Foxall Russell, Narang Priyanka, Glaysher Bridget, Hub Elin, Teal Emma, Coles Mark C, Ashton-Key Margaret, Beers Stephen A, Cragg Mark S
Antibody and Vaccine Group, Centre for Cancer Immunology, School of Cancer Sciences, University of Southampton Faculty of Medicine, Southampton, United Kingdom.
Centre for Immunology and Infection, University of York, York, United Kingdom.
Front Immunol. 2021 Feb 8;11:605231. doi: 10.3389/fimmu.2020.605231. eCollection 2020.
Diffuse large cell B cell lymphoma (DLBCL) accounts for approximately 30%-40% of all non-Hodgkin lymphoma (NHL) cases. Current first line DLBCL treatment results in long-term remission in more than 60% of cases. However, those patients with primary refractory disease or early relapse exhibit poor prognosis, highlighting a requirement for alternative therapies. Our aim was to develop a novel model of DLBCL that facilitates testing of current and novel therapies by replicating key components of the tumor microenvironment (TME) in a three-dimensional (3D) culture system that would enable primary DLBCL cell survival and study . The TME is a complex ecosystem, comprising malignant and non-malignant cells, including cancer-associated fibroblasts (CAF) and tumor-associated macrophages (TAM) whose reciprocal crosstalk drives tumor initiation and growth while fostering an immunosuppressive milieu enabling its persistence. The requirement to recapitulate, at least to some degree, this complex, interactive network is exemplified by the rapid cell death of primary DLBCL cells removed from their TME and cultured alone . Building on previously described methodologies to generate lymphoid-like fibroblasts from adipocyte derived stem cells (ADSC), we confirmed lymphocytes, specifically B cells, interacted with this ADSC-derived stroma, in the presence or absence of monocyte-derived macrophages (MDM), in both two-dimensional (2D) cultures and a 3D collagen-based spheroid system. Furthermore, we demonstrated that DLBCL cells cultured in this system interact with its constituent components, resulting in their improved viability as compared to 2D monocultures. We then assessed the utility of this system as a platform to study therapeutics in the context of antibody-directed phagocytosis, using rituximab as a model immunotherapeutic antibody. Overall, we describe a novel 3D spheroid co-culture system comprising key components of the DLBCL TME with the potential to serve as a testbed for novel therapeutics, targeting key cellular constituents of the TME, such as CAF and/or TAM.
弥漫性大细胞B细胞淋巴瘤（DLBCL）约占所有非霍奇金淋巴瘤（NHL）病例的30%-40%。目前DLBCL的一线治疗使超过60%的病例实现长期缓解。然而，那些原发性难治性疾病或早期复发的患者预后较差，这凸显了对替代疗法的需求。我们的目标是开发一种新型的DLBCL模型，通过在三维（3D）培养系统中复制肿瘤微环境（TME）的关键成分，促进对当前和新型疗法的测试，该系统能够使原发性DLBCL细胞存活并进行研究。TME是一个复杂的生态系统，由恶性和非恶性细胞组成，包括癌症相关成纤维细胞（CAF）和肿瘤相关巨噬细胞（TAM），它们之间的相互作用驱动肿瘤的起始和生长，同时营造一个免疫抑制环境以使其持续存在。从TME中取出并单独培养的原发性DLBCL细胞迅速死亡，这体现了至少在一定程度上重现这种复杂的交互网络的必要性。基于先前描述的从脂肪来源干细胞（ADSC）生成类淋巴成纤维细胞的方法，我们证实淋巴细胞，特别是B细胞，在二维（2D）培养和基于3D胶原蛋白的球体系统中，无论有无单核细胞衍生巨噬细胞（MDM），都与这种ADSC衍生的基质相互作用。此外，我们证明在该系统中培养的DLBCL细胞与其组成成分相互作用，与2D单培养相比，其活力得到改善。然后，我们以利妥昔单抗作为模型免疫治疗抗体，评估了该系统作为研究抗体介导吞噬作用背景下治疗方法的平台的效用。总体而言，我们描述了一种新型的3D球体共培养系统，该系统包含DLBCL TME的关键成分，有可能作为新型疗法的试验平台，针对TME的关键细胞成分，如CAF和/或TAM。
