Project description:We have employed single cell RNA sequencing using the 10x Genomics platform to study the molecular mechanisms underlying resistance to treatment. In clinically relevant mouse models of G3 medulloblastoma, CT-guided fractionated radiotherapy was given to induce the clonal selection of radioresistant subpopulations. Comparison of recurrent and treatment naïve tumors revealed a prognostic gene expression signature reflective of radio-resistance.
Project description:Medulloblastoma (MB) is the most common malignant paediatric brain tumour, with Group 3 disease representing the most aggressive and therapy-refractory subtype. Despite intensive craniospinal radiotherapy, outcomes remain poor and the molecular basis of radioresistance is unclear. Here we identify CALCOCO2 as a key determinant of Group 3 MB aggressiveness and radiotherapy resistance. CALCOCO2 is markedly upregulated in Group 3 tumours, where high expression correlates with adverse clinical outcome. We show that CALCOCO2 is transcriptionally regulated by the developmental factor OTX2, defining a previously unrecognised axis that sustains a progenitor-like, transformation-permissive state. Functionally, nuclear CALCOCO2 promotes efficient repair of radiation-induced DNA double-strand breaks by engaging POLθ-dependent alternative end joining. Disruption of the CALCOCO2–POLθ pathway impairs DNA repair, sensitizes tumour cells to radiotherapy and suppresses tumour growth in vitro and in vivo. These findings uncover a developmentally wired DNA repair dependency in Group 3 MB and highlight POLθ inhibition as a rational strategy to overcome therapeutic resistance.
Project description:Group 3 medulloblastoma is the most aggressive among the four medulloblastoma subgroups, with poor prognosis and early metastasis making treatment particularly challenging. The lack of an immune-competent mouse model has hindered effective in vivo testing of potential drugs and mechanisms. Given the prevalent MYC overexpression in Group 3 medulloblastoma, we developed a Group 3-like medulloblastoma mouse model in C57BL/6J mice by overexpressing MYC in the cerebellum of newborn pups. This model provides a valuable tool for studying Group 3 medulloblastoma more effectively.
Project description:Group 3 medulloblastoma is the most aggressive among the four medulloblastoma subgroups, with poor prognosis and early metastasis making treatment particularly challenging. The lack of an immune-competent mouse model has hindered effective in vivo testing of potential drugs and mechanisms. Given the prevalent MYC overexpression in Group 3 medulloblastoma, we developed a Group 3-like medulloblastoma mouse model in C57BL/6J mice by overexpressing MYC in the cerebellum of newborn pups. This model provides a valuable tool for studying Group 3 medulloblastoma more effectively.
Project description:PRTG+ve cells show high self renewability in Group 3 medulloblastoma tumors. To access the proteins differentially expressed in this subset of cells, we sorted PRTG+ve and PRTG-ve cells by surface staining with Anti-PRTG antibody from Gr3 medulloblastoma xenografts.
Project description:Investigate the DNA binding pattern as well as transcriptional consequences of ZIC1 and its medulloblastoma mutants in group 3 medulloblastoma cell lines and granule neuron progenitors
Project description:Investigate the DNA binding pattern as well as transcriptional consequences of ZIC1 and its medulloblastoma mutants in group 3 medulloblastoma cell lines and granule neuron progenitors
Project description:We evaluated changes in active and repressive histone modifications following the silencing of OTX2 in Group 3 medulloblastoma tumorspheres