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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Research Article Cell biology Oncology

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 6

CXCR4 inhibition decreases leukemia cell invasiveness both in vitro and in vivo.

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CXCR4 inhibition decreases leukemia cell invasiveness both in vitro and ...
(A) CXCR4-specific-antagonist treatment of KOPTK1 cells expressing WT-PFKP, NLS-PFKP, or PFKP-RXL significantly decreases cell invasiveness in vitro. Invasion assays were performed on cells with and without treatment with either motixafortide (4 μM) or plerixafor (10 μM). Boyden chambers coated with Matrigel and CXCL12 (100 ng/mL) in the bottom chambers were used for cell invasion assay as in other figures. (B) CXCR4 knockdown with shCXCR4-1 or shCXCR4-2 significantly decreases the invasive capability of DND41 cells expressing WT-, NLS-, or S679E-PFKP. shCon is nontargeting control shRNA. (C) Flow cytometry analysis of KOPTK1 cells expressing WT-, NLS-PFKP, or PFKP-RXL in peripheral blood, bone marrow, spleen, or liver of immunodeficient mice by gating on human CD45 3 weeks after the mice were tail vein injected with KOPTK1 cells and treated with CXCR4 antagonist (plerixafor) daily. Tissues were dissociated by collagenase treatment followed by passing through a 70-μm strainer. Percentage of cells represents KOPTK1 cells over total gated cells. Each symbol represents data for an individual mouse. (D) Immunohistochemical staining with an anti–human CD45 antibody of spleens from mice that received KOPTK1 cells expressing WT-, NLS-PFKP, or PFKP-RXL. Tumor-bearing mice were treated with plerixafor (lower) or vehicle control (upper). Arrows indicate blood vessels. Scale bar: 100 μm. (E) Kaplan-Meier survival curves of tumor-bearing mice that received KOPTK1 cells expressing WT-, NLS-PFKP, or PFKP-RXL. Tumor-bearing mice were treated with CXCR4 antagonist (plerixafor) daily, 7 days after the implantation of KOPTK1 cells. n = 3 (A), 4 (B), and 6 (C and D) mice/group for WT-PFKP and NLS-PFKP, n = 5 mice/group for PFKP-RXL; n = 5 mice/group (E). Data represent mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 by 1-way ANOVA (A–C). NS, not significant.