Expression of somatostatin receptors 2 and 5 in circulating tumour cells from patients with neuroendocrine tumours

Alexa Childs, Clare Vesely, Leah Ensell, Helen Lowe, Tu Vinh Luong, Martyn E Caplin, Christos Toumpanakis, Christina Thirlwell, John A Hartley, Tim Meyer, Alexa Childs, Clare Vesely, Leah Ensell, Helen Lowe, Tu Vinh Luong, Martyn E Caplin, Christos Toumpanakis, Christina Thirlwell, John A Hartley, Tim Meyer

Abstract

Background: Neuroendocrine tumours (NET) overexpress somatostatin receptors (SSTR) that can be targeted for therapy. Somatostatin receptor expression is routinely measured by molecular imaging but the resolution is insufficient to define heterogeneity. We hypothesised that SSTR expression could be measured on circulating tumour cells (CTCs) and used to investigate heterogeneity of expression and track changes during therapy.

Methods: MCF-7 cells were transfected with SSTR2 or 5 and spiked into donor blood for analysis by CellSearch. Optimum anti-SSTR antibody concentration and exposure time were determined, and flow cytometry was used to evaluate assay sensitivity. For clinical evaluation, blood was analysed by CellSearch, and SSTR2/5 immunohistochemistry was performed on matched tissue samples.

Results: Flow cytometry confirmed CellSearch was sensitive and that detection of SSTR was unaffected by the presence of somatostatin analogue up to a concentration of 100 ng ml-l. Thirty-one NET patients were recruited: grade; G1 (29%), G2 (45%), G3 (13%), primary site; midgut (58%), pancreatic (39%). Overall, 87% had SSTR-positive tumours according to somatostatin receptor scintigraphy or 68-Ga-DOTATE PET/CT. Circulating tumour cells were detected in 21 out of 31 patients (68%), of which 33% had evidence of heterogeneous expression of either SSTR2 (n=5) or SSTR5 (n=2).

Conclusions: Somatostatin receptors 2 and 5 are detectable on CTCs from NET patients and may be a useful biomarker for evaluating SSTR-targeted therapies and this is being prospectively evaluated in the Phase IV CALMNET trial (NCT02075606).

Conflict of interest statement

TM has received research funding for this work from Ipsen, MEC has received grant funding and consultancy fees from Ipsen.

Figures

Figure 1
Figure 1
CellTracks Analyzer II images. (A) SSTR transfected MCF-7-spiked blood samples. (i) SSTR2-positive tumour cell. (ii) SSTR2-negative tumour cell. (iii) SSTR5-positive tumour cell. (iv) SSTR5-negative tumour cell. (B) Clinical validation in patient samples. (i) SSTR2-positive CTC in patient 21. (ii) SSTR2-negative CTC in patient 21. (iii) SSTR5-positive CTC in patient 7. (iv) SSTR5-negative CTC in patient 7.
Figure 2
Figure 2
Immunohistochemistry for SSTR2 and SSTR5. (A) SSTR2 staining score 0. (B) SSTR2 staining score 2. (C) SSTR2 staining score 3. (D) SSTR5 staining score 0. (E) SSTR5 staining score 2. (F) SSTR5 staining score 3.

References

    1. Caplin ME, Pavel M, Ćwikła JB, Phan AT, Raderer M, Sedláčková E, Cadiot G, Wolin EM, Capdevila J, Wall L, Rindi G, Langley A, Martinez S, Blumberg J, Ruszniewski P CLARINET Investigators (2014) Lanreotide in metastatic enteropancreatic neuroendocrine tumors. N Engl J Med 371: 224–233.
    1. de Herder WW, Hofland LJ, van der Lely AJ, SWJ Lamberts (2003) Somatostatin receptors in gastroentero-pancreatic neuroendocrine tumours. Endocr Relat Cancer 10: 451–458.
    1. Fazio N, Cinieri S, Lorizzo K, Squadroni M, Orlando L, Spada F, Maiello E, Bodei L, Paganelli G, Delle Fave G, De Braud F (2010) Biological targeted therapies in patients with advanced enteropancreatic neuroendocrine carcinomas. Cancer Treat Rev 36(Suppl 3): S87–S94.
    1. Flores LM, Kindelberger DW, Ligon AH, Capelletti M, Fiorentino M, Loda M, Cibas ES, Jänne PA, Krop IE (2010) Improving the yield of circulating tumour cells facilitates molecular characterisation and recognition of discordant HER2 amplification in breast cancer. Br J Cancer 102: 1495–1502.
    1. Gabriel M, Decristoforo C, Kendler D, Dobrozemsky G, Heute D, Uprimny C, Kovacs P, Guggenberg Von E, Bale R, Virgolini IJ (2007) 68Ga-DOTA-Tyr3-octreotide PET in neuroendocrine tumors: comparison with somatostatin receptor scintigraphy and CT. J Nucl Med 48: 508–518.
    1. Hofman MS, Hicks RJ (2012) Changing paradigms with molecular imaging of neuroendocrine tumors. Discov Med 14: 71–81.
    1. Kaemmerer D, Peter L, Lupp A, Schulz S, Sänger J, Baum RP, Prasad V, Hommann M (2012) Comparing of IRS and Her2 as immunohistochemical scoring schemes in gastroenteropancreatic neuroendocrine tumors. Int J Clin Exp Pathol 5: 187–194.
    1. Kaemmerer D, Peter L, Lupp A, Schulz S, Sänger J, Prasad V, Kulkarni H, Haugvik S-P, Hommann M, Baum RP (2011) Molecular imaging with 68Ga-SSTR PET/CT and correlation to immunohistochemistry of somatostatin receptors in neuroendocrine tumours. Eur J Nucl Med Mol Imaging 38: 1659–1668.
    1. Kaemmerer D, Wirtz RM, Fischer EK, Hommann M, Sänger J, Prasad V, Specht E, Baum RP, Schulz S, Lupp A (2015) Analysis of somatostatin receptor 2A immunohistochemistry, RT-qPCR, and in vivo PET/CT data in patients with pancreatic neuroendocrine neoplasm. Pancreas 44: 648–654.
    1. Khan MS, Kirkwood A, Tsigani T, Garcia-Hernandez J, Hartley JA, Caplin ME, Meyer T (2013. a) Circulating tumor cells as prognostic markers in neuroendocrine tumors. J Clin Oncol 31: 365–372.
    1. Khan MS, Kirkwood AA, Tsigani T, Lowe H, Goldstein R, Hartley JA, Caplin ME, Meyer T (2015) Early changes in circulating tumor cells are associated with response and survival following treatment of metastatic neuroendocrine neoplasms. Clin Cancer Res 22: 79–85, clincanres.1008.2015.
    1. Khan MS, Luong TV, Watkins J, Toumpanakis C, Caplin ME, Meyer T (2013. b) A comparison of Ki-67 and mitotic count as prognostic markers for metastatic pancreatic and midgut neuroendocrine neoplasms. Br J Cancer 108: 1838–1845.
    1. Khan MS, Tsigani T, Rashid M, Rabouhans JS, Yu D, Luong TV, Caplin M, Meyer T (2011. a) Circulating tumor cells and EpCAM expression in neuroendocrine tumors. Clin Cancer Res 17: 337–345.
    1. Khan MS, Tsigani T, Rashid M, Rabouhans JS, Yu D, Luong TV, Caplin M, Meyer T (2011. b) Circulating tumor cells and EpCAM expression in neuroendocrine tumors. Clin Cancer Res 17: 337–345.
    1. Krenning EP, Valkema R, Kwekkeboom DJ, de Herder WW, van Eijck CHJ, de Jong M, Pauwels S, Reubi JC (2005) Molecular imaging as in vivo molecular pathology for gastroenteropancreatic neuroendocrine tumors: implications for follow-up after therapy. J Nucl Med 46(Suppl 1): 76S–82S.
    1. Mazel M, Jacot W, Pantel K, Bartkowiak K, Topart D, Cayrefourcq L, Rossille D, Maudelonde T, Fest T, Alix-Panabières C (2015) Frequent expression of PD-L1 on circulating breast cancer cells. Mol Oncol 9: 1773–1782.
    1. Mehta S, de Reuver PR, Gill P, Andrici J, D'Urso L, Mittal A, Pavlakis N, Clarke S, Samra JS, Gill AJ (2015) Somatostatin receptor SSTR-2a expression is a stronger predictor for survival than Ki-67 in pancreatic neuroendocrine tumors. Medicine (Baltimore) 94: e1281.
    1. Miederer M, Seidl S, Buck A, Scheidhauer K, Wester H-J, Schwaiger M, Perren A (2009) Correlation of immunohistopathological expression of somatostatin receptor 2 with standardised uptake values in 68Ga-DOTATOC PET/CT. Eur J Nucl Med Mol Imaging 36: 48–52.
    1. Munzone E, Nolé F, Goldhirsch A, Botteri E, Esposito A, Zorzino L, Curigliano G, Minchella I, Adamoli L, Cassatella MC, Casadio C, Sandri M-T (2010) Changes of HER2 status in circulating tumor cells compared with the primary tumor during treatment for advanced breast cancer. Clin Breast Cancer 10: 392–397.
    1. Nasir A, Stridsberg M, Strosberg J, Su P-H, Livingston S, Malik HA, Kelley ST, Centeno BA, Coppola D, Malafa ME, Yeatman TJ, Kvols LK (2006) Somatostatin receptor profiling in hepatic metastases from small intestinal and pancreatic neuroendocrine neoplasms: immunohistochemical approach with potential clinical utility. Cancer Control 13: 52–60.
    1. Pentheroudakis G, Batistatou A, Kalogeras KT, Kronenwett R, Wirtz RM, Bournakis E, Eleftheraki AG, Pectasides D, Bobos M, Papaspirou I, Kamina S, Gogas H, Koutras AK, Pavlidis N, Fountzilas G (2011) Prognostic utility of β-tubulin isotype III and correlations with other molecular and clinicopathological variables in patients with early breast cancer: a translational Hellenic Cooperative Oncology Group (HeCOG) study. Breast Cancer Res Treat 127: 179–193.
    1. Pestrin M, Bessi S, Galardi F, Truglia M, Biggeri A, Biagioni C, Cappadona S, Biganzoli L, Giannini A, Di Leo A (2009) Correlation of HER2 status between primary tumors and corresponding circulating tumor cells in advanced breast cancer patients. Breast Cancer Res Treat 118: 523–530.
    1. Punnoose EA, Atwal SK, Spoerke JM, Savage H, Pandita A, Yeh R-F, Pirzkall A, Fine BM, Amler LC, Chen DS, Lackner MR (2010) Molecular biomarker analyses using circulating tumor cells. PLoS One 5: e12517.
    1. Reubi JC (2011) Peptide receptors as molecular targets for cancer diagnosis and therapy. Endocr Rev 24: 389–427.
    1. Rindi G, Klöppel G, Alhman H, Caplin M, Couvelard A, de Herder WW, Erikssson B, Falchetti A, Falconi M, Komminoth P, Körner M, Lopes JM, McNicol A-M, Nilsson O, Perren A, Scarpa A, Scoazec JY, Wiedenmann B all other Frascati Consensus Conference participantsEuropean Neuroendocrine Tumor Society (ENETS) (2006) TNM staging of foregut (neuro)endocrine tumors: a consensus proposal including a grading system. Virchows Arch 449: 395–401.
    1. Rinke A, Müller H-H, Schade-Brittinger C, Klose K-J, Barth P, Wied M, Mayer C, Aminossadati B, Pape U-F, Bläker M, Harder J, Arnold C, Gress T, Arnold R PROMID Study Group (2009) Placebo-controlled, double-blind, prospective, randomized study on the effect of octreotide LAR in the control of tumor growth in patients with metastatic neuroendocrine midgut tumors: a report from the PROMID Study Group. J Clin Oncol 27: 4656–4663.
    1. SOMATULINE® DEPOT (lanreotide) INJECTION (2015) Available at .
    1. Volante M, Brizzi MP, Faggiano A, La Rosa S, Rapa I, Ferrero A, Mansueto G, Righi L, Garancini S, Capella C, De Rosa G, Dogliotti L, Colao A, Papotti M (2007) Somatostatin receptor type 2A immunohistochemistry in neuroendocrine tumors: a proposal of scoring system correlated with somatostatin receptor scintigraphy. Mod Pathol 20: 1172–1182.
    1. Yao JC, Hassan M, Phan A, Dagohoy C, Leary C, Mares JE, Abdalla EK, Fleming JB, Vauthey J-N, Rashid A, Evans DB (2008) One hundred years after ‘carcinoid': epidemiology of and prognostic factors for neuroendocrine tumors in 35 825 cases in the United States. J Clin Oncol 26: 3063–3072.

Source: PubMed

Sponzoři a spolupracovníci

Zdravotní podmínky

Drogové intervence

3
Předplatit