Autophagy Cholesterol Axis Remodeling Supports Malignant Progression and Chemoresistance in Glioma

Shojaei, S.; Barzegar Behrooz, A.; Naghibzadeh, K.; Basso, J.; Alizadeh, J.; Dehesh, T.; Saberi, R.; Bhushan, B.; Eshraghi, M.; Rosa, S. C.; Clark, C.; Tomczyk, M. M.; Cole, L.; Hatch, G.; Dolinsky, V. W.; Yathindranath, V.; Miller, D.; Pascoe, C.; Dhingra, S.; Srivastava, A.; Ravandi, A.; Vitorino, R.; Pecic, S.; Azarpira, N.; Aghaei, M.; Ghavami, S.

doi:10.64898/2026.01.06.697885

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bioRxiv · 10.64898/2026.01.06.697885

Autophagy Cholesterol Axis Remodeling Supports Malignant Progression and Chemoresistance in Glioma

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Abstract

Glioma progression and resistance to temozolomide (TMZ) remain major clinical challenges. Here, we investigated whether dysregulated autophagy and cholesterol metabolism are coordinately remodeled during glioma progression and TMZ resistance. Tissue microarray analysis of astrocytoma and glioblastoma specimens revealed progressive autophagosome accumulation, reflected by increased LC3{beta} puncta, coupled with impaired autophagic flux compared with adjacent normal brain tissue. These alterations intensified with tumor grade and were associated with upregulation of farnesyl diphosphate synthase (FDPS), linking malignant progression to cholesterol pathway remodeling. TMZ-resistant (R) glioblastoma cells exhibited epithelial-to-mesenchymal transition, mitotic quiescence, and mitochondrial remodeling consistent with a therapy-tolerant phenotype. Bioenergetic profiling demonstrated reduced respiratory reserve, diminished ATP-linked respiration, and elevated proton leak, indicating constrained metabolic flexibility. In parallel, impaired autophagy flux was associated with suppression of de novo cholesterol synthesis and transcriptional downregulation of SREBP-2 and LDL-R. Comprehensive lipidomic profiling revealed marked cholesterol metabolic reprogramming in R cells, characterized by accumulation of specific cholesteryl esters, including CE 22:5, CE 22:6, CE 22:4, and CE 20:4, despite reduced cholesterol biosynthesis. Pharmacologic inhibition of the mevalonate pathway with simvastatin significantly altered cholesteryl ester profiles but failed to restore autophagy flux or sensitize R cells to TMZ-induced apoptosis, even under combined TMZ-simvastatin treatment. Lay AbstractAs gliomas progress from astrocytoma to glioblastoma, autophagy becomes dysregulated and cholesterol metabolism is rewired. This coordinated remodeling supports tumor survival, metabolic plasticity, and resistance to temozolomide therapy. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=133 SRC="FIGDIR/small/697885v2_ufig1.gif" ALT="Figure 1000"> View larger version (79K): org.highwire.dtl.DTLVardef@1183dd2org.highwire.dtl.DTLVardef@82e20dorg.highwire.dtl.DTLVardef@c6c8dforg.highwire.dtl.DTLVardef@adb427_HPS_FORMAT_FIGEXP M_FIG C_FIG HighlightsAutophagy flux blockade intensifies during progression from astrocytoma to glioblastoma Dysregulated autophagy is coupled to altered cholesterol metabolism in malignant gliomas TMZ-resistant glioblastoma cells undergo epithelial-to-mesenchymal transition and mitotic quiescence Resistant cells exhibit constrained bioenergetic capacity and mitochondrial remodeling Impaired autophagy suppresses de novo cholesterol synthesis and lipid recycling Lipidomics reveals accumulation of long-chain cholesteryl esters in TMZ-resistant cells Statin-based cholesterol inhibition fails to resensitize glioblastoma cells to temozolomide

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Shojaei, S., Barzegar Behrooz, A., Naghibzadeh, K., Basso, J., Alizadeh, J., Dehesh, T., Saberi, R., Bhushan, B., Eshraghi, M., Rosa, S. C., Clark, C., Tomczyk, M. M., Cole, L., Hatch, G., Dolinsky, V. W., Yathindranath, V., Miller, D., Pascoe, C., Dhingra, S., Srivastava, A., Ravandi, A., Vitorino, R., Pecic, S., Azarpira, N., Aghaei, M., Ghavami, S.. 2026-01-07. Autophagy Cholesterol Axis Remodeling Supports Malignant Progression and Chemoresistance in Glioma. https://doi.org/10.64898/2026.01.06.697885

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