AML-158 Phase Separation Mediates NUP98 Fusion Oncoprotein Leukemic Transformation.

作者：Nicole L. Michmerhuizen, Bappaditya Chandra, H. Shirnekhi, Swarnendu Tripathi, B. Pioso, D. Baggett, Diana M. Mitrea, Ilaria Lacobucci, Michael R. H. White, Jingjing Chen, Cheon‐Gil Park, Huiyun Wu, S. Pounds, A. Medyukhina, K. Khairy, Qingsong Gao, Chunxu Qu, S. Gorman, Simran Bawa, Carolyn Maslanka, Swati Kinger, Priyanka Dogra, Danika Di Giacomo, C. Mecucci, J. Klco, C. Mullighan, R. Kriwacki · 发表于：Clinical Lymphoma, Myeloma & Leukemia · 年份：2022 · DOI：10.1016/s2152-2650(22)01234-4 · 被引用次数：34 · 研究领域：Medicine

CONTEXT Rearrangements of the nucleoporin 98 gene (NUP98) define a high-risk subset of childhood acute myeloid leukemia (AML). The resulting fusion oncoproteins (FOs) involve the N-terminal, intrinsically disordered region of NUP98, and the C-terminal portion of one of more than 30 identified fusion partners. Approximately one third of fusion partners have DNA-binding homeodomains, and the remaining partners have other domains involved in gene regulation. OBJECTIVE NUP98 FOs have long been known to localize in nuclear puncta. Here, we investigated whether these puncta form by liquid-liquid phase separation (LLPS) and how they might contribute to cell transformation. DESIGN We first focused on the NUP98::HOXA9 (NHA9) FO, in which the HOXA9 fusion partner includes a DNA-binding homeodomain. We expressed GFP-tagged NHA9 in HEK293T cells and characterized the resulting FO-associated nuclear puncta. We also investigated lentiviral FO expression in mouse hematopoietic stem and progenitor cells (HSPCs), studying localization using confocal imaging, self-renewal using colony forming unit assays, and gene expression using RNA sequencing. We next mutated the phenylalanine glycine (FG) repeats of NUP98 to disrupt interactions with other FOs and interacting proteins or mutated the homeodomain of HOXA9 to disrupt DNA binding. Finally, we investigated the applicability of our findings to other NUP98 FOs with and without homeodomains. RESULTS NHA9 localizes in nuclear puncta in HEK293...