PO.TB04.02 · 肿瘤生物学
开发下一代用于临床前肿瘤学的定制化人类免疫系统小鼠
Developing the next generation of customized human immune system mice for preclinical oncology
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
人源化免疫系统（HIS）小鼠可移植人细胞系来源或患者来源的肿瘤，已成为肿瘤学中细胞疗法和生物制剂的临床前及IND申报开发的重要工具。然而，鉴于可人源化的小鼠品系和免疫移植方案的多样性，选择合适的HIS模型仍具有挑战性。我们首先比较了移植脐带血来源CD34⁺造血干细胞（HSCs）或外周血单个核细胞（PBMCs）的重度免疫缺陷小鼠的免疫谱、临床症状、体重和生存情况。PBMC移植小鼠因早期移植物抗宿主病（GvHD）而表现出较差的生存，并表现出具有部分耗竭表型（Lag-3⁺、TIM-3⁺）的人T细胞扩增。相比之下，CD34移植小鼠在30周内未出现健康恶化，并发育出包含T、B、NK和髓系细胞的完整人类免疫系统。这些结果表明CD34+ HSC移植的HIS小鼠是总体上更优的药物评估模型，因为它提供了更长的、无GvHD的治疗窗口和更完整的免疫系统。接下来，我们采用了一种通用的、无需辐照的、基于化学消融的CD34⁺ HSC移植方案，以比较多种重度免疫缺陷品系间的免疫人源化程度，包括基础模型（NOG、NCG、BNDG、BRG）和下一代品系（NOG-EXL，其过表达人GM-CSF和IL-3；以及FcResolv NOG，其缺乏小鼠Fcγ受体）。对于每个品系，我们测量了生存率、总体人源化率（人类免疫细胞占总免疫细胞的百分比）以及血液中人类免疫亚群的频率。最后，我们展示了当无法获得过表达这些细胞因子的转基因品系时，如何通过将一种或多种人细胞因子进行流体动力学基因递送至HIS小鼠来增强特定免疫群体。总之，我们的发现为选择具有最合适免疫重建谱的人源化免疫系统小鼠模型（基于候选药物的作用机制评估任何候选药物）提供了知识基础。
查看英文原文 English abstract
Humanized immune system (HIS) mice, which can be engrafted with human cell line-derived or patient-derived tumors, have become essential tools for preclinical and IND-enabling development of cell therapies and biologics in oncology. However, selecting the appropriate HIS model remains challenging given the diversity of humanizable mouse strains and immune-engraftment protocols. We first compared immune profiles, clinical symptoms, body weight, and survival in severely immunodeficient mice engrafted with either cord blood-derived CD34⁺ hematopoietic stem cells (HSCs) or peripheral blood mononuclear cells (PBMCs). PBMC-engrafted mice demonstrated poor survival associated with early graft-versus-host disease (GvHD) and exhibited amplification of human T cells with a partially exhausted phenotype (Lag-3⁺, TIM-3⁺). In contrast, CD34-engrafted mice showed no health deterioration over 30 weeks and developed a complete human immune system comprising T, B, NK, and myeloid cells.These results place the CD34+ HSC engrafted HIS mice as overall superior model for drug assessment as it allows a longer, GvHD-free treatment window and a more complete immune system. We next implemented a universal, irradiation-free, chemoablation-based CD34⁺ HSC-engraftment protocol to compare the extent of immune humanization across several severely immunodeficient strains, including foundational models (NOG, NCG, BNDG, BRG, and next-generation strains (NOG-EXL, which overexpresses human GM-CSF and IL-3; and FcResolv NOG, which lacks murine Fcgamma receptors). For each strain, we measured survival, overall humanization rate (percentage of human among total immune cells), and human immune-subset frequencies in blood. Finally, we demonstrate how hydrodynamic gene delivery of one or more human cytokines into HIS mice can boost certain immune populations when a transgenic strain overexpressing these cytokines is unavailable. Altogether, our findings provide a knowledge base for the selection of the humanized immune system mouse model with the most suitable immune reconstitution profile for assessing any drug candidate based on its mechanisms of action.
利益披露 Disclosure
A. Wetzel, None..
E. Bayon, None..
S. Tabruyn, None..
D. Georgess, None.