Single-cell profiling of pediatric T-cell acute lymphoblastic leukemia: Impact of PTEN exon 7 mutation on PI3K/Akt and JAK-STAT signaling pathways.
Cell Line, Tumor
Child
Exons
/ genetics
Gene Expression Regulation, Neoplastic
/ genetics
Humans
Interleukin-7
/ genetics
Interleukin-7 Receptor alpha Subunit
/ genetics
Janus Kinases
/ antagonists & inhibitors
Mutation
/ genetics
PTEN Phosphohydrolase
/ genetics
Phosphatidylinositol 3-Kinases
/ genetics
Precursor T-Cell Lymphoblastic Leukemia-Lymphoma
/ diagnosis
Proteomics
/ methods
STAT5 Transcription Factor
/ genetics
Signal Transduction
/ genetics
Single-Cell Analysis
/ methods
T-Lymphocytes
/ metabolism
PTEN
acute lymphoblastic leukemia
cell signaling
childhood
interleukin 7
phoshoflow
Journal
Cytometry. Part B, Clinical cytometry
ISSN: 1552-4957
Titre abrégé: Cytometry B Clin Cytom
Pays: United States
ID NLM: 101235690
Informations de publication
Date de publication:
11 2020
11 2020
Historique:
received:
03
10
2019
revised:
16
04
2020
accepted:
17
04
2020
pubmed:
2
6
2020
medline:
20
8
2021
entrez:
2
6
2020
Statut:
ppublish
Résumé
The PI3K/Akt/mTOR (PI3K) signaling pathway has a crucial role in T-cell acute lymphoblastic leukemias (T-ALLs). Although loss-of-function of phosphatase and tensin homolog (PTEN) is a common event in pediatric T-ALLs, the exact role of this tumor suppressor in T-ALL development has yet to be defined. Here, we report an optimized cytometric method for accurate proteomic profiling of T-ALL leukemic blasts at single-cell level. We determined the expression of PI3K and JAK-STAT signaling components in both primary and immortalized T-ALL cells as well as in normal T cells. We observed that PTEN exon 7 mutated T-ALL cells retain a distinct PI3K activation; in particular, these cells show higher pAkt levels and a lower pS6 expression. Interestingly, we demonstrated for the first time that PTEN exon 7 mutated T-ALL are nonresponsive to IL7 in vitro as assessed by lack of pSTAT5 activation, although they do express IL7R. Phosphoflow analysis represents a fast, reliable, and accurate method to study the signaling profile of T-ALL. PTEN exon 7 mutated T-ALL cells are nonresponsive to IL7 in vitro suggesting that they may activate other mechanisms to support their viability and proliferation such as a higher constitutive PI3K/Akt signaling. Further investigations are necessary to elucidate the significance of this peculiar signaling behavior. Our observations should be taken into account in future studies aiming at molecular targeting of PI3K and/or JAK/STAT pathways for pharmacological intervention in T-ALL.
Sections du résumé
BACKGROUND
The PI3K/Akt/mTOR (PI3K) signaling pathway has a crucial role in T-cell acute lymphoblastic leukemias (T-ALLs). Although loss-of-function of phosphatase and tensin homolog (PTEN) is a common event in pediatric T-ALLs, the exact role of this tumor suppressor in T-ALL development has yet to be defined.
METHODS
Here, we report an optimized cytometric method for accurate proteomic profiling of T-ALL leukemic blasts at single-cell level. We determined the expression of PI3K and JAK-STAT signaling components in both primary and immortalized T-ALL cells as well as in normal T cells.
RESULTS
We observed that PTEN exon 7 mutated T-ALL cells retain a distinct PI3K activation; in particular, these cells show higher pAkt levels and a lower pS6 expression. Interestingly, we demonstrated for the first time that PTEN exon 7 mutated T-ALL are nonresponsive to IL7 in vitro as assessed by lack of pSTAT5 activation, although they do express IL7R.
CONCLUSIONS
Phosphoflow analysis represents a fast, reliable, and accurate method to study the signaling profile of T-ALL. PTEN exon 7 mutated T-ALL cells are nonresponsive to IL7 in vitro suggesting that they may activate other mechanisms to support their viability and proliferation such as a higher constitutive PI3K/Akt signaling. Further investigations are necessary to elucidate the significance of this peculiar signaling behavior. Our observations should be taken into account in future studies aiming at molecular targeting of PI3K and/or JAK/STAT pathways for pharmacological intervention in T-ALL.
Identifiants
pubmed: 32479694
doi: 10.1002/cyto.b.21882
doi:
Substances chimiques
IL7 protein, human
0
IL7R protein, human
0
Interleukin-7
0
Interleukin-7 Receptor alpha Subunit
0
STAT5 Transcription Factor
0
Janus Kinases
EC 2.7.10.2
PTEN Phosphohydrolase
EC 3.1.3.67
PTEN protein, human
EC 3.1.3.67
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
491-503Informations de copyright
© 2020 International Clinical Cytometry Society.
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