Identifying new biomarkers of aggressive Group 3 and SHH medulloblastoma using 3D hydrogel models, single cell RNA sequencing and 3D OrbiSIMS imaging

Acta Neuropathologica Communications

Research

Franziska Linke¹  James E. C. Johnson¹  Anbarasu Lourdusamy¹  Beth Coyle¹  Daniel Lea²  Andrew C. Peet³  Christopher D. Bennett³  Ian D. Kerr⁴  Catherine L. R. Merry⁵  Anna M. Grabowska⁵  Snow Stolnik⁶  Morgan R. Alexander⁶  Stefanie Kern⁶  David J. Scurr⁶  Rian L. Griffiths⁶  Steven C. Clifford⁷ 

[1] Children’s Brain Tumour Research Centre, School of Medicine, Biodiscovery Institute, University of Nottingham, Nottingham, UK;Digital Research Service, University of Nottingham, Nottingham, UK;Institute of Cancer and Genomic Sciences, University of Birmingham, Birmingham, UK;Birmingham Children’s Hospital, Birmingham, UK;School of Life Sciences, University of Nottingham, Nottingham, UK;School of Medicine, Biodiscovery Institute, University of Nottingham, Nottingham, UK;School of Pharmacy, University of Nottingham, Nottingham, UK;Wolfson Childhood Cancer Research Centre, Translational & Clinical Research Institute, Newcastle University Centre for Cancer, NE1 7RU, Newcastle Upon Tyne, UK;

关键词: Medulloblastoma;    Group 3;    SHH;    Tricarboxylic acid (TCA) cycle;    Small leucine-rich proteoglycans (SLRPs);   

DOI  :  10.1186/s40478-022-01496-4

 received in 2022-10-12, accepted in 2022-12-19,  发布年份 2022

来源: Springer

【 摘 要 】

The most common malignant brain tumour in children, medulloblastoma (MB), is subdivided into four clinically relevant molecular subgroups, although targeted therapy options informed by understanding of different cellular features are lacking. Here, by comparing the most aggressive subgroup (Group 3) with the intermediate (SHH) subgroup, we identify crucial differences in tumour heterogeneity, including unique metabolism-driven subpopulations in Group 3 and matrix-producing subpopulations in SHH. To analyse tumour heterogeneity, we profiled individual tumour nodules at the cellular level in 3D MB hydrogel models, which recapitulate subgroup specific phenotypes, by single cell RNA sequencing (scRNAseq) and 3D OrbiTrap Secondary Ion Mass Spectrometry (3D OrbiSIMS) imaging. In addition to identifying known metabolites characteristic of MB, we observed intra- and internodular heterogeneity and identified subgroup-specific tumour subpopulations. We showed that extracellular matrix factors and adhesion pathways defined unique SHH subpopulations, and made up a distinct shell-like structure of sulphur-containing species, comprising a combination of small leucine-rich proteoglycans (SLRPs) including the collagen organiser lumican. In contrast, the Group 3 tumour model was characterized by multiple subpopulations with greatly enhanced oxidative phosphorylation and tricarboxylic acid (TCA) cycle activity. Extensive TCA cycle metabolite measurements revealed very high levels of succinate and fumarate with malate levels almost undetectable particularly in Group 3 tumour models. In patients, high fumarate levels (NMR spectroscopy) alongside activated stress response pathways and high Nuclear Factor Erythroid 2-Related Factor 2 (NRF2; gene expression analyses) were associated with poorer survival. Based on these findings we predicted and confirmed that NRF2 inhibition increased sensitivity to vincristine in a long-term 3D drug treatment assay of Group 3 MB. Thus, by combining scRNAseq and 3D OrbiSIMS in a relevant model system we were able to define MB subgroup heterogeneity at the single cell level and elucidate new druggable biomarkers for aggressive Group 3 and low-risk SHH MB.

【 授权许可】

CC BY   
© The Author(s) 2023

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