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. 2024 Jan;38(1):193-197.
doi: 10.1038/s41375-023-02096-4. Epub 2023 Nov 25.

A positive feedback loop regulation between NOTCH1 and USP11 in T-cell leukemia

Igor Fijalkowski #  1   2   3 , Jin Wang #  4 , Qi Jin  5   6 , Jolien Van Laere  1   2   3 , Valentina Serafin  7   8 , John D Crispino  6 , Panagiotis Ntziachristos  9   10   11
Affiliations

A positive feedback loop regulation between NOTCH1 and USP11 in T-cell leukemia

Igor Fijalkowski et al. Leukemia. 2024 Jan.

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No abstract available

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Conflict of interest statement

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1. NOTCH1 and USP11 expression levels are positively correlated in T-ALL.
A Volcano plot showing Pearson correlation between 52 USPs and NOTCH1 mRNA levels in T-ALL patient samples analyzed from the Pediatric Cancer Genome Project data portal (PeCan, St. Jude, Memphis). B Pearson correlation between NOTCH1 and USP11 mRNA levels in T-ALL patient samples (source: PeCan). C RPKM values for NOTCH1 and USP11 in 176 blood cancer cell lines were obtained from https://software.broadinstitute.org/morpheus/, using the CCLE RNA sequencing data. These include 17 T-ALL cell lines, and all other cell lines were analyzed against these T-ALL cell lines. A two-tailed unpaired t-test was conducted using the RPKM values (****, P < 0.0001). D Pearson correlation between NOTCH1 and USP11 mRNA levels in 17 T-ALL cell lines analyzed from CCLE RNA sequencing data. E Reverse-phase protein array (RPPA) for USP11 and intracellular NOTCH1 (N1IC) protein levels (n = 61). F Tracks showing NOTCH1, BRD4, and the activating histone marks H3K27Ac ChIP-Seq signal enrichment in T-ALL cells (CUTLL1) at the USP11 locus. G Heatmap representation of significant gene expression changes upon treatment of CUTLL1 cells with gamma-secretase inhibitor (γSI) followed by drug wash-off for 320’ (wash-off) [7]. Classical NOTCH1 targets (e.g., HES1, MYC, NRAPR) and the deubiquitinases USP7 and USP11 follow the intracellular NOTCH1 levels. H JURKAT T-ALL cells were treated with γSI (1 µM) for 24 h. RT-qPCR analysis of USP11 and other NOTCH1 targets was shown (*P < 0.05). I RT-qPCR analysis of Usp11 and Notch1 in normal mouse thymocytes and spleen cells isolated from N1ΔE induced mouse T-ALL model (*P < 0.05, ****P < 0.00001).
Fig. 2
Fig. 2. USP11 is a NOTCH1 deubiquitinase.
A 293 T cells were transfected with the indicated plasmids for 48 h. Immunoblot studies after USP11 immunoprecipitation showing NOTCH1, USP11, and Actin. USP11 WT: catalytically active USP11; USP11 CD: catalytically inactive USP11. B 293 T cells were transfected with FLAG-tagged NOTCH1 and HA-tagged USP11 expressing constructs for 48 h coupled to immunoprecipitation using HA and IgG (control) antibodies. Immunoblot results for the detection of NOTCH1 and USP11 are shown. C Immunoblots for 293 T cells transfected with FLAG-tagged NOTCH1 only or FLAG-tagged NOTCH1 and HA-tagged USP11 for 48 h coupled to immunoprecipitation using FLAG and IgG (control) antibodies. Immunoblot results for the detection of NOTCH1 and USP11 are shown. In A-C, 10% of the lysate volume used in the IP reaction was loaded as input to indicate the protein size and amount in whole cell lysates. D Immunoblots studies for NOTCH1, USP7, and USP11 following isolation of whole-cell extracts and gel filtration chromatography in JURKAT cells. Fractions containing high to low molecular weight complexes (left to right) were run on a gradient 4–15% polyacrylamide gel. E 293T cells were transfected with FLAG-NOTCH1, ubiquitin, and FBXW7 associated with USP11 WT or USP11 CD as indicated. Immunoblots studies following immunoprecipitation of Flag-NOTCH1 in 293 T cells at denaturing condition. USP11 WT: catalytically active USP11; USP11 CD: catalytically inactive USP11. F JURKAT (left), or CUTLL1 (right) cells were transduced with indicated lentiviral vectors. Immunoblot studies show protein levels of NOTCH1, USP11, Actin, and HSC70. G Growth study over 4 days of wild-type (WT) and catalytically inactive (CD) USP11-expressing CUTLL1 cells transduced with control or shUSP11.2 lentivirus (n = 3 biological replicates, *P ≤ 0.05). H Immunoblot detection of USP11 and GAPDH protein levels (day 0) in cells from (G).
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References

    1. Ntziachristos P, Lim JS, Sage J, Aifantis I. From fly wings to targeted cancer therapies: a centennial for notch signaling. Cancer Cell. 2014;25:318–34. doi: 10.1016/j.ccr.2014.02.018. - DOI - PMC - PubMed
    1. Kliza K, Husnjak K. Resolving the complexity of ubiquitin networks. Front Mol Biosci. 2020;7:21. doi: 10.3389/fmolb.2020.00021. - DOI - PMC - PubMed
    1. Lim R, Sugino T, Nolte H, Andrade J, Zimmermann B, Shi C, et al. Deubiquitinase USP10 regulates Notch signaling in the endothelium. Science. 2019;364:188–93. doi: 10.1126/science.aat0778. - DOI - PubMed
    1. Jin Q, Martinez CA, Arcipowski KM, Zhu Y, Gutierrez-Diaz BT, Wang KK, et al. USP7 cooperates with NOTCH1 to drive the oncogenic transcriptional program in T-cell leukemia. Clin Cancer Res. 2019;25:222–39. doi: 10.1158/1078-0432.CCR-18-1740. - DOI - PMC - PubMed
    1. Jin Q, Gutierrez Diaz B, Pieters T, Zhou Y, Narang S, Fijalkwoski I, et al. Oncogenic deubiquitination controls tyrosine kinase signaling and therapy response in acute lymphoblastic leukemia. Sci Adv. 2022;8:eabq8437. doi: 10.1126/sciadv.abq8437. - DOI - PMC - PubMed

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