Download PDF
J Neurol Surg B Skull Base 2022; 83(01): 087-098
DOI: 10.1055/s-0040-1716694
Original Article

Advancement of PI3 Kinase Inhibitor Combination Therapies for PI3K-Aberrant Chordoma

Molly E. Heft Neal‡   *
1   Department of Otolaryngology—Head and Neck Surgery, University of Michigan, Ann Arbor, Michigan, United States
,
Nicole L. Michmerhuizen‡   *
1   Department of Otolaryngology—Head and Neck Surgery, University of Michigan, Ann Arbor, Michigan, United States
2   Department of Pharmacology, University of Michigan Medical School, Ann Arbor, Michigan, United States
,
Kevin J. Kovatch
1   Department of Otolaryngology—Head and Neck Surgery, University of Michigan, Ann Arbor, Michigan, United States
,
John Henry J. Owen
1   Department of Otolaryngology—Head and Neck Surgery, University of Michigan, Ann Arbor, Michigan, United States
,
Jingyi Zhai
3   Department of Biostatistics, School of Public Heath, University of Michigan, Ann Arbor, Michigan, United States
,
Hui Jiang
3   Department of Biostatistics, School of Public Heath, University of Michigan, Ann Arbor, Michigan, United States
4   Rogel Cancer Center, University of Michigan Medical School, Ann Arbor, Michigan, United States
,
Erin L. McKean
1   Department of Otolaryngology—Head and Neck Surgery, University of Michigan, Ann Arbor, Michigan, United States
,
Mark E.P. Prince‡   *
1   Department of Otolaryngology—Head and Neck Surgery, University of Michigan, Ann Arbor, Michigan, United States
,
J. Chad Brenner   *
1   Department of Otolaryngology—Head and Neck Surgery, University of Michigan, Ann Arbor, Michigan, United States
2   Department of Pharmacology, University of Michigan Medical School, Ann Arbor, Michigan, United States
4   Rogel Cancer Center, University of Michigan Medical School, Ann Arbor, Michigan, United States
› Author Affiliations Funding M.E.H.N. received funding from AAO-HNSF Resident Research Award 513933 and NIH grant T32 DC005356. J.C.B. received funding from NIH Grants P30-CA046592 and the American Cancer Society. Funding sources had no role in study design, collection/analysis/interpretation of data, writing of decision for submission of this article.
› Further Information
    Permissions and Reprints

Abstract

Objectives Targeted inhibitors of the PI3 kinase (PI3K) pathway have shown promising but incomplete antitumor activity in preclinical chordoma models. The aim of this study is to advance methodology for a high-throughput drug screen using chordoma models to identify new combination therapies for chordoma.

Study Design Present work is an in vitro study.

Setting The study conducted at an academic research laboratory.

Materials and Methods An in vitro study on automated high-throughput screening of chordoma cells was performed using a library of 1,406 drugs as both mono- and combination therapies with PI3K inhibitors. Combination indices were determined for dual therapies and synergistic outliers were identified as potential therapeutic agents. T (brachyury) siRNA knockdown in combination with PI3K pathway inhibition was also assessed.

Results Fifty-nine combination therapies were identified as having potential therapeutic efficacy. Effective combinations included PI3K inhibitors with GSK1838705A (ALK/IGF-1R inhibitor), LY2874455 (VEGFR/FGFR inhibitor), El1 (selective Ezh2 inhibitor), and (-)-p-bromotetramisole oxalate (alkaline phosphatase inhibitor). The top ranking targets identified included ALK, PDGFR, VEGFR, aurora kinase, and BCL-2. T (brachyury) inhibition produced significant reduction in cell viability and growth; however PI3K inhibition in combination with T (brachyury) knockdown did not result in further reduction in growth and viability in vitro.

Conclusion High throughput with in vitro combination screening is feasible with chordoma cells and allows for rapid identification of synergistic dual-therapies. Potential combination therapies and targetable pathways were identified. T (brachyury) knockdown produced significant reduction in cell viability, but did not show additional benefit with PI3K pathway inhibition in this model. Further in vitro and in vivo validation of these therapeutic combinations is warranted.

Keywords

chordoma - brachyury - PI3K - high throughput

Note

This study was presented at AAO-HNSF Annual Meeting, 2019.


* Authors contributed equally.


Supplementary Material



Publication History

Received: 05 May 2020

Accepted: 19 July 2020

Article published online:
12 October 2020

© 2020. Thieme. All rights reserved.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 

  • References

  • 1 Smoll NR, Gautschi OP, Radovanovic I, Schaller K, Weber DC. Incidence and relative survival of chordomas: the standardized mortality ratio and the impact of chordomas on a population. Cancer 2013; 119 (11) 2029-2037
    CrossrefPubMedGoogle Scholar
  • 2 Colia V, Stacchiotti S. Medical treatment of advanced chordomas. Eur J Cancer 2017; 83: 220-228
    CrossrefPubMedGoogle Scholar
  • 3 Azzarelli A, Quagliuolo V, Cerasoli S. et al. Chordoma: natural history and treatment results in 33 cases. J Surg Oncol 1988; 37 (03) 185-191
    CrossrefPubMedGoogle Scholar
  • 4 Meng T, Jin J, Jiang C. et al. Molecular targeted therapy in the treatment of chordoma: a systematic review. Front Oncol 2019; 9: 30
    CrossrefPubMedGoogle Scholar
  • 5 McMaster ML, Goldstein AM, Bromley CM, Ishibe N, Parry DM. Chordoma: incidence and survival patterns in the United States, 1973-1995. Cancer Causes Control 2001; 12 (01) 1-11
    CrossrefPubMedGoogle Scholar
  • 6 Tarpey PS, Behjati S, Young MD. et al. The driver landscape of sporadic chordoma. Nat Commun 2017; 8 (01) 890
    CrossrefPubMedGoogle Scholar
  • 7 Tauziéde-Espariat A, Bresson D, Polivka M. et al. Prognostic and therapeutic markers in chordomas: a study of 287 tumors. J Neuropathol Exp Neurol 2016; 75 (02) 111-120
    CrossrefPubMedGoogle Scholar
  • 8 Choy E, MacConaill LE, Cote GM. et al. Genotyping cancer-associated genes in chordoma identifies mutations in oncogenes and areas of chromosomal loss involving CDKN2A, PTEN, and SMARCB1. PLoS One 2014; 9 (07) e101283
    CrossrefPubMedGoogle Scholar
  • 9 Otani R, Mukasa A, Shin M. et al. Brachyury gene copy number gain and activation of the PI3K/Akt pathway: association with upregulation of oncogenic Brachyury expression in skull base chordoma. J Neurosurg 2018; 128 (05) 1428-1437
    CrossrefPubMedGoogle Scholar
  • 10 Presneau N, Shalaby A, Idowu B. et al. Potential therapeutic targets for chordoma: PI3K/AKT/TSC1/TSC2/mTOR pathway. Br J Cancer 2009; 100 (09) 1406-1414
    CrossrefPubMedGoogle Scholar
  • 11 Schwab J, Antonescu C, Boland P. et al. Combination of PI3K/mTOR inhibition demonstrates efficacy in human chordoma. Anticancer Res 2009; 29 (06) 1867-1871
    PubMedGoogle Scholar
  • 12 Michmerhuizen NL, Owen JH, Heft Neal ME. et al. Rationale for the advancement of PI3K pathway inhibitors for personalized chordoma therapy. J Neurooncol 2020; 147 (01) 25-35
    CrossrefPubMedGoogle Scholar
  • 13 Tamborini E, Virdis E, Negri T. et al. Analysis of receptor tyrosine kinases (RTKs) and downstream pathways in chordomas. Neuro-oncol 2010; 12 (08) 776-789
    CrossrefPubMedGoogle Scholar
  • 14 Sharifnia T, Wawer MJ, Chen T. et al. Small-molecule targeting of brachyury transcription factor addiction in chordoma. Nat Med 2019; 25 (02) 292-300
    CrossrefPubMedGoogle Scholar
  • 15 Jambhekar NA, Rekhi B, Thorat K, Dikshit R, Agrawal M, Puri A. Revisiting chordoma with brachyury, a “new age” marker: analysis of a validation study on 51 cases. Arch Pathol Lab Med 2010; 134 (08) 1181-1187
    CrossrefPubMedGoogle Scholar
  • 16 Vujovic S, Henderson S, Presneau N. et al. Brachyury, a crucial regulator of notochordal development, is a novel biomarker for chordomas. J Pathol 2006; 209 (02) 157-165
    CrossrefPubMedGoogle Scholar
  • 17 Wasserman JK, Gravel D, Purgina B. Chordoma of the head and neck: a review. Head Neck Pathol 2018; 12 (02) 261-268
    CrossrefPubMedGoogle Scholar
  • 18 Kitamura Y, Sasaki H, Yoshida K. Genetic aberrations and molecular biology of skull base chordoma and chondrosarcoma. Brain Tumor Pathol 2017; 34 (02) 78-90
    CrossrefPubMedGoogle Scholar
  • 19 Sun HI, Guduk M, Gucyetmez B, Yapicier O, Pamir MN. Chordoma: immunohistochemical analysis of brachury. Turk Neurosurg 2018; 28 (02) 174-178
    PubMedGoogle Scholar
  • 20 Sa JK, Lee IH, Hong SD, Kong DS, Nam DH. Genomic and transcriptomic characterization of skull base chordoma. Oncotarget 2017; 8 (01) 1321-1328
    CrossrefPubMedGoogle Scholar
  • 21 Hu Y, Mintz A, Shah SR, Quinones-Hinojosa A, Hsu W. The FGFR/MEK/ERK/brachyury pathway is critical for chordoma cell growth and survival. Carcinogenesis 2014; 35 (07) 1491-1499
    CrossrefPubMedGoogle Scholar
  • 22 Jian J, Zhong N, Jiang D. et al. The embryonic transcription factor Brachyury confers chordoma chemoresistance via upregulating CA9. Am J Transl Res 2018; 10 (03) 936-947
    PubMedGoogle Scholar
  • 23 Hsu W, Mohyeldin A, Shah SR. et al. Generation of chordoma cell line JHC7 and the identification of Brachyury as a novel molecular target. J Neurosurg 2011; 115 (04) 760-769
    CrossrefPubMedGoogle Scholar
  • 24 Owen JH, Komarck CM, Wang AC. et al. UM-Chor1: establishment and characterization of the first validated clival chordoma cell line. J Neurosurg 2018; 128 (03) 701-709
    CrossrefPubMedGoogle Scholar
  • 25 Brenner JC, Graham MP, Kumar B. et al. Genotyping of 73 UM-SCC head and neck squamous cell carcinoma cell lines. Head Neck 2010; 32 (04) 417-426
    CrossrefPubMedGoogle Scholar
  • 26 Michmerhuizen NL, Leonard E, Matovina C. et al. Rationale for using irreversible epidermal growth factor receptor inhibitors in combination with phosphatidylinositol 3-kinase inhibitors for advanced head and neck squamous cell carcinoma. Mol Pharmacol 2019; 95 (05) 528-536
    CrossrefPubMedGoogle Scholar
  • 27 Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001; 25 (04) 402-408
    CrossrefPubMedGoogle Scholar
  • 28 Birkeland AC, Yanik M, Tillman BN. et al. Identification of targetable ERBB2 aberrations in head and neck squamous cell carcinoma. JAMA Otolaryngol Head Neck Surg 2016; 142 (06) 559-567
    CrossrefPubMedGoogle Scholar
  • 29 Akhavan-Sigari R, Gaab MR, Rohde V, Abili M, Ostertag H. Expression of PDGFR-α, EGFR and c-MET in spinal chordoma: a series of 52 patients. Anticancer Res 2014; 34 (02) 623-630
    PubMedGoogle Scholar
  • 30 Tamborini E, Miselli F, Negri T. et al. Molecular and biochemical analyses of platelet-derived growth factor receptor (PDGFR) B, PDGFRA, and KIT receptors in chordomas. Clin Cancer Res 2006; 12 (23) 6920-6928
    CrossrefPubMedGoogle Scholar
  • 31 Lee DH, Zhang Y, Kassam AB. et al. Combined PDGFR and HDAC inhibition overcomes PTEN disruption in chordoma. PLoS One 2015; 10 (08) e0134426
    CrossrefPubMedGoogle Scholar
  • 32 Casali PG, Messina A, Stacchiotti S. et al. Imatinib mesylate in chordoma. Cancer 2004; 101 (09) 2086-2097
    CrossrefPubMedGoogle Scholar
  • 33 Stacchiotti S, Casali PG. Systemic therapy options for unresectable and metastatic chordomas. Curr Oncol Rep 2011; 13 (04) 323-330
    CrossrefPubMedGoogle Scholar
  • 34 Stacchiotti S, Longhi A, Ferraresi V. et al. Phase II study of imatinib in advanced chordoma. J Clin Oncol 2012; 30 (09) 914-920
    CrossrefPubMedGoogle Scholar
  • 35 Li X, Ji Z, Ma Y, Qiu X, Fan Q, Ma B. Expression of hypoxia-inducible factor-1α, vascular endothelial growth factor and matrix metalloproteinase-2 in sacral chordomas. Oncol Lett 2012; 3 (06) 1268-1274
    CrossrefPubMedGoogle Scholar
  • 36 Morimoto Y, Tamura R, Ohara K. et al. Prognostic significance of VEGF receptors expression on the tumor cells in skull base chordoma. J Neurooncol 2019; 144 (01) 65-77
    CrossrefPubMedGoogle Scholar
  • 37 Bompas E, Le Cesne A, Tresch-Bruneel E. et al. Sorafenib in patients with locally advanced and metastatic chordomas: a phase II trial of the French Sarcoma Group (GSF/GETO). Ann Oncol 2015; 26 (10) 2168-2173
    CrossrefPubMedGoogle Scholar
  • 38 George S, Merriam P, Maki RG. et al. Multicenter phase II trial of sunitinib in the treatment of nongastrointestinal stromal tumor sarcomas. J Clin Oncol 2009; 27 (19) 3154-3160
    CrossrefPubMedGoogle Scholar
  • 39 Lebellec L, Bertucci F, Tresch-Bruneel E. et al. Circulating vascular endothelial growth factor (VEGF) as predictive factor of progression-free survival in patients with advanced chordoma receiving sorafenib: an analysis from a phase II trial of the french sarcoma group (GSF/GETO). Oncotarget 2016; 7 (45) 73984-73994
    CrossrefPubMedGoogle Scholar
  • 40 Lebellec L, Chauffert B, Blay JY. et al. Advanced chordoma treated by first-line molecular targeted therapies: Outcomes and prognostic factors. A retrospective study of the French Sarcoma Group (GSF/GETO) and the Association des Neuro-Oncologues d'Expression Française (ANOCEF). Eur J Cancer 2017; 79: 119-128
    CrossrefPubMedGoogle Scholar
  • 41 Lipplaa A, Dijkstra S, Gelderblom H. Efficacy of pazopanib and sunitinib in advanced axial chordoma: a single reference centre case series. Clin Sarcoma Res 2016; 6: 19
    CrossrefPubMedGoogle Scholar
  • 42 Svoboda RM, Mackay D, Welsch MJ, Anderson BE. Multiple cutaneous metastatic chordomas from the sacrum. J Am Acad Dermatol 2012; 66 (06) e246-e247
    CrossrefPubMedGoogle Scholar
  • 43 Ribeiro MFSA, de Sousa MC, Hanna SA. et al. Tumor reduction with pazopanib in a patient with recurrent lumbar chordoma. Case Rep Oncol Med 2018; 2018: 4290131
    PubMedGoogle Scholar
  • 44 Sommer J, Itani DM, Homlar KC. et al. Methylthioadenosine phosphorylase and activated insulin-like growth factor-1 receptor/insulin receptor: potential therapeutic targets in chordoma. J Pathol 2010; 220 (05) 608-617
    CrossrefPubMedGoogle Scholar
  • 45 Weinberger PM, Yu Z, Kowalski D. et al. Differential expression of epidermal growth factor receptor, c-Met, and HER2/neu in chordoma compared with 17 other malignancies. Arch Otolaryngol Head Neck Surg 2005; 131 (08) 707-711
    CrossrefPubMedGoogle Scholar
  • 46 Magnaghi P, Salom B, Cozzi L. et al. Afatinib is a new therapeutic approach in chordoma with a unique ability to target EGFR and brachyury. Mol Cancer Ther 2018; 17 (03) 603-613
    CrossrefPubMedGoogle Scholar
  • 47 Heery CR, Singh BH, Rauckhorst M. et al. Phase I trial of a yeast-based therapeutic cancer vaccine (GI-6301) targeting the transcription factor brachyury. Cancer Immunol Res 2015; 3 (11) 1248-1256
    CrossrefPubMedGoogle Scholar
  • 48 Fruman DA, Bismuth G. Fine tuning the immune response with PI3K. Immunol Rev 2009; 228 (01) 253-272
    CrossrefPubMedGoogle Scholar
  • 49 Liu X, Xu Y, Zhou Q. et al. PI3K in cancer: its structure, activation modes and role in shaping tumor microenvironment. Future Oncol 2018; 14 (07) 665-674
    CrossrefPubMedGoogle Scholar
  • 50 Di Veroli GY, Fornari C, Wang D. et al. Combenefit: an interactive platform for the analysis and visualization of drug combinations. Bioinformatics 2016; 32 (18) 2866-2868
    CrossrefPubMedGoogle Scholar