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Quantitative characterization of in vitro bystander effect of antibody-drug conjugates

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Abstract

Antibody–drug conjugates (ADCs) are designed to target antigen expressing (Ag+) cells in a tumor. Once processed by the Ag+ cells, ADCs can release cytotoxic drug molecules that can diffuse out of Ag+ cells into the neighboring antigen-negative (Ag−) cells to induce their cytotoxicity. This additional efficacy of ADCs on Ag− cells in the presence of Ag+ cells is known as the ‘bystander effect’. Although the importance of this phenomena is widely acknowledged for effective killing of a heterogeneous tumor, the rate and extent of the bystander killing in a heterogeneous system is not quantitatively understood yet. Thus, the objectives of this manuscript were to: (1) synthesize and characterize a tool ADC Trastuzumab-vc-MMAE that is capable of exhibiting bystander effect, (2) quantify the time course of the bystander effect for the tool ADC using in vitro co-culture systems created using mixture of various HER2-expressing cell lines, and (3) develop a pharmacodynamic (PD) model that is capable of characterizing the bystander effect of ADCs. Co-culture studies conducted using GFP labelled MCF7 cells as Ag− cells and N87, BT474, and SKBR3 as Ag+ cells revealed that the bystander effect of ADC increases with increasing fraction of Ag+ cells in a co-culture system, and with increased expression level of target on Ag+ cells. A notable lag time after ADC incubation was also observed prior to significant bystander killing of Ag− cells. Based on our results we hypothesize that there may be other determinants apart from the antigen expression level that can also influence the ability of Ag+ cells to demonstrate the bystander effect in a co-culture system. The co-culture analysis also suggested that the bystander effect of the ADC can dissipate over the period of time as the population of Ag+ cells declines. A novel PD model was developed to mathematically characterize the bystander effect of ADCs by combining two different cell distribution models to represent the population of Ag+ and Ag− cells in a co-culture system. This PD model can be integrated with the systems PK model for ADCs in the future to generate a quantitative framework that is capable of supporting the discovery and development of novel ADCs with optimal bystander killing capabilities.

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Acknowledgments

This work was supported by NIH Grant GM114179 to D.K.S., and the Centre for Protein Therapeutics at the State University of New York at Buffalo. S.S. was supported by a post-doctoral fellowship from Hoffman La-Roche, Switzerland to D.K.S. Authors would also like to thank Dr. Wojciech Krzyzanski, University at Buffalo, for his helpful discussion on mathematical modeling section of the manuscript.

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  1. Department of Pharmaceutical Sciences, School of Pharmacy and Pharmaceutical Sciences, The State University of New York at Buffalo, 455 Kapoor Hall, Buffalo, NY, 14214-8033, USA

    Aman P. Singh, Sharad Sharma & Dhaval K. Shah

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  1. Aman P. Singh
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Correspondence to Dhaval K. Shah.

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Singh, A.P., Sharma, S. & Shah, D.K. Quantitative characterization of in vitro bystander effect of antibody-drug conjugates. J Pharmacokinet Pharmacodyn 43, 567–582 (2016). https://doi.org/10.1007/s10928-016-9495-8

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  • DOI: https://doi.org/10.1007/s10928-016-9495-8

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