Letters to the Editor 1835

OPEN A critical role of periostin in B-cell acute lymphoblastic leukemia

Leukemia (2017) 31, 1835–1837; doi:10.1038/leu.2017.149 actively regulate leukemia cell growth and migration.1 The extra- cellular matrix, which is produced and shaped by both leukemia and Microenvironment plays important roles in leukemia initiation, stromal cells, mediates the interactions between leukemia cells and progression and metastasis. While leukemia cells affect stromal cells their microenvironment. However, the functions of individual through cell–cell contact and secreted factors, stromal cells also extracellular matrix components remain poorly understood.

Figure 1. For caption see next page.

Accepted article preview online 22 May 2017; advance online publication, 16 June 2017

© 2017 Macmillan Publishers Limited, part of Springer Nature. Leukemia (2017) 1816 – 1842 Letters to the Editor 1836 Periostin (POSTN), also known as osteoblast-specific factor 2, We first evaluated the expression of POSTN in the bone marrow of was originally identified in MC3T3-E1 osteoblast cells as an healthy individuals and B-ALL patients. Immunohistochemistry assays adhesion molecule.2,3 As a matricellular protein, POSTN is highly showed that the bone marrow from B-ALL patients exhibited expressed in collagen-rich fibrous connective tissues at sites of elevated expression of POSTN (Figure 1a). We further analyzed the injury or tissue inflammation, as well as by tumor stromal cells. It B-ALL mRNA microarray data in the Oncomine database. The data plays a critical role in tissue remodeling, inflammatory responses from the Zhang leukemia database,15 which includes bone marrow and tumor metastasis by regulating the cross talk between cells from 483 B-ALL patients and 8 healthy individuals, reveal that POSTN and the extracellular matrix.4–8 We previously showed that POSTN mRNA is expressed at significantly higher levels in the bone marrow promotes the metastasis of colon cancer cells to the liver by of B-ALL patients than that of healthy individuals (Supplementary enhancing cancer cell survival and angiogenesis.9 Malanchi et al.10 Figure S1). These results demonstrate that POSTN expression is found that fibroblast-derived POSTN promotes metastatic coloni- elevated in the bone marrow of B-ALL patients, suggesting that zation in the lungs by establishing a metastatic niche for POSTN is involved in B-ALL development and progression. disseminated breast cancer cells. Bissell and colleagues demon- We then examined POSTN expression in animal models of B-ALL. strated that POSTN surrounding the tip cells of the neovasculature In SCID mice transplanted with human B-ALL cells (a NALM-6 cell awakens the dormant disseminated breast tumor cells and line), immunohistochemistry analysis of the bone marrow showed augments metastatic tumor outgrowth.11 Recently, we further that POSTN expression significantly increased after 4 weeks of B-ALL showed that POSTN is a novel component of immunosuppressive cell transplantation (Figure 1b). This was further confirmed by premetastatic niches and is exploited by breast tumor cells to quantitative real-time PCR and immunoblot analysis of POSTN facilitate lung metastasis.12 These studies suggest that POSTN mRNA and protein levels, respectively (Figure 1c). In another model, plays critical roles in various tumor microenvironments, including we injected L1210 cells, a mouse B-ALL cell line, into the tail vein of the cancer stem cell niche,6,10 the perivascular niche11 and the Balb/c mice and examined POSTN expression in the bone marrow of premetastatic niche12 during tumor metastasis, and may represent these mice 3 weeks later. Consistent with the SCID mouse model, a novel diagnostic and therapeutic target for tumor metastasis. both the protein and mRNA levels of POSTN were elevated in Balb/c While the expression pattern and function of POSTN in the mice injected with L1210 cells (Supplementary Figure S2). We development and metastasis of solid tumors have been exten- further isolated peripheral blood mononuclear cells from the sively investigated, not much is known about its roles in peripheral blood of a B-ALL patient and injected these cells into hematological malignancies. B-cell acute lymphoblastic leukemia the tail vein of NOD-SCID mice. Four weeks later, the protein and (B-ALL) is a subtype of malignant hematological neoplasm driven mRNA expression of POSTN in the bone marrow of the mice bearing by dysregulated proliferation of lymphoid progenitor cells and human B-ALL cells was also significantly increased (Supplementary occurs in both adults and children.13,14 In this study, we Figure S3). Taken together, our study of B-ALL patients and mouse investigated the expression and function of POSTN in B-ALL. models of B-ALL suggests that elevated expression of POSTN is a

Figure 2. POSTN deletion decreases B-ALL leukemia burden in mice. (a) Bioluminescent imaging of representative wild-type and POSTN-deficient mice at day 21 after injection (i.v.) with L1210-GFP/Luc cells. Three independent experiments were performed, with n = 3(left),3(middle)and4–6 (right) mice per group. (b) Bioluminescent imaging of limbs, liver and spleen of one representative wild-type and one representative POSTN- deficient mouse at day 21 after injection (i.v.) with L1210-GFP/Luc cells. (c) Hematoxylin and eosin staining and immunohistochemistry analysis of GFP and Ki67 expression in the liver of wild-type and POSTN-deficient mice injected with L1210-GFP/Luc cells in a. Scale bars, 50 μm.

Figure 1. POSTN is highly expressed in the bone marrow of human patients and mice with B-ALL. (a) The expression of POSTN in the bone marrow of 20 B-ALL patients and 8 healthy individuals was assessed by immunohistochemistry. Four representative cases of each group were shown. (b, c) SCID mice were injected (i.v.) with 1 × 107 NALM-6 cells. Four weeks later, POSTN expression in the bone marrow was examined by immunohistochemistry (b), quantitative real-time PCR and immunoblot (c). Normal bone marrow from non-engrafted SCID mice was used as control. n = 4 mice per group. Scale bars, 50 μm. **Po0.01.

Leukemia (2017) 1816 – 1842 © 2017 Macmillan Publishers Limited, part of Springer Nature. Letters to the Editor 1837 common feature of B-ALL and that POSTN may play an important the data interpretation and conceptual advice. CX, GF and GO designed the role in the pathogenesis of B-ALL. experiments, interpreted the data and wrote the manuscript. We speculated that the deletion of POSTN may decrease leukemia burden in mouse models of B-ALL. To test this, we injected L1210- ZMa1,5, X Zhao1,5, J Huang2,3,5, X Jia1, M Deng1, D Cui1,ZDu4, GFP/Luc cells, which were engineered to express both the green GFu1, G Ouyang1 and C Xiao1 fluorescent protein (GFP) and firefly luciferase (Luc), into the tail vein 1State Key Laboratory of Cellular Stress Biology, Innovation Center for of wild-type and POSTN-deficient mice. Three weeks later, leukemia Cell Signaling Network, School of Life Sciences, Xiamen University, burden was evaluated by live imaging of bioluminescence. While Xiamen, Fujian, China; wild-type mice showed significant leukemia burdens, the accumula- 2Translational Medicine Research Center, School of Pharmaceutical tion of leukemia cells in POSTN-deficient mice was much less than in Science, Xiamen University, Xiamen, Fujian, China; their wild-type counterparts (Figure 2a; Supplementary Figure S4a). 3Department of Hematology, the Affiliated Hospital of Putian Moreover, POSTN-deficient mice showed a significantly decreased University, Putian, Fujian, China and leukemia dissemination to the limbs, liver and spleen (Figure 2b; 4Department of Translational Science, Amgen Asia R&D Center, Supplementary Figures S4b and c). Immunohistochemistry analysis Shanghai, China showed that a greater number of GFP-labeled B-ALL cells and Ki67- E-mail: [email protected] or [email protected] or cxiao@xmu. positive cells were observed in the liver of wild-type mice than that edu.cn of POSTN-deficient mice (Figure 2c; Supplementary Figures S4d and 5These authors contributed equally to this work. e). These results demonstrate that POSTN deletion significantly decreased B-ALL cell proliferation and accumulation in the bone marrow and impaired leukemia cell dissemination, and suggest that REFERENCES POSTN plays critical roles in the microenvironment to promote the 1 Ayala F, Dewar R, Kieran M, Kalluri R. Contribution of bone microenvironment to proliferation, accumulation and migration of leukemia cells. leukemogenesis and leukemia progression. Leukemia 2009; 23:2233–2241. We isolated B-ALL cells and bone marrow-derived mesenchymal 2 Takeshita S, Kikuno R, Tezuka K, Amann E. 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This work was supported by grants from the National Nature Science Foundation of This work is licensed under a Creative Commons Attribution- China (grants 81572598, 81372841 and 31171339 to GO, and 31570911 to GF), the NonCommercial-NoDerivs 4.0 International License. The images or Fundamental Research Funds for the Central Universities of China (grant other third party material in this article are included in the article’s Creative Commons 20720150060 to GO and 20720150065 to GF) and the 100 Talents Program of Fujian license, unless indicated otherwise in the credit line; if the material is not included under Province, China (K08008 to CX). the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http:// creativecommons.org/licenses/by-nc-nd/4.0/ AUTHOR CONTRIBUTIONS ZM, XZ, JH, XJ and MD performed the experiments and generated data. JH performed diagnostic interpretation of patient samples. ZD and DC assisted in © The Author(s) 2017

Supplementary Information accompanies this paper on the Leukemia website (http://www.nature.com/leu)

© 2017 Macmillan Publishers Limited, part of Springer Nature. Leukemia (2017) 1816 – 1842