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Synergistic cytotoxicity of a prostate cancer-specific immunotoxin in combination with the BH3 mimetic ABT-737

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Abstract

In many tumors, including prostate cancer, anti-apoptotic members of the Bcl-2 family are overexpressed and cause cell death resistance, which is a typical hallmark of cancer. Different therapeutic approaches, therefore, aim to restore the death mechanisms for enhanced apoptosis. Our recombinant immunotoxin D7(VL-VH)-PE40 is composed of the scFv D7(VL-VH) against the prostate-specific membrane antigen (PSMA) on the surface of prostate cancer cells and of the cytotoxic domain of the bacterial toxin Pseudomonas Exotoxin A (PE40). Since Pseudomonas Exotoxin A-based immunotoxins are known to preferentially inhibit the expression of the anti-apoptotic protein Mcl-1, the rationale was to test our immunotoxin in combination with the BH3 mimetic ABT-737, which specifically inhibits Bcl-2, Bcl-xl, and Bcl-w for enhanced induction of apoptosis in prostate cancer cells. The immunotoxin showed high and specific binding and cytotoxicity against PSMA expressing prostate cancer cells marked by a direct inhibition of Mcl-1. The combination of the immunotoxin with a subtoxic concentration of ABT-737 caused additive or even synergistic effects, which were based on an enhanced apoptosis induction as detected by poly(ADP-ribose) polymerase (PARP) and Caspase-3 cleavage in Western blot. Our study shows that the combination therapy of immunotoxin plus ABT-737 is a promising approach for the future treatment of advanced prostate cancer to improve therapeutic efficacy and to reduce adverse side effects.

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Abbreviations

ABT-737:

4-[4-[[2-(4-chlorophenyl)phenyl]methyl]piperazin-1-yl]-N-[4-[[(2R)-4-(dimethylamino)-1-phenylsulfanylbutan-2-yl]amino]-3-nitrophenyl] sulfonylbenzamide

Bak:

Bcl-2 antagonist/killer

Bax:

Bcl-2-associated X protein

BCA:

Bicinchoninic acid

Bcl-2:

B cell lymphoma 2

Bcl-w:

Bcl-2 like 2

Bcl-xl:

B cell lymphoma extra-large

BH3:

Bcl-2 homology domain 3

CI:

Combination Index

c-myc:

Avian myelocytomatosis virus oncogene cellular homolog

ECL:

Enhanced chemiluminescence

eEF-2:

Eukaryotic elongation factor-2

Mcl-1:

Myeloid cell leukemia sequence 1

PARP:

Poly(ADP-ribose) polymerase

PE:

Pseudomonas aeruginosa Exotoxin A

PSMA:

Prostate-specific membrane antigen

WST-1:

Water-soluble tetrazolium salt

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Acknowledgements

This study was funded by the Wilhelm Sander-Stiftung (Grant No. 2016.089.01 to Philipp Wolf).

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Authors and Affiliations

  1. Department of Urology, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Breisacher Str. 66, 79106, Freiburg, Germany

    Theresa Noll, Susanne Schultze-Seemann, Irina Kuckuck, Marta Michalska & Philipp Wolf

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  1. Theresa Noll
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  2. Susanne Schultze-Seemann
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  3. Irina Kuckuck
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  4. Marta Michalska
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  5. Philipp Wolf
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Correspondence to Philipp Wolf.

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Noll, T., Schultze-Seemann, S., Kuckuck, I. et al. Synergistic cytotoxicity of a prostate cancer-specific immunotoxin in combination with the BH3 mimetic ABT-737. Cancer Immunol Immunother 67, 413–422 (2018). https://doi.org/10.1007/s00262-017-2097-5

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  • DOI: https://doi.org/10.1007/s00262-017-2097-5

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