Abstract
BH3-mimetics are small molecule inhibitors that neutralize the function of anti-apoptotic BCL-2 family members. BH3-mimetics have recently gained a lot of popularity in oncology because of their success in cancer treatment. However, BH3-mimetics might have a broader clinical application. Here, we established an ex vivo flow cytometric assay allowing the comparison of the impact of BH3-mimetics (ABT-199, ABT-263, WEHI-539, and S63845) on leukocyte populations of both, healthy human subjects and C57BL/6 J wild type mice. BH3-mimetics were added to freshly drawn blood that was diluted 1/2 in cell medium, and BH3-mimetics-mediated impact on leukocyte count was assessed by flow cytometry. Our results demonstrate that responses towards 1μM of BH3-mimetics can be identical as well as considerably different in leukocytes of humans and mice. For instance, the inhibition of BCL-2 by ABT-199 caused cell death in all types of lymphocytes in mice but was exclusively specific for B cells in humans. Moreover, inhibition of BCL-XL by WEHI-539 affected solely mouse leukocytes while targeting MCL-1 by S63845 resulted in efficient induction of cell death in human neutrophils but not in their mouse counterparts. Our ex vivo assay enables initial identification of analogies and differences between human and mouse leukocytes in response towards BH3-mimetics.
Highlights
Excessive cell death or evasion from apoptosis is associated with the development of autoimmune disorders or cancer[1]
We present an ex vivo flow cytometry assay that allows quantitative assessment of human and mouse leukocyte viabilities in whole blood samples in response to BCL-2 homology domain 3 (BH3)-mimetics
We aimed to establish an assay to test the effects of BH3-mimetics in human and mouse specimens under non-invasive conditions, which are close to physiological conditions
Summary
Excessive cell death or evasion from apoptosis is associated with the development of autoimmune disorders or cancer[1]. Upregulation of certain anti-apoptotic BCL-2 family members, such as BCL-2, B cell lymphoma-extra-large (BCL-XL), and myeloid cell leukemia (MCL-1), are known to support development and survival of cancerous cells as well as evasion from cancer therapy[6]. To overcome such resistance towards apoptosis, small molecule inhibitors called BCL-2 homology domain 3 (BH3) -mimetics have been developed to neutralize anti-apoptotic BCL-2 family members in order to restore normal apoptotic signaling in cancerous cells[7]. We established an assay valid for both human and mouse derived blood samples, allowing an initial comparison of the response of human and mouse leukocytes towards BH3-mimetics
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