Structural characterisation of high affinity Siglec-2 (CD22) ligands in complex with whole Burkitt's lymphoma (BL) Daudi cells by NMR spectroscopy

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Author(s)
Madge, Paul D
Maggioni, Andrea
Pascolutti, Mauro
Amin, Moein
Waespy, Mario
Bellette, Bernadette
Thomson, Robin J
Kelm, Sorge
von Itzstein, Mark
Haselhorst, Thomas
Griffith University Author(s)
Year published
2016
Metadata
Show full item recordAbstract
Siglec-2 undergoes constitutive endocytosis and is a drug target for autoimmune diseases and B cellderived
malignancies, including hairy cell leukaemia, marginal zone lymphoma, chronic lymphocytic
leukaemia and non-Hodgkin’s lymphoma (NHL). An alternative to current antibody-based therapies
is the use of liposomal nanoparticles loaded with cytotoxic drugs and decorated with Siglec-2 ligands.
We have recently designed the first Siglec-2 ligands (9-biphenylcarboxamido-4-meta-nitrophenylcarboxamido-
Neu5Acα2Me, 9-BPC-4-mNPC-Neu5Acα2Me) with simultaneous modifications at
C-4 and C-9 position. In the current study we have used ...
View more >Siglec-2 undergoes constitutive endocytosis and is a drug target for autoimmune diseases and B cellderived malignancies, including hairy cell leukaemia, marginal zone lymphoma, chronic lymphocytic leukaemia and non-Hodgkin’s lymphoma (NHL). An alternative to current antibody-based therapies is the use of liposomal nanoparticles loaded with cytotoxic drugs and decorated with Siglec-2 ligands. We have recently designed the first Siglec-2 ligands (9-biphenylcarboxamido-4-meta-nitrophenylcarboxamido- Neu5Acα2Me, 9-BPC-4-mNPC-Neu5Acα2Me) with simultaneous modifications at C-4 and C-9 position. In the current study we have used Saturation Transfer Difference (STD) NMR spectroscopy to monitor the binding of 9-BPC-4-mNPC-Neu5Acα2Me to Siglec-2 present on intact Burkitt’s lymphoma Daudi cells. Pre-treatment of cells with periodate resulted in significantly higher STD NMR signal intensities for 9-BPC-4-mNPC-Neu5Acα2Me as the cells were more susceptible to ligand binding because cis-binding on the cell surface was removed. Quantification of STD NMR effects led to a cell-derived binding epitope of 9-BPC-4-mNPC-Neu5Acα2Me that facilitated the design and synthesis of C-2, C-3, C-4 and C-9 tetra-substituted Siglec-2 ligands showing an 88-fold higher affinity compared to 9-BPC-Neu5Acα2Me. This is the first time a NMR-based binding study of high affinity Siglec-2 (CD22) ligands in complex with whole Burkitt’s lymphoma Daudi cells has been described that might open new avenues in developing tailored therapeutics and personalised medicine.
View less >
View more >Siglec-2 undergoes constitutive endocytosis and is a drug target for autoimmune diseases and B cellderived malignancies, including hairy cell leukaemia, marginal zone lymphoma, chronic lymphocytic leukaemia and non-Hodgkin’s lymphoma (NHL). An alternative to current antibody-based therapies is the use of liposomal nanoparticles loaded with cytotoxic drugs and decorated with Siglec-2 ligands. We have recently designed the first Siglec-2 ligands (9-biphenylcarboxamido-4-meta-nitrophenylcarboxamido- Neu5Acα2Me, 9-BPC-4-mNPC-Neu5Acα2Me) with simultaneous modifications at C-4 and C-9 position. In the current study we have used Saturation Transfer Difference (STD) NMR spectroscopy to monitor the binding of 9-BPC-4-mNPC-Neu5Acα2Me to Siglec-2 present on intact Burkitt’s lymphoma Daudi cells. Pre-treatment of cells with periodate resulted in significantly higher STD NMR signal intensities for 9-BPC-4-mNPC-Neu5Acα2Me as the cells were more susceptible to ligand binding because cis-binding on the cell surface was removed. Quantification of STD NMR effects led to a cell-derived binding epitope of 9-BPC-4-mNPC-Neu5Acα2Me that facilitated the design and synthesis of C-2, C-3, C-4 and C-9 tetra-substituted Siglec-2 ligands showing an 88-fold higher affinity compared to 9-BPC-Neu5Acα2Me. This is the first time a NMR-based binding study of high affinity Siglec-2 (CD22) ligands in complex with whole Burkitt’s lymphoma Daudi cells has been described that might open new avenues in developing tailored therapeutics and personalised medicine.
View less >
Journal Title
Scientific Reports
Volume
6
Copyright Statement
© The Author(s). 2016. This work is licensed under a Creative Commons Attribution 4.0 International License. The images
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Subject
Biochemistry and Cell Biology not elsewhere classified