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A genetic strategy involving a glycosyltransferase promoter and a lipid translocating enzyme to eliminate cancer cells

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Abstract

The most common therapeutic strategy for the treatment of cancer uses antimetabolites, which block uncontrolled division of cancer cells and kill them. However, such antimetabolites also kill normal cells, thus yielding detrimental side effects. This emphasizes the need for an alternative therapy, which would have little or no side effects. Our approach involves designing genetic means to alter surface lipid determinants that induce phagocytosis of cancer cells. The specific target of this strategy has been the enzyme activity termed aminophospholipid translocase (APLT) or flippase that causes translocation of phosphatidylserine (PS) from the outer to the inner leaflet of the plasma membrane in viable cells. Efforts to identify the enigmatic, plasma membrane APLT of mammalian cells have led investigators to some P-type ATPases, which have often proven to be the APLT of internal membranes rather than the plasma membrane. By measuring kinetic parameters for the plasma membrane APLT activity, we have shown that the P-type ATPase Atp8a1 is the plasma membrane APLT of the tumorigenic N18 cells, but not the non-tumorigenic HN2 (hippocampal neuron × N18) cells. Targeted knockdown of this enzyme causes PS externalization in the N18 cells, which would trigger phagocytic removal of these cells. But how would we specifically express the mutants or antisense Atp8a1 in the cancer cells? This has brought us to a glycosyltransferase, GnT-V, which is highly expressed in the transformed cells. By using the GnT-V promoter to drive a luciferase reporter gene we have demonstrated a dramatic increase in luciferase expression selectively in tumor cells. The described strategy could be tested for the removal of cancer cells without the use of antimetabolites that often kill normal cells.

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Acknowledgment

FJ and KL were supported by funds from the New York Louis Stokes Alliance for Minority Participation Program, MAGNET/AGEP, and NIH minority supplements. The authors gratefully acknowledge the gifts of GnT-V antibody from Drs. Naoyuki Taniguchi and Eiji Miyoshi (Osaka University Medical School, Japan) and Atp8a1 antibody from Dr. Xiao-Song Xie (University of Texas Southwestern Medical Center). This project was supported by a grant from the NIH (CA77803-03).

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Author notes

  1. Farah Jayman

    Present address: Walter Reed Army Medical Center, Washington, DC, 20307, USA

Authors and Affiliations

  1. Department of Chemistry, Neuroscience Program, City University of New York at the College of Staten Island, Staten Island, NY, 10314, USA

    Tomasz Sobocki, Priya Ranjan Debata & Probal Banerjee

  2. CUNY Doctoral Program in Biochemistry, City University of New York at the College of Staten Island, Staten Island, NY, 10314, USA

    Kelly Levano & Farah Jayman

  3. Department of Medicine, SUNY Downstate Medical Center, Brooklyn, NY, 10203, USA

    Malgorzata B. Sobocka

  4. Department of Chemistry and the CSI/IBR Center for Developmental Neuroscience, The City University of New York at The College of Staten Island, Staten Island, NY, USA

    Probal Banerjee

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  1. Kelly Levano
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Correspondence to Probal Banerjee.

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Tomasz Sobocki, Farah Jayman contributed equally.

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Levano, K., Sobocki, T., Jayman, F. et al. A genetic strategy involving a glycosyltransferase promoter and a lipid translocating enzyme to eliminate cancer cells. Glycoconj J 26, 739–748 (2009). https://doi.org/10.1007/s10719-009-9233-1

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