PT  - JOURNAL ARTICLE
AU  - Rebecca Caeser
AU  - Miriam Di Re
AU  - Joanna A Krupka
AU  - Jie Gao
AU  - Maribel Lara-Chica
AU  - João M.L Dias
AU  - Susanna L Cooke
AU  - Rachel Fenner
AU  - Zelvera Usheva
AU  - Hendrik Runge
AU  - Philip A Beer
AU  - Hesham Eldaly
AU  - Hyo-Kyung Pak
AU  - Chan-Sik Park
AU  - George Vassiliou
AU  - Brian J.P Huntly
AU  - Annalisa Mupo
AU  - Rachael JM Bashford-Rogers
AU  - Daniel J Hodson
TI  - Genetic modification of primary human B cells generates translationally-relevant models of high-grade lymphoma
AID  - 10.1101/618835
DP  - 2019 Jan 01
TA  - bioRxiv
PG  - 618835
4099  - http://biorxiv.org/content/early/2019/04/26/618835.short
4100  - http://biorxiv.org/content/early/2019/04/26/618835.full
AB  - 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 utilize 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 center 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 tumors 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.