Biomarker Predictive Ibrutinib Response Using Profiled ABC-DLBCL Patient Derived Xenografts

Background. Activated B cell-like (ABC) is a subtype of DLBCL difficult to treat. Activation of B-cell receptor (BCR) signaling pathway, including CD79 transmembrane proteins (A/B subunits) and Bruton's tyrosine kinase (BTK), are required for the disease maintenance. The MYD88 (an adapter for Toll-like receptors in a separate pathway from BCR pathway) activating mutation, L265P, has been suggested to activate BCR pathway (29% of ABC-DLBCL) and thus drives the disease. These DLBCLs seem responsive to ibrutinib (a BTK inhibitor) in the clinics^1,2. Majority of MYD88-L265P mutant DLBCL patients also have CD79B mutation (e.g. Y197N), i.e. double mutants (21% of total ABC-DLBCL), hinting collaborative oncogenesis, while some are MYD88-L265P single mutant with wild type CD79B. It remains unclear whether the single and double mutant ABC-DLBCL differ in their molecular pathogenesis as well as response to ibrutinib.

Method. We created DLBCL patient derived xenografts (PDX) as experimental system mimicking patients in order to investigate the molecular pathogenesis/response to ibrutinib. We first transcriptome-sequenced the established PDXs. We categorized them into ABC/GCB subtypes along with mutation status of CD79B/MYD88 per transcriptome profiles. We then assessed these PDXs for their response to ibrutinib and trying to correlate antitumor activity to molecular signature.

Results. We have created 20 NHL-PDX, with 5 of them transcriptome-sequenced. Clustering analysis per expression profile of 14-genes have classified all these 5 into subclass ABC-DLBCL. Among the 5, one is MYD88-L265P/CD79B-Y197N double mutants (LY2298), one MYD88-L265P single mutant (LY0257), and three wild types (LY2345, LY2214, LY2266). Among the 3 models tested so far, LY2214 (wildtype) and LY0257 (single mutation) did not respond to ibrutinib, but LY2298 (double mutations) did. These results, although small sample sizes, seem to suggest that double mutations are required for the response to ibrutinib, while single mutant and wild type does not. These observation seem to be consistent with the implication from recent clinical study¹. We are currently profile and test the remaining models to further confirm these observations. We will also further elucidate the downstream signaling along with the drug response.

Conclusion. Our preliminary results confirmed that ibrutinib sensitivity correlates with the concomitant double mutation of CD79B and MYD88.

References

1. Wilson, W.H., et al. Targeting B cell receptor signaling with ibrutinib in diffuse large B cell lymphoma. Nature medicine (2015).

2. Aalipour, A. & Advani, R.H. Bruton's tyrosine kinase inhibitors and their clinical potential in the treatment of B-cell malignancies: focus on ibrutinib. Therapeutic advances in hematology5, 121-133 (2014).