Developing a 3D B Cell Lymphoma Culture System to Model Antibody Therapy

Abstract

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 in vitro 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 ex vivo. 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 in vitro. 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 ex-vivo 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.

Development and Characterization of 3D Hybrid Spheroids for the Investigation of the Crosstalk Between B-Cell Non-Hodgkin Lymphomas and Mesenchymal Stromal Cells

“… 35 Recently, Foxall et al described a collagen scaffolds-based spheroid co-culture system comprising DLBCL cells, cancer-associated fibroblasts (CAF), and tumor-associated macrophages (TAM). 36 Notably, both studies demonstrated that DLBCL cells interact with their constituent components, resulting in their improved viability compared to 2D mono-cultures. Finally, An et al successfully developed a canine 3D hybrid model by co-culturing the lymphoma cells, and lymph node-derived primary stromal cells.…”

A novel 3D culture model recapitulates primary FL B-cell features and promotes their survival

“… 20 However, modeling the interactions of the TME with primary lymphoma B cells in 3D cocultures remains a challenge. In particular, 3D cocultures have been successful only with primary DLBCL cells 21 but not with primary FL cells.…”

Tumor Microenvironment and Hydrogel-Based 3D Cancer Models for In Vitro Testing Immunotherapies

“…Therefore, the importance of developing more reliable in vitro systems taking into account the complex reciprocal interactions occurring in vivo between malignant and non-malignant cells is evident. Recently, some platforms prepared the groundwork for the investigation of novel agents (e.g., immunotherapeutic antibodies) aimed at targeting the key cellular components of the TME (e.g., CAFs or TAMs) in a clinically relevant context [ 173 ].…”