC已被证明可以进化促炎的表型并成为关键 通常在老年人中观察到的骨髓（BM）髓鞘增加的驱动因素。这部分是通过 造血细胞和髓样祖细胞数量的PC依赖性膨胀 与老年人口中髓样白血病的发病率升高有关。最近，我们有 确定了与年轻胸腺中PB/PC种群有关的性别差异，其中3 与雄性相比，几个月大的雌性小鼠的这些细胞数量明显更高。有趣的是 足够，这些您的PBS/PC表达了您的T细胞选择所需的关键蛋白质及其数量 与整体你的细胞性显着相关。与BM相似，我们观察到您的PB/PC的变化 种群在衰老的背景下。尤其是，雄性小鼠会累积您的PBS/PC，而数字 在女性中保持恒定。这与BM和Spleen（SPL）形成鲜明对比，PBS/PC随着年龄的增长而增加 性别。因此，了解PBS/PCS年龄如何使我们能够更好地评估其在适应性免疫中的作用 老年人的反应以及其不依赖AB的功能不断发展的曲目。最终，这是5月 导致开发改变PB/PC行为并促进老年人的HealthSpan的疗法 个人。目前，很难识别出长寿命或老化的浆细胞。而各种细胞 已经证明表面标记可以识别浆细胞，这些决定因素不一定与 寿命但相当成熟。另外，细胞标记技术依赖于上游B细胞的有效标记 并在随后生成的PBS/PC中标记保留率。在某些情况下，检测这些标签需要 细胞通透性因此消除了进一步研究生活，老化的PBS/PC的能力。 AIM 1专注于 使用荧光记者开发可诱导的系统以不可磨灭的标记或时间戳浆细胞 因此，促进了评估种群动态和潜在分离各个年龄的细胞的能力。 AIM 2意志 证明该记者系统的可行性是专门确定差异人群周转和/或 女性与雄性浆细胞的维持。