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.
Highlights
Diffuse large cell B cell lymphoma (DLBCL) is the most common type of non-Hodgkin lymphoma (NHL); accounting for approximately 30%–40% of cases [1]
We demonstrated that normal and malignant human B cells interact with adipocyte derived stem cells (ADSC)-derived human lymphoid-like fibroblasts, in the presence or absence of human monocyte-derived macrophages (MDM), in both 2D and 3D spheroid co-cultures
Positive for alpha-smooth muscle actin (a-SMA), podoplanin and VCAM-1, and CD20+ DLBCL cells were observed in close proximity to CD68+ macrophages (Figure 1A)
Summary
Diffuse large cell B cell lymphoma (DLBCL) is the most common type of non-Hodgkin lymphoma (NHL); accounting for approximately 30%–40% of cases [1]. It is subcategorized into germinal-center B cell-like (GCB) and activated B cell-like (ABC) entities, based on cell-of-origin and gene expression profiling (GEP) [2], with ABC DLBCL associated with substantially worse outcomes with current treatments [3]. Despite growing knowledge of the under-pinning mutations and oncogenic drivers in DLBCL, new targeted therapeutics have so far failed to deliver improved clinical outcomes, highlighting a need for better models to study DLBCL
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