Top

Open Access 17-04-2024 | Cytokines | Research

Rebound growth of BRAF mutant pediatric glioma cells after MAPKi withdrawal is associated with MAPK reactivation and secretion of microglia-recruiting cytokines

Authors: Daniela Kocher, Lei Cao, Romain Guiho, Melanie Langhammer, Yun-Lu Lai, Pauline Becker, Hiba Hamdi, Dennis Friedel, Florian Selt, David Vonhören, Julia Zaman, Gintvile Valinciute, Sonja Herter, Daniel Picard, Johanna Rettenmeier, Kendra K. Maass, Kristian W. Pajtler, Marc Remke, Andreas von Deimling, Stefan Pusch, Stefan M. Pfister, Ina Oehme, David T.W. Jones, Sebastian Halbach, Tilman Brummer, Juan Pedro Martinez-Barbera, Olaf Witt, Till Milde, Romain Sigaud

Published in: Journal of Neuro-Oncology

Abstract

Introduction

Patients with pediatric low-grade gliomas (pLGGs), the most common primary brain tumors in children, can often benefit from MAPK inhibitor (MAPKi) treatment. However, rapid tumor regrowth, also referred to as rebound growth, may occur once treatment is stopped, constituting a significant clinical challenge.

Methods

Four patient-derived pediatric glioma models were investigated to model rebound growth in vitro based on viable cell counts in response to MAPKi treatment and withdrawal. A multi-omics dataset (RNA sequencing and LC-MS/MS based phospho-/proteomics) was generated to investigate possible rebound-driving mechanisms. Following in vitro validation, putative rebound-driving mechanisms were validated in vivo using the BT-40 orthotopic xenograft model.

Results

Of the tested models, only a BRAF^V600E-driven model (BT-40, with additional CDKN2A/Bdel) showed rebound growth upon MAPKi withdrawal. Using this model, we identified a rapid reactivation of the MAPK pathway upon MAPKi withdrawal in vitro, also confirmed in vivo. Furthermore, transient overactivation of key MAPK molecules at transcriptional (e.g. FOS) and phosphorylation (e.g. pMEK) levels, was observed in vitro. Additionally, we detected increased expression and secretion of cytokines (CCL2, CX3CL1, CXCL10 and CCL7) upon MAPKi treatment, maintained during early withdrawal. While increased cytokine expression did not have tumor cell intrinsic effects, presence of these cytokines in conditioned media led to increased attraction of microglia cells in vitro.

Conclusion

Taken together, these data indicate rapid MAPK reactivation upon MAPKi withdrawal as a tumor cell intrinsic rebound-driving mechanism. Furthermore, increased secretion of microglia-recruiting cytokines may play a role in treatment response and rebound growth upon withdrawal, warranting further evaluation.

Available only for authorised users

Ostrom QT, Price M, Ryan K et al (2022) CBTRUS Statistical Report: Pediatric Brain Tumor Foundation Childhood and adolescent primary brain and other Central Nervous System tumors diagnosed in the United States in 2014–2018. Neuro Oncol 24:iii1–iii38. https://doi.org/10.1093/neuonc/noac161CrossRefPubMedPubMedCentral

Louis DN, Perry A, Wesseling P et al (2021) The 2021 WHO classification of tumors of the central nervous system: a summary. Neuro Oncol 23:1231–1251. https://doi.org/10.1093/neuonc/noab106CrossRefPubMedPubMedCentral

Milde T, Rodriguez FJ, Barnholtz-Sloan JS et al (2021) Reimagining pilocytic astrocytomas in the context of pediatric low-grade gliomas. Neuro Oncol. https://doi.org/10.1093/neuonc/noab138CrossRefPubMedPubMedCentral

Armstrong GT, Conklin HM, Huang S et al (2011) Survival and long-term health and cognitive outcomes after low-grade glioma. Neuro Oncol 13:223–234. https://doi.org/10.1093/neuonc/noq178CrossRefPubMed

Jones DTW, Bandopadhayay P, Jabado N (2019) The power of human cancer genetics as revealed by low-grade Gliomas. Annu Rev Genet 53:483–503. https://doi.org/10.1146/annurev-genet-120417-031642CrossRefPubMed

Manoharan N, Liu KX, Mueller S et al (2023) Pediatric low-grade glioma: targeted therapeutics and clinical trials in the molecular era. Neoplasia (United States) 36:100857. https://doi.org/10.1016/j.neo.2022.100857CrossRef

Karoulia Z, Gavathiotis E, Poulikakos PI (2017) New perspectives for targeting RAF kinase in human cancer. Nat Rev Cancer 17:676–691CrossRefPubMedPubMedCentral

Nobre L, Zapotocky M, Ramaswamy V et al (2020) Outcomes of BRAF V600E pediatric Gliomas treated with targeted BRAF inhibition. JCO Precis Oncol 561–571. https://doi.org/10.1200/po.19.00298

Hargrave DR, Bouffet E, Tabori U et al (2019) Efficacy and safety of Dabrafenib in Pediatric patients with BRAF V600 mutation-positive relapsed or refractory low-Grade Glioma: results from a phase I/IIa study. Clin Cancer Res 25:7303–7311. https://doi.org/10.1158/1078-0432.CCR-19-2177CrossRefPubMed

10.

Barbato MI, Nashed J, Bradford D et al (2023) FDA approval Summary: Dabrafenib in combination with trametinib for BRAF V600E mutation-positive low-grade glioma. Clin Cancer Res 30:OF1–OF6. https://doi.org/10.1158/1078-0432.CCR-23-1503CrossRef

11.

Bouffet E, Hansford JR, Garrè ML et al (2023) Dabrafenib plus Trametinib in Pediatric Glioma with BRAF V600 mutations. N Engl J Med 389:1108–1120. https://doi.org/10.1056/NEJMoa2303815CrossRefPubMed

12.

Tsai JW, Choi JJ, Ouaalam H et al (2023) Integrated response analysis of pediatric low-grade gliomas during and after targeted therapy treatment. Neuro-Oncology Adv 5. https://doi.org/10.1093/noajnl/vdac182

13.

Selt F, van Tilburg CM, Bison B et al (2020) Response to trametinib treatment in progressive pediatric low-grade glioma patients. J Neurooncol 149:499–510. https://doi.org/10.1007/s11060-020-03640-3CrossRefPubMedPubMedCentral

14.

Selt F, Hohloch J, Hielscher T et al (2017) Establishment and application of a novel patient-derived KIAA1549: BRAF-driven pediatric pilocytic astrocytoma model for preclinical drug testing. Oncotarget 8:11460–11479. https://doi.org/10.18632/oncotarget.14004CrossRefPubMed

15.

Selt F, Sigaud R, Valinciute G et al (2023) BH3 mimetics targeting BCL-XL impact the senescent compartment of pilocytic astrocytoma. Neuro Oncol 25:735–747. https://doi.org/10.1093/neuonc/noac199CrossRefPubMed

16.

Selt F, El Damaty A, Schuhmann MU et al (2023) Generation of patient-derived pediatric pilocytic astrocytoma in-vitro models using SV40 large T: evaluation of a modeling workflow. J Neurooncol 165:467–478. https://doi.org/10.1007/s11060-023-04500-6CrossRefPubMedPubMedCentral

17.

Valinciute G, Ecker J, Selt F et al (2023) Class I HDAC inhibitor entinostat synergizes with PLK1 inhibitors in MYC-amplified medulloblastoma cells. J Neurooncol 163:143–158. https://doi.org/10.1007/s11060-023-04319-1CrossRefPubMedPubMedCentral

18.

Milde T, Oehme I, Korshunov A et al (2010) HDAC5 and HDAC9 in medulloblastoma: novel markers for risk stratification and role in tumor cell growth. Clin Cancer Res 16:3240–3252. https://doi.org/10.1158/1078-0432.CCR-10-0395CrossRefPubMed

19.

Kolb EA, Gorlick R, Houghton PJ et al (2010) Initial testing (stage 1) of AZD6244 (ARRY-142886) by the pediatric preclinical testing program. Pediatr Blood Cancer 55:668–677. https://doi.org/10.1002/pbc.22576CrossRefPubMedPubMedCentral

20.

Ram A, Murphy D, DeCuzzi N et al (2023) A guide to ERK dynamics, part 2: downstream decoding. Biochem J 480:1909–1928. https://doi.org/10.1042/BCJ20230277CrossRefPubMed

21.

Murphy LO, Smith S, Chen RH et al (2002) Molecular, interpretation of ERK signal duration by immediate early gene products. Nat Cell Biol 4:556–564. https://doi.org/10.1038/ncb822CrossRefPubMed

22.

He M, Dong H, Huang Y et al (2016) Astrocyte-derived CCL2 is associated with m1 activation and recruitment of cultured microglial cells. Cell Physiol Biochem 38:859–870. https://doi.org/10.1159/000443040CrossRefPubMed

23.

Hulshof S, van Haastert ES, Kuipers HF et al (2003) CX3CL1 and CX3CR1 expression in human brain tissue: Noninflammatory Control versus multiple sclerosis. J Neuropathol Exp Neurol 62:899–907. https://doi.org/10.1093/jnen/62.9.899CrossRefPubMed

24.

Held-Feindt J, Hattermann K, Müerköster SS et al (2010) CX3CR1 promotes recruitment of human glioma-infiltrating microglia/macrophages (GIMs). Exp Cell Res 316:1553–1566. https://doi.org/10.1016/j.yexcr.2010.02.018CrossRefPubMed

25.

Milde T, Fangusaro J, Fisher MJ et al (2023) Optimizing preclinical pediatric low-grade glioma models for meaningful clinical translation. Neuro Oncol 25:1920–1931. https://doi.org/10.1093/neuonc/noad125CrossRefPubMed

26.

Buhl JL, Selt F, Hielscher T et al (2019) The senescence-associated secretory phenotype mediates Oncogene-induced senescence in Pediatric Pilocytic Astrocytoma. Clin Cancer Res 25:1851–1866. https://doi.org/10.1158/1078-0432.CCR-18-1965CrossRefPubMed

27.

Ahuja D, Sáenz-Robles MT, Pipas JM (2005) SV40 large T antigen targets multiple cellular pathways to elicit cellular transformation. Oncogene 24:7729–7745CrossRefPubMed

28.

Usta D, Sigaud R, Buhl JL et al (2020) A cell-based MAPK reporter assay reveals synergistic MAPK pathway activity suppression by MAPK inhibitor combination in BRAF-driven pediatric low-grade glioma cells. Mol Cancer Ther 19:1736–1750. https://doi.org/10.1158/1535-7163.MCT-19-1021CrossRefPubMed

29.

Soto-Gamez A, Quax WJ, Demaria M (2019) Regulation of Survival Networks in senescent cells: from mechanisms to interventions. J Mol Biol 431:2629–2643CrossRefPubMed

30.

Lassaletta A, Zapotocky M, Mistry M et al (2017) Therapeutic and prognostic implications of BRAF V600E in Pediatric Low-Grade Gliomas. J Clin Oncol 35:2934–2941. https://doi.org/10.1200/JCO.2016.71.8726CrossRefPubMedPubMedCentral

31.

Leung GP, Feng T, Sigoillot FD et al (2019) Hyperactivation of MAPK signaling is deleterious to RAS/RAF-mutant Melanoma. Mol Cancer Res 17:199–211. https://doi.org/10.1158/1541-7786.MCR-18-0327CrossRefPubMed

32.

Chiou LW, Chan CH, Jhuang YL et al (2023) DNA replication stress and mitotic catastrophe mediate sotorasib addiction in KRASG12C-mutant cancer. J Biomed Sci 30:50. https://doi.org/10.1186/s12929-023-00940-4CrossRefPubMedPubMedCentral

33.

Farnsworth DA, Inoue Y, Johnson FD et al (2022) MEK inhibitor resistance in lung adenocarcinoma is associated with addiction to sustained ERK suppression. npj Precis Oncol 6:1–15. https://doi.org/10.1038/s41698-022-00328-xCrossRef

34.

Pratilas CA, Taylor BS, Ye Q et al (2009) V600EBRAF is associated with disabled feedback inhibition of RAF-MEK signaling and elevated transcriptional output of the pathway. Proc Natl Acad Sci U S A 106:4519–4524. https://doi.org/10.1073/pnas.0900780106CrossRefPubMedPubMedCentral

35.

Ito T, Young MJ, Li R et al (2021) Paralog knockout profiling identifies DUSP4 and DUSP6 as a digenic dependence in MAPK pathway-driven cancers. Nat Genet 53:1664–1672. https://doi.org/10.1038/s41588-021-00967-zCrossRefPubMed

36.

Ecker V, Brandmeier L, Stumpf M et al (2023) Negative feedback regulation of MAPK signaling is an important driver of chronic lymphocytic leukemia progression. Cell Rep 42:113017. https://doi.org/10.1016/j.celrep.2023.113017CrossRefPubMed

37.

Corrales E, Levit-Zerdoun E, Metzger P et al (2022) PI3K/AKT signaling allows for MAPK/ERK pathway independency mediating dedifferentiation-driven treatment resistance in melanoma. Cell Commun Signal 20:187. https://doi.org/10.1186/s12964-022-00989-yCrossRefPubMedPubMedCentral

38.

Obenauf AC, Zou Y, Ji AL et al (2015) Therapy-induced tumour secretomes promote resistance and tumour progression. Nature 520:368–372. https://doi.org/10.1038/nature14336CrossRefPubMedPubMedCentral

39.

Shi H, Hugo W, Kong X et al (2014) Acquired resistance and clonal evolution in melanoma during BRAF inhibitor therapy. Cancer Discov 4:80–93. https://doi.org/10.1158/2159-8290.CD-13-0642CrossRefPubMed

40.

Shi H, Kong X, Ribas A, Lo RS (2011) Combinatorial treatments that overcome PDGFRβ-driven resistance of melanoma cells to V600EB-RAF inhibition. Cancer Res 71:5067–5074. https://doi.org/10.1158/0008-5472.CAN-11-0140CrossRefPubMedPubMedCentral

41.

Steinbrunn T, Stühmer T, Sayehli C et al (2012) Combined targeting of MEK/MAPK and PI3K/Akt signalling in multiple myeloma. Br J Haematol 159:430–440. https://doi.org/10.1111/bjh.12039CrossRefPubMed

42.

Schreck KC, Allen AN, Wang J, Pratilas CA (2020) Combination MEK and mTOR inhibitor therapy is active in models of glioblastoma. https://doi.org/10.1093/noajnl/vdaa138. Neuro-Oncology Adv 2:

43.

Arnold A, Yuan M, Price A et al (2019) Synergistic activity of mTORC1/2 kinase and MEK inhibitors suppresses pediatric low-grade glioma tumorigenicity and vascularity. Neuro Oncol. https://doi.org/10.1093/neuonc/noz230CrossRefPubMedPubMedCentral

44.

Liu Y, Cai Y, Liu L et al (2018) Crucial biological functions of CCL7 in cancer. https://doi.org/10.7717/peerj.4928. PeerJ 2018:

45.

Liu M, Guo S, Stiles JK (2011) The emerging role of CXCL10 in cancer. Oncol Lett 2:583–589CrossRefPubMedPubMedCentral

46.

Xu M, Wang Y, Xia R et al (2021) Role of the CCL2-CCR2 signalling axis in cancer: mechanisms and therapeutic targeting. Cell Prolif. 54

47.

Conroy MJ, Lysaght J (2020) CX3CL1 Signaling in the Tumor Microenvironment. Advances in Experimental Medicine and Biology. Springer, pp 1–12

48.

Clarner T, Janssen K, Nellessen L et al (2015) CXCL10 triggers early microglial activation in the Cuprizone Model. 194:3400–3413

49.

Wetzel K, Menten P, Opdënakker G et al (2001) Transduction of human MCP-3 by a parvoviral vector induces leukocyte infiltration and reduces growth of human cervical carcinoma cell xenografts. J Gene Med 3:326–337. https://doi.org/10.1002/jgm.191CrossRefPubMed

50.

Tsou CL, Peters W, Si Y et al (2007) Critical roles for CCR2 and MCP-3 in monocyte mobilization from bone marrow and recruitment to inflammatory sites. J Clin Invest 117:902–909. https://doi.org/10.1172/JCI29919CrossRefPubMedPubMedCentral

51.

Yoneyama H, Narumi S, Zhang Y et al (2002) Pivotal role of dendritic cell-derived CXCL10 in the retention of T helper cell 1 lymphocytes in secondary lymph nodes. J Exp Med 195:1257–1266. https://doi.org/10.1084/jem.20011983CrossRefPubMedPubMedCentral

52.

Chen R, Keoni C, Waker CA et al (2019) KIAA1549-BRAF expression establishes a permissive Tumor Microenvironment through NFκB-Mediated CCL2 production. Neoplasia (United States) 21:52–60. https://doi.org/10.1016/j.neo.2018.11.007CrossRef

53.

Sigaud R, Albert TK, Hess C et al (2023) MAPK inhibitor sensitivity scores predict sensitivity driven by the immune infiltration in pediatric low-grade gliomas. Nat Commun 14. https://doi.org/10.1038/s41467-023-40235-8

54.

Krug K, Mertins P, Zhang B et al (2019) A curated resource for Phosphosite-specific signature analysis. Mol Cell Proteom 18:576–593. https://doi.org/10.1074/mcp.TIR118.000943CrossRef

55.

Wagle M-C, Kirouac D, Klijn C et al (2018) A transcriptional MAPK pathway activity score (MPAS) is a clinically relevant biomarker in multiple cancer types. npj Precis Oncol 2:7. https://doi.org/10.1038/s41698-018-0051-4CrossRefPubMedPubMedCentral

56.

Maxwell MJ, Arnold A, Sweeney H et al (2021) Unbiased proteomic and phosphoproteomic analysis identifies response signatures and novel susceptibilities after combined MEK and mTOR inhibition in BRAFV600E mutant glioma. Mol Cell Proteom 20. https://doi.org/10.1016/j.mcpro.2021.100123

Title: Rebound growth of BRAF mutant pediatric glioma cells after MAPKi withdrawal is associated with MAPK reactivation and secretion of microglia-recruiting cytokines
Authors: Daniela Kocher
Lei Cao
Romain Guiho
Melanie Langhammer
Yun-Lu Lai
Pauline Becker
Hiba Hamdi
Dennis Friedel
Florian Selt
David Vonhören
Julia Zaman
Gintvile Valinciute
Sonja Herter
Daniel Picard
Johanna Rettenmeier
Kendra K. Maass
Kristian W. Pajtler
Marc Remke
Andreas von Deimling
Stefan Pusch
Stefan M. Pfister
Ina Oehme
David T.W. Jones
Sebastian Halbach
Tilman Brummer
Juan Pedro Martinez-Barbera
Olaf Witt
Till Milde
Romain Sigaud
Publication date: 17-04-2024
Publisher: Springer US
Keywords: Cytokines
Glioma
Glioma
Glioma
Published in: Journal of Neuro-Oncology
Print ISSN: 0167-594X
Electronic ISSN: 1573-7373
DOI: https://doi.org/10.1007/s11060-024-04672-9

At a glance: The STEP trials

Springer Medicine

Rebound growth of BRAF mutant pediatric glioma cells after MAPKi withdrawal is associated with MAPK reactivation and secretion of microglia-recruiting cytokines

Abstract

Introduction

Methods

Results

Conclusion

At a glance: The STEP trials

Springer Medicine

Abstract

Introduction

Methods

Results

Conclusion

Please log in to get access to this content