Nuclear PFKP promotes CXCR4-dependent infiltration by T cell acute lymphoblastic leukemia
Xueliang Gao, … , Xue-Zhong Yu, Haizhen Wang
Xueliang Gao, … , Xue-Zhong Yu, Haizhen Wang
Published July 13, 2021
Citation Information: J Clin Invest. 2021;131(16):e143119. https://doi.org/10.1172/JCI143119.
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Nuclear PFKP promotes CXCR4-dependent infiltration by T cell acute lymphoblastic leukemia

Abstract

PFKP (phosphofructokinase, platelet), the major isoform of PFK1 expressed in T cell acute lymphoblastic leukemia (T-ALL), is predominantly expressed in the cytoplasm to carry out its glycolytic function. Our study showed that PFKP is a nucleocytoplasmic shuttling protein with functional nuclear export and nuclear localization sequences (NLSs). Cyclin D3/CDK6 facilitated PFKP nuclear translocation by dimerization and by exposing the NLS of PFKP to induce the interaction between PFKP and importin 9. Nuclear PFKP stimulated the expression of C-X-C chemokine receptor type 4 (CXCR4), a chemokine receptor regulating leukemia homing/infiltration, to promote T-ALL cell invasion, which depended on the activity of c-Myc. In vivo experiments showed that nuclear PFKP promoted leukemia homing/infiltration into the bone marrow, spleen, and liver, which could be blocked with CXCR4 antagonists. Immunohistochemical staining of tissues from a clinically well-annotated cohort of T cell lymphoma/leukemia patients showed nuclear PFKP localization in invasive cancers, but not in nonmalignant T lymph node or reactive hyperplasia. The presence of nuclear PFKP in these specimens correlated with poor survival in patients with T cell malignancy, suggesting the potential utility of nuclear PFKP as a diagnostic marker.

Authors

Xueliang Gao, Shenghui Qin, Yongxia Wu, Chen Chu, Baishan Jiang, Roger H. Johnson, Dong Kuang, Jie Zhang, Xi Wang, Anand Mehta, Kenneth D. Tew, Gustavo W. Leone, Xue-Zhong Yu, Haizhen Wang

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Figure 7

CXCR4 inhibition decreases primary T-ALL invasion.

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CXCR4 inhibition decreases primary T-ALL invasion. (A) Immunoblotting (l...
(A) Immunoblotting (left) with quantification (right) shows nuclear PFKP expression in primary T-ALL cells from different patients. The 3 primary cultures are designated DFAT-24836, DFAT-28537, and CBAT-93917. (B) CXCR4 expression levels in primary T-ALL cells from different patients were analyzed with flow cytometry. (C) Invasive capability of primary T-ALL cells from different patients was analyzed. Boyden chambers coated with Matrigel and CXCL12 (100 ng/mL) in the bottom chambers as a stimulus for 24 hours were used for in vitro invasion assays here and Supplemental Figure 7. (D) Nuclear PFKP expression in primary T-ALL cells (DFAT-24836) treated with the CDK4/6 inhibitor palbociclib (Palbo, 1 μM), CDK6 degrader (CDK6D; BSJ-03-123, 10 μM), or importin inhibitor importazole (Impor, 40 μM) for 24 hours. (E) CXCR4 expression analyzed with flow cytometry in primary T-ALL cells (DFAT-24836) treated with palbociclib (Palbo, 1 μM), BSJ-03-123 (CDK6D, 10 μM), importazole (Impor, 40 μM), or c-Myc inhibitor (10058-F4, 50 μM) for 24 hours. (F) Invasion assay of primary T-ALL cells (DFAT-24836) treated with palbociclib (Palbo, 1 μM), CXCR4 antagonist (plerixafor, 10 μM), or c-Myc inhibitor (10058-F4, 50 μM) for 24 hours. (G) Analysis of primary T-ALL cells (DFAT-24836 and DFAT-28537) in peripheral blood, bone marrow, spleen, or liver of NOG mice using flow cytometry with gating on human CD45 after the mice were tail vein injected with primary T-ALL cells and then treated with plerixafor daily for 29 days. Percentage of cells represents primary T-ALL over total gated cells. Each symbol represents data for an individual mouse. n = 5 mice/group. n = 3 (A, B, D, and E) and 4 (C and F). Data represent mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 by 1-way ANOVA (A–C and E–G). NS, not significant.