Skip to main content
Top
Published in: Pediatric Surgery International 1/2023

01-12-2023 | Neuroblastoma | Original Article

Expression of programmed cell death-1 on neuroblastoma cells in TH-MYCN transgenic mice

Authors: Yuta Takeuchi, Seiichiro Inoue, Akio Odaka

Published in: Pediatric Surgery International | Issue 1/2023

Login to get access

Abstract

Background

Immunotherapy may improve the poor prognosis of high-risk neuroblastoma. Programmed cell death-1 (PD-1) is expressed in several cancers. The tyrosine hydroxylase MYCN (TH-MYCN) transgenic mouse model is widely used in neuroblastoma research, but detailed information on its immunological background is lacking. Therefore, we studied the immunological tumor microenvironment and tumor cell surface antigen expression in homozygote and hemizygote mice and effects of antibody therapy against PD-1.

Methods

CD4, CD8, CD11b, and CD11c expression in immune cells from retroperitoneal lymph nodes and spleen was analyzed by flow cytometry. Tumor cell surface antigen expression was confirmed, and data from homozygote and hemizygote mice were compared. Effects of anti-PD-1 antibody were evaluated.

Results

CD4-, CD8-, CD11b-, and CD11c-positive cells were not significantly different in homozygote and hemizygote mice, and CD11b- and CD11c-positive cells were identified in the tumor microenvironment in both. Tumor cells expressed PD-1, and anti-PD-1 antibody had anti-tumor effects and significantly reduced the percentage of living tumor cells in cultures after 2 h.

Conclusion

The immunological background is similar in homozygote and hemizygote TH-MYCN transgenic mice, and both have PD-1–positive tumor cells. Anti-PD-1 antibody suppresses tumor growth. This mouse model may be a useful for studying immunotherapy of neuroblastoma.
Literature
2.
go back to reference Whittle SB, Smith V, Doherty E, Zhao S, McCarty S, Zage PE (2017) Overview and recent advances in the treatment of neuroblastoma. Expert Rev Anticancer Ther 17(4):369–386CrossRefPubMed Whittle SB, Smith V, Doherty E, Zhao S, McCarty S, Zage PE (2017) Overview and recent advances in the treatment of neuroblastoma. Expert Rev Anticancer Ther 17(4):369–386CrossRefPubMed
3.
go back to reference Newman EA, Nuchtern JG (2016) Recent biologic and genetic advances in neuroblastoma: implications for diagnostic, risk stratification, and treatment strategies. Semin Pediatr Surg 25(5):257–264CrossRefPubMed Newman EA, Nuchtern JG (2016) Recent biologic and genetic advances in neuroblastoma: implications for diagnostic, risk stratification, and treatment strategies. Semin Pediatr Surg 25(5):257–264CrossRefPubMed
4.
go back to reference Morandi F, Sabatini F, Podesta M, Airoldi I (2021) Immunotherapeutic strategies for neuroblastoma: present past and future. Vaccines (Basel) 9(1):43CrossRefPubMed Morandi F, Sabatini F, Podesta M, Airoldi I (2021) Immunotherapeutic strategies for neuroblastoma: present past and future. Vaccines (Basel) 9(1):43CrossRefPubMed
6.
go back to reference Keir ME, Liang SC, Guleria I, Latchman YE, Qipo A, Albacker LA, Koulmanda M, Freeman GJ, Sayegh MH, Sharpe AH (2006) Tissue expression of PD-L1 mediates peripheral T cell tolerance. J Exp Med 203(4):883–895CrossRefPubMedPubMedCentral Keir ME, Liang SC, Guleria I, Latchman YE, Qipo A, Albacker LA, Koulmanda M, Freeman GJ, Sayegh MH, Sharpe AH (2006) Tissue expression of PD-L1 mediates peripheral T cell tolerance. J Exp Med 203(4):883–895CrossRefPubMedPubMedCentral
8.
go back to reference Webb ER, Lanati S, Wareham C, Easton A, Dunn SN, Inzhelevskaya T, Sadler FM, James S, Ashton-Key M, Cragg MS, Beers SA, Gray JC (2020) Immune characterization of pre-clinical murine models of neuroblastoma. Sci Rep 10(1):16695CrossRefPubMedPubMedCentral Webb ER, Lanati S, Wareham C, Easton A, Dunn SN, Inzhelevskaya T, Sadler FM, James S, Ashton-Key M, Cragg MS, Beers SA, Gray JC (2020) Immune characterization of pre-clinical murine models of neuroblastoma. Sci Rep 10(1):16695CrossRefPubMedPubMedCentral
9.
go back to reference Eissler N, Mao Y, Brodin D, Reutersward P, Andersson Svahn H, Johnsen JI, Kiessling R, Kogner P (2016) Regulation of myeloid cells by activated T cells determines the efficacy of PD-1 blockade. Oncoimmunology 5(12):e1232222CrossRefPubMedPubMedCentral Eissler N, Mao Y, Brodin D, Reutersward P, Andersson Svahn H, Johnsen JI, Kiessling R, Kogner P (2016) Regulation of myeloid cells by activated T cells determines the efficacy of PD-1 blockade. Oncoimmunology 5(12):e1232222CrossRefPubMedPubMedCentral
10.
go back to reference Ardavin C (2003) Origin, precursors and differentiation of mouse dendritic cells. Nat Rev Immunol 3(7):582–590CrossRefPubMed Ardavin C (2003) Origin, precursors and differentiation of mouse dendritic cells. Nat Rev Immunol 3(7):582–590CrossRefPubMed
11.
go back to reference Asselin-Paturel C, Brizard G, Pin JJ, Briere F, Trinchieri G (2003) Mouse strain differences in plasmacytoid dendritic cell frequency and function revealed by a novel monoclonal antibody. J Immunol 171(12):6466–6477CrossRefPubMed Asselin-Paturel C, Brizard G, Pin JJ, Briere F, Trinchieri G (2003) Mouse strain differences in plasmacytoid dendritic cell frequency and function revealed by a novel monoclonal antibody. J Immunol 171(12):6466–6477CrossRefPubMed
13.
go back to reference Kroesen M, Brok IC, Reijnen D, van Hout-Kuijer MA, Zeelenberg IS, Den Brok MH, Hoogerbrugge PM, Adema GJ (2015) Intra-adrenal murine TH-MYCN neuroblastoma tumors grow more aggressive and exhibit a distinct tumor microenvironment relative to their subcutaneous equivalents. Cancer Immunol Immunother 64(5):563–572CrossRefPubMedPubMedCentral Kroesen M, Brok IC, Reijnen D, van Hout-Kuijer MA, Zeelenberg IS, Den Brok MH, Hoogerbrugge PM, Adema GJ (2015) Intra-adrenal murine TH-MYCN neuroblastoma tumors grow more aggressive and exhibit a distinct tumor microenvironment relative to their subcutaneous equivalents. Cancer Immunol Immunother 64(5):563–572CrossRefPubMedPubMedCentral
14.
go back to reference Asgharzadeh S, Salo JA, Ji L, Oberthuer A, Fischer M, Berthold F, Hadjidaniel M, Liu CW, Metelitsa LS, Pique-Regi R, Wakamatsu P, Villablanca JG, Kreissman SG, Matthay KK, Shimada H, London WB, Sposto R, Seeger RC (2012) Clinical significance of tumor-associated inflammatory cells in metastatic neuroblastoma. J Clin Oncol 30(28):3525–3532CrossRefPubMedPubMedCentral Asgharzadeh S, Salo JA, Ji L, Oberthuer A, Fischer M, Berthold F, Hadjidaniel M, Liu CW, Metelitsa LS, Pique-Regi R, Wakamatsu P, Villablanca JG, Kreissman SG, Matthay KK, Shimada H, London WB, Sposto R, Seeger RC (2012) Clinical significance of tumor-associated inflammatory cells in metastatic neuroblastoma. J Clin Oncol 30(28):3525–3532CrossRefPubMedPubMedCentral
15.
go back to reference Fox BA, Schendel DJ, Butterfield LH, Aamdal S, Allison JP, Ascierto PA, Atkins MB, Bartunkova J, Bergmann L, Berinstein N, Bonorino CC, Borden E, Bramson JL, Britten CM, Cao X, Carson WE, Chang AE, Characiejus D, Choudhury AR, Coukos G, de Gruijl T, Dillman RO, Dolstra H, Dranoff G, Durrant LG, Finke JH, Galon J, Gollob JA, Gouttefangeas C, Grizzi F, Guida M, Hakansson L, Hege K, Herberman RB, Hodi FS, Hoos A, Huber C, Hwu P, Imai K, Jaffee EM, Janetzki S, June CH, Kalinski P, Kaufman HL, Kawakami K, Kawakami Y, Keilholtz U, Khleif SN, Kiessling R, Kotlan B, Kroemer G, Lapointe R, Levitsky HI, Lotze MT, Maccalli C, Maio M, Marschner JP, Mastrangelo MJ, Masucci G, Melero I, Melief C, Murphy WJ, Nelson B, Nicolini A, Nishimura MI, Odunsi K, Ohashi PS, O’Donnell-Tormey J, Old LJ, Ottensmeier C, Papamichail M, Parmiani G, Pawelec G, Proietti E, Qin S, Rees R, Ribas A, Ridolfi R, Ritter G, Rivoltini L, Romero PJ, Salem ML, Scheper RJ, Seliger B, Sharma P, Shiku H, Singh-Jasuja H, Song W, Straten PT, Tahara H, Tian Z, van Der Burg SH, von Hoegen P, Wang E, Welters MJ, Winter H, Withington T, Wolchok JD, Xiao W, Zitvogel L, Zwierzina H, Marincola FM, Gajewski TF, Wigginton JM, Disis ML (2011) Defining the critical hurdles in cancer immunotherapy. J Transl Med 9:214CrossRefPubMedPubMedCentral Fox BA, Schendel DJ, Butterfield LH, Aamdal S, Allison JP, Ascierto PA, Atkins MB, Bartunkova J, Bergmann L, Berinstein N, Bonorino CC, Borden E, Bramson JL, Britten CM, Cao X, Carson WE, Chang AE, Characiejus D, Choudhury AR, Coukos G, de Gruijl T, Dillman RO, Dolstra H, Dranoff G, Durrant LG, Finke JH, Galon J, Gollob JA, Gouttefangeas C, Grizzi F, Guida M, Hakansson L, Hege K, Herberman RB, Hodi FS, Hoos A, Huber C, Hwu P, Imai K, Jaffee EM, Janetzki S, June CH, Kalinski P, Kaufman HL, Kawakami K, Kawakami Y, Keilholtz U, Khleif SN, Kiessling R, Kotlan B, Kroemer G, Lapointe R, Levitsky HI, Lotze MT, Maccalli C, Maio M, Marschner JP, Mastrangelo MJ, Masucci G, Melero I, Melief C, Murphy WJ, Nelson B, Nicolini A, Nishimura MI, Odunsi K, Ohashi PS, O’Donnell-Tormey J, Old LJ, Ottensmeier C, Papamichail M, Parmiani G, Pawelec G, Proietti E, Qin S, Rees R, Ribas A, Ridolfi R, Ritter G, Rivoltini L, Romero PJ, Salem ML, Scheper RJ, Seliger B, Sharma P, Shiku H, Singh-Jasuja H, Song W, Straten PT, Tahara H, Tian Z, van Der Burg SH, von Hoegen P, Wang E, Welters MJ, Winter H, Withington T, Wolchok JD, Xiao W, Zitvogel L, Zwierzina H, Marincola FM, Gajewski TF, Wigginton JM, Disis ML (2011) Defining the critical hurdles in cancer immunotherapy. J Transl Med 9:214CrossRefPubMedPubMedCentral
16.
go back to reference Bibby MC (2004) Orthotopic models of cancer for preclinical drug evaluation: advantages and disadvantages. Eur J Cancer 40(6):852–857CrossRefPubMed Bibby MC (2004) Orthotopic models of cancer for preclinical drug evaluation: advantages and disadvantages. Eur J Cancer 40(6):852–857CrossRefPubMed
17.
go back to reference Youn J-I, Nagaraj S, Collazo M, Gabrilovich DI (2009) Subsets of myeloid-derived suppressor cells in tumor bearing mice. PMC 181(8):5791–5802 Youn J-I, Nagaraj S, Collazo M, Gabrilovich DI (2009) Subsets of myeloid-derived suppressor cells in tumor bearing mice. PMC 181(8):5791–5802
18.
go back to reference Weiss WA, Aldape K, Mohapatra G, Feuerstein BG, Bishop JM (1997) Targeted expression of MYCN causes neuroblastoma in transgenic mice. EMBO J 16(11):2985–2995CrossRefPubMedPubMedCentral Weiss WA, Aldape K, Mohapatra G, Feuerstein BG, Bishop JM (1997) Targeted expression of MYCN causes neuroblastoma in transgenic mice. EMBO J 16(11):2985–2995CrossRefPubMedPubMedCentral
19.
go back to reference Terrile M, Bryan K, Vaughan L, Hallsworth A, Webber H, Chesler L, Stallings RL (2011) miRNA expression profiling of the murine TH-MYCN neuroblastoma model reveals similarities with human tumors and identifies novel candidate miRNAs. PLoS ONE 6(12):e28356CrossRefPubMedPubMedCentral Terrile M, Bryan K, Vaughan L, Hallsworth A, Webber H, Chesler L, Stallings RL (2011) miRNA expression profiling of the murine TH-MYCN neuroblastoma model reveals similarities with human tumors and identifies novel candidate miRNAs. PLoS ONE 6(12):e28356CrossRefPubMedPubMedCentral
20.
go back to reference Hansford LM, Thomas WD, Keating JM, Burkhart CA, Peaston AE, Norris MD, Haber M, Armati PJ, Weiss WA, Marshall GM (2004) Mechanisms of embryonal tumor initiation distinct roles for MycN expression and MYCN amplification. PNAS 101(34):12669CrossRef Hansford LM, Thomas WD, Keating JM, Burkhart CA, Peaston AE, Norris MD, Haber M, Armati PJ, Weiss WA, Marshall GM (2004) Mechanisms of embryonal tumor initiation distinct roles for MycN expression and MYCN amplification. PNAS 101(34):12669CrossRef
21.
go back to reference Moore HC, Wood KM, Jackson MS, Lastowska MA, Hall D, Imrie H, Redfern CP, Lovat PE, Ponthan F, O’Toole K, Lunec J, Tweddle DA (2008) Histological profile of tumours from MYCN transgenic mice. J Clin Pathol 61(10):1098–1103CrossRefPubMed Moore HC, Wood KM, Jackson MS, Lastowska MA, Hall D, Imrie H, Redfern CP, Lovat PE, Ponthan F, O’Toole K, Lunec J, Tweddle DA (2008) Histological profile of tumours from MYCN transgenic mice. J Clin Pathol 61(10):1098–1103CrossRefPubMed
22.
go back to reference Rasmuson A, Segerstrom L, Nethander M, Finnman J, Elfman LH, Javanmardi N, Nilsson S, Johnsen JI, Martinsson T, Kogner P (2012) Tumor development, growth characteristics and spectrum of genetic aberrations in the TH-MYCN mouse model of neuroblastoma. PLoS ONE 7(12):e51297CrossRefPubMedPubMedCentral Rasmuson A, Segerstrom L, Nethander M, Finnman J, Elfman LH, Javanmardi N, Nilsson S, Johnsen JI, Martinsson T, Kogner P (2012) Tumor development, growth characteristics and spectrum of genetic aberrations in the TH-MYCN mouse model of neuroblastoma. PLoS ONE 7(12):e51297CrossRefPubMedPubMedCentral
23.
go back to reference Kawakubo N, Harada Y, Ishii M, Souzaki R, Kinoshita Y, Tajiri T, Taguchi T, Yonemitsu Y (2018) Natural antibody against neuroblastoma of TH-MYCN transgenic mice does not correlate with spontaneous regression. Biochem Biophys Res Commun 503(3):1666–1673CrossRefPubMed Kawakubo N, Harada Y, Ishii M, Souzaki R, Kinoshita Y, Tajiri T, Taguchi T, Yonemitsu Y (2018) Natural antibody against neuroblastoma of TH-MYCN transgenic mice does not correlate with spontaneous regression. Biochem Biophys Res Commun 503(3):1666–1673CrossRefPubMed
24.
go back to reference Mao Y, Eissler N, Blanc KL, Johnsen JI, Kogner P, Kiessling R (2016) Targeting suppressive myeloid cells potentiates checkpoint inhibitors to control spontaneous neuroblastoma. Clin Cancer Res 22(15):3849–3859CrossRefPubMed Mao Y, Eissler N, Blanc KL, Johnsen JI, Kogner P, Kiessling R (2016) Targeting suppressive myeloid cells potentiates checkpoint inhibitors to control spontaneous neuroblastoma. Clin Cancer Res 22(15):3849–3859CrossRefPubMed
26.
go back to reference Wang X, Yang X, Zhang C, Wang Y, Cheng T, Duan L, Tong Z, Tan S, Zhang H, Saw PE, Gu Y, Wang J, Zhang Y, Shang L, Liu Y, Jiang S, Yan B, Li R, Yang Y, Yu J, Chen Y, Gao GF, Ye Q, Gao S (2020) Tumor cell-intrinsic PD-1 receptor is a tumor suppressor and mediates resistance to PD-1 blockade therapy. Proc Natl Acad Sci U S A 117(12):6640–6650CrossRefPubMedPubMedCentral Wang X, Yang X, Zhang C, Wang Y, Cheng T, Duan L, Tong Z, Tan S, Zhang H, Saw PE, Gu Y, Wang J, Zhang Y, Shang L, Liu Y, Jiang S, Yan B, Li R, Yang Y, Yu J, Chen Y, Gao GF, Ye Q, Gao S (2020) Tumor cell-intrinsic PD-1 receptor is a tumor suppressor and mediates resistance to PD-1 blockade therapy. Proc Natl Acad Sci U S A 117(12):6640–6650CrossRefPubMedPubMedCentral
28.
go back to reference Majzner RG, Simon JS, Grosso JF, Martinez D, Pawel BR, Santi M, Merchant MS, Geoerger B, Hezam I, Marty V, Vielh P, Daugaard M, Sorensen PH, Mackall CL, Maris JM (2017) Assessment of programmed death-ligand 1 expression and tumor-associated immune cells in pediatric cancer tissues. Cancer 123(19):3807–3815CrossRefPubMed Majzner RG, Simon JS, Grosso JF, Martinez D, Pawel BR, Santi M, Merchant MS, Geoerger B, Hezam I, Marty V, Vielh P, Daugaard M, Sorensen PH, Mackall CL, Maris JM (2017) Assessment of programmed death-ligand 1 expression and tumor-associated immune cells in pediatric cancer tissues. Cancer 123(19):3807–3815CrossRefPubMed
29.
go back to reference Aoki T, Hino M, Koh K, Kyushiki M, Kishimoto H, Arakawa Y, Hanada R, Kawashima H, Kurihara J, Shimojo N, Motohashi S (2016) Low frequency of programmed death ligand 1 expression in pediatric cancers. Pediatr Blood Cancer 63(8):1461–1464CrossRefPubMedPubMedCentral Aoki T, Hino M, Koh K, Kyushiki M, Kishimoto H, Arakawa Y, Hanada R, Kawashima H, Kurihara J, Shimojo N, Motohashi S (2016) Low frequency of programmed death ligand 1 expression in pediatric cancers. Pediatr Blood Cancer 63(8):1461–1464CrossRefPubMedPubMedCentral
30.
go back to reference Moreno-Vicente J, Beers SA, Gray JC (2019) PD-1/PD-L1 blockade in paediatric cancers: what does the future hold? Cancer Lett 457:74–85CrossRefPubMed Moreno-Vicente J, Beers SA, Gray JC (2019) PD-1/PD-L1 blockade in paediatric cancers: what does the future hold? Cancer Lett 457:74–85CrossRefPubMed
31.
go back to reference Davis KL, Fox E, Merchant MS, Reid JM, Kudgus RA, Liu X, Minard CG, Voss S, Berg SL, Weigel BJ, Mackall CL (2020) Nivolumab in children and young adults with relapsed or refractory solid tumours or lymphoma (ADVL1412): a multicentre, open-label, single-arm, phase 1–2 trial. Lancet Oncol 21(4):541–550CrossRefPubMedPubMedCentral Davis KL, Fox E, Merchant MS, Reid JM, Kudgus RA, Liu X, Minard CG, Voss S, Berg SL, Weigel BJ, Mackall CL (2020) Nivolumab in children and young adults with relapsed or refractory solid tumours or lymphoma (ADVL1412): a multicentre, open-label, single-arm, phase 1–2 trial. Lancet Oncol 21(4):541–550CrossRefPubMedPubMedCentral
32.
go back to reference Ehlert K, Hansjuergens I, Zinke A, Otto S, Siebert N, Henze G, Lode H (2020) Nivolumab and dinutuximab beta in two patients with refractory neuroblastoma. J Immunother Cancer 8(1):540CrossRef Ehlert K, Hansjuergens I, Zinke A, Otto S, Siebert N, Henze G, Lode H (2020) Nivolumab and dinutuximab beta in two patients with refractory neuroblastoma. J Immunother Cancer 8(1):540CrossRef
33.
go back to reference Kleffel S, Posch C, Barthel SR, Mueller H, Schlapbach C, Guenova E, Elco CP, Lee N, Juneja VR, Zhan Q, Lian CG, Thomi R, Hoetzenecker W, Cozzio A, Dummer R, Mihm MC Jr, Flaherty KT, Frank MH, Murphy GF, Sharpe AH, Kupper TS, Schatton T (2015) Melanoma cell-intrinsic pd-1 receptor functions promote tumor growth. Cell 162(6):1242–1256CrossRefPubMedPubMedCentral Kleffel S, Posch C, Barthel SR, Mueller H, Schlapbach C, Guenova E, Elco CP, Lee N, Juneja VR, Zhan Q, Lian CG, Thomi R, Hoetzenecker W, Cozzio A, Dummer R, Mihm MC Jr, Flaherty KT, Frank MH, Murphy GF, Sharpe AH, Kupper TS, Schatton T (2015) Melanoma cell-intrinsic pd-1 receptor functions promote tumor growth. Cell 162(6):1242–1256CrossRefPubMedPubMedCentral
34.
go back to reference Li H, Li X, Liu S, Guo L, Zhang B, Zhang J, Ye Q (2017) Programmed cell death-1 (PD-1) checkpoint blockade in combination with a mammalian target of rapamycin inhibitor restrains hepatocellular carcinoma growth induced by hepatoma cell-intrinsic PD-1. Hepatology 66(6):1920–1933CrossRefPubMed Li H, Li X, Liu S, Guo L, Zhang B, Zhang J, Ye Q (2017) Programmed cell death-1 (PD-1) checkpoint blockade in combination with a mammalian target of rapamycin inhibitor restrains hepatocellular carcinoma growth induced by hepatoma cell-intrinsic PD-1. Hepatology 66(6):1920–1933CrossRefPubMed
35.
go back to reference Du S, McCall N, Park K, Guan Q, Fontina P, Ertel A, Zhan T, Dicker AP, Lu B (2018) Blockade of tumor-expressed pd-1 promotes lung cancer growth. Oncoimmunology 7(4):e1408747CrossRefPubMedPubMedCentral Du S, McCall N, Park K, Guan Q, Fontina P, Ertel A, Zhan T, Dicker AP, Lu B (2018) Blockade of tumor-expressed pd-1 promotes lung cancer growth. Oncoimmunology 7(4):e1408747CrossRefPubMedPubMedCentral
Metadata
Title
Expression of programmed cell death-1 on neuroblastoma cells in TH-MYCN transgenic mice
Authors
Yuta Takeuchi
Seiichiro Inoue
Akio Odaka
Publication date
01-12-2023
Publisher
Springer Berlin Heidelberg
Published in
Pediatric Surgery International / Issue 1/2023
Print ISSN: 0179-0358
Electronic ISSN: 1437-9813
DOI
https://doi.org/10.1007/s00383-022-05292-y

Other articles of this Issue 1/2023

Pediatric Surgery International 1/2023 Go to the issue