Pediatric Acute Myeloid Leukaemia (AML), has seen limited therapeutic advancement over the last two decades. Notably, cryptic translocations involving the NUP98 gene have emerged as key contributors to high-risk, chemoresistant cases. Within this context, LNP1 stands out as a "tdark" fusion partner. LNP1's obscure physiological functions and its specific role in AML onset and progression present a knowledge gap that hinders improved therapeutic strategies.
By integrating advanced structural and molecular techniques with functional assays, our aim is to decode the multifaceted interactions and roles that LNP1 undertakes, especially when fused with NUP98. We aim to:
Aim 1: Leveraging cryo-electron microscopy, we will characterize LNP1 and NUP98-LNP1 molecular structure. Its cellular localization will be determined using immunofluorescence microscopy. Overexpression studies will discern the impact of LNP1 and NUP98-LNP1 in hematopoietic cells.
Aim 2: A combination of lentiviral transductions, Co-IP, CHIP-seq and RNA-seq will be employed. This comprehensive experimental set up will uncover protein-protein/DNA interactions, transcriptional alterations, and broader cellular implications resulting from the NUP98-LNP1 fusion.
Aim 3: To decipher the LNP1 functional domains relevant to malignant transformation, we will deploy an incremental lentiviral truncation library targeting NUP98-LNP1. Assessing effects of specific domain truncations on NUP98-LNP1 oncogenic attributes will spotlight key functional segments. Experiments will be conducted on CD34+ cells from cord blood.
We anticipate gaining critical insights into LNP1 molecular structure, delving into its cellular dynamics, and understanding its pathogenic implications. We are determined to illuminate the molecular mechanisms and the key domains by which NUP98-LNP1 induces malignant transformation.
Our findings will fill the current knowledge gap surrounding LNP1 in NUP98-rearranged AML, paving the way for new therapies.