Long non-coding RNA NEAT1 drives nuclear condensates regulating gene expression and ferroptosis vulnerability in pancreatic cancer cells

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive malignancy with poor patient outcomes due to development of tumor cell drug resistance impacting treatment efficacy. Shifts in tumor cell transcriptional subtypes from well-differentiated classical to poorly differentiated basal-like cells can drive PDAC chemotherapeutic resistance. Such transcriptomic changes have been linked to cellular metabolic states, but how coordination of transcriptional and metabolic programs is mediated at the molecular level remains unclear. The stress-responsive long noncoding RNA NEAT1 , an essential constituent of nuclear condensates called paraspeckles, has been associated with classical PDAC and was previously linked to regulation of cellular metabolism. In this study, we utilized a high-throughput drug screening approach in NEAT1 -deficient cancer cells to uncover functional metabolic targets in these cells, identifying ferroptosis via GPX4 inhibition as a major vulnerability of cells lacking paraspeckles. We find that the paraspeckle-forming isoform of NEAT1 is central to ferroptosis suppression, and we extend these results into PDAC cell line models with genetic suppression or activation of NEAT1 . We find that NEAT1 -nucleated paraspeckles underlie the expression program tied to the classical PDAC subtype and promote ferroptosis resistance. We pinpoint the paraspeckle-suppressive RNA-binding protein QKI, associated with basal-like PDAC, as a positive regulator of ferroptosis susceptibility, possibly through its suppression of paraspeckles. These results link NEAT1 and regulation of paraspeckle condensates to subtype-specific ferroptosis susceptibility in PDAC and offer novel avenues for targeting metabolic vulnerabilities of pancreatic cancer cells on the basis of transcriptional subtype.