Effective Targeting of Melanoma Cells by Combination of Mcl-1 and Bcl-2/Bcl-xL/Bcl-w Inhibitors
<p>Combinations of BH3 mimetics decrease cell viability and induce apoptosis in melanoma cells at 24 h. Melanoma cell lines A-375, Mel-HO, MeWo, and SK-Mel-23 were seeded in 24-well plates and were treated with S63845 (S63, 1 µM) as well as with ABT-199, ABT-263, or ABT-737 (0.01, 0.1, 1 µM) as indicated. (<b>A</b>) After 24 h, cell viability was determined via calcein-AM staining and flow cytometry. Values represent the percentage of cells with high calcein staining (viable cells). Effects on cell viability are displayed as a percentage of non-treated controls (100%). (<b>B</b>) After 24 h, apoptotic cells were identified as sub-G1 cells in cell cycle analyses via flow cytometry after propidium-iodide staining. (<b>A</b>,<b>B</b>) At least two series of experiments were performed, each one consisting of independent triplicate values. Mean values of all individual values (at least 6) are shown here. Statistical significance is indicated by asterisks (*; <span class="html-italic">p</span> < 0.05) and was calculated for the single treatments as compared to non-treated control cells. The combination treatments were compared both to the respective single treatments with ABT (light bars) and to S63845 alone. Largely comparable findings were obtained after 48 h treatments (<a href="#app1-ijms-25-03453" class="html-app">Supplementary Figure S2</a>). Example flow cytometry readings of cells treated for 48 h with combinations of ABTs and S63845 (1 µM concentrations) are shown as overlays vs. controls. Non-viable and viable cell populations as well as cell cycle phases G1 (gap 1), S (synthesis), G2 (gap 2), and sub-G1 cells are indicated.</p> "> Figure 2
<p>Early induction of apoptosis plays a leading role. Mel-HO and SK-Mel-23 cells were seeded in 24-well plates treated with ABT-199, ABT-263, ABT-737, and/or S63845 (1 µM concentrations; ---, no ABTs were added). (<b>A</b>) Cell death analysis by Annexin V/PI staining and flow cytometry was performed at 6 h, 12 h, and 24 h. Early apoptotic cells were determined as AnnV(+)/PI(−), while late apoptotic or necrotic cells corresponded to AnnV(+)/PI(+) staining. Mean values (in %) and SDs were calculated of three series of experiments, each one consisting of double values (six values in a group). Statistically significant changes observed in combination treatments, as compared to the respective single treatment with ABT and to S63845 alone, are indicated for the two cell death fractions (*; <span class="html-italic">p</span> < 0.01). (<b>B</b>) Representative flow cytometry histograms of combination-treated cells and control cells are shown. Readings are separated into four quadrants (according to PI and AnnV positivity). Thus, the lower left quadrant corresponds to AnnV(−)/PI(−) cells (viable cells), the lower right quadrant corresponds to AnnV(+)/PI(−) cells (early apoptosis), and the upper right quadrant corresponds to AnnV(+)/PI(+) cells (late apoptosis).</p> "> Figure 3
<p>Combinations of BH3 mimetics induce mitochondrial apoptosis in vitro. The four cell lines were treated with S63845 (S63), ABT-199, ABT-263, ABT-737, and combinations (1 µM concentrations; ---, no ABTs were added). (<b>A</b>) Mitochondrial membrane potential (MMP) was determined at 4 h and at 24 h via TMRM<sup>+</sup> staining and flow cytometry. Values represent the percentage of cells with low MMP. Mean values and SDs (in %) were calculated for two series of experiments, each one consisting of independent triplicate values (six values in a group). Statistical significance (* <span class="html-italic">p</span> < 0.05) was calculated for combination treatments as compared to the respective single treatments (ABTs alone and S63845 alone). (<b>B</b>) Examples of flow cytometry readings are shown as overlays of combination treatments (dark graphs) vs. the controls (open graphs). Cell populations with low MMP are indicated. (<b>C</b>) For microscopic visualization of low MMP, Mel-HO and SK-Mel-23 cells were stained with JC-1 and counterstained with Hoechst-33342 at 6 h and at 12 h of treatment. Cell nuclei are stained in blue, while mitochondria with high (normal) MMP are stained in red. Green staining indicates cytosolic JC1 localization upon release of JC1 from mitochondria with low MMP. Two independent experiments revealed highly comparable results.</p> "> Figure 4
<p>Reactive oxygen species are produced after combination treatment. A-375, Mel-HO, MeWo, and SK-Mel-23 cells were treated with ABT-199, ABT-263, ABT-737, S63845 (S63), and combinations (1 µM concentrations; ---, no ABTs were added). (<b>A</b>) Cellular levels of ROS were determined at 4 h and at 24 h via H<sub>2</sub>DCF-DA staining and flow cytometry. Values represent the percentage of cells with high ROS. Mean values and SDs (in %) were calculated for two independent experiments, each one consisting of triplicate values (six values in a group). Statistical significance (* <span class="html-italic">p</span> < 0.05) was calculated for combination treatments as compared to the respective single treatments (ABTs alone and S63845 alone). (<b>B</b>) Examples of flow cytometry readings are shown on the right side for the combination treatments (dark graphs) vs. the controls (open graphs). Cell populations with high ROS are indicated.</p> "> Figure 5
<p>Pro-apoptotic pathways are efficiently activated only after combination treatment. Mel-HO and SK-Mel-23 cells were treated for 8 h with ABT-199, ABT-263, ABT-737, S63845, and combinations (1 µM concentrations). Total protein extracts were analyzed by Western blotting for cleaved caspase-3, total caspase-8, and caspase-9 (Csp). Further, processing of PARP from 116 to 89 kD and phosphorylation of histone H2AX (γ-H2AX) were analyzed. Equal protein amounts (30 µg per lane) were separated by SDS-PAGE, and consistent blotting was proven by Ponceau staining as well as via evaluation of GAPDH expression. Molecular weights are indicated in kD. Each two independent series of protein extracts and Western blots revealed highly comparable results.</p> "> Figure 6
<p>Cell death induced by BH3 mimetic combinations in melanoma cells is caspase-dependent in vitro. Mel-HO and SK-Mel-23 cells were treated with combinations of S63845 with ABT-199, ABT-263, and ABT-737 (1 µM concentrations). When indicated, the pan-caspase inhibitor QVD-Oph (QVD, 5 µM) was applied at 1 h before other treatments started. Cell viability (calcein staining; (<b>A</b>)), apoptosis induction (cell cycle analysis; (<b>B</b>)), loss of MMP (TMRM<sup>+</sup> staining; (<b>C</b>)) and production of ROS (H<sub>2</sub>DCF-DA staining; (<b>D</b>)) were determined via flow cytometry at 24 h. Mean values and SDs (in %) were calculated from two series of experiments, each one consisting of independent triplicate values. Statistical significance (* <span class="html-italic">p</span> < 0.05) was calculated for QVD treatments as compared to ABT/S63845 combinations without QVD (at least 6 individual values in a group). On the right side, example flow cytometry readings of ABT/S63845 combination treatments +/− QVD are shown as overlays. Viable and non-viable cell populations (<b>A</b>), sub-G1, G1, S and G2 cell populations (<b>B</b>), cell populations with low MMP (<b>C</b>), and cell populations with high ROS levels (<b>D</b>) are indicated.</p> "> Figure 7
<p>Apoptosis proteins Mcl-1 and XIAP are downregulated by combination treatments. Mel-HO and SK-Mel-23 cells were treated with ABT-199, ABT-263, ABT-737, S63845, and combinations (1 µM concentrations, treatment time: 8 h). Total protein extracts were analyzed by Western blotting for expression of Mcl-1 (41 kD), Bcl-2 (26 kD), Bcl-w (18 kD), Bcl-x<sub>L</sub> (30 kD), and XIAP (53 kD). Equal protein amounts (30 µg per lane) were separated by SDS-PAGE, and consistent blotting was proven via Ponceau staining as well as through evaluation of GAPDH expression. Molecular weights are indicated in kD. Two independent series of protein extracts and Western blots revealed highly comparable results.</p> ">
Abstract
:1. Introduction
2. Results
2.1. Significant Decrease in Cell Viability Only after Combination of BH3 Mimetics
2.2. Decreased Cell Viability Correlates with Induction of Apoptosis
2.3. Activation of Mitochondrial Apoptosis Pathways
2.4. Crucial Role of Caspase-Mediated Pathways
2.5. Downregulation of Anti-Apoptotic Factors
3. Discussion
4. Materials and Methods
4.1. Cell Culture and Treatment
4.2. Quantification of Cell Viability and Apoptosis
4.3. Assays for Mitochondrial Membrane Potential and Reactive Oxygen Species (ROS)
4.4. Western Blotting
4.5. Statistical Analyses
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
- Schadendorf, D.; van Akkooi, A.C.J.; Berking, C.; Griewank, K.G.; Gutzmer, R.; Hauschild, A.; Stang, A.; Roesch, A.; Ugurel, S. Melanoma. Lancet 2018, 392, 971–984. [Google Scholar] [CrossRef] [PubMed]
- Garbe, C.; Amaral, T.; Peris, K.; Hauschild, A.; Arenberger, P.; Basset-Seguin, N.; Bastholt, L.; Bataille, V.; Del Marmol, V.; Dreno, B.; et al. European consensus-based interdisciplinary guideline for melanoma. Part 1: Diagnostics: Update 2022. Eur. J. Cancer 2022, 170, 236–255. [Google Scholar] [CrossRef] [PubMed]
- Davies, H.; Bignell, G.R.; Cox, C.; Stephens, P.; Edkins, S.; Clegg, S.; Teague, J.; Woffendin, H.; Garnett, M.J.; Bottomley, W.; et al. Mutations of the BRAF gene in human cancer. Nature 2002, 417, 949–954. [Google Scholar] [CrossRef] [PubMed]
- Postow, M.A.; Callahan, M.K.; Wolchok, J.D. Immune Checkpoint Blockade in Cancer Therapy. J. Clin. Oncol. 2015, 33, 1974–1982. [Google Scholar] [CrossRef] [PubMed]
- Menzies, A.M.; Long, G.V. Systemic treatment for BRAF-mutant melanoma: Where do we go next? Lancet Oncol. 2014, 15, e371–e381. [Google Scholar] [CrossRef] [PubMed]
- Garbe, C.; Amaral, T.; Peris, K.; Hauschild, A.; Arenberger, P.; Basset-Seguin, N.; Bastholt, L.; Bataille, V.; Del Marmol, V.; Dreno, B.; et al. European consensus-based interdisciplinary guideline for melanoma. Part 2: Treatment—Update 2022. Eur. J. Cancer 2022, 170, 256–284. [Google Scholar] [CrossRef]
- Wolchok, J.D.; Chiarion-Sileni, V.; Gonzalez, R.; Grob, J.J.; Rutkowski, P.; Lao, C.D.; Cowey, C.L.; Schadendorf, D.; Wagstaff, J.; Dummer, R.; et al. Long-Term Outcomes With Nivolumab Plus Ipilimumab or Nivolumab Alone Versus Ipilimumab in Patients With Advanced Melanoma. J. Clin. Oncol. 2022, 40, 127–137. [Google Scholar] [CrossRef] [PubMed]
- Hughes, T.; Klairmont, M.; Sharfman, W.H.; Kaufman, H.L. Interleukin-2, Ipilimumab, and Anti-PD-1: Clinical management and the evolving role of immunotherapy for the treatment of patients with metastatic melanoma. Cancer Biol. Ther. 2021, 22, 513–526. [Google Scholar] [CrossRef]
- Johnpulle, R.A.; Johnson, D.B.; Sosman, J.A. Molecular Targeted Therapy Approaches for BRAF Wild-Type Melanoma. Curr. Oncol. Rep. 2016, 18, 6. [Google Scholar] [CrossRef]
- Dimitriou, F.; Long, G.V.; Menzies, A.M. Novel adjuvant options for cutaneous melanoma. Ann. Oncol. 2021, 32, 854–865. [Google Scholar] [CrossRef]
- Carlino, M.S.; Larkin, J.; Long, G.V. Immune checkpoint inhibitors in melanoma. Lancet 2021, 398, 1002–1014. [Google Scholar] [CrossRef]
- Hanahan, D.; Weinberg, R.A. Hallmarks of cancer: The next generation. Cell 2011, 144, 646–674. [Google Scholar] [CrossRef]
- Eberle, J.; Fecker, L.F.; Forschner, T.; Ulrich, C.; Rowert-Huber, J.; Stockfleth, E. Apoptosis pathways as promising targets for skin cancer therapy. Brit. J. Dermatol. 2007, 156, 18–24. [Google Scholar] [CrossRef]
- Krammer, P.H.; Arnold, R.; Lavrik, I.N. Life and death in peripheral T cells. Nat. Rev. Immunol. 2007, 7, 532–542. [Google Scholar] [CrossRef] [PubMed]
- Fischer, U.; Janicke, R.U.; Schulze-Osthoff, K. Many cuts to ruin: A comprehensive update of caspase substrates. Cell Death Differ. 2003, 10, 76–100. [Google Scholar] [CrossRef] [PubMed]
- Soldani, C.; Scovassi, A.I. Poly(ADP-ribose) polymerase-1 cleavage during apoptosis: An update. Apoptosis 2002, 7, 321–328. [Google Scholar] [CrossRef]
- Rogakou, E.P.; Nieves-Neira, W.; Boon, C.; Pommier, Y.; Bonner, W.M. Initiation of DNA fragmentation during apoptosis induces phosphorylation of H2AX histone at serine 139. J. Biol. Chem. 2000, 275, 9390–9395. [Google Scholar] [CrossRef] [PubMed]
- Lu, C.; Zhu, F.; Cho, Y.Y.; Tang, F.; Zykova, T.; Ma, W.Y.; Bode, A.M.; Dong, Z. Cell apoptosis: Requirement of H2AX in DNA ladder formation, but not for the activation of caspase-3. Mol. Cell 2006, 23, 121–132. [Google Scholar] [CrossRef]
- Singh, R.; Letai, A.; Sarosiek, K. Regulation of apoptosis in health and disease: The balancing act of BCL-2 family proteins. Nat. Rev. Mol. Cell Biol. 2019, 20, 175–193. [Google Scholar] [CrossRef] [PubMed]
- Zhivkova, V.; Kiecker, F.; Langer, P.; Eberle, J. Crucial role of reactive oxygen species (ROS) for the proapoptotic effects of indirubin derivative DKP-073 in melanoma cells. Mol. Carcinog. 2019, 58, 258–269. [Google Scholar] [CrossRef]
- Quast, S.A.; Berger, A.; Eberle, J. ROS-dependent phosphorylation of Bax by wortmannin sensitizes melanoma cells for TRAIL-induced apoptosis. Cell Death Dis. 2013, 4, e839. [Google Scholar] [CrossRef]
- Chipuk, J.E.; Moldoveanu, T.; Llambi, F.; Parsons, M.J.; Green, D.R. The BCL-2 family reunion. Mol. Cell 2010, 37, 299–310. [Google Scholar] [CrossRef]
- Raisova, M.; Hossini, A.M.; Eberle, J.; Riebeling, C.; Wieder, T.; Sturm, I.; Daniel, P.T.; Orfanos, C.E.; Geilen, C.C. The Bax/Bcl-2 ratio determines the susceptibility of human melanoma cells to CD95/Fas-mediated apoptosis. J. Investig. Dermatol. 2001, 117, 333–340. [Google Scholar] [CrossRef]
- Jansen, B.; Schlagbauer-Wadl, H.; Brown, B.D.; Bryan, R.N.; van Elsas, A.; Muller, M.; Wolff, K.; Eichler, H.G.; Pehamberger, H. bcl-2 antisense therapy chemosensitizes human melanoma in SCID mice. Nat. Med. 1998, 4, 232–234. [Google Scholar] [CrossRef]
- Eberle, J. Countering TRAIL Resistance in Melanoma. Cancers 2019, 11, 656. [Google Scholar] [CrossRef]
- Hanada, M.; Delia, D.; Aiello, A.; Stadtmauer, E.; Reed, J.C. bcl-2 gene hypomethylation and high-level expression in B-cell chronic lymphocytic leukemia. Blood 1993, 82, 1820–1828. [Google Scholar] [CrossRef] [PubMed]
- Krajewski, S.; Krajewska, M.; Shabaik, A.; Wang, H.G.; Irie, S.; Fong, L.; Reed, J.C. Immunohistochemical analysis of in vivo patterns of Bcl-X expression. Cancer Res. 1994, 54, 5501–5507. [Google Scholar] [PubMed]
- Kozopas, K.M.; Yang, T.; Buchan, H.L.; Zhou, P.; Craig, R.W. MCL1, a gene expressed in programmed myeloid cell differentiation, has sequence similarity to BCL2. Proc. Natl. Acad. Sci. USA 1993, 90, 3516–3520. [Google Scholar] [CrossRef]
- Diepstraten, S.T.; Anderson, M.A.; Czabotar, P.E.; Lessene, G.; Strasser, A.; Kelly, G.L. The manipulation of apoptosis for cancer therapy using BH3-mimetic drugs. Nat. Rev. Cancer 2022, 22, 45–64. [Google Scholar] [CrossRef]
- Leiter, U.; Schmid, R.M.; Kaskel, P.; Peter, R.U.; Krahn, G. Antiapoptotic bcl-2 and bcl-xL in advanced malignant melanoma. Arch. Dermatol. Res. 2000, 292, 225–232. [Google Scholar] [CrossRef]
- Trisciuoglio, D.; Desideri, M.; Ciuffreda, L.; Mottolese, M.; Ribatti, D.; Vacca, A.; Del Rosso, M.; Marcocci, L.; Zupi, G.; Del Bufalo, D. Bcl-2 overexpression in melanoma cells increases tumor progression-associated properties and in vivo tumor growth. J. Cell. Physiol. 2005, 205, 414–421. [Google Scholar] [CrossRef]
- Beroukhim, R.; Mermel, C.H.; Porter, D.; Wei, G.; Raychaudhuri, S.; Donovan, J.; Barretina, J.; Boehm, J.S.; Dobson, J.; Urashima, M.; et al. The landscape of somatic copy-number alteration across human cancers. Nature 2010, 463, 899–905. [Google Scholar] [CrossRef]
- Lee, E.F.; Harris, T.J.; Tran, S.; Evangelista, M.; Arulananda, S.; John, T.; Ramnac, C.; Hobbs, C.; Zhu, H.; Gunasingh, G.; et al. BCL-XL and MCL-1 are the key BCL-2 family proteins in melanoma cell survival. Cell Death Dis. 2019, 10, 342. [Google Scholar] [CrossRef]
- Hartman, M.L.; Czyz, M. BCL-w: Apoptotic and non-apoptotic role in health and disease. Cell Death Dis. 2020, 11, 260. [Google Scholar] [CrossRef]
- Kim, Y.J.; Tsang, T.; Anderson, G.R.; Posimo, J.M.; Brady, D.C. Inhibition of BCL2 Family Members Increases the Efficacy of Copper Chelation in BRAFV600E-Driven Melanoma. Cancer Res. 2020, 80, 1387–1400. [Google Scholar] [CrossRef]
- Sarif, Z.; Tolksdorf, B.; Fechner, H.; Eberle, J. Mcl-1 targeting strategies unlock the proapoptotic potential of TRAIL in melanoma cells. Mol. Carcinog. 2020, 59, 1256–1268. [Google Scholar] [CrossRef]
- Placzek, W.J.; Wei, J.; Kitada, S.; Zhai, D.; Reed, J.C.; Pellecchia, M. A survey of the anti-apoptotic Bcl-2 subfamily expression in cancer types provides a platform to predict the efficacy of Bcl-2 antagonists in cancer therapy. Cell Death Dis. 2010, 1, e40. [Google Scholar] [CrossRef]
- Merino, D.; Kelly, G.L.; Lessene, G.; Wei, A.H.; Roberts, A.W.; Strasser, A. BH3-Mimetic Drugs: Blazing the Trail for New Cancer Medicines. Cancer Cell 2018, 34, 879–891. [Google Scholar] [CrossRef]
- Oltersdorf, T.; Elmore, S.W.; Shoemaker, A.R.; Armstrong, R.C.; Augeri, D.J.; Belli, B.A.; Bruncko, M.; Deckwerth, T.L.; Dinges, J.; Hajduk, P.J.; et al. An inhibitor of Bcl-2 family proteins induces regression of solid tumours. Nature 2005, 435, 677–681. [Google Scholar] [CrossRef]
- Tse, C.; Shoemaker, A.R.; Adickes, J.; Anderson, M.G.; Chen, J.; Jin, S.; Johnson, E.F.; Marsh, K.C.; Mitten, M.J.; Nimmer, P.; et al. ABT-263: A potent and orally bioavailable Bcl-2 family inhibitor. Cancer Res. 2008, 68, 3421–3428. [Google Scholar] [CrossRef]
- Souers, A.J.; Leverson, J.D.; Boghaert, E.R.; Ackler, S.L.; Catron, N.D.; Chen, J.; Dayton, B.D.; Ding, H.; Enschede, S.H.; Fairbrother, W.J.; et al. ABT-199, a potent and selective BCL-2 inhibitor, achieves antitumor activity while sparing platelets. Nat. Med. 2013, 19, 202–208. [Google Scholar] [CrossRef]
- Kotschy, A.; Szlavik, Z.; Murray, J.; Davidson, J.; Maragno, A.L.; Le Toumelin-Braizat, G.; Chanrion, M.; Kelly, G.L.; Gong, J.N.; Moujalled, D.M.; et al. The MCL1 inhibitor S63845 is tolerable and effective in diverse cancer models. Nature 2016, 538, 477–482. [Google Scholar] [CrossRef]
- Peng, Z.; Gillissen, B.; Richter, A.; Sinnberg, T.; Schlaak, M.S.; Eberle, J. Enhanced Apoptosis and Loss of Cell Viability in Melanoma Cells by Combined Inhibition of ERK and Mcl-1 Is Related to Loss of Mitochondrial Membrane Potential, Caspase Activation and Upregulation of Proapoptotic Bcl-2 Proteins. Int. J. Mol. Sci. 2023, 24, 4961. [Google Scholar] [CrossRef]
- Torrens-Mas, M.; Cordani, M.; Mullappilly, N.; Pacchiana, R.; Riganti, C.; Palmieri, M.; Pons, D.G.; Roca, P.; Oliver, J.; Donadelli, M. Mutant p53 induces SIRT3/MnSOD axis to moderate ROS production in melanoma cells. Arch. Biochem. Biophys. 2020, 679, 108219. [Google Scholar] [CrossRef]
- Tantawy, S.I.; Sarkar, A.; Hubner, S.; Tan, Z.; Wierda, W.G.; Eldeib, A.; Zhang, S.; Kornblau, S.; Gandhi, V. Mechanisms of MCL-1 Protein Stability Induced by MCL-1 Antagonists in B-Cell Malignancies. Clin. Cancer Res. 2023, 29, 446–457. [Google Scholar] [CrossRef]
- Herrant, M.; Jacquel, A.; Marchetti, S.; Belhacène, N.; Colosetti, P.; Luciano, F.; Auberger, P. Cleavage of Mcl-1 by caspases impaired its ability to counteract Bim-induced apoptosis. Oncogene 2004, 23, 7863–7873. [Google Scholar] [CrossRef]
- Kaloni, D.; Diepstraten, S.T.; Strasser, A.; Kelly, G.L. BCL-2 protein family: Attractive targets for cancer therapy. Apoptosis 2023, 28, 20–38. [Google Scholar] [CrossRef]
- Wilson, W.H.; O’Connor, O.A.; Czuczman, M.S.; LaCasce, A.S.; Gerecitano, J.F.; Leonard, J.P.; Tulpule, A.; Dunleavy, K.; Xiong, H.; Chiu, Y.L.; et al. Navitoclax, a targeted high-affinity inhibitor of BCL-2, in lymphoid malignancies: A phase 1 dose-escalation study of safety, pharmacokinetics, pharmacodynamics, and antitumour activity. Lancet Oncol. 2010, 11, 1149–1159. [Google Scholar] [CrossRef]
- Mason, K.D.; Carpinelli, M.R.; Fletcher, J.I.; Collinge, J.E.; Hilton, A.A.; Ellis, S.; Kelly, P.N.; Ekert, P.G.; Metcalf, D.; Roberts, A.W.; et al. Programmed anuclear cell death delimits platelet life span. Cell 2007, 128, 1173–1186. [Google Scholar] [CrossRef] [PubMed]
- Deeks, E.D. Venetoclax: First Global Approval. Drugs 2016, 76, 979–987. [Google Scholar] [CrossRef]
- Lok, S.W.; Whittle, J.R.; Vaillant, F.; Teh, C.E.; Lo, L.L.; Policheni, A.N.; Bergin, A.R.T.; Desai, J.; Ftouni, S.; Gandolfo, L.C.; et al. A Phase Ib Dose-Escalation and Expansion Study of the BCL2 Inhibitor Venetoclax Combined with Tamoxifen in ER and BCL2-Positive Metastatic Breast Cancer. Cancer Discov. 2019, 9, 354–369. [Google Scholar] [CrossRef] [PubMed]
- Lochmann, T.L.; Floros, K.V.; Naseri, M.; Powell, K.M.; Cook, W.; March, R.J.; Stein, G.T.; Greninger, P.; Maves, Y.K.; Saunders, L.R.; et al. Venetoclax Is Effective in Small-Cell Lung Cancers with High BCL-2 Expression. Clin. Cancer Res. 2018, 24, 360–369. [Google Scholar] [CrossRef] [PubMed]
- Place, A.E.; Goldsmith, K.; Bourquin, J.P.; Loh, M.L.; Gore, L.; Morgenstern, D.A.; Sanzgiri, Y.; Hoffman, D.; Zhou, Y.; Ross, J.A.; et al. Accelerating drug development in pediatric cancer: A novel Phase I study design of venetoclax in relapsed/refractory malignancies. Future Oncol. 2018, 14, 2115–2129. [Google Scholar] [CrossRef]
- Kehr, S.; Vogler, M. It’s time to die: BH3 mimetics in solid tumors. Biochim. Biophys. Acta Mol. Cell Res. 2021, 1868, 118987. [Google Scholar] [CrossRef] [PubMed]
- Mukherjee, N.; Skees, J.; Todd, K.J.; West, D.A.; Lambert, K.A.; Robinson, W.A.; Amato, C.M.; Couts, K.L.; Van Gulick, R.; MacBeth, M.; et al. MCL1 inhibitors S63845/MIK665 plus Navitoclax synergistically kill difficult-to-treat melanoma cells. Cell Death Dis. 2020, 11, 443. [Google Scholar] [CrossRef] [PubMed]
- Miller, L.A.; Goldstein, N.B.; Johannes, W.U.; Walton, C.H.; Fujita, M.; Norris, D.A.; Shellman, Y.G. BH3 Mimetic ABT-737 and a Proteasome Inhibitor Synergistically Kill Melanomas through Noxa-Dependent Apoptosis. J. Investig. Dermatol. 2009, 129, 964–971. [Google Scholar] [CrossRef]
- Wertz, I.E.; Kusam, S.; Lam, C.; Okamoto, T.; Sandoval, W.; Anderson, D.J.; Helgason, E.; Ernst, J.A.; Eby, M.; Liu, J.; et al. Sensitivity to antitubulin chemotherapeutics is regulated by MCL1 and FBW7. Nature 2011, 471, 110–114. [Google Scholar] [CrossRef]
- Brennan, M.S.; Chang, C.; Tai, L.; Lessene, G.; Strasser, A.; Dewson, G.; Kelly, G.L.; Herold, M.J. Humanized Mcl-1 mice enable accurate preclinical evaluation of MCL-1 inhibitors destined for clinical use. Blood 2018, 132, 1573–1583. [Google Scholar] [CrossRef]
- Dastur, A.; Choi, A.; Costa, C.; Yin, X.Q.; Williams, A.; McClanaghan, J.; Greenberg, M.; Roderick, J.; Patel, N.U.; Boisvert, J.; et al. NOTCH1 Represses MCL-1 Levels in GSI-resistant T-ALL, Making them Susceptible to ABT-263. Clin. Cancer Res. 2019, 25, 312–324. [Google Scholar] [CrossRef]
- Guieze, R.; Liu, V.M.; Rosebrock, D.; Jourdain, A.A.; Hernandez-Sanchez, M.; Martinez Zurita, A.; Sun, J.; Ten Hacken, E.; Baranowski, K.; Thompson, P.A.; et al. Mitochondrial Reprogramming Underlies Resistance to BCL-2 Inhibition in Lymphoid Malignancies. Cancer Cell 2019, 36, 369–384.E13. [Google Scholar] [CrossRef]
- Faber, A.C.; Farago, A.F.; Costa, C.; Dastur, A.; Gomez-Caraballo, M.; Robbins, R.; Wagner, B.L.; Rideout, W.M., 3rd; Jakubik, C.T.; Ham, J.; et al. Assessment of ABT-263 activity across a cancer cell line collection leads to a potent combination therapy for small-cell lung cancer. Proc. Natl. Acad. Sci. USA 2015, 112, E1288–E1296. [Google Scholar] [CrossRef]
- Williams, M.M.; Lee, L.; Hicks, D.J.; Joly, M.M.; Elion, D.; Rahman, B.; McKernan, C.; Sanchez, V.; Balko, J.M.; Stricker, T.; et al. Key Survival Factor, Mcl-1, Correlates with Sensitivity to Combined Bcl-2/Bcl-xL Blockade. Mol. Cancer Res. 2017, 15, 259–268. [Google Scholar] [CrossRef] [PubMed]
- Wang, H.; Guo, M.; Wei, H.; Chen, Y. Targeting MCL-1 in cancer: Current status and perspectives. J. Hematol. Oncol. 2021, 14, 67. [Google Scholar] [CrossRef] [PubMed]
- Szlavik, Z.; Ondi, L.; Csekei, M.; Paczal, A.; Szabo, Z.B.; Radics, G.; Murray, J.; Davidson, J.; Chen, I.; Davis, B.; et al. Structure-Guided Discovery of a Selective Mcl-1 Inhibitor with Cellular Activity. J. Med. Chem. 2019, 62, 6913–6924. [Google Scholar] [CrossRef] [PubMed]
- Senichkin, V.V.; Streletskaia, A.Y.; Gorbunova, A.S.; Zhivotovsky, B.; Kopeina, G.S. Saga of Mcl-1: Regulation from transcription to degradation. Cell Death Differ. 2020, 27, 405–419. [Google Scholar] [CrossRef] [PubMed]
- Tseng, H.Y.; Dreyer, J.; Emran, A.A.; Gunatilake, D.; Pirozyan, M.; Cullinane, C.; Dutton-Regester, K.; Rizos, H.; Hayward, N.K.; McArthur, G.; et al. Co-targeting bromodomain and extra-terminal proteins and MCL1 induces synergistic cell death in melanoma. Int. J. Cancer 2020, 147, 2176–2189. [Google Scholar] [CrossRef] [PubMed]
- Weeden, C.E.; Ah-Cann, C.; Holik, A.Z.; Pasquet, J.; Garnier, J.M.; Merino, D.; Lessene, G.; Asselin-Labat, M.L. Dual inhibition of BCL-XL and MCL-1 is required to induce tumour regression in lung squamous cell carcinomas sensitive to FGFR inhibition. Oncogene 2018, 37, 4475–4488. [Google Scholar] [CrossRef] [PubMed]
- Kehr, S.; Haydn, T.; Bierbrauer, A.; Irmer, B.; Vogler, M.; Fulda, S. Targeting BCL-2 proteins in pediatric cancer: Dual inhibition of BCL-X(L) and MCL-1 leads to rapid induction of intrinsic apoptosis. Cancer Lett. 2020, 482, 19–32. [Google Scholar] [CrossRef] [PubMed]
- Luo, M.J.; Palmieri, M.; Riffkin, C.D.; Sakthianandeswaren, A.; Djajawi, T.M.; Hirokawa, Y.; Shuttleworth, V.; Segal, D.H.; White, C.A.; Nhu, D.; et al. Defining the susceptibility of colorectal cancers to BH3-mimetic compounds. Cell Death Dis. 2020, 11, 735. [Google Scholar] [CrossRef]
- Mukherjee, N.; Amato, C.M.; Skees, J.; Todd, K.J.; Lambert, K.A.; Robinson, W.A.; Van Gulick, R.; Weight, R.M.; Dart, C.R.; Tobin, R.P.; et al. Simultaneously Inhibiting BCL2 and MCL1 Is a Therapeutic Option for Patients with Advanced Melanoma. Cancers 2020, 12, 2182. [Google Scholar] [CrossRef]
- Griffioen, M.S.; de Leeuw, D.C.; Janssen, J.; Smit, L. Targeting Acute Myeloid Leukemia with Venetoclax; Biomarkers for Sensitivity and Rationale for Venetoclax-Based Combination Therapies. Cancers 2022, 14, 3456. [Google Scholar] [CrossRef] [PubMed]
- Bierbrauer, A.; Jacob, M.; Vogler, M.; Fulda, S. A direct comparison of selective BH3-mimetics reveals BCL-X(L), BCL-2 and MCL-1 as promising therapeutic targets in neuroblastoma. Br. J. Cancer 2020, 122, 1544–1551. [Google Scholar] [CrossRef] [PubMed]
- Yu, R.; Lu, Y.; Yu, R.; Xie, J.; Zhou, S. Synergistic Effects of TW-37 and ABT-263 on Renal Cell Carcinoma Cells. Cancer Manag. Res. 2021, 13, 953–963. [Google Scholar] [CrossRef] [PubMed]
- Mukherjee, N.; Strosnider, A.; Vagher, B.; Lambert, K.A.; Slaven, S.; Robinson, W.A.; Amato, C.M.; Couts, K.L.; Bemis, J.G.T.; Turner, J.A.; et al. BH3 mimetics induce apoptosis independent of DRP-1 in melanoma. Cell Death Dis. 2018, 9, 907. [Google Scholar] [CrossRef]
- Hartman, M.L.; Gajos-Michniewicz, A.; Talaj, J.A.; Mielczarek-Lewandowska, A.; Czyz, M. BH3 mimetics potentiate pro-apoptotic activity of encorafenib in BRAF(V600E) melanoma cells. Cancer Lett. 2021, 499, 122–136. [Google Scholar] [CrossRef] [PubMed]
- Fofaria, N.M.; Frederick, D.T.; Sullivan, R.J.; Flaherty, K.T.; Srivastava, S.K. Overexpression of Mcl-1 confers resistance to BRAFV600E inhibitors alone and in combination with MEK1/2 inhibitors in melanoma. Oncotarget 2015, 6, 40535–40556. [Google Scholar] [CrossRef]
- Haasler, L.; Kondadi, A.K.; Tsigaras, T.; von Montfort, C.; Graf, P.; Stahl, W.; Brenneisen, P. The BH3 mimetic (±) gossypol induces ROS-independent apoptosis and mitochondrial dysfunction in human A375 melanoma cells in vitro. Arch. Toxicol. 2021, 95, 1349–1365. [Google Scholar] [CrossRef]
- Jiang, C.C.; Wroblewski, D.; Yang, F.; Hersey, P.; Zhang, X.D. Human melanoma cells under endoplasmic reticulum stress are more susceptible to apoptosis induced by the BH3 mimetic obatoclax. Neoplasia 2009, 11, 945–955. [Google Scholar] [CrossRef]
- Redza-Dutordoir, M.; Averill-Bates, D.A. Activation of apoptosis signalling pathways by reactive oxygen species. Biochim. Biophys. Acta 2016, 1863, 2977–2992. [Google Scholar] [CrossRef] [PubMed]
- Franke, J.C.; Plotz, M.; Prokop, A.; Geilen, C.C.; Schmalz, H.G.; Eberle, J. New caspase-independent but ROS-dependent apoptosis pathways are targeted in melanoma cells by an iron-containing cytosine analogue. Biochem. Pharmacol. 2010, 79, 575–586. [Google Scholar] [CrossRef] [PubMed]
- Verhaegen, M.; Bauer, J.A.; Martin de la Vega, C.; Wang, G.; Wolter, K.G.; Brenner, J.C.; Nikolovska-Coleska, Z.; Bengtson, A.; Nair, R.; Elder, J.T.; et al. A novel BH3 mimetic reveals a mitogen-activated protein kinase-dependent mechanism of melanoma cell death controlled by p53 and reactive oxygen species. Cancer Res. 2006, 66, 11348–11359. [Google Scholar] [CrossRef]
- Howard, A.N.; Bridges, K.A.; Meyn, R.E.; Chandra, J. ABT-737, a BH3 mimetic, induces glutathione depletion and oxidative stress. Cancer Chemother. Pharmacol. 2009, 65, 41–54. [Google Scholar] [CrossRef]
- Winkler, M.; Friedrich, J.; Boedicker, C.; Dolgikh, N. Co-targeting MCL-1 and ERK1/2 kinase induces mitochondrial apoptosis in rhabdomyosarcoma cells. Transl. Oncol. 2022, 16, 101313. [Google Scholar] [CrossRef] [PubMed]
- Li, Z.; He, S.; Look, A.T. The MCL1-specific inhibitor S63845 acts synergistically with venetoclax/ABT-199 to induce apoptosis in T-cell acute lymphoblastic leukemia cells. Leukemia 2019, 33, 262–266. [Google Scholar] [CrossRef]
- Alcon, C.; Gomez Tejeda Zanudo, J.; Albert, R.; Wagle, N.; Scaltriti, M.; Letai, A.; Samitier, J.; Montero, J. ER+ Breast Cancer Strongly Depends on MCL-1 and BCL-xL Anti-Apoptotic Proteins. Cells 2021, 10, 1659. [Google Scholar] [CrossRef]
- Keuling, A.M.; Felton, K.E.; Parker, A.A.; Akbari, M.; Andrew, S.E.; Tron, V.A. RNA silencing of Mcl-1 enhances ABT-737-mediated apoptosis in melanoma: Role for a caspase-8-dependent pathway. PLoS ONE 2009, 4, e6651. [Google Scholar] [CrossRef]
- Bauer, D.; Werth, F.; Nguyen, H.A.; Kiecker, F.; Eberle, J. Critical role of reactive oxygen species (ROS) for synergistic enhancement of apoptosis by vemurafenib and the potassium channel inhibitor TRAM-34 in melanoma cells. Cell Death Dis. 2017, 8, e2594. [Google Scholar] [CrossRef]
- Berger, A.; Quast, S.A.; Plotz, M.; Kammermeier, A.; Eberle, J. Sensitization of melanoma cells for TRAIL-induced apoptosis by BMS-345541 correlates with altered phosphorylation and activation of Bax. Cell Death Dis. 2013, 4, e477. [Google Scholar] [CrossRef] [PubMed]
- Zhang, X.D.; Gillespie, S.K.; Borrow, J.M.; Hersey, P. The histone deacetylase inhibitor suberic bishydroxamate: A potential sensitizer of melanoma to TNF-related apoptosis-inducing ligand (TRAIL) induced apoptosis. Biochem. Pharmacol. 2003, 66, 1537–1545. [Google Scholar] [CrossRef] [PubMed]
- Giard, D.J.; Aaronson, S.A.; Todaro, G.J.; Arnstein, P.; Kersey, J.H.; Dosik, H.; Parks, W.P. In vitro cultivation of human tumors: Establishment of cell lines derived from a series of solid tumors. J. Natl. Cancer Inst. 1973, 51, 1417–1423. [Google Scholar] [CrossRef]
- Holzmann, B.; Lehmann, J.M.; Ziegler-Heitbrock, H.W.; Funke, I.; Riethmuller, G.; Johnson, J.P. Glycoprotein P3.58, associated with tumor progression in malignant melanoma, is a novel leukocyte activation antigen. Int. J. Cancer 1988, 41, 542–547. [Google Scholar] [CrossRef] [PubMed]
- Bean, M.A.; Bloom, B.R.; Herberman, R.B.; Old, L.J.; Oettgen, H.F.; Klein, G.; Terry, W.D. Cell-mediated cytotoxicity for bladder carcinoma: Evaluation of a workshop. Cancer Res. 1975, 35, 2902–2913. [Google Scholar] [PubMed]
- Carey, T.E.; Takahashi, T.; Resnick, L.A.; Oettgen, H.F.; Old, L.J. Cell surface antigens of human malignant melanoma: Mixed hemadsorption assays for humoral immunity to cultured autologous melanoma cells. Proc. Natl. Acad. Sci. USA 1976, 73, 3278–3282. [Google Scholar] [CrossRef] [PubMed]
- Chou, T.C. Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacol. Rev. 2006, 58, 621–681. [Google Scholar] [CrossRef]
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Peng, Z.; Gillissen, B.; Richter, A.; Sinnberg, T.; Schlaak, M.S.; Eberle, J. Effective Targeting of Melanoma Cells by Combination of Mcl-1 and Bcl-2/Bcl-xL/Bcl-w Inhibitors. Int. J. Mol. Sci. 2024, 25, 3453. https://doi.org/10.3390/ijms25063453
Peng Z, Gillissen B, Richter A, Sinnberg T, Schlaak MS, Eberle J. Effective Targeting of Melanoma Cells by Combination of Mcl-1 and Bcl-2/Bcl-xL/Bcl-w Inhibitors. International Journal of Molecular Sciences. 2024; 25(6):3453. https://doi.org/10.3390/ijms25063453
Chicago/Turabian StylePeng, Zhe, Bernhard Gillissen, Antje Richter, Tobias Sinnberg, Max S. Schlaak, and Jürgen Eberle. 2024. "Effective Targeting of Melanoma Cells by Combination of Mcl-1 and Bcl-2/Bcl-xL/Bcl-w Inhibitors" International Journal of Molecular Sciences 25, no. 6: 3453. https://doi.org/10.3390/ijms25063453