Targeting NUP214 Eradicates Leukemia Stem Cells By Inducing Ferroptosis

Abstract

Acute myeloid leukemia (AML) arises from and are maintained by leukemia stem cell (LSCs), which are also critical causes for drug resistance and relapse of AML. The critical genes involved in LSC maintenance remain largely unknown. Here, we performed a pooled negative-selection screen with a customized small hairpin RNA (shRNA) library against 1191 known pan-essential genes of AML blast cells. We found that several Nucleoporins (NUPs) genes were essential for LSC survival, and NUP214 was highly expressed in LSCs. Knock-out of NUP214 completely eradicated LSCs in MLL-AF9 and HOXA9-MEIS1 AML mouse models, but also comprised HSC maintenance and function. Haploinsufficiency of NUP214 resulted in the increased death of LSCs (~4% to ~33%), reduced LSC numbers, and extended survival of AML mice. In contrast, the haploinsufficiency of NUP214 had a negligible effect on normal hematopoietic stem cells (HSCs) and hematopoiesis. Haploinsufficiency of NUP214 substantially induced lipid peroxidation and resulted in ferroptosis of LSCs, attributed to the remarkably increased transcription of lipoxygenase-15 (ALOX15) and heme oxygenase-1 (HMOX1), evidenced by the results of RNA-seq, proteome, and functional experiments. Immunofluorescence revealed that NUP214 was a mobile nucleoporin and largely located inside the nucleoplasm in LSCs, while NUP214 was mainly located in the nuclear periphery. Co-immunoprecipitation and Cleavage Under Targets and Tagmentation (CUT&Tag) showed that NUP214 directly interacted with transcriptional coactivator Sub1 in LSCs, binding the gene locus of HMOX1 and ALOX15, eventually inhibiting their transcription. Collectively, our findings reveal the essential roles of NUP214 in LSC maintenance and provide a valuable target for AML therapy.