免疫细胞迁移是免疫细胞能够发现入侵病原体、控制组织损伤和消除原发性肿瘤的关键特征之一。嵌合抗原受体（CAR）T 细胞疗法是对抗各种癌症的一种新策略。它在治疗血液肿瘤方面取得了成功，但在实体瘤方面仍面临许多挑战。在这项工作中，我们评估了致密胶原基水凝胶中 T 细胞和 CAR-T 细胞的三维 (3D) 迁移能力。量化三维 (3D) 细胞迁移需要可能不易获得的显微镜技术。因此，我们引入了一种简单的数学模型，旨在从二维 (2D) 细胞轨迹推断细胞的 3D 轨迹。我们开发了一种基于 3D 代理的模型 (ABM)，通过逆变换模拟迁移方向的时间变化抽样方法。然后，我们提出了一种优化程序，可以随着时间的推移准确定向细胞迁移，以从 2D 实验细胞轨迹重现细胞迁移。利用该模型，我们模拟了微流体装置中在具有不同浓度 I 型胶原的水凝胶下进行的 T 细胞和 CAR-T 细胞的细胞迁移测定，并使用光片显微镜验证了我们的 3D 细胞迁移预测。我们的研究结果表明 CAR-T 细胞与 T 细胞相比，迁移对胶原蛋白浓度的增加更敏感，从而导致其侵袭性更显着降低。此外，我们的计算模型揭示了 T 细胞和 CAR-T 细胞之间 3D 运动模式的显着差异。 T 细胞表现出 3D 迁移行为，而 CAR-T 细胞主要在 XY 平面内移动，在 Z 方向上移动有限。然而，在引入 CXCL12 化学梯度后，CAR-T 细胞呈现出与 T 细胞非常相似的迁移模式。该框架表明 3D 轨迹的 2D 投影可能无法准确代表真实的迁移模式。此外，它还提供了一种从 2D 实验数据估计 3D 迁移模式的工具，可以通过自动量化算法轻松获得。这种方法有助于减少实验室对复杂且昂贵的显微镜设备的需求，以及生成和分析 3D 实验数据所涉及的计算负担。版权所有 © 2024 作者。由 Elsevier B.V. 出版。保留所有权利。

Immune cell migration is one of the key features that enable immune cells to find invading pathogens, control tissue damage, and eliminate primary developing tumors. Chimeric antigen receptor (CAR) T-cell therapy is a novel strategy in the battle against various cancers. It has been successful in treating hematological tumors, yet it still faces many challenges in the case of solid tumors. In this work, we evaluate the three-dimensional (3D) migration capacity of T and CAR-T cells within dense collagen-based hydrogels. Quantifying three-dimensional (3D) cell migration requires microscopy techniques that may not be readily accessible. Thus, we introduce a straightforward mathematical model designed to infer 3D trajectories of cells from two-dimensional (2D) cell trajectories.We develop a 3D agent-based model (ABM) that simulates the temporal changes in the direction of migration with an inverse transform sampling method. Then, we propose an optimization procedure to accurately orient cell migration over time to reproduce cell migration from 2D experimental cell trajectories. With this model, we simulate cell migration assays of T and CAR-T cells in microfluidic devices conducted under hydrogels with different concentrations of type I collagen and validate our 3D cell migration predictions with light-sheet microscopy.Our findings indicate that CAR-T cell migration is more sensitive to collagen concentration increases than T cells, resulting in a more pronounced reduction in their invasiveness. Moreover, our computational model reveals significant differences in 3D movement patterns between T and CAR-T cells. T cells exhibit migratory behavior in 3D whereas that CAR-T cells predominantly move within the XY plane, with limited movement in the Z direction. However, upon the introduction of a CXCL12 chemical gradient, CAR-T cells present migration patterns that closely resemble those of T cells.This framework demonstrates that 2D projections of 3D trajectories may not accurately represent real migration patterns. Moreover, it offers a tool to estimate 3D migration patterns from 2D experimental data, which can be easily obtained with automatic quantification algorithms. This approach helps reduce the need for sophisticated and expensive microscopy equipment required in laboratories, as well as the computational burden involved in producing and analyzing 3D experimental data.Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.