PO.CL05.13 · 临床研究
用于评估B细胞CAR-T疗法疗效和安全性的人源化SLE小鼠模型
Humanized SLE mouse model for evaluating B-cell CAR-T therapy efficacy and safety
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
系统性红斑狼疮（SLE）是一种复杂的自身免疫性疾病，以B细胞激活失调、自身抗体产生和全身性炎症为特征。疗法开发受限于物种特异性的免疫差异、传统小鼠对人类病理建模不佳以及临床预测价值低。同样，免疫相关不良事件（IRAE），如细胞因子释放综合征（CRS），仍是CAR-T等基于细胞的癌症免疫治疗的一大挑战。为解决这些问题，我们开发了一种人源化临床前SLE小鼠模型，该模型能够反映人类免疫应答，并作为评估靶向B细胞的CAR-T疗法及免疫毒性的平台。SLE表型由咪喹莫特（IMQ）诱导，这是一种Toll样受体7激动剂，可激活浆细胞样树突状细胞和B细胞。在CD34⁺人源化小鼠中，IMQ诱导了人源化的SLE样表型，使人类免疫细胞能够对TLR7刺激作出应答，从而紧密模拟人类疾病。CAR-T细胞通过偶联抗CD5抗体的脂质纳米颗粒（LNP）在体内递送，以靶向CD5⁺ T细胞。LNP内化递送编码CD19特异性CAR的DNA，转染的T细胞通过流式细胞术和数字微滴PCR进行表征。LNP药代动力学通过LC-MS/MS分析，组织分布通过CAR基因检测进行评估。通过针对细胞因子（IFN-gamma、TNF-alpha、IL-2、IL-4、IL-6、IL-10）和趋化因子（MCP-1、CCL5）的多重检测监测T细胞激活和CRS。通过抗PEG ELISA和抗CAR MSD检测评估抗药抗体应答。CAR-T疗效通过血清抗dsDNA抗体和尿脂质运载蛋白-2（SLE活动的关键生物标志物）进行评估。总之，这一整合了分析组套的人源化SLE模型提供了一个具有生理相关性的平台，用于评估治疗疗效和免疫毒性，加速自身免疫性疾病和癌症免疫治疗中人类靶向疗法的临床前开发。
查看英文原文 English abstract
Systemic lupus erythematosus (SLE) is a complex autoimmune disease marked by dysregulated B-cell activation, autoantibody production, and systemic inflammation. Therapy development is limited by species-specific immune differences, poor modeling of human pathology in conventional mice, and low clinical predictive value. Similarly, immune-related adverse events (IRAEs), such as cytokine release syndrome (CRS), remain a major challenge in cell-based cancer immunotherapies like CAR-T. To address these issues, we developed a humanized preclinical SLE mouse model that mirrors human immune responses and serves as a platform to evaluate B-cell-targeted CAR-T therapies and immune toxicity.The SLE phenotype was induced by imiquimod (IMQ), a Toll-like receptor 7 agonist that activates plasmacytoid dendritic cells and B cells. In CD34⁺ humanized mice, IMQ induced a humanized SLE-like phenotype, allowing human immune cells to respond to TLR7 stimulation and closely modeling human disease.CAR-T cells were delivered in vivo using lipid nanoparticles (LNPs) conjugated with anti-CD5 antibodies to target CD5⁺ T cells. LNP internalization delivers DNA encoding a CD19-specific CAR, and transfected T cells were characterized by flow cytometry and digital droplet PCR. LNP pharmacokinetics were analyzed via LC-MS/MS, and tissue distribution was assessed by CAR gene detection.T-cell activation and CRS were monitored via multiplex assays for cytokines (IFN-gamma, TNF-alpha, IL-2, IL-4, IL-6, IL-10) and chemokines (MCP-1, CCL5). Anti-drug antibody responses were evaluated by anti-PEG ELISA and anti-CAR MSD assays. CAR-T efficacy was assessed using serum anti-dsDNA antibodies and urinary lipocalin-2, key biomarkers of SLE activity.In summary, this humanized SLE model with integrated analytical panels provides a physiologically relevant platform for evaluating therapeutic efficacy and immune toxicity, accelerating preclinical development of human-targeted therapies for autoimmune disease and cancer immunotherapy.
利益披露 Disclosure
R. Zhang, None..
T. Ho, None..
B. Peng, None..
H. Qu, None..
Q. Jiang, None..
B. Wei, None..
Q. Lin, None.