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Article ; Online: The effect of HYPE knock-out on the AMPylome of human OSU-CLL leukemia cells.

Fatima, Narjis / Best, O Giles / Belov, Larissa / Christopherson, Richard I

2024 Volume 65, Issue 2, Page(s) 242–249

Abstract: In humans, AMPylation of cellular proteins is carried out by Huntingtin yeast-interacting protein E (HYPE), activated under conditions of endoplasmic reticulum stress, such as in cancer cells. Extracts of the human chronic lymphocytic leukemia cell line, ...

Abstract	In humans, AMPylation of cellular proteins is carried out by Huntingtin yeast-interacting protein E (HYPE), activated under conditions of endoplasmic reticulum stress, such as in cancer cells. Extracts of the human chronic lymphocytic leukemia cell line, OSU-CLL, were fractionated using immuno-precipitation with antibodies against adenosine-phosphate and then AMP-Tyr. The proteins isolated were modified with AMP, the 'AMPylome.' AMP-labelled peptides isolated from HYPE wild-type (WT) and HYPE knock-out (KO) cells were identified using tandem mass spectrometry. A total of 213 proteins were identified from WT cell extracts, while only 23 of these were pulled down from KO cells, consistent with the presence of another AMPylator, besides HYPE. The KO cells were more sensitive to fludarabine nucleoside (2-FaraA) than WT cells. Ingenuity Pathway Analysis of the AMPylated proteins identified in WT cells clustered actin binding proteins of the cytoskeleton, and proteins of the RHO GTPase pathway that would jointly stimulate cell proliferation.
MeSH term(s)	Humans ; Leukemia, Lymphocytic, Chronic, B-Cell/metabolism ; Cell Line ; Endoplasmic Reticulum Stress ; Adenosine Monophosphate/metabolism ; Vidarabine
Chemical Substances	Adenosine Monophosphate (415SHH325A) ; Vidarabine (FA2DM6879K)
Language	English
Publishing date	2024-01-24
Publishing country	United States
Document type	Journal Article
ZDB-ID	1042374-6
ISSN	1029-2403 ; 1042-8194
ISSN (online)	1029-2403
ISSN	1042-8194
DOI	10.1080/10428194.2023.2275530
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Article ; Online: Adenylated proteins in mouse B16-F10 melanoma cells cluster in functional categories: a new paradigm for cellular regulation?

Fatima, Narjis / Alomari, Munther / Belov, Larissa / Shen, Yandong / Christopherson, Richard I

Nucleosides, nucleotides & nucleic acids

2021 Volume 41, Issue 3, Page(s) 255–263

Abstract: In mammals, AMPylation of cellular proteins is carried out by Huntingtin yeast-interacting protein E, and pseudokinase SelO. Lysates from mouse B16-F10 melanoma cells have been fractionated by immuno-precipitation using magnetic Dynabeads coated with ... ...

Abstract	In mammals, AMPylation of cellular proteins is carried out by Huntingtin yeast-interacting protein E, and pseudokinase SelO. Lysates from mouse B16-F10 melanoma cells have been fractionated by immuno-precipitation using magnetic Dynabeads coated with antibodies against both adenosine 5'-monophosphate in phosphate ester linkage to tyrosine, and adenosine-phosphate. Proteins pulled down with both these antibodies were subject to post-translational modification, most likely AMPylation. Using tandem mass spectrometry, analysis of these protein fractions identified 333 proteins that could be pulled down by both antibodies. Many of these proteins clustered in 13 functional Ingenuity Pathway Analysis categories of 4 or more adenylated proteins including some from the cytoskeleton, and some involved with initiating the unfolded protein response.Supplemental data for this article is available online at https://doi.org/10.1080/15257770.2021.1995608 .
MeSH term(s)	Adenosine Monophosphate/metabolism ; Animals ; Mammals/metabolism ; Melanoma ; Mice ; Protein Processing, Post-Translational
Chemical Substances	Adenosine Monophosphate (415SHH325A)
Language	English
Publishing date	2021-11-05
Publishing country	United States
Document type	Journal Article
ZDB-ID	2008956-9
ISSN	1532-2335 ; 1525-7770
ISSN (online)	1532-2335
ISSN	1525-7770
DOI	10.1080/15257770.2021.1995608
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Article ; Online: Fludarabine nucleoside induces major changes in the p53 interactome in human B-lymphoid cancer cell lines.

Almazi, Juhura G / Alomari, Munther / Belov, Larissa / Best, O Giles / Shen, Yandong / Graham, Mark E / Mulligan, Stephen P / Christopherson, Richard I

Nucleosides, nucleotides & nucleic acids

2021 Volume 41, Issue 3, Page(s) 314–320

Abstract: Triple combination FCR (fludarabine, cyclophosphamide and rituximab) is often used as front-line treatment for chronic lymphocytic leukemia (CLL) and non-Hodgkin's lymphoma. Results from our laboratory indicate that 2-FaraAMP (fludarabine) has multiple ... ...

Abstract	Triple combination FCR (fludarabine, cyclophosphamide and rituximab) is often used as front-line treatment for chronic lymphocytic leukemia (CLL) and non-Hodgkin's lymphoma. Results from our laboratory indicate that 2-FaraAMP (fludarabine) has multiple mechanisms of cytotoxicity that include accumulation of isoforms and phosphorylated derivatives of p53, and induction of the unfolded protein response (UPR). Using protein pull-downs with Dynabeads coated with p53 antibody, we have found that 2-FaraA (fludarabine nucleoside) induces major changes in the p53 interactome in human Raji lymphoma and IM9 multiple myeloma cells. These changes are likely driven by DNA strand breaks induced by 2-FaraA that activate protein kinases such as ATM, ATR and Chk1.
MeSH term(s)	Antineoplastic Agents/pharmacology ; Antineoplastic Agents/therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; Cell Line ; Cyclophosphamide ; Humans ; Neoplasms/drug therapy ; Nucleosides ; Tumor Suppressor Protein p53/genetics ; Tumor Suppressor Protein p53/metabolism ; Vidarabine/analogs & derivatives ; Vidarabine/pharmacology
Chemical Substances	Antineoplastic Agents ; Nucleosides ; Tumor Suppressor Protein p53 ; Cyclophosphamide (8N3DW7272P) ; Vidarabine (FA2DM6879K) ; fludarabine (P2K93U8740)
Language	English
Publishing date	2021-12-10
Publishing country	United States
Document type	Journal Article
ZDB-ID	2008956-9
ISSN	1532-2335 ; 1525-7770
ISSN (online)	1532-2335
ISSN	1525-7770
DOI	10.1080/15257770.2021.2013500
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Article: Extensive surface protein profiles of extracellular vesicles from cancer cells may provide diagnostic signatures from blood samples.

Belov, Larissa / Matic, Kieran J / Hallal, Susannah / Best, O Giles / Mulligan, Stephen P / Christopherson, Richard I

Journal of extracellular vesicles

2016 Volume 5, Page(s) 25355

Abstract: Extracellular vesicles (EV) are membranous particles (30-1,000 nm in diameter) secreted by cells. Important biological functions have been attributed to 2 subsets of EV, the exosomes (bud from endosomal membranes) and the microvesicles (MV; bud from ... ...

Abstract	Extracellular vesicles (EV) are membranous particles (30-1,000 nm in diameter) secreted by cells. Important biological functions have been attributed to 2 subsets of EV, the exosomes (bud from endosomal membranes) and the microvesicles (MV; bud from plasma membranes). Since both types of particles contain surface proteins derived from their cell of origin, their detection in blood may enable diagnosis and prognosis of disease. We have used an antibody microarray (DotScan) to compare the surface protein profiles of live cancer cells with those of their EV, based on their binding patterns to immobilized antibodies. Initially, EV derived from the cancer cell lines, LIM1215 (colorectal cancer) and MEC1 (B-cell chronic lymphocytic leukaemia; CLL), were used for assay optimization. Biotinylated antibodies specific for EpCAM (CD326) and CD19, respectively, were used to detect captured particles by enhanced chemiluminescence. Subsequently, this approach was used to profile CD19(+) EV from the plasma of CLL patients. These EV expressed a subset (~40%) of the proteins detected on CLL cells from the same patients: moderate or high levels of CD5, CD19, CD31, CD44, CD55, CD62L, CD82, HLA-A,B,C, HLA-DR; low levels of CD21, CD49c, CD63. None of these proteins was detected on EV from the plasma of age- and gender-matched healthy individuals.
Language	English
Publishing date	2016-04-15
Publishing country	Sweden
Document type	Journal Article
ZDB-ID	2683797-3
ISSN	2001-3078
ISSN	2001-3078
DOI	10.3402/jev.v5.25355
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Article ; Online: Cell surface markers in colorectal cancer prognosis.

Belov, Larissa / Zhou, Jerry / Christopherson, Richard I

International journal of molecular sciences

2010 Volume 12, Issue 1, Page(s) 78–113

Abstract: The classification of colorectal cancers (CRC) is currently based largely on histologically determined tumour characteristics, such as differentiation status and tumour stage, i.e., depth of tumour invasion, involvement of regional lymph nodes and the ... ...

Abstract	The classification of colorectal cancers (CRC) is currently based largely on histologically determined tumour characteristics, such as differentiation status and tumour stage, i.e., depth of tumour invasion, involvement of regional lymph nodes and the occurrence of metastatic spread to other organs. These are the conventional prognostic factors for patient survival and often determine the requirement for adjuvant therapy after surgical resection of the primary tumour. However, patients with the same CRC stage can have very different disease-related outcomes. For some, surgical removal of early-stage tumours leads to full recovery, while for others, disease recurrence and metastasis may occur regardless of adjuvant therapy. It is therefore important to understand the molecular processes that lead to disease progression and metastasis and to find more reliable prognostic markers and novel targets for therapy. This review focuses on cell surface proteins that correlate with tumour progression, metastasis and patient outcome, and discusses some of the challenges in finding prognostic protein markers in CRC.
MeSH term(s)	Biomarkers, Tumor/metabolism ; Colorectal Neoplasms/diagnosis ; Colorectal Neoplasms/metabolism ; Disease Progression ; Humans ; Membrane Proteins/metabolism ; Neoplasm Metastasis ; Neoplastic Stem Cells/metabolism ; Prognosis ; Proteomics ; Survival Analysis ; Tumor Microenvironment
Chemical Substances	Biomarkers, Tumor ; Membrane Proteins
Language	English
Publishing date	2010-12-28
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms12010078
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Article ; Online: Cell Surface Markers in Colorectal Cancer Prognosis

Richard I. Christopherson / Larissa Belov / Jerry Zhou

International Journal of Molecular Sciences, Vol 12, Iss 1, Pp 78-

2010 Volume 113

Abstract	The classification of colorectal cancers (CRC) is currently based largely on histologically determined tumour characteristics, such as differentiation status and tumour stage, i.e., depth of tumour invasion, involvement of regional lymph nodes and the occurrence of metastatic spread to other organs. These are the conventional prognostic factors for patient survival and often determine the requirement for adjuvant therapy after surgical resection of the primary tumour. However, patients with the same CRC stage can have very different disease-related outcomes. For some, surgical removal of early-stage tumours leads to full recovery, while for others, disease recurrence and metastasis may occur regardless of adjuvant therapy. It is therefore important to understand the molecular processes that lead to disease progression and metastasis and to find more reliable prognostic markers and novel targets for therapy. This review focuses on cell surface proteins that correlate with tumour progression, metastasis and patient outcome, and discusses some of the challenges in finding prognostic protein markers in CRC.
Keywords	colorectal cancer ; biomarkers ; proteomics ; prognostic ; Biology (General) ; QH301-705.5 ; Chemistry ; QD1-999
Subject code	610 ; 616
Language	English
Publishing date	2010-12-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
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Article ; Online: Profiles of surface mosaics on chronic lymphocytic leukemias distinguish stable and progressive subtypes.

Huang, Pauline Y / Kohnke, Philippa / Belov, Larissa / Best, O Giles / Mulligan, Stephen Peter / Christopherson, Richard I

Journal of pharmacy & pharmaceutical sciences : a publication of the Canadian Society for Pharmaceutical Sciences, Societe canadienne des sciences pharmaceutiques

2013 Volume 16, Issue 2, Page(s) 231–237

Abstract: Purpose: Chronic lymphocytic leukemia (CLL) is a heterogeneous disease, some patients may survive for many years, while 20-30% of patients progress and may die within several years. Currently, there is not a single procedure that enables accurate ... ...

Abstract	Purpose: Chronic lymphocytic leukemia (CLL) is a heterogeneous disease, some patients may survive for many years, while 20-30% of patients progress and may die within several years. Currently, there is not a single procedure that enables accurate prognosis and triaging of those patients who need immediate and aggressive treatment. All CLL cells are characterised by the expression of the B-cell antigens CD19, CD20, CD21, CD22 and CD23, with aberrant expression of the T-cell antigen CD5. Methods: We have developed a CD antibody microarray (DotScan) containing 182 immobilised CD antibodies that has been used to obtain extensive surface profiles of CLL cells obtained from 96 patients. Results: Of these 182 antigens, 27 were significantly differentially expressed between stable, stable-progressive and progressive CLL. Some of these antigens are not expressed on normal B-cells and may be targets for therapeutic antibodies against CLL. Unsupervised hierarchical clustering of the surface profiles from 96 patients showed that those with progressive CLL could be distinguished based solely upon this 'disease signature'. The sensitivity (proportion of actual positives correctly identified) was 67.9%, the specificity (proportion of negatives correctly identified) was 77.5%, and the accuracy was 71.9%. Conclusions: Considerable effort by a number of research groups has resulted in identification of individual markers for progressive CLL, but their collective use is yet to provide a test that identifies CLL patients at risk. Data presented here provide a basis for development of a simple test using an antibody microarray.
MeSH term(s)	Antibodies/immunology ; Antigens, CD/immunology ; Disease Progression ; Humans ; Leukemia, Lymphocytic, Chronic, B-Cell/immunology ; Microarray Analysis
Chemical Substances	Antibodies ; Antigens, CD
Language	English
Publishing date	2013-08-13
Publishing country	Canada
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1482-1826
ISSN (online)	1482-1826
DOI	10.18433/j3f01c
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Article: All-trans retinoic acid induces different immunophenotypic changes on human HL60 and NB4 myeloid leukaemias.

Barber, Nicole / Belov, Larissa / Christopherson, Richard I

Leukemia research

2008 Volume 32, Issue 2, Page(s) 315–322

Abstract: All-trans retinoic acid (ATRA) is used to treat patients with acute promyelocytic leukaemia (APL), inducing APL cells to differentiate into abnormal neutrophils. To investigate the possible relationship between the chromosome translocation t(15;17) found ...

Abstract	All-trans retinoic acid (ATRA) is used to treat patients with acute promyelocytic leukaemia (APL), inducing APL cells to differentiate into abnormal neutrophils. To investigate the possible relationship between the chromosome translocation t(15;17) found in APL and ATRA treatment, the human myeloid leukaemia cell lines HL60 and NB4, that are PML-RARalpha negative and positive, respectively, were treated with ATRA and immunophenotyped using a CD antibody microarray. For HL60 cells, ATRA induced major increases in descending order of CD38, CD11b, CD45RO, CD11c, CD54 and CD36 with repression of CD117 and CD44. For NB4 cells, ATRA induced major increases in descending order of CD11c, CD54, CD11a, CD11b, CD53, CD65, CD138, CD66c and T-cell receptor alpha/beta (TCRalpha/beta), with repression of CD38 and CD9. The induction of a number of these CD antigens is consistent with the known differentiation of these leukaemias to abnormal neutrophils. Approximately half of the antigens up-regulated by ATRA on NB4 cells were adhesion molecules, including CD11a, CD11b, CD11c, CD54, CD66c and CD138, consistent with the increased adhesiveness of leukaemia cells observed for APL patients treated with ATRA. On HL60 cells, ATRA induced expression of CD38, CD43 and CD45RO and repressed CD117, while the converse was true on NB4 cells that contain chimeric PML-RARalpha. For NB4 cells, ATRA induced some remarkable increases in CD antigens not seen for HL60: CD14 (16.6-fold), CD32 (27.8), CD53 (20.5), CD65 (139), CD66c (79.7), CD126 (15.1), and CD138 (57.6). The expression of these antigens may be regulated by PML-RARalpha in the presence of ATRA. Such CD antigens could be targets for synergistic treatment of APL with therapeutic antibodies following ATRA treatment.
MeSH term(s)	Antigens, CD/drug effects ; Antineoplastic Agents/pharmacology ; Gene Expression/drug effects ; HL-60 Cells ; Humans ; Immunophenotyping ; Leukemia, Myeloid, Acute/metabolism ; Protein Array Analysis ; Tretinoin/pharmacology
Chemical Substances	Antigens, CD ; Antineoplastic Agents ; Tretinoin (5688UTC01R)
Language	English
Publishing date	2008-02
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	752396-8
ISSN	1873-5835 ; 0145-2126
ISSN (online)	1873-5835
ISSN	0145-2126
DOI	10.1016/j.leukres.2007.04.013
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Zs.A 1321: Show issues

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Article ; Online: Surface profiles of live colorectal cancer cells and tumor infiltrating lymphocytes from surgical samples correspond to prognostic categories.

Zhou, Jerry / Belov, Larissa / Chapuis, Pierre / Chan, Charles / Armstrong, Nicola / Kaufman, Kimberley L / Solomon, Michael J / Clarke, Stephen J / Christopherson, Richard I

Journal of immunological methods

2015 Volume 416, Page(s) 59–68

Abstract: Extensive surface profiles of colorectal cancer (CRC) cells and tumor infiltrating lymphocytes (TIL) have been obtained from 45 surgical resection samples. Live cells were captured on an antibody microarray and stained with fluorescently-labeled ... ...

Abstract	Extensive surface profiles of colorectal cancer (CRC) cells and tumor infiltrating lymphocytes (TIL) have been obtained from 45 surgical resection samples. Live cells were captured on an antibody microarray and stained with fluorescently-labeled antibodies. Minimal panels of 11 CRC antigens (CD13, CD24, CD26, CD49d, CD138, CD166, CA-125, CA19-9, EGFR, Galectin-4 and HLA-DR) and 11 T-cell antigens (CD10, CD11b, CD11c, CD25, CD31, CD95, CD151, CD181, Galectin-4, CA19-9, TSP-1) provide signatures for relapse and survival. Hierarchical clustering of profiles from CRC cells and TIL identified groups of patients for survival, systemic relapse and death. The groups from CRC and TIL profiles for systemic relapse showed 79.2% concordance, enabling prediction of relapse after surgery. The results demonstrate communication between CRC cells and TIL.
MeSH term(s)	Aged ; Antigens, CD/immunology ; Colorectal Neoplasms/immunology ; Colorectal Neoplasms/pathology ; Epitopes, T-Lymphocyte/immunology ; Female ; HLA-DR Antigens/immunology ; Humans ; Lymphocytes, Tumor-Infiltrating/immunology ; Lymphocytes, Tumor-Infiltrating/pathology ; Male ; Neoplasm Recurrence, Local/immunology ; Neoplasm Recurrence, Local/pathology ; Prognosis
Chemical Substances	Antigens, CD ; Epitopes, T-Lymphocyte ; HLA-DR Antigens
Language	English
Publishing date	2015-01
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	120142-6
ISSN	1872-7905 ; 0022-1759
ISSN (online)	1872-7905
ISSN	0022-1759
DOI	10.1016/j.jim.2014.11.001
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Zs.A 795: Show issues

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Article ; Online: Surface profiles for subclassification of chronic lymphocytic leukemia.

Huang, Pauline Y / Best, O Giles / Belov, Larissa / Mulligan, Stephen P / Christopherson, Richard I

Leukemia & lymphoma

2012 Volume 53, Issue 6, Page(s) 1046–1056

Abstract: Chronic lymphocytic leukemia (CLL) has a variable clinical course. Some patients have stable disease while others progress and require treatment. Levels of several cluster of differentiation (CD) antigens are known to correlate with prognosis and may be ... ...

Abstract	Chronic lymphocytic leukemia (CLL) has a variable clinical course. Some patients have stable disease while others progress and require treatment. Levels of several cluster of differentiation (CD) antigens are known to correlate with prognosis and may be used to stratify patients according to risk. In this review, we summarize current information on surface CD antigens found on CLL, their pathological significance and their detection using CD antibody microarrays. The use of extensive immunophenotypic patterns or surface profiles as disease signatures for CLL subclassification, prognosis and patient management is discussed with a focus on triaging patients with CLL with progressive disease.
MeSH term(s)	Antigens, CD/genetics ; Antigens, CD/metabolism ; Antigens, Surface/genetics ; Antigens, Surface/metabolism ; Biomarkers, Tumor/genetics ; Biomarkers, Tumor/metabolism ; Disease Progression ; Gene Expression Profiling ; Humans ; Leukemia, Lymphocytic, Chronic, B-Cell/classification ; Leukemia, Lymphocytic, Chronic, B-Cell/diagnosis ; Leukemia, Lymphocytic, Chronic, B-Cell/genetics ; Leukemia, Lymphocytic, Chronic, B-Cell/pathology ; Microarray Analysis ; Models, Biological ; Neoplasm Staging/methods ; Prognosis
Chemical Substances	Antigens, CD ; Antigens, Surface ; Biomarkers, Tumor
Language	English
Publishing date	2012-04-03
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	1042374-6
ISSN	1029-2403 ; 1042-8194
ISSN (online)	1029-2403
ISSN	1042-8194
DOI	10.3109/10428194.2011.631370
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