PiggyBac transposon tools for recessive screening identify B-cell lymphoma drivers in mice

Abstract

B-cell lymphoma (BCL) is the most common hematologic malignancy. While sequencing studies gave insights into BCL genetics, identification of non-mutated cancer genes remains challenging. Here, we describe PiggyBac transposon tools and mouse models for recessive screening and show their application to study clonal B-cell lymphomagenesis. In a genome-wide screen, we discover BCL genes related to diverse molecular processes, including signaling, transcriptional regulation, chromatin regulation, or RNA metabolism. Cross-species analyses show the efficiency of the screen to pinpoint human cancer drivers altered by non-genetic mechanisms, including clinically relevant genes dysregulated epigenetically, transcriptionally, or post-transcriptionally in human BCL. We also describe a CRISPR/Cas9-based in vivo platform for BCL functional genomics, and validate discovered genes, such as Rfx7 , a transcription factor, and Phip , a chromatin regulator, which suppress lymphomagenesis in mice. Our study gives comprehensive insights into the molecular landscapes of BCL and underlines the power of genome-scale screening to inform biology.

H3K9me3-mediated epigenetic regulation of senescence in mice predicts outcome of lymphoma patients

“…f Circular workflow of functional interrogations in mouse models of cancer harboring defined genetic lesions, investigation of genetically unmanipulated mouse tumors in a clinical trial-like setting in vivo, and cross-species application of the genetic determinants of novel biological functions and intervention-evoked dynamic state switches learned therein in corresponding human cancer patient cohorts. exploration of functional lymphoma features in Eµ-myc mice and the subsequent validation of these findings in human DLBCL 23,27,28,40,[54][55][56] , we certainly acknowledge limitations of this transgenic model as a reflection of DLBCL pathogenesis, particularly in light of numerous mouse models developed to more faithfully recapitulate GCB-or ABC-subtype features of human DLBCL [57][58][59][60][61][62][63][64][65][66][67][68] . However, while those models elegantly provide examples for distinct routes into GCB-or ABC-skewed diffuse large B-cell lymphomagenesis, they are, in turn, selectively composed of complexity-reduced genetics out of the overwhelming heterogeneity human DLBCL exhibit as a cardinal property.…”

Structural basis of seamless excision and specific targeting by piggyBac transposase

“…pB has proved to be extremely versatile and the lack of a DNA footprint left behind after its transposition is a unique and valuable property [13][14][15][16] . It is used in non-viral vectors for transgenesis 17,18 , gene therapy 7,19 , insertional mutagenesis 20 , and genetic screens [21][22][23] . It has also found application in novel therapeutic strategies including CAR T-cell engineering [24][25][26] , CRISPR/Cas-mediated gene therapy [27][28][29] , and human induced pluripotent stem cells (iPSC) engineering [30][31][32] .…”

Genetically Engineered Mouse Models of Liver Tumorigenesis Reveal a Wide Histological Spectrum of Neoplastic and Non-Neoplastic Liver Lesions

“…All samples were obtained from the Comparative Experimental Pathology (CEP) at the Institute of Pathology, Technical University Munich (TUM), one of the largest core facilities in Europe for comparative pathology. An adequate level of genetic modification/deletion of each mouse and model has been assured by the collaboration partners [31,32]. Animals were initially provided to our collaboration partners by the Welcome Trust Sanger Institute, Genome Campus, Hinxton, Cambridge, CB10 1SA, UK.…”