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Phosphoproteome of glioblastoma cells after FG1059 treatment

ABSTRACT: We found that RPL22L1 isoform switch is regulated by SRSF4 protein and identified a small molecule compound FG1059, that inhibits this process and decreases tumor growth. To confirm that FG1059 affects RPL22L1 due to the inhibition of SRSF protein phosphorylation we studied the phosphoproteome of patient-derived GBM neurospheres treated with FG1059 for 0, 3, 6 and 12 hours.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Brain, Primary Cell, Cell Culture

DISEASE(S): Glioblastoma

SUBMITTER: Georgij Arapidi

LAB HEAD: Georgij Arapidi

PROVIDER: PXD035855 | Pride | 2023-10-24

REPOSITORIES: Pride

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Alternative RNA splicing modulates ribosomal composition and determines the spatial phenotype of glioblastoma cells.

Larionova Tatyana D TD Bastola Soniya S Aksinina Tatiana E TE Anufrieva Ksenia S KS Wang Jia J Shender Victoria O VO Andreev Dmitriy E DE Kovalenko Tatiana F TF Arapidi Georgij P GP Shnaider Polina V PV Kazakova Anastasia N AN Latyshev Yaroslav A YA Tatarskiy Victor V VV Shtil Alexander A AA Moreau Pascale P Giraud Francis F Li Chaoxi C Wang Yichan Y Rubtsova Maria P MP Dontsova Olga A OA Condro Michael M Ellingson Benjamin M BM Shakhparonov Mikhail I MI Kornblum Harley I HI Nakano Ichiro I Pavlyukov Marat S MS

Nature cell biology 20221003 10

Glioblastoma (GBM) is characterized by exceptionally high intratumoral heterogeneity. However, the molecular mechanisms underlying the origin of different GBM cell populations remain unclear. Here, we found that the compositions of ribosomes of GBM cells in the tumour core and edge differ due to alternative RNA splicing. The acidic pH in the core switches before messenger RNA splicing of the ribosomal gene RPL22L1 towards the RPL22L1b isoform. This allows cells to survive acidosis, increases ste ...[more]

PMID: 36192632

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LC-MS/MS identification of proteins that were co-purified with Fc-tagged isoforms of RPL22L1

Project description:Glioblastoma (GBM) is characterized by an exceptionally high intratumoral heterogeneity. However, the molecular mechanisms underlying the origin of different GBM cell populations remain unclear. Here we found that the composition of ribosomes of GBM cells in the tumor core and edge differ due to alternative RNA splicing. The acidic pH in the core switches pre-mRNA splicing of the ribosomal gene RPL22L1 toward the RPL22L1b isoform. To elucidate the functions of RPL22L1 isoforms we identified proteins that interact with RPL22L1a and RPL22L1b. First, we generated GBM cells with stable expression of Fc-tagged RPL22L1 isoforms. Fc-RPL22L1a and Fc-RPL22L1b together with their binding partners were isolated from neurospheres using magnetic beads and analyzed by LC-MS/MS.

2023-10-24 | PXD035767 | Pride

LC-MS/MS identification of proteins that were bound to recombinant His-tagged isoforms of RPL22L1

Project description:Glioblastoma (GBM) is characterized by an exceptionally high intratumoral heterogeneity. However, the molecular mechanisms underlying the origin of different GBM cell populations remain unclear. Here we found that the composition of ribosomes of GBM cells in the tumor core and edge differ due to alternative RNA splicing. The acidic pH in the core switches pre-mRNA splicing of the ribosomal gene RPL22L1 toward the RPL22L1b isoform. To elucidate the functions of RPL22L1 isoforms we identified proteins that interact with RPL22L1a and RPL22L1b. First, each isoform was expressed in E. coli and immobilized on magnetic beads. The beads were incubated with GBM cell lysates and proteins bound to the beads were identified using LC-MS/MS.

2023-10-24 | PXD035849 | Pride

Generation of CNS neural stem cells and PNS derivatives from neural crest derived peripheral stem cells

Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series

2014-11-01 | E-GEOD-57003 | biostudies-arrayexpress

RAS/MAPK activation drives resistance to Smo inhibition, metastasis and tumor evolution in Shh pathway-dependent tumors

Project description:Aberrant Shh signaling promotes tumor growth in diverse human cancers. The importance of Shh signaling is particularly evident in medulloblastoma and basal cell carcinoma (BCC), where inhibitors targeting the Shh pathway component Smoothened (Smo) show great therapeutic promise. However, the emergence of drug resistance limits long-term efficacy and the mechanisms of resistance remain poorly understood. Using new culturing techniques, we established a cohort of Shh pathway-driven medulloblastoma cell lines derived from Ptch+/- mice. Using this new model, we identify activation of the RAS/MAPK pathway circumvents Shh pathway-dependency, drives tumor growth and enhances metastatic behavior.Together these findings reveal a critical role of RAS/MAPK pathway in drug resistance and tumor evolution of Shh pathway-dependent tumors. We performed gene expression profile analysis of SMB cells, in vivo primary MB, as well as cerebellum of normal P6 and adult mice. We also compared gene expression profiles of SMB cells to those of growth factor treated and RAS transformed cells. 24 mouse samples were collected for RNA extraction and hybridization on Affymetrix microarrays, including 3 normal cerebellar as control, 3 primary tumor samples and 18 cell samples with different treatment conditions. We first compared gene expression profiles of SMB cells to those of in vivo primary MB and normal cerebellum. We then compared gene expression profiles of SMB cells to those of growth factor treated and RAS transformed cells.

2015-07-22 | E-GEOD-69359 | biostudies-arrayexpress

Transcription profiling of mouse secondary neurospheres cultured from cells taken from mouse forebrains was compared between FoxO3+/+ (wildtype) and FoxO3-/- (null mutant) mice from the FVB/N background

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2009-12-05 | E-GEOD-18326 | biostudies-arrayexpress

Gene Expression Profiles of Adult Human Olfactory Bulb and Embryonic Neural Stem Cells

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2012-01-30 | E-GEOD-35390 | biostudies-arrayexpress

Generation of CNS neural stem cells and PNS derivatives from neural crest derived peripheral stem cells [Dataset 2]

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2014-11-01 | E-GEOD-57001 | biostudies-arrayexpress

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