BRAF Modulates Lipid Use and Accumulation
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Plasmids and Gene Constructs
2.2. Stable Gene Expression, Cell Lines, and Culture Conditions
2.3. Mass Spectrometry, Metabolomics, and Lipidomics
2.4. Metabolic Flux Assay
2.5. Immunofluorescence and Microscopy
2.6. Melanoma Patient Samples
2.7. Sanger Sequencing and Quantitative Real-Time PCR
2.8. Western Immunoblotting
2.9. Statistical Analyses
3. Results
3.1. BRAF Expression and Mutation Modulates Metabolic Profiles
3.2. Cells Expressing BRAF V600E Do Not Exhibit Warburg-Like Metabolism
3.3. BRAF V600E Expression Promotes Formation of Tunneling Nanotube (TNT)-like Protrusions Which Preferentially Accumulate Lipids
3.4. Expression of BRAF V600E Enriches for Immunomodulatory Profiles
3.5. Circulating Plasma Lipids Are Increased in Melanoma Patients That Do Not Respond to MAPK Inhibitor Therapy
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Liu, F.; Yang, X.; Geng, M.; Huang, M. Targeting ERK, an Achilles’ Heel of the MAPK pathway, in cancer therapy. Acta Pharm. Sin. B 2018, 8, 552–562. [Google Scholar] [CrossRef] [PubMed]
- Yaeger, R.; Corcoran, R.B. Targeting Alterations in the RAF–MEK Pathway. Cancer Discov. 2019, 9, 329–341. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Gardner, A.M.; Vaillancourt, R.R.; Lange-Carter, C.A.; Johnson, G.L. MEK-1 phosphorylation by MEK kinase, Raf, and mitogen-activated protein kinase: Analysis of phosphopeptides and regulation of activity. Mol. Biol. Cell 1994, 5, 193–201. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Graves, J.D.; Campbell, J.S.; Krebs, E.G. Protein Serine/Threonine Kinases of the MAPK Cascade. Ann. N. Y. Acad. Sci. 1995, 766, 320–343. [Google Scholar] [CrossRef]
- Ahn, N.G. The MAP kinase cascade. Discovery of a new signal transduction pathway. Mol. Cell. Biochem. 1993, 127–128, 201–209. [Google Scholar] [CrossRef]
- 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]
- Bollag, G.; Hirth, P.; Tsai, J.; Zhang, J.; Ibrahim, P.N.; Cho, H.; Spevak, W.; Zhang, C.; Zhang, Y.; Habets, G.; et al. Clinical efficacy of a RAF inhibitor needs broad target blockade in BRAF-mutant melanoma. Nature 2010, 467, 596–599. [Google Scholar] [CrossRef]
- Wan, P.T.; Garnett, M.J.; Roe, S.M.; Lee, S.; Niculescu-Duvaz, D.; Good, V.M.; Jones, C.M.; Marshall, C.J.; Springer, C.J.; Barford, D.; et al. Mechanism of activation of the RAF-ERK signaling pathway by oncogenic mutations of B-RAF. Cell 2004, 116, 855–867. [Google Scholar] [CrossRef] [Green Version]
- Yao, Z.; Torres, N.M.; Tao, A.; Gao, Y.; Luo, L.; Li, Q.; De Stanchina, E.; Abdel-Wahab, O.; Solit, D.B.; Poulikakos, P.I.; et al. BRAF Mutants Evade ERK-Dependent Feedback by Different Mechanisms that Determine Their Sensitivity to Pharmacologic Inhibition. Cancer Cell 2015, 28, 370–383. [Google Scholar] [CrossRef] [Green Version]
- Garnett, M.J.; Rana, S.; Paterson, H.; Barford, D.; Marais, R. Wild-Type and Mutant B-RAF Activate C-RAF through Distinct Mechanisms Involving Heterodimerization. Mol. Cell 2005, 20, 963–969. [Google Scholar] [CrossRef]
- Haling, J.R.; Sudhamsu, J.; Yen, I.; Sideris, S.; Sandoval, W.; Phung, W.; Bravo, B.J.; Giannetti, A.M.; Peck, A.; Masselot, A.; et al. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell 2014, 26, 402–413. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Tiacci, E.; Trifonov, V.; Schiavoni, G.; Holmes, A.; Kern, W.; Martelli, M.P.; Pucciarini, A.; Bigerna, B.; Pacini, R.; Wells, V.A.; et al. BRAFMutations in Hairy-Cell Leukemia. N. Engl. J. Med. 2011, 364, 2305–2315. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Blombery, P.A.; Wong, S.Q.; Hewitt, C.A.; Dobrovic, A.; Maxwell, E.L.; Juneja, S.; Grigoriadis, G.; Westerman, D.A. Detection of BRAF mutations in patients with hairy cell leukemia and related lymphoproliferative disorders. Haematologica 2012, 97, 780–783. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Pratilas, C.A.; Xing, F.; Solit, D.B. Targeting Oncogenic BRAF in Human Cancer. Curr. Top. Microbiol. Immunol. 2010, 355, 83–98. [Google Scholar] [CrossRef] [Green Version]
- Hélias-Rodzewicz, Z.; Funck-Brentano, E.; Baudoux, L.; Jung, C.K.; Zimmermann, U.; Marin, C.; Clérici, T.; Le Gall, C.; Peschaud, F.; Taly, V.; et al. Variations of BRAF mutant allele percentage in melanomas. BMC Cancer 2015, 15, 497. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Dasgupta, T.; Haas-Kogan, D.A. The Combination of Novel Targeted Molecular Agents and Radiation in the Treatment of Pediatric Gliomas. Front. Oncol. 2013, 3, 110. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Molteni, R.; Biavasco, R.; Stefanoni, D.; Nemkov, T.; Domínguez-Andrés, J.; Arts, R.J.; Merelli, I.; Mazza, D.; Zambrano, S.; Panigada, M.; et al. Oncogene-induced maladaptive activation of trained immunity in the pathogenesis and treatment of Erdheim-Chester disease. Blood 2021, 138, 1554–1569. [Google Scholar] [CrossRef]
- Paton, E.L.; Turner, J.A.; Schlaepfer, I.R. Overcoming Resistance to Therapies Targeting the MAPK Pathway in BRAF-Mutated Tumours. J. Oncol. 2020, 2020, 1079827. [Google Scholar] [CrossRef]
- Hardeman, K.; Peng, C.; Paudel, B.; Meyer, C.; Luong, T.; Tyson, D.; Young, J.D.; Quaranta, V.; Fessel, J.P. Dependence on Glycolysis Sensitizes BRAF-mutated Melanomas For Increased Response To Targeted BRAF Inhibition. Sci. Rep. 2017, 7, 42604. [Google Scholar] [CrossRef] [Green Version]
- Parmenter, T.J.; Kleinschmidt, M.; Kinross, K.M.; Bond, S.T.; Li, J.; Kaadige, M.R.; Rao, A.; Sheppard, K.; Hugo, W.; Pupo, G.M.; et al. Response of BRAF-Mutant Melanoma to BRAF Inhibition Is Mediated by a Network of Transcriptional Regulators of Glycolysis. Cancer Discov. 2014, 4, 423–433. [Google Scholar] [CrossRef] [Green Version]
- Haq, R.; Shoag, J.; Andreu-Perez, P.; Yokoyama, S.; Edelman, H.; Rowe, G.C.; Frederick, D.T.; Hurley, A.D.; Nellore, A.; Kung, A.; et al. Oncogenic BRAF Regulates Oxidative Metabolism via PGC1α and MITF. Cancer Cell 2013, 23, 302–315. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Talebi, A.; Dehairs, J.; Rambow, F.; Rogiers, A.; Nittner, D.; Derua, R.; Vanderhoydonc, F.; Duarte, J.A.G.; Bosisio, F.; Eynde, K.V.D.; et al. Sustained SREBP-1-dependent lipogenesis as a key mediator of resistance to BRAF-targeted therapy. Nat. Commun. 2018, 9, 2500. [Google Scholar] [CrossRef] [PubMed]
- Aloia, A.; Müllhaupt, D.; Chabbert, C.D.; Eberhart, T.; Flückiger-Mangual, S.; Vukolic, A.; Eichhoff, O.; Irmisch, A.; Alexander, L.T.; Scibona, E.; et al. A Fatty Acid Oxidation-dependent Metabolic Shift Regulates the Adaptation of BRAF-mutated Melanoma to MAPK Inhibitors. Clin. Cancer Res. 2019, 25, 6852–6867. [Google Scholar] [CrossRef] [PubMed]
- Turner, J.A.; Bemis, J.G.T.; Bagby, S.M.; Capasso, A.; Yacob, B.W.; Chimed, T.-S.; Van Gulick, R.; Lee, H.; Tobin, R.; Tentler, J.J.; et al. BRAF fusions identified in melanomas have variable treatment responses and phenotypes. Oncogene 2019, 38, 1296–1308. [Google Scholar] [CrossRef] [PubMed]
- Zarini, S.; Hankin, J.A.; Murphy, R.C.; Gijón, M.A. Lysophospholipid acyltransferases and eicosanoid biosynthesis in zebrafish myeloid cells. Prostaglandins Other Lipid Mediat. 2014, 113–115, 52–61. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Nemkov, T.; Reisz, J.A.; Gehrke, S.; Hansen, K.C.; D’Alessandro, A. High-Throughput Metabolomics: Isocratic and Gradient Mass Spectrometry-Based Methods. Methods Mol. Biol. 2019, 1978, 13–26. [Google Scholar] [CrossRef]
- Reisz, J.A.; Zheng, C.; D’Alessandro, A.; Nemkov, T. Untargeted and Semi-targeted Lipid Analysis of Biological Samples Using Mass Spectrometry-Based Metabolomics. Methods Mol. Biol. 2019, 1978, 121–135. [Google Scholar] [CrossRef]
- Garnett, M.J.; Marais, R. Guilty as charged: B-RAF is a human oncogene. Cancer Cell 2004, 6, 313–319. [Google Scholar] [CrossRef] [Green Version]
- Lv, X.; Wang, D.; Ma, Y.; Long, Z. Analysis of the oncogene BRAF mutation and the correlation of the expression of wild-type BRAF and CREB1 in endometriosis. Int. J. Mol. Med. 2018, 41, 1349–1356. [Google Scholar] [CrossRef] [Green Version]
- Senol, A.D.; Pepe, A.; Grudina, C.; Sassoon, N.; Reiko, U.; Bousset, L.; Melki, R.; Piel, J.; Gugger, M.; Zurzolo, C. Effect of tolytoxin on tunneling nanotube formation and function. Sci. Rep. 2019, 9, 5741. [Google Scholar] [CrossRef]
- Kimura, S.; Hase, K.; Ohno, H. The molecular basis of induction and formation of tunneling nanotubes. Cell Tissue Res. 2013, 352, 67–76. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Liu, J.J.; Green, P.; Mann, J.J.; Rapoport, S.I.; Sublette, M.E. Pathways of polyunsaturated fatty acid utilization: Implications for brain function in neuropsychiatric health and disease. Brain Res. 2015, 1597, 220–246. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Levy, B.D.; Clish, C.; Schmidt, B.A.; Gronert, K.; Serhan, C.N. Lipid mediator class switching during acute inflammation: Signals in resolution. Nat. Immunol. 2001, 2, 612–619. [Google Scholar] [CrossRef] [PubMed]
- Matloubian, M.; Lo, C.G.; Cinamon, G.; Lesneski, M.J.; Xu, Y.; Brinkmann, V.; Allende, M.L.; Proia, R.; Cyster, J.G. Lymphocyte egress from thymus and peripheral lymphoid organs is dependent on S1P receptor 1. Nature 2004, 427, 355–360. [Google Scholar] [CrossRef] [PubMed]
- Hsu, P.P.; Sabatini, D.M. Cancer Cell Metabolism: Warburg and Beyond. Cell 2008, 134, 703–707. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Liberti, M.V.; Locasale, J.W. The Warburg Effect: How Does it Benefit Cancer Cells? Trends Biochem. Sci. 2016, 41, 211–218. [Google Scholar] [CrossRef] [Green Version]
- Warburg, O. On the Origin of Cancer Cells. Science 1956, 123, 309–314. [Google Scholar] [CrossRef]
- Warburg, O.; Wind, F.; Negelein, E. The metabolism of tumors in the body. J. Gen. Physiol. 1927, 8, 519–530. [Google Scholar] [CrossRef] [Green Version]
- Knudson, A.G. Two genetic hits (more or less) to cancer. Nat. Rev. Cancer 2001, 1, 157–162. [Google Scholar] [CrossRef]
- Dhomen, N.; Reis-Filho, J.S.; Dias, S.D.R.; Hayward, R.; Savage, K.; Delmas, V.; LaRue, L.; Pritchard, C.; Marais, R. Oncogenic Braf Induces Melanocyte Senescence and Melanoma in Mice. Cancer Cell 2009, 15, 294–303. [Google Scholar] [CrossRef]
- Pollock, P.; Harper, U.L.; Hansen, K.S.; Yudt, L.M.; Stark, M.; Robbins, C.M.; Moses, T.Y.; Hostetter, G.; Wagner, U.; Kakareka, J.; et al. High frequency of BRAF mutations in nevi. Nat. Genet. 2003, 33, 19–20. [Google Scholar] [CrossRef] [PubMed]
- Yazdi, A.S.; Palmedo, G.; Flaig, M.J.; Puchta, U.; Reckwerth, A.; Rütten, A.; Mentzel, T.; Hügel, H.; Hantschke, M.; Schmid-Wendtner, M.-H.; et al. Mutations of the BRAF Gene in Benign and Malignant Melanocytic Lesions. J. Investig. Dermatol. 2003, 121, 1160–1162. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Falini, B.; Martelli, M.P.; Tiacci, E. BRAF V600E mutation in hairy cell leukemia: From bench to bedside. Blood 2016, 128, 1918–1927. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Dupont, M.; Souriant, S.; Lugo-Villarino, G.; Maridonneau-Parini, I.; Vérollet, C. Tunneling Nanotubes: Intimate Communication between Myeloid Cells. Front. Immunol. 2018, 9, 43. [Google Scholar] [CrossRef] [Green Version]
- Tomić, S.; Joksimović, B.; Bekić, M.; Vasiljević, M.; Milanović, M.; Čolić, M.; Vučević, D. Prostaglanin-E2 Potentiates the Suppressive Functions of Human Mononuclear Myeloid-Derived Suppressor Cells and Increases Their Capacity to Expand IL-10-Producing Regulatory T Cell Subsets. Front. Immunol. 2019, 10, 475. [Google Scholar] [CrossRef] [Green Version]
- Mathew, D.; Kremer, K.N.; Strauch, P.; Tigyi, G.; Pelanda, R.; Torres, R.M. LPA5 Is an Inhibitory Receptor That Suppresses CD8 T-Cell Cytotoxic Function via Disruption of Early TCR Signaling. Front. Immunol. 2019, 10, 1159. [Google Scholar] [CrossRef] [Green Version]
- Ridley, A.J.; Hall, A. The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factors. Cell 1992, 70, 389–399. [Google Scholar] [CrossRef]
- Pyne, N.J.; Pyne, S. Sphingosine 1-phosphate and cancer. Nat. Rev. Cancer 2010, 10, 489–503. [Google Scholar] [CrossRef] [Green Version]
- Schlaepfer, I.R.; Rider, L.; Rodrigues, L.U.; Gijón, M.A.; Pac, C.T.; Romero, L.; Cimic, A.; Sirintrapun, S.J.; Glodé, L.M.; Eckel, R.H.; et al. Lipid Catabolism via CPT1 as a Therapeutic Target for Prostate Cancer. Mol. Cancer Ther. 2014, 13, 2361–2371. [Google Scholar] [CrossRef] [Green Version]
Patient No. | Sex | BRAF Status | Stage | Treatment | Response |
---|---|---|---|---|---|
1 | F | V600E | IV | Dabrafenib + Trametinib | R |
2 | M | V600E | IV | Dabrafenib + Trametinib | R |
3 | M | V600E | IV | Dabrafenib + Trametinib | R |
4 | F | V600E | IV | Dabrafenib + Trametinib | R |
5 | M | V600E | IV | Dabrafenib + Trametinib | R |
6 | M | V600E | IV | Dabrafenib + Trametinib | R |
7 | F | V600E | IV | Dabrafenib + Trametinib | NR |
8 | F | V600E | III | Vemurafenib | NR |
9 | F | V600E | IV | Vemurafenib | NR |
10 | M | V600E | IV | Vemurafenib/Dabrafenib + Trametinib | NR/NR |
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Turner, J.A.; Paton, E.L.; Van Gulick, R.; Stefanoni, D.; Cendali, F.; Reisz, J.; Tobin, R.P.; McCarter, M.; D’Alessandro, A.; Torres, R.M.; et al. BRAF Modulates Lipid Use and Accumulation. Cancers 2022, 14, 2110. https://doi.org/10.3390/cancers14092110
Turner JA, Paton EL, Van Gulick R, Stefanoni D, Cendali F, Reisz J, Tobin RP, McCarter M, D’Alessandro A, Torres RM, et al. BRAF Modulates Lipid Use and Accumulation. Cancers. 2022; 14(9):2110. https://doi.org/10.3390/cancers14092110
Chicago/Turabian StyleTurner, Jacqueline A., Emily L. Paton, Robert Van Gulick, Davide Stefanoni, Francesca Cendali, Julie Reisz, Richard P. Tobin, Martin McCarter, Angelo D’Alessandro, Raul M. Torres, and et al. 2022. "BRAF Modulates Lipid Use and Accumulation" Cancers 14, no. 9: 2110. https://doi.org/10.3390/cancers14092110