RT Journal Article
SR Electronic
T1 Genetic modification of primary human B cells generates translationally-relevant models of high-grade lymphoma
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 618835
DO 10.1101/618835
A1 Rebecca Caeser
A1 Miriam Di Re
A1 Joanna A Krupka
A1 Jie Gao
A1 Maribel Lara-Chica
A1 João M.L Dias
A1 Susanna L Cooke
A1 Rachel Fenner
A1 Zelvera Usheva
A1 Hendrik Runge
A1 Philip A Beer
A1 Hesham Eldaly
A1 Hyo-Kyung Pak
A1 Chan-Sik Park
A1 George Vassiliou
A1 Brian J.P Huntly
A1 Annalisa Mupo
A1 Rachael JM Bashford-Rogers
A1 Daniel J Hodson
YR 2019
UL http://biorxiv.org/content/early/2019/04/26/618835.abstract
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.