Reactivating PTEN to impair glioma stem cells by inhibiting cytosolic iron-sulfur assembly pathway

Glioblastoma (GBM), the most lethal primary brain tumor, harbors glioma stem cells (GSCs) that not only initiate and maintain malignant phenotypes but also enhance therapeutic resistance. Although frequently mutated in GBMs, the function and regulation of PTEN in PTEN-intact GSCs are unknown. Here we found that PTEN directly interacts with MMS19 and competitively disrupts MMS19-based cytosolic iron-sulfur (Fe-S) cluster assembly (CIA) machinery in the differentiated glioma cells (DGCs). Interrogation of GSCs, when compared with their matched DGCs, revealed that PTEN is specifically succinated at cysteine (C) 211 in GSCs. Isotope tracing coupled with mass spectrometry analysis confirmed that fumarate, generated by adenylosuccinate lyase (ADSL) in de novo purine synthesis pathway which is highly activated in GSCs, promotes PTEN C211 succination. This modification abrogates the interaction between PTEN and MMS19, thereby reactivating CIA machinery pathway in GSCs. Functionally, inhibiting ..., Mice
To identify the potential tumor formation ability of GSCs and DGCs, luciferase-expressing cells (1 × 104) were injected intracranially into 4-week-old female athymic old nude mice (BALB/cNj-Foxn1nu/Gpt, Strain NO. D000521, GemPharmatech) as previously described (25). Each group contained five mice. Mice were fed autoclaved food and water and maintained in a specific pathogen-free facility. Tumor volumes were monitored by detecting the flux activity using a bioluminescence imaging system at different time points.
To teste whether inhibiting PTEN C211sc sensitizes GSCs to TMZ treatment, PTEN-depleted-MGG8 or MES28 GSCs expressing with or without WT Flag-PTEN or Flag-PTEN C211S were transfected with firefly luciferase. 1 × 104 cells were injected intracranially into 4-week-old female athymic old nude mice. Each group contained five mice. 7 days after the injection, mice were treated with PBS or TMZ (20 mg/kg) by intraperitoneal injection. To teste whether inhibiting PTEN C211sc sensit..., , # Reactivating PTEN to impair glioma stem cells by inhibiting cytosolic iron-sulfur assembly pathway

[https://doi.org/10.5061/dryad.2v6wwpzvs ](https://doi.org/10.5061/dryad.2v6wwpzvs)

In this study, we have shown that PTEN to be a substrate of fumarate, it is possible that other substrates could be succinated contributing to glioma progression. In addition, to evaluate the clinical applicability of NAC supplementation in glioma treatment, the GSCs-derived xenograft transplantation mouse model was used to evaluate tumor growth in vivo. While more mouse models, like PDX mouse model, will make the results more solid. Moreover, PTEN C211sc expression in clinical samples are needed to further evaluate the correlation between PTEN C211sc expression and TMZ or radiation therapy sensitivity of glioma patients.

### Description of the data and file structure

Datasets included:

1\. Uncropped blots

Each blots contains the information of ce...