Related Experiment Video
Updated: Aug 28, 2025
Flow Cytometry to Estimate Leukemia Stem Cells in Primary Acute Myeloid Leukemia and in Patient-derived-xenografts, at Diagnosis and Follow Up
Published on: March 26, 2018
Ex Vivo Expansion of Phenotypic and Transcriptomic Chronic Myeloid Leukemia Stem Cells
Sweta B Patel1, Valeriya Kuznetsova2, Victoria R Matkins2
1Department of Medicine, Division of Hematology/Oncology, O'Neal Comprehensive Cancer Center, University of Alabama, Birmingham, AL; Division of Hematology, University of Colorado Anschutz Medical Campus, Aurora, CO.
Abstract:
Despite decades of research, standard therapies remain ineffective for most leukemias, pushing toward an essential unmet need for targeted drug screens. Moreover, preclinical drug testing is an important consideration for success of clinical trials without affecting non-transformed stem cells. Using the transgenic chronic myeloid leukemia (CML) mouse model, we determine that leukemic stem cells (LSCs) are transcriptionally heterogenous with a preexistent drug-insensitive signature. To test targeting of potentially important pathways, we establish ex vivo expanded LSCs that have long-term engraftment and give rise to multilineage hematopoiesis. Expanded LSCs share transcriptomic signatures with primary LSCs including enrichment in Wnt, JAK-STAT, MAPK, mTOR and transforming growth factor β signaling pathways. Drug testing on expanded LSCs show that transforming growth factor β and Wnt inhibitors had significant effects on the viability of LSCs, but not leukemia-exposed healthy HSCs. This platform allows testing of multiple drugs at the same time to identify vulnerabilities of LSCs.
Insights
Targeted drug screens are crucial for leukemia treatment. Researchers identified specific drug-insensitive signatures in leukemic stem cells (LSCs) and found Wnt and TGF-β inhibitors effective against LSCs but not healthy stem cells.
Area of Science:
- Hematology
- Cancer Biology
- Pharmacology
Background:
- Standard leukemia therapies are often ineffective, highlighting the need for novel therapeutic strategies.
- Preclinical drug testing is essential for successful clinical trials, particularly for targeting leukemic stem cells (LSCs) without harming healthy stem cells.
Purpose of the Study:
- To investigate the transcriptional heterogeneity of LSCs and identify potential drug targets.
- To develop and validate an ex vivo platform for drug screening against LSCs.
Main Methods:
- Utilized a transgenic chronic myeloid leukemia (CML) mouse model.
- Established ex vivo expanded LSCs with long-term engraftment and multilineage hematopoietic potential.
- Performed transcriptomic analysis to identify enriched signaling pathways in LSCs.
- Conducted drug testing on expanded LSCs and healthy hematopoietic stem cells (HSCs).
Main Results:
- Identified transcriptional heterogeneity and a pre-existing drug-insensitive signature in LSCs.
- Expanded LSCs shared transcriptomic signatures with primary LSCs, including enrichment in Wnt, JAK-STAT, MAPK, mTOR, and transforming growth factor β (TGF-β) pathways.
- Wnt and TGF-β inhibitors demonstrated significant efficacy against LSC viability, while sparing healthy HSCs.
Conclusions:
- Leukemic stem cells exhibit distinct transcriptional profiles that influence drug sensitivity.
- An ex vivo drug screening platform using expanded LSCs can effectively identify targeted therapies that spare healthy stem cells.
- Targeting Wnt and TGF-β signaling pathways represents a promising therapeutic strategy for leukemia.
More Related Videos
09:34A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations
Published on: October 25, 2018
06:33Author Spotlight: Analyzing Bone Marrow Microenvironment in Murine Hematological Malignancies
Published on: November 10, 2023