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Endocrine
Published in: Endocrine 3/2018

01-12-2018 | Original Article

Effect of combined treatment with a pan-PI3K inhibitor or an isoform-specific PI3K inhibitor and everolimus on cell proliferation in GH-secreting pituitary tumour in an experimental setting

Authors: Claudia Pivonello, Roberta Patalano, Domenico Solari, Renata S. Auriemma, Federico Frio, Francesca Vitulli, Ludovica F. S. Grasso, Marialuisa Di Cera, Maria Cristina De Martino, Luigi M. Cavallo, Paolo Cappabianca, Annamaria Colao, Rosario Pivonello

Published in: Endocrine | Issue 3/2018

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Abstract

Purpose

PI3K/Akt/mTOR pathway activation is common in GH-secreting pituitary tumours, and a target for treatment with mTOR inhibitors, including everolimus (EVE). The current study aimed to evaluate the efficacy of two PI3K inhibitors (PI3Ki), NVP-BKM120 and NVP-BYL719, alone and in combination with EVE in rat GH-secreting pituitary tumour cell line (GH3) and human GH-secreting pituitary tumour cell cultures.

Methods

In GH3 cell line and in six GH-secreting tumour cell cultures, the effects of PI3Ki and EVE, as single agents and in combination, were tested on cell viability and colony survival, by MTT and clonogenic assay, respectively, whereas western blot was performed to evaluate the underlying intracellular signalling pathways.

Results

PI3Ki and EVE showed a dose-dependent inhibition of cell viability in GH3 cell line, with PI3Ki displaying a synergistic effect when combined with EVE. PI3Ki and EVE inhibited colony survival in GH3 cell line with no further improvement in combination. In GH-secreting pituitary tumour cell cultures PI3Ki are effective in inhibiting cell viability increasing the slight and non significant inhibition induced by EVE as single agent, generally showing a synergistic effect. Despite in both GH3 cell line and GH-secreting pituitary tumour cell cultures combination of PI3Ki enhanced EVE effect, the study of intracellular signalling pathways revealed a different regulation of PI3K/Akt/mTOR and MAPK between the two models.

Conclusions

The results of the current study demonstrated that PI3Ki, especially in combination with EVE, are effective in inhibiting cell proliferation, therefore representing a promising therapeutic tool for the treatment of aggressive GH-secreting pituitary tumours, not responsive to standard medical therapies.
Literature
1.
S. Melmed, A. Colao, A. Barkan, M. Molitch, A.B. Grossman, D. Kleinberg et al., Guidelines for acromegaly management: an update. J. Clin. Endocrinol. Metab. 94, 1509–17 (2009).CrossRef
2.
R.N. Clayton, Cardiovascular function in acromegaly. Endocr. Rev. 24, 272–7 (2003)CrossRef
3.
R. Pivonello, R.S. Auriemma, L.F. Grasso, C. Pivonello, C. Simeoli, R. Patalano et al., Complications of acromegaly: cardiovascular, respiratory and metabolic comorbidities. Pituitary 20, 46–62 (2017).CrossRef
4.
A. Colao, D. Ferone, P. Marzullo, G. Lombardi, Systemic complications of acromegaly: epidemiology, pathogenesis, and management. Endocr. Rev. 25, 102–52 (2004)CrossRef
5.
L. Katznelson, E.R. Laws Jr., S. Melmed, M.E. Molitch, M.H. Murad, A. Utz et al., Acromegaly: an endocrine society clinical practice guideline. J. Clin. Endocrinol. Metab. 99, 3933–51 (2014)CrossRef
6.
A. Giustina, A. Barkan, F.F. Casanueva, F. Cavagnini, L. Frohman, K. Ho et al., Criteria for cure of acromegaly: a consensus statement. J. Clin. Endocrinol. Metab. 85, 526–9 (2000)PubMed
7.
E.R. Laws, G. Lanzino. Transsphenoidal Surgery. (Saunders - Elsevier, Philadelphia, 2010)
8.
D. Solari, L.M. Cavallo, P. Cappabianca, Surgical approach to pituitary tumors. Handb. Clin. Neurol. 124, 291–301 (2014)CrossRef
9.
A. Colao, R. Pivonello, C. Di Somma, S. Savastano, L.F. Grasso, G. Lombardi, Medical therapy of pituitary adenomas: effects on tumor shrinkage. Rev. Endocr. Metab. Disord. 10, 111–23 (2009)CrossRef
10.
A. Giustina, M.R. Ambrosio, P. Beck Peccoz, F. Bogazzi, S. Cannavo, L. De Marinis et al., Use of Pegvisomant in acromegaly. An Italian Society of Endocrinology guideline. J. Endocrinol. Invest. 37, 1017–30 (2014)CrossRef
11.
A. Colao, R.S. Auriemma, G. Lombardi, R. Pivonello, Resistance to somatostatin analogs in acromegaly. Endocr. Rev. 32, 247–71 (2011)CrossRef
12.
M.R. Gadelha, L.E. Wildemberg, M.D. Bronstein, F. Gatto, D. Ferone, Somatostatin receptor ligands in the treatment of acromegaly. Pituitary 20, 100–8 (2017)CrossRef
13.
C. Beauregard, U. Truong, J. Hardy, O. Serri, Long-term outcome and mortality after transsphenoidal adenomectomy for acromegaly. Clin. Endocrinol. 58, 86–91 (2003)CrossRef
14.
G. Minniti, M.L. Jaffrain-Rea, V. Esposito, A. Santoro, G. Tamburrano, G. Cantore, Evolving criteria for post-operative biochemical remission of acromegaly: can we achieve a definitive cure? An audit of surgical results on a large series and a review of the literature. Endocr. Relat. Cancer 10, 611–9 (2003)CrossRef
15.
F. Di Nicolantonio, S. Arena, J. Tabernero, S. Grosso, F. Molinari, T. Macarulla et al., Deregulation of the PI3K and KRAS signaling pathways in human cancer cells determines their response to everolimus. J. Clin. Invest. 120, 2858–66 (2010)CrossRef
16.
Y. Lin, X. Jiang, Y. Shen, M. Li, H. Ma, M. Xing et al., Frequent mutations and amplifications of the PIK3CA gene in pituitary tumors. Endocr. Relat. Cancer 16, 301–10 (2009)CrossRef
17.
K.E. O’Reilly, F. Rojo, Q.B. She, D. Solit, G.B. Mills, D. Smith et al., mTOR inhibition induces upstream receptor tyrosine kinase signaling and activates Akt. Cancer Res. 66, 1500–8 (2006)CrossRef
18.
I. Vivanco, C.L. Sawyers, The phosphatidylinositol 3-Kinase AKT pathway in human cancer. Nat. Rev. Cancer 2, 489–501 (2002)CrossRef
19.
D.A. Altomare, J.R. Testa, Perturbations of the AKT signaling pathway in human cancer. Oncogene 24, 7455–64 (2005)CrossRef
20.
K.H. Khan, T.A. Yap, L. Yan, D. Cunningham, Targeting the PI3K-AKT-mTOR signaling network in cancer. Chin. J. Cancer 32, 253–65 (2013)CrossRef
21.
C. Pivonello, M. Negri, M.C. De Martino, M. Napolitano, C. de Angelis, D.P. Provvisiero et al., The dual targeting of insulin and insulin-like growth factor 1 receptor enhances the mTOR inhibitor-mediated antitumor efficacy in hepatocellular carcinoma. Oncotarget 7, 9718–31 (2016)CrossRef
22.
L. Zhao, P.K. Vogt, Class I PI3K in oncogenic cellular transformation. Oncogene 27, 5486–96 (2008)CrossRef
23.
T.L. Yuan, L.C. Cantley, PI3K pathway alterations in cancer: variations on a theme. Oncogene 27, 5497–510 (2008)CrossRef
24.
A. Carracedo, L. Ma, J. Teruya-Feldstein, F. Rojo, L. Salmena, A. Alimonti et al., Inhibition of mTORC1 leads to MAPK pathway activation through a PI3K-dependent feedback loop in human cancer. J. Clin. Investig. 118, 3065–74 (2008)PubMed
25.
E. Monsalves, K. Juraschka, T. Tateno, S. Agnihotri, S.L. Asa, S. Ezzat et al., The PI3K/AKT/mTOR pathway in the pathophysiology and treatment of pituitary adenomas. Endocr. Relat. Cancer 21, R331–44 (2014)CrossRef
26.
S. Jean, A.A. Kiger, Classes of phosphoinositide 3-kinases at a glance. J. Cell. Sci. 127, 923–8 (2014)CrossRef
27.
M. Chanal, P. Chevallier, V. Raverot, G. Fonteneau, K. Lucia, J.L. Monteserin Garcia et al., Differential effects of PI3K and dual PI3K/mTOR inhibition in rat prolactin-secreting pituitary tumors. Mol. Cancer Ther. 15, 1261–70 (2016)CrossRef
28.
C.B. Murat, P.B. Braga, M.A. Fortes, M.D. Bronstein, M.L. Correa-Giannella, R.R. Giorgi, Mutation and genomic amplification of the PIK3CA proto-oncogene in pituitary adenomas. Braz. J. Med. Biol. Res. 45, 851–5 (2012)CrossRef
29.
M. Cakir, A.B. Grossman, Targeting MAPK (Ras/ERK) and PI3K/Akt pathways in pituitary tumorigenesis. Expert. Opin. Ther. Targets 13, 1121–34 (2009)CrossRef
30.
D. Dworakowska, E. Wlodek, C.A. Leontiou, S. Igreja, M. Cakir, M. Teng et al., Activation of RAF/MEK/ERK and PI3K/AKT/mTOR pathways in pituitary adenomas and their effects on downstream effectors. Endocr. Relat. Cancer 16, 1329–38 (2009)CrossRef
31.
M. Musat, M. Korbonits, B. Kola, N. Borboli, M.R. Hanson, A.M. Nanzer et al., Enhanced protein kinase B/Akt signalling in pituitary tumours. Endocr. Relat. Cancer 12, 423–33 (2005)CrossRef
32.
B. Svejda, M. Kidd, A. Kazberouk, B. Lawrence, R. Pfragner, I.M. Modlin, Limitations in small intestinal neuroendocrine tumor therapy by mTor kinase inhibition reflect growth factor-mediated PI3K feedback loop activation via ERK1/2 and AKT. Cancer 117, 4141–54 (2011)CrossRef
33.
D.A. Cantrell, Phosphoinositide 3-kinase signalling pathways. J. Cell. Sci. 114, 1439–45 (2001)PubMed
34.
B.T. Hennessy, D.L. Smith, P.T. Ram, Y. Lu, G.B. Mills, Exploiting the PI3K/AKT pathway for cancer drug discovery. Nat. Rev. Drug. Discov. 4, 988–1004 (2005)CrossRef
35.
X. Bai, Y. Jiang, Key factors in mTOR regulation. Cell. Mol. Life Sci. 67, 239–53 (2010)CrossRef
36.
R. Chen, J. Duan, L. Li, Q. Ma, Q. Sun, J. Ma et al., mTOR promotes pituitary tumor development through activation of PTTG1. Oncogene 36, 979–88 (2017)CrossRef
37.
S.M. Maira, S. Pecchi, A. Huang, M. Burger, M. Knapp, D. Sterker et al., Identification and characterization of NVP-BKM120, an orally available pan-class I PI3-kinase inhibitor. Mol. Cancer Ther. 11, 317–28 (2012)CrossRef
38.
P. Furet, V. Guagnano, R.A. Fairhurst, P. Imbach-Weese, I. Bruce, M. Knapp et al., Discovery of NVP-BYL719 a potent and selective phosphatidylinositol-3 kinase alpha inhibitor selected for clinical evaluation. Bioorg. Med. Chem. Lett. 23, 3741–8 (2013)CrossRef
39.
R.J. Motzer, B. Escudier, S. Oudard, T.E. Hutson, C. Porta, S. Bracarda et al., Efficacy of everolimus in advanced renal cell carcinoma: a double-blind, randomised, placebo-controlled phase III trial. Lancet 372, 449–56 (2008)CrossRef
40.
A.X. Zhu, M. Kudo, E. Assenat, S. Cattan, Y.K. Kang, H.Y. Lim et al., Effect of everolimus on survival in advanced hepatocellular carcinoma after failure of sorafenib: the EVOLVE-1 randomized clinical trial. JAMA 312, 57–67 (2014)CrossRef
41.
N. Wagle, B.C. Grabiner, E.M. Van Allen, A. Amin-Mansour, A. Taylor-Weiner, M. Rosenberg et al., Response and acquired resistance to everolimus in anaplastic thyroid cancer. N. Engl. J. Med. 371, 1426–33 (2014)CrossRef
42.
M.E. Pavel, J.D. Hainsworth, E. Baudin, M. Peeters, D. Horsch, R.E. Winkler et al., Everolimus plus octreotide long-acting repeatable for the treatment of advanced neuroendocrine tumours associated with carcinoid syndrome (RADIANT-2): a randomised, placebo-controlled, phase 3 study. Lancet 378, 2005–12 (2011)CrossRef
43.
J.C. Yao, M.H. Shah, T. Ito, C.L. Bohas, E.M. Wolin, E. Van Cutsem et al., Everolimus for advanced pancreatic neuroendocrine tumors. N. Engl. J. Med. 364, 514–23 (2011)CrossRef
44.
A. Gorshtein, H. Rubinfeld, E. Kendler, M. Theodoropoulou, V. Cerovac, G.K. Stalla et al., Mammalian target of rapamycin inhibitors rapamycin and RAD001 (everolimus) induce anti-proliferative effects in GH-secreting pituitary tumor cells in vitro. Endocr. Relat. Cancer 16, 1017–27 (2009)CrossRef
45.
S. Sukumari-Ramesh, N. Singh, K.M. Dhandapani, J.R. Vender, mTOR inhibition reduces cellular proliferation and sensitizes pituitary adenoma cells to ionizing radiation. Surg. Neurol. Int. 2, 22 (2011)CrossRef
46.
E. Rozengurt, H.P. Soares, J. Sinnet-Smith, Suppression of feedback loops mediated by PI3K/mTOR induces multiple overactivation of compensatory pathways: an unintended consequence leading to drug resistance. Mol. Cancer Ther. 13, 2477–88 (2014)CrossRef
47.
G. Brabant, A. von zur Muhlen, C. Wuster, M.B. Ranke, J. Kratzsch, W. Kiess et al., Serum insulin-like growth factor I reference values for an automated chemiluminescence immunoassay system: results from a multicenter study. Horm. Res. 60, 53–60 (2003)PubMed
48.
C. Pivonello, P. Rousaki, M. Negri, M. Sarnataro, M. Napolitano, F.Z. Marino et al., Effects of the single and combined treatment with dopamine agonist, somatostatin analog and mTOR inhibitors in a human lung carcinoid cell line: an in vitro study. Endocrine 56, 603–20 (2017)CrossRef
49.
C. Desbois-Mouthon, A. Baron, M.J. Blivet-Van Eggelpoel, L. Fartoux, C. Venot, F. Bladt et al., Insulin-like growth factor-1 receptor inhibition induces a resistance mechanism via the epidermal growth factor receptor/HER3/AKT signaling pathway: rational basis for cotargeting insulin-like growth factor-1 receptor and epidermal growth factor receptor in hepatocellular carcinoma. Clin. Cancer Res. 15, 5445–56 (2009)CrossRef
50.
D.A. Donoho, N. Bose, G. Zada, J.D. Carmichael, Management of aggressive growth hormone secreting pituitary adenomas. Pituitary 20, 169–78 (2017)CrossRef
51.
E.A. Sajjad, G. Zielinski, M. Maksymowicz, L. Hutnik, T. Bednarczuk, P. Wlodarski, mTOR is frequently active in GH-secreting pituitary adenomas without influencing their morphopathological features. Endocr. Pathol. 24, 11–9 (2013)CrossRef
52.
M. Lee, T. Wiedemann, C. Gross, I. Leinhauser, F. Roncaroli, R. Braren et al., Targeting PI3K/mTOR signaling displays potent antitumor efficacy against nonfunctioning pituitary adenomas. Clin. Cancer Res. 21, 3204–15 (2015)CrossRef
53.
M. Lee, M. Theodoropoulou, J. Graw, F. Roncaroli, M.C. Zatelli, N.S. Pellegata, Levels of p27 sensitize to dual PI3K/mTOR inhibition. Mol. Cancer Ther. 10, 1450–9 (2011)CrossRef
54.
H. Rubinfeld, I. Shimon, PI3K/Akt/mTOR and Raf/MEK/ERK signaling pathways perturbations in non-functioning pituitary adenomas. Endocrine 42, 285–91 (2012)CrossRef
55.
J.D. Valentino, J. Li, Y.Y. Zaytseva, W.C. Mustain, V.A. Elliott, J.T. Kim et al., Cotargeting the PI3K and RAS pathways for the treatment of neuroendocrine tumors. Clin. Cancer Res. 20, 1212–22 (2014)CrossRef
56.
S. Nolting, J. Rentsch, H. Freitag, K. Detjen, F. Briest, M. Mobs et al., The selective PI3Kalpha inhibitor BYL719 as a novel therapeutic option for neuroendocrine tumors: results from multiple cell line models. PLoS ONE 12, e0182852 (2017)CrossRef
57.
Y. Hu, R. Guo, J. Wei, Y. Zhou, W. Ji, J. Liu et al., Effects of PI3K inhibitor NVP-BKM120 on overcoming drug resistance and eliminating cancer stem cells in human breast cancer cells. Cell Death Dis. 6, e2020 (2015)CrossRef
58.
Y. Zheng, J. Yang, J. Qian, L. Zhang, Y. Lu, H. Li et al., Novel phosphatidylinositol 3-kinase inhibitor NVP-BKM120 induces apoptosis in myeloma cells and shows synergistic anti-myeloma activity with dexamethasone. J. Mol. Med. 90, 695–706 (2012)CrossRef
59.
E. Musi, G. Ambrosini, E. de Stanchina, G.K. Schwartz, The phosphoinositide 3-kinase alpha selective inhibitor BYL719 enhances the effect of the protein kinase C inhibitor AEB071 in GNAQ/GNA11-mutant uveal melanoma cells. Mol. Cancer Ther. 13, 1044–53 (2014)CrossRef
60.
H. Ren, H. Guo, A. Thakur, S. Zhang, T. Wang, Y. Liang et al., Blockade efficacy of MEK/ERK-dependent autophagy enhances PI3K/Akt inhibitor NVP-BKM120’s therapeutic effectiveness in lung cancer cells. Oncotarget 7, 67277–87 (2016)PubMedPubMedCentral
61.
J.C. Bendell, J. Rodon, H.A. Burris, M. de Jonge, J. Verweij, D. Birle et al., Phase I, dose-escalation study of BKM120, an oral pan-Class I PI3K inhibitor, in patients with advanced solid tumors. J. Clin. Oncol. 30, 282–90 (2012)CrossRef
62.
S.S. De Buck, A. Jakab, M. Boehm, D. Bootle, D. Juric, C. Quadt et al., Population pharmacokinetics and pharmacodynamics of BYL719, a phosphoinositide 3-kinase antagonist, in adult patients with advanced solid malignancies. Br. J. Clin. Pharmacol. 78, 543–55 (2014)CrossRef
63.
M.C. Zatelli, M. Minoia, C. Filieri, F. Tagliati, M. Buratto, M.R. Ambrosio et al., Effect of everolimus on cell viability in nonfunctioning pituitary adenomas. J. Clin. Endocrinol. Metab. 95, 968–76 (2010)CrossRef
64.
V. Cerovac, J. Monteserin-Garcia, H. Rubinfeld, M. Buchfelder, M. Losa, T. Florio et al., The somatostatin analogue octreotide confers sensitivity to rapamycin treatment on pituitary tumor cells. Cancer Res. 70, 666–74 (2010)CrossRef
65.
R. Loewith, E. Jacinto, S. Wullschleger, A. Lorberg, J.L. Crespo, D. Bonenfant et al., Two TOR complexes, only one of which is rapamycin sensitive, have distinct roles in cell growth control. Mol. Cell 10, 457–68 (2002)CrossRef
66.
S.C. Hanna, S.A. Heathcote, W.Y. Kim, mTOR pathway in renal cell carcinoma. Expert. Rev. AntiCancer Ther. 8, 283–92 (2008)CrossRef
67.
A. O’Donnell, S. Faivre, H.A. Burris 3rd, D. Rea, V. Papadimitrakopoulou, N. Shand et al., Phase I pharmacokinetic and pharmacodynamic study of the oral mammalian target of rapamycin inhibitor everolimus in patients with advanced solid tumors. J. Clin. Oncol.: Off. J. Am. Soc. Clin. Oncol. 26, 1588–95 (2008)CrossRef
68.
C. Di Pasquale, E. Gentilin, S. Falletta, M. Bellio, M. Buratto, E. Degli Uberti et al. , PI3K/Akt/mTOR pathway involvement in regulating growth hormone secretion in a rat pituitary adenoma cell line. Endocrine 60, 308–316 (2018).CrossRef
69.
R.J. Shaw, L.C. Cantley, Ras, PI(3)K and mTOR signalling controls tumour cell growth. Nature 441, 424–30 (2006)CrossRef
70.
M. Breuleux, M. Klopfenstein, C. Stephan, C.A. Doughty, L. Barys, S.M. Maira et al., Increased AKT S473 phosphorylation after mTORC1 inhibition is rictor dependent and does not predict tumor cell response to PI3K/mTOR inhibition. Mol. Cancer Ther. 8, 742–53 (2009)CrossRef
Metadata
Title
Effect of combined treatment with a pan-PI3K inhibitor or an isoform-specific PI3K inhibitor and everolimus on cell proliferation in GH-secreting pituitary tumour in an experimental setting
Authors
Claudia Pivonello
Roberta Patalano
Domenico Solari
Renata S. Auriemma
Federico Frio
Francesca Vitulli
Ludovica F. S. Grasso
Marialuisa Di Cera
Maria Cristina De Martino
Luigi M. Cavallo
Paolo Cappabianca
Annamaria Colao
Rosario Pivonello
Publication date
01-12-2018
Publisher
Springer US
Published in
Endocrine / Issue 3/2018
Print ISSN: 1355-008X
Electronic ISSN: 1559-0100
DOI
https://doi.org/10.1007/s12020-018-1677-2

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