High Expression of NTRK1 in ETV6::RUNX1 Positive Acute Lymphoblastic Leukaemia Drives Factor Independence and Sensitivity to Larotrectinib

Luke Quinlan; Elyse C Page; Jacqueline Rehn; Barbara J McClure; Michael P Osborn; Andrew S Moore; Rishi S Kotecha; Michelle J Henderson; Michelle Martin; Frank Alvaro; David T Yeung; Deborah L White; Susan L Heatley

doi:10.1002/pbc.31983

High Expression of NTRK1 in ETV6::RUNX1 Positive Acute Lymphoblastic Leukaemia Drives Factor Independence and Sensitivity to Larotrectinib

Pediatr Blood Cancer. 2025 Nov;72(11):e31983. doi: 10.1002/pbc.31983. Epub 2025 Aug 28.

Authors

Luke Quinlan^{1

2}, Elyse C Page^{1

2}, Jacqueline Rehn^{1

2}, Barbara J McClure^{1

2}, Michael P Osborn^{2

3

4

5}, Andrew S Moore^{4

6

7}, Rishi S Kotecha^{8

9

10}, Michelle J Henderson^{11

12}, Michelle Martin¹³, Frank Alvaro¹⁴, David T Yeung^{1

2

5

15}, Deborah L White^{1

2

4

15}, Susan L Heatley^{1

2

4}

Affiliations

¹ South Australian Health and Medical Research Institute (SAHMRI), Blood Cancer Program, Precision Cancer Medicine Theme, Adelaide, South Australia, Australia.
² Faculty of Health and Medical Sciences, University of Adelaide, Adelaide, South Australia, Australia.
³ Women's and Children's Hospital, Adelaide, South Australia, Australia.
⁴ Australian and New Zealand Children's Haematology/Oncology Group (ANZCHOG), Clayton, Victoria, Australia.
⁵ Royal Adelaide Hospital, Adelaide, South Australia, Australia.
⁶ Oncology Service, Children's Health Queensland Hospital and Health Service, Brisbane, Queensland, Australia.
⁷ Child Health Research Centre, The University of Queensland, Brisbane, Australia.
⁸ The Kids Research Institute Australia, Leukaemia Translational Research Laboratory, WA Kids Cancer Centre, University of Western Australia, Perth, Western Australia, Australia.
⁹ Perth Children's Hospital, Perth, Western Australia, Australia.
¹⁰ Curtin Medical School, Curtin University, Perth, Western Australia, Australia.
¹¹ Children's Cancer Institute, Sydney, New South Wales, Australia.
¹² School of Clinical Medicine, UNSW Medicine and Health, University of New South Wales Sydney, New South Wales, Australia.
¹³ Monash Children's Cancer Centre, Melbourne, Victoria, Australia.
¹⁴ John Hunter Children's Hospita, Newcastle, New South Wales, Australia.
¹⁵ Australasian Leukaemia and Lymphoma Group (ALLG), Melbourne, Victoria, Australia.

PMID: 40873269
DOI: 10.1002/pbc.31983

Abstract

Background: ETV6::RUNX1 is one of the most common recurrent genomic abnormalities in acute lymphoblastic leukaemia (ALL) and is associated with a good prognosis. High expression of NTRK1, encoding tropomyosin receptor kinase A (TrkA), confers a poor prognosis in other malignancies and may contribute to therapy resistance in patients with ETV6::RUNX1 B-ALL.

Method: Relapse-free survival was estimated by the Kaplan-Meier and log-rank analyses. ETV6::RUNX1 and NTRK1 were isolated from patient cDNA and transduced into IL-3-dependent Ba/F3 cells. Proliferation was assessed via CellTiter-Glo-2.0 with the addition of the TrkA agonist, nerve growth factor (NGF). Sensitivity to the TRK inhibitor larotrectinib was assessed via Annexin V/7AAD staining and flow cytometry. Changes in TrkA signalling through effectors, ERK and AKT, were evaluated by western blotting.

Results: Patients with ETV6::RUNX1 had increased NTRK1 expression in comparison with other B-ALL cases (p < 0.0001), with those overexpressing NTRK1 exhibiting a trend towards increased relapse. in vitro experiments revealed that only Ba/F3 ETV6::RUNX1+NTRK1 cells demonstrated IL-3 independence, indicative of leukaemic transformation (vs parental Ba/F3, p < 0.0001). These cells were sensitive to larotrectinib (LD₅₀ 161 nM) and significantly decreased phosphorylation of ERK and AKT (vs NGF p = 0.0004; p = 0.007, respectively).

Conclusion: NTRK1 overexpression has not previously been reported in B-ALL and was associated with increased relapse in patients with ETV6::RUNX1 and NTKR1 overexpression. The in vitro success of larotrectinib treatment warrants further in vivo investigation and may be a viable therapeutic addition for patients with ETV6::RUNX1 B-ALL and NTRK1 overexpression.

Keywords: ALL; ALL relapse; Cancer genetics; Hematology/Oncology; Pediatric hematology/oncology; molecular diagnosis & therapy.

MeSH terms

Adolescent
Child
Child, Preschool
Core Binding Factor Alpha 2 Subunit* / genetics
Core Binding Factor Alpha 2 Subunit* / metabolism
Drug Resistance, Neoplasm*
ETS Translocation Variant 6 Protein
Female
Humans
Male
Oncogene Proteins, Fusion* / genetics
Oncogene Proteins, Fusion* / metabolism
Precursor Cell Lymphoblastic Leukemia-Lymphoma* / drug therapy
Precursor Cell Lymphoblastic Leukemia-Lymphoma* / genetics
Precursor Cell Lymphoblastic Leukemia-Lymphoma* / metabolism
Precursor Cell Lymphoblastic Leukemia-Lymphoma* / pathology
Prognosis
Proto-Oncogene Proteins c-ets* / genetics
Proto-Oncogene Proteins c-ets* / metabolism
Pyrazoles* / pharmacology
Pyrazoles* / therapeutic use
Pyrimidines* / pharmacology
Pyrimidines* / therapeutic use
Receptor, trkA* / biosynthesis
Receptor, trkA* / genetics
Receptor, trkA* / metabolism
Repressor Proteins* / genetics
Repressor Proteins* / metabolism
Tumor Cells, Cultured

Substances

Receptor, trkA
Core Binding Factor Alpha 2 Subunit
NTRK1 protein, human
larotrectinib
Pyrazoles
Pyrimidines
RUNX1 protein, human
Oncogene Proteins, Fusion
ETS Translocation Variant 6 Protein
Proto-Oncogene Proteins c-ets
Repressor Proteins

Abstract

MeSH terms

Substances

Grants and funding