Synergistic inhibition of SCR1- and ERBB2-driven brain metastatic breast cancer cells

Abstract

Aim: Metastasis to the brain has become a major limitation to the life expectancy and quality of life for many patients with breast cancer. Unfortunately, other than radiation and palliative treatments with trastuzumab, and pertuzumab, no effective therapy for brain metastases is currently available. This study seeks to identify novel gene targets and pharmaceutical Intervention against breast cancer brain metastasis.

Methods: The detailed methods applied to this study, including comparative RNA sequencing and bioinformatics analysis of sequence data, ingenuity pathway analysis, protein-protein interaction analysis, high throughput screening of clinical and pre-clinical drugs, cell viability and proliferation assay, toxicity and apoptosis assay using fluorescence-activated cell sorting, real-time PCR, western blotting, statistical analysis of data.

Results: The study reveals critical roles for SRC, ERBB2, PIK3CA, and GABA in the proliferation and survival of breast cancer brain metastatic (BBM) cells and showed that SRC- and ERBB2-mediated activation of PIK3-AKT/mTOR signaling regulates BBM cell survival. Selective inhibition of these candidate genes alone or in combination induces robust apoptosis in BBM cells

Conclusion: The findings of this study provide a rationale for further preclinical evaluation of SRC-targeting regimens in combination with ERBB2 inhibitors and/or GABA agonists to target breast cancer brain metastasis.

Keywords

Breast cancer / brain metastasis / human epidermal growth factor receptor 2 / ERBB2 / SRC / astrocytes / PI3KCA

References

[1] Swain SM,Miles D,Quah C.Incidence of central nervous system metastases in patients with HER2-positive metastatic breast cancer treated with pertuzumab, trastuzumab, and docetaxel: results from the randomized phase III study CLEOPATRA..Ann Oncol2014;25:1116-21 PMCID:PMC4037862

[2] Brufsky AM,Rugo HS,Tan-Chiu E.Central nervous system metastases in patients with HER2-positive metastatic breast cancer: incidence, treatment, and survival in patients from registHER..Clin Cancer Res2011;17:4834-43

[3] Moliterni A,Stella GC,Bianchi GV.Long-term results from INT-HER study: retrospective evaluation of adjuvant trastuzumab in unselected HER2-positive breast cancer patients-single institution experience..J Clin Oncol2014;32:

[4] Ow SGW,Chieng WS,Lee SC.Clinical outcome of HER2-positive breast cancer patients after failure on adjuvant trastuzumab: the potential of the time to relapse..Clin Oncol2014;26:174-5

[5] Ramakrishna N,Lin NU.Recommendations on disease management for patients with advanced human epidermal growth factor receptor 2-positive breast cancer and brain metastases: ASCO clinical practice guideline update summary..J Oncol Pract2018;14:505-7

[6] Ramakrishna N,Chandarlapaty S,Davidson NE.Recommendations on disease management for patients with advanced human epidermal growth factor receptor 2-positive breast cancer and brain metastases: American Society of Clinical Oncology clinical practice guideline..J Clin Oncol2014;32:2100-8 PMCID:PMC6366342

[7] Steeg PS,Smith QR.Brain metastases as preventive and therapeutic targets..Nat Rev Cancer2011;11:352-63

[8] Stemmler HJ.Central nervous system metastases in HER-2-overexpressing metastatic breast cancer: a treatment challenge..Oncologist2008;13:739-50

[9] Lin NU,Liu MC,Come SE.Phase II trial of lapatinib for brain metastases in patients with human epidermal growth factor receptor 2-positive breast cancer..J Clin Oncol2008;26:1993-9 PMCID:PMC4524351

[10] Lin NU,Paul D,Christodoulou C.Multicenter phase II study of lapatinib in patients with brain metastases from HER2-positive breast cancer..Clin Cancer Res2009;15:1452-9

[11] Yap TA,Adam J,Spicer J.Phase I trial of the irreversible EGFR and HER2 kinase inhibitor BIBW 2992 in patients with advanced solid tumors..J Clin Oncol2010;28:3965-72

[12] Da Silva L,Smart CE,Waddell N.HER3 and downstream pathways are involved in colonization of brain metastases from breast cancer..Breast Cancer Res2010;12:R46 PMCID:PMC2949633

[13] Kodack DP,Ferraro GB,Jain RK.Emerging strategies for treating brain metastases from breast cancer..Cancer Cell2015;27:163-75 PMCID:PMC4325273

[14] Lin NU,Younger WJ,Freedman RA.Phase II trial of carboplatin (C) and bevacizumab (BEV) in patients (pts) with breast cancer brain metastases (BCBM)..J Clin Oncol2013;31:

[15] Lu YS,Lin CH,Yeh DC.Bevacizumab, etoposide, and cisplatin (BEEP) in brain metastases of breast cancer progressing from radiotherapy: results of the first stage of a multicenter phase II study..J Clin Oncol2012;30:

[16] Bartsch R,Preusser M.Breast cancer brain metastases responding to primary systemic therapy with T-DM1..J Neurooncol2014;116:205-6

[17] Krop IE,Gonzalez-Martin A,Ferrero JM.Trastuzumab emtansine versus treatment of physician's choice for pretreated HER2-positive advanced breast cancer (TH3RESA): a randomised, open-label, phase 3 trial..Lancet Oncol2014;15:689-99

[18] Verma S,Gianni L,Welslau M.Trastuzumab emtansine for HER2-positive advanced breast cancer..N Engl J Med2012;367:1783-91 PMCID:PMC5125250

[19] Beadnell TC,Rose MM,Danysh BP.Src-mediated regulation of the PI3K pathway in advanced papillary and anaplastic thyroid cancer..Oncogenesis2018;7:23 PMCID:PMC5833015

[20] Rexer BN,Rinehart C,Granja-Ingram ND.Phosphoproteomic mass spectrometry profiling links Src family kinases to escape from HER2 tyrosine kinase inhibition..Oncogene2011;30:4163-74 PMCID:PMC3204390

[21] Zhang SY,Zhang L,Lowery FJ.Src family kinases as novel therapeutic targets to treat breast cancer brain metastases..Cancer Res2013;73:5764-74 PMCID:PMC3781592

[22] Du Y,Zheng H,Li S.GABA exists as a negative regulator of cell proliferation in spermaogonial stem cells..Cell Mol Biol Lett2013;18:149-62 PMCID:PMC6275874

[23] Young SZ.GABA's control of stem and cancer cell proliferation in adult neural and peripheral niches..Physiology (Bethesda)2009;24:171-85 PMCID:PMC2931807

[24] Young SZ.GABA's control of stem and cancer cell proliferation in adult neural and peripheral niches..Physiology2009;24:171-85 PMCID:PMC2931807

[25] Choy C,Neman J,Duenas MJ.Cooperation of neurotrophin receptor TrkB and Her2 in breast cancer cells facilitates brain metastases..Breast Cancer Res2017;19:51 PMCID:PMC5406906

[26] Opdam FL,Beijnen JH.Lapatinib for advanced or metastatic breast cancer..Oncologist2012;17:536-42 PMCID:PMC3336826

[27] Freedman RA,Ottesen RA,He Y.Use of adjuvant trastuzumab in women with human epidermal growth factor receptor 2 (HER2)-positive breast cancer by race/ethnicity and education within the National Comprehensive Cancer Network..Cancer2013;119:839-46 PMCID:PMC3565006

[28] Collins I,Mileshkin L.Adjuvant trastuzumab in HER2-positive breast cancer..N Engl J Med2012;366:664; author reply 664-6

[29] Chen L,Yao L,Zuo WJ.Characterization of PIK3CA and PIK3R1 somatic mutations in Chinese breast cancer patients..Nat Commun2018;9:1357 PMCID:PMC5893593

[30] Jensen JD,Laenkholm AV,Jensen MB.PIK3CA mutations, PTEN, and pHER2 expression and impact on outcome in HER2-positive early-stage breast cancer patients treated with adjuvant chemotherapy and trastuzumab..Ann Oncol2012;23:2034-42

[31] Zardavas D,Milne RL,Fountzilas G.Tumor PIK3CA genotype and prognosis in early-stage breast cancer: a pooled analysis of individual patient data..J Clin Oncol2018;36:981-90

[32] Paplomata E.The PI3K/AKT/mTOR pathway in breast cancer: targets, trials and biomarkers..Ther Adv Med Oncol2014;6:154-66 PMCID:PMC4107712

[33] Abraham J.PI3K/AKT/mTOR pathway inhibitors: the ideal combination partners for breast cancer therapies?.Expert Rev Anticancer Ther2015;15:51-68

[34] Bahrami A,Shahidsales S,Hasanzadeh M.The therapeutic potential of PI3K/Akt/mTOR inhibitors in breast cancer: rational and progress..J Cell Biochem2018;119:213-22

[35] Chia S,Joy AA,Gorr M.Novel agents and associated toxicities of inhibitors of the pi3k/Akt/mtor pathway for the treatment of breast cancer..Curr Oncol2015;22:33-48 PMCID:PMC4324342

[36] Ghayad SE.Inhibitors of the PI3K/Akt/mTOR pathway: new hope for breast cancer patients..Recent Pat Anticancer Drug Discov2010;5:29-57

[37] Lee JJ,Yap YS.PI3K/Akt/mTOR inhibitors in breast cancer..Cancer Biol Med2015;12:342-54 PMCID:PMC4706528

[38] Bos PD,Nadal C,Gomis RR.Genes that mediate breast cancer metastasis to the brain..Nature2009;459:1005-9 PMCID:PMC2698953

[39] Ulasov IV,Pytel P,Liu F.Clinical significance of KISS1 protein expression for brain invasion and metastasis..Cancer2012;118:2096-105 PMCID:PMC3674482

[40] Neman J,Wilczynski S,Choy C.Human breast cancer metastases to the brain display GABAergic properties in the neural niche..Proc Natl Acad Sci U S A2014;111:984-9 PMCID:PMC3903266

[41] Smits A,Elsir T,Nister M.GABA-A channel subunit expression in human glioma correlates with tumor histology and clinical outcome..PLoS One2012;7:e37041 PMCID:PMC3355166

[42] Quail DF.The microenvironmental landscape of brain tumors..Cancer Cell2017;31:326-41 PMCID:PMC5424263

[43] Pontes A,Hu W.Novel functions of GABA signaling in adult neurogenesis..Front Biol (Beijing)2013;8: PMCID:PMC3839947

[44] Berg DA,Song H.Neurotransmitter-mediated control of neurogenesis in the adult vertebrate brain..Development2013;140:2548-61 PMCID:PMC3666382

[45] Pohl SG,Agostino M,Kumar AP.Wnt signaling in triple-negative breast cancer..Oncogenesis2017;6:e310 PMCID:PMC5520491

[46] Bilir B,Moreno CS.Wnt signaling blockage inhibits cell proliferation and migration, and induces apoptosis in triple-negative breast cancer cells..J Transl Med2013;11:280 PMCID:PMC4228255

[47] King TD,Li YH.The wnt/beta-catenin signaling pathway: a potential therapeutic target in the treatment of triple negative breast cancer..J Cell Biochem2012;113:13-8