Session: 621. Lymphomas: Translational – Molecular and Genetic: Poster II
Hematology Disease Topics & Pathways:
Lymphoid Leukemias, CLL, Lymphomas, Non-Hodgkin lymphoma, B Cell lymphoma, Diseases, Aggressive lymphoma, Lymphoid Malignancies
Methods: We created the triple transgenic iMYC-TCL1 murine model of temporally controlled MYC overexpression in B cells of the Eμ-TCL1 CLL model by crossing it to the R26StopFLMYC and CD19.cre-ERT2 murine models (R26StopFLMYChet. CD19.cre-ERT2het. Eμ-TCL1hemi). The R26StopFLMYC allows for the conditional overexpression of both MYC and an inactive human CD2 marker. Cre recombination was induced by administration of Tamoxifen (TAM, 110mg/kg) or vehicle (Veh, corn oil) via oral gavage (QD x 5 days). Mice were monitored monthly by flow cytometry for circulating CLL cells (cCLL; Cd19+Cd5+) and enrolled into disease load groups (<5% and >20% of cCLL) and treated with TAM or Veh. Limiting cell RNAseq (lcRNAseq) was performed on circulating malignant B cell populations (Cd19+Cd5+, Cd19+Cd5+hCD2+, and Cd19+hCD2+) for evaluation of BCR gene expression and global DEGs.
Results: Induction of MYC in pre-leukemic mice (<5% cCLL) (n=9 TAM, n=6 Veh) resulted in rapid disease expansion with 5/9 TAM vs 0/6 Veh mice achieving >25% circulating disease after 3.2 months (p<0.03). The MYC induced population had increased cell size (FSC) by flow cytometry. Since the clonal relationship of RT to CLL is a key prognostic factor, we assessed MYC induction's clonal dynamics by analyzing malignant B cell populations in TAM-treated mice using FACS and lcRNAseq (n=4). Clonal relationship between Cd19+Cd5+ and Cd19+Cd5+hCD2+ was established in all TAM treated mice with light chain CDR3 sequences of the dominant Cd19+Cd5+hCD2+ clone emerging from a pre-existing Cd19+Cd5+ clone. Furthermore, the Cd19+Cd5+hCD2+ population shows enrichment of E2F target genes (NES: 1.29, p < 0.05) and decreased interferon gamma response genes (NES: -1.56, p < 0.05) when compared to CLL-like B cells (Cd19+Cd5+) as has been previously shown in RT vs CLL tumors. Intestinal tumors composed of Cd19+Cd5+ and Cd19+Cd5-hCD2+ cells were seen in 4/9 TAM treated mice and was a phenotype seen only within this group.
Next, to mimic RT development with MYC hyperactivation post-CLL development we induced the iMYC-TCL1 model at >20% cCLL (n=14, 7 per treatment group). Induction led to a rapid expansion of Cd19+Cd5+hCD2+ cells infiltrating the spleen, lymph nodes, and marrow of mice, as early as 4 weeks post-TAM. Splenic B cells in TAM treated mice at ERC were predominantly Cd19+Cd5+hCD2+ (mean: 72% vs 5% Cd19+Cd5+, p<0.01). MYC induced, Cd19+Cd5+hCD2+, cells show increased Pdl1 expression relative to Cd19+Cd5+hCD2- cells. Significant median survival differences were observed (TAM: 80 vs Veh: 98 days post-induction, p<0.05). Histopathologic analyses are currently ongoing.
To assess the impact of MYC induction on the tumor microenvironment, we analyzed circulating T cell profiles in post-leukemic TAM and Veh treated mice. Veh treated mice had predominantly memory CD8+ T cells, while TAM treated mice showed reduced T cell abundance but increased CD8+ T cell diversity (memory, effector, naïve).
Conclusion: Induction prior to CLL development leads to the development and expansion of large B cells, clonally related to the CLL B cell population, and akin to RT, upregulate E2F target genes. Notably, GI tumors were observed only in the preleukemic TAM-treated mice which demonstrates the impact of disease burden prior to induction in iMYC-TCL1 mice. MYC induction post-CLL development further accelerates clonal disease expansion, upregulates Pdl1, skews the T cell repertoire, and shortens overall survival. B cell-specific overexpression of MYC in the iMYC-TCL1 model imparts features of RT and serves as a RT disease model and therapeutic development platform.
Disclosures: Woyach: Pharmacyclics: Consultancy, Research Funding; Newave: Consultancy; Merck: Consultancy; Loxo Lilly: Consultancy; Janssen: Research Funding; Genentech, Inc.: Consultancy; BeiGene: Consultancy; AstraZeneca: Consultancy; AbbVie: Research Funding; Schrodinger: Research Funding; Morphosys: Research Funding.
See more of: Oral and Poster Abstracts