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Aberrant Wnt Signaling Pathway in the Hematopoietic Stem/Progenitor Compartment in Experimental Leukemic Animal

  • Sukalpa Chattopadhyay
  • Malay Chaklader
  • Sujata Law
Research Article

Abstract

The evolutionarily conserved Wnt signaling pathway regulates physiological hematopoiesis, a process of formation of blood cells and has been shown to play crucial role in the development of both myeloid and lymphoid malignancies. The Wnt signaling pathway can be broadly divided into canonical and non-canonical pathways. In the present study, we investigated the pathobiology of leukemia by studying the expression profile of Wnt proteins, receptors, key signaling intermediates and endogenous Wnt antagonist involved in canonical and non-canonical pathways in the bone marrow (BM) hematopoietic stem/progenitor cell (HSPC) compartment of experimental leukemic mice. Cell adhesion molecule N-Cadherin and leukemic BM microenvironment with reference to Wnt were also studied. We used ENU, a potent carcinogen, to induce leukemia in wild type Swiss albino mice and malignant transformation was cofirmed by peripheral blood and BM studies. Flow cytometric expression analysis revealed profound up-regulation of canonical Wnt3a/β-catenin/CyclinD1 signaling axis along with N-Cadherin whereas down-regulation of non-canonical Wnt5a/Ca2+/CaMKII signaling axis in the leukemic HSPC compartment. Subsequent use of anti-Wnt3a antibody in the in vitro clonogenicity assay uncovered that anti-Wnt3a antibody preferentially inhibited the growth and number of the primitive leukemic hematopoietic CFU-GEMM and BFU-E colonies. Stromal cells derived from the leukemic BM also exhibited aberrant Wnt3a and Wnt5a protein expression. Taken together, alteration of canonical and non-canonical Wnt signaling pathways in the HSPC compartment along with classical Wnt protein expression pattern in the leukemic stromal microenvironment resulted in progression of leukemia.

Keywords

ENU Leukemia N-Cadherin Wnt signaling Wnt3a Wnt5a 

Abbreviations

BM

Bone marrow

HSPC

Hematopoietic stem/progenitor cell

ENU

N-ethyl-N-nitrosourea

MPO

Myeloperoxidase

NSE

Nonspecific-esterase

Ctrl

Control

Leuk

Leukemia

CFU-GEMM

Colony-forming unit of granulocyte/erythrocyte/macrophage/megakaryocyte

BFU-E

Burst-forming unit of erythroid cells

CFU-GM

Colony-forming unit of granulocyte/macrophage

CFU-G

Colony-forming unit of granulocyte

CFU-E

Colony-forming unit of erythroid cells

Notes

Acknowledgements

Authors are thankful to the Director, Calcutta School of Tropical Medicine for supporting the work. We are also thankful to Leica Microsystem and Dept. of Biological Sciences, Presidency University for providing facilities of confocal microscope.

Funding

This work was supported by the INSPIRE Fellowship, Dept. of Science and Technology, Govt. of India [Fellowship Number: DST/INSPIRE Fellowship/2013/283(IF130354)]; and the Council for Scientific and Industrial Research (CSIR), Govt. of India [Grant Number: No. 37(1429)/10/EMRII].

Compliance with ethical standards

Conflict of Interest

Authors unanimously declare no potential conflict of interest.

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Copyright information

© The International CCN Society 2018

Authors and Affiliations

  • Sukalpa Chattopadhyay
    • 1
  • Malay Chaklader
    • 1
    • 2
  • Sujata Law
    • 1
  1. 1.Stem Cell Research and Application Unit, Department of Biochemistry and Medical BiotechnologyCalcutta School of Tropical MedicineKolkataIndia
  2. 2.Tissue Engineering Laboratory, Skeletal Biology and Engineering Research Center, Prometheus, Division of Skeletal Tissue EngineeringKU LeuvenLeuvenBelgium

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