Genetic modification of primary human B cells generates translationally-relevant models of high-grade lymphoma

Rebecca Caeser, Miriam Di Re, Joanna A Krupka, Jie Gao, Maribel Lara-Chica, João M.L Dias, Susanna L Cooke, Rachel Fenner, Zelvera Usheva, Hendrik Runge, Philip A Beer, Hesham Eldaly, Chan-Sik Park, George Vassiliou, Brian J.P Huntly, Annalisa Mupo, Rachael JM Bashford-Rogers & Daniel J Hodson

May 06, 2019

Received Date: 15th April 19

Sequencing studies of Diffuse Large B Cell Lymphoma (DLBCL) have identified hundreds of recurrently altered genes. However, it remains largely unknown whether and how these mutations may contribute to lymphomagenesis, either individually or in combination. Existing strategies to address this problem predominantly utilise cell lines, which are limited by their initial characteristics and subsequent adaptions to prolonged in vitro culture. Here, we describe a novel co-culture system that enables the ex vivo expansion and viral transduction of primary human germinal centre B cells. The incorporation of CRISPR/Cas9 technology enables high-throughput functional interrogation of genes recurrently mutated in DLBCL. Using a backbone of BCL2 with either BCL6 or MYC we have identified co-operating oncogenes that promote growth and survival, or even full transformation into synthetically engineered models of DLBCL. The resulting tumours can be expanded and sequentially transplanted in vivo, providing a scalable platform to test putative cancer genes and for the creation of mutation-directed, bespoke lymphoma models.

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This is an abstract of a preprint hosted on an independent third party site. It has not been peer reviewed but is currently under consideration at Nature Communications.

Nature Communications

Nature Research, Springer Nature