FLT3 is genetically essential for ITD-mutated leukemic stem cells but dispensable for human hematopoietic stem cells

J Joana L. Araújo (1Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada) E Elvin Wagenblast V Veronique Voisin J Jessica McLeod O Olga I. Gan (1Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada) S Suraj Bansal (1Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada) L Liqing Jin A Amanda Mitchell (1Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada) B Blaise Gratton (1Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada) S Sarah Cutting (1Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada) A Andrea Arruda (Princess Margaret Cancer Centre, University Health Network) M Monica Doedens (1Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada) A Anthea Travas (10Department of Medical Oncology and Hematology, Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada) D Dennis Kim (1Princess Margaret Cancer Centre, University Health Network, University of Toronto, Toronto, Canada) J Jose-Mario Capo-Chichi (2University Health Network, Advanced Molecular Diagnostics Laboratory, Toronto, Canada) S Sagi Abelson M Mark D. Minden (Princess Margaret Cancer Centre, University Health Network) J Jean C. Y. Wang (1Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada) M Manuel A. Sobrinho-Simões (2Department of Hematology, Centro Hospitalar Universitário de São João, Porto, Portugal) P Perpétua Pinto-do-Ó (4Instituto de Investigação e Inovação em Saúde, University of Porto, Porto, Portugal) E Eric Lechman (2University of Pittsburgh, Pittsburgh, United States) J John E. Dick

Abstract

Abstract Leukemic stem cells (LSCs) fuel acute myeloid leukemia (AML) growth and relapse, but therapies tailored toward eradicating LSCs without harming normal hematopoietic stem cells (HSCs) are lacking. FMS-like tyrosine kinase 3 (FLT3) is considered an important therapeutic target due to frequent mutation in AML and association with relapse. However, there has been limited clinical success with FLT3 drug targeting, suggesting either that FLT3 is not a vulnerability in LSC or that more potent inhibition is required, a scenario where HSC toxicity could become limiting. We tested these possibilities by ablating FLT3 using CRISPR/Cas9-mediated FLT3 knockout (FLT3-KO) in human LSCs and HSCs followed by functional xenograft assays. FLT3-KO in LSCs from FLT3–internal tandem duplication (ITD)-mutated but not FLT3–wild-type AMLs resulted in short-term leukemic grafts of FLT3-KO edited cells that disappeared by 12 weeks. By contrast, FLT3-KO in HSCs from the fetal liver, cord blood, and adult bone marrow did not impair multilineage hematopoiesis in primary and secondary xenografts. Our study establishes FLT3 as an ideal therapeutic target where ITD-positive LSCs are eradicated upon FLT3 deletion whereas HSCs are spared. These findings support the development of more potent FLT3-targeting drugs or gene-editing approaches for LSC eradication to improve clinical outcomes.

Article Details

Journal Blood
Volume / Issue Vol. 145, Issue 20
Published May 15, 2025
Pages 2361-2373
ISSN 0006-4971
Publisher Elsevier BV

Journal Info

Blood

Elsevier BV

ISSN: 0006-4971 Health Sciences

Authors (22)

J

Joana L. Araújo

1Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada

E

Elvin Wagenblast

V

Veronique Voisin

J

Jessica McLeod

O

Olga I. Gan

1Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada

S

Suraj Bansal

1Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada

L

Liqing Jin

A

Amanda Mitchell

1Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada

B

Blaise Gratton

1Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada

S

Sarah Cutting

1Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada

A

Andrea Arruda

Princess Margaret Cancer Centre, University Health Network

M

Monica Doedens

1Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada

A

Anthea Travas

10Department of Medical Oncology and Hematology, Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada

D

Dennis Kim

1Princess Margaret Cancer Centre, University Health Network, University of Toronto, Toronto, Canada

J

Jose-Mario Capo-Chichi

2University Health Network, Advanced Molecular Diagnostics Laboratory, Toronto, Canada

S

Sagi Abelson

M

Mark D. Minden

Princess Margaret Cancer Centre, University Health Network

J

Jean C. Y. Wang

1Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada

M

Manuel A. Sobrinho-Simões

2Department of Hematology, Centro Hospitalar Universitário de São João, Porto, Portugal

P

Perpétua Pinto-do-Ó

4Instituto de Investigação e Inovação em Saúde, University of Porto, Porto, Portugal

E

Eric Lechman

2University of Pittsburgh, Pittsburgh, United States

J

John E. Dick