In Vivo Screening Unveils Pervasive RNA-Binding Protein Dependencies in Leukemic Stem Cells and Identifies ELAVL1 as a Therapeutic Target.
Abstract
Acute myeloid leukemia (AML) is fueled by leukemic stem cells (LSC) whose determinants are challenging to discern from hematopoietic stem cells (HSC) or uncover by approaches focused on general cell properties. We have identified a set of RNA-binding proteins (RBP) selectively enriched in human AML LSCs. Using an in vivo two-step CRISPR-Cas9 screen to assay stem cell functionality, we found 32 RBPs essential for LSCs in MLL-AF9;NrasG12D AML. Loss-of-function approaches targeting key hit RBP ELAVL1 compromised LSC-driven in vivo leukemic reconstitution, and selectively depleted primitive malignant versus healthy cells. Integrative multiomics revealed differentiation, splicing, and mitochondrial metabolism as key features defining the leukemic ELAVL1-mRNA interactome with mitochondrial import protein, TOMM34, being a direct ELAVL1-stabilized target whose repression impairs AML propagation. Altogether, using a stem cell-adapted in vivo CRISPR screen, this work demonstrates pervasive reliance on RBPs as regulators of LSCs and highlights their potential as therapeutic targets in AML. LSC-targeted therapies remain a significant unmet need in AML. We developed a stem-cell-adapted in vivo CRISPR screen to identify key LSC drivers. We uncover widespread RNA-binding protein dependencies in LSCs, including ELAVL1, which we identify as a novel therapeutic vulnerability through its regulation of mitochondrial metabolism. This article is highlighted in the In This Issue feature, p. 171.
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Linked Datasets (1)
A two-step in vivo CRISPR screen unveils pervasive RNA binding protein dependencies for leukemic stem cells and identifies ELAVL1 as a therapeutic target
Homo sapiens24 data files
| File | Type | Size |
|---|---|---|
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| 1-090191-shSCR-Rep1_S1_L001_R1_001.fastq.gz | RNA-Seq | 2.3 GB |
| 1-090191-shSCR-Rep1_S1_L002_R1_001.fastq.gz | RNA-Seq | 2.3 GB |
| 1-090191-shSCR-Rep1_S1_L002_R1_001.fastq.gz | RNA-Seq | 2.3 GB |
| 2-090191-shSCR-Rep2_S2_L001_R1_001.fastq.gz | RNA-Seq | 1.6 GB |
| 2-090191-shSCR-Rep2_S2_L001_R1_001.fastq.gz | RNA-Seq | 1.6 GB |
| 2-090191-shSCR-Rep2_S2_L002_R1_001.fastq.gz | RNA-Seq | 1.6 GB |
| 2-090191-shSCR-Rep2_S2_L002_R1_001.fastq.gz | RNA-Seq | 1.6 GB |
| 3-090191-shSCR-Rep3_S3_L001_R1_001.fastq.gz | RNA-Seq | 1.5 GB |
| 3-090191-shSCR-Rep3_S3_L001_R1_001.fastq.gz | RNA-Seq | 1.5 GB |
| 3-090191-shSCR-Rep3_S3_L002_R1_001.fastq.gz | RNA-Seq | 1.5 GB |
| 3-090191-shSCR-Rep3_S3_L002_R1_001.fastq.gz | RNA-Seq | 1.5 GB |
| 4-090191-shELAVL1-7-Rep1_S4_L001_R1_001.fastq.gz | RNA-Seq | 1.5 GB |
| 4-090191-shELAVL1-7-Rep1_S4_L001_R1_001.fastq.gz | RNA-Seq | 1.5 GB |
| 4-090191-shELAVL1-7-Rep1_S4_L002_R1_001.fastq.gz | RNA-Seq | 1.5 GB |
| 4-090191-shELAVL1-7-Rep1_S4_L002_R1_001.fastq.gz | RNA-Seq | 1.5 GB |
| 5-090191-shELAVL1-7-Rep2_S5_L001_R1_001.fastq.gz | RNA-Seq | 1.2 GB |
| 5-090191-shELAVL1-7-Rep2_S5_L001_R1_001.fastq.gz | RNA-Seq | 1.2 GB |
| 5-090191-shELAVL1-7-Rep2_S5_L002_R1_001.fastq.gz | RNA-Seq | 1.2 GB |
| 5-090191-shELAVL1-7-Rep2_S5_L002_R1_001.fastq.gz | RNA-Seq | 1.2 GB |
| 6-090191-shELAVL1-7-Rep3_S6_L001_R1_001.fastq.gz | RNA-Seq | 1.4 GB |
| 6-090191-shELAVL1-7-Rep3_S6_L001_R1_001.fastq.gz | RNA-Seq | 1.4 GB |
| 6-090191-shELAVL1-7-Rep3_S6_L002_R1_001.fastq.gz | RNA-Seq | 1.4 GB |
| 6-090191-shELAVL1-7-Rep3_S6_L002_R1_001.fastq.gz | RNA-Seq | 1.4 GB |
Potentially Related Datasets (3)
These accessions were text-mined from the PMC full text. They may be referenced for comparison, cited from other studies, or otherwise mentioned without being primary data for this paper.
In vivo CRISPR screening unveils RNA binding protein dependencies for leukemic stem cells and identifies ELAVL1 as a potential therapeutic …
In vivo CRISPR screening unveils RNA binding protein dependencies for leukemic stem cells and identifies ELAVL1 as a potential therapeutic …