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Identification and Targeting of Thomsen–Friedenreich and IL1RAP Antigens on Chronic Myeloid Leukemia Stem Cells Using Bi-Specific Antibodies

INTRODUCTION: Quiescent leukemia stem cells (LSCs) play a major role in therapeutic resistance and disease progression of chronic myeloid leukemia (CML). LSCs belong to the primitive population; CD34+CD38-Lin-, which does not distinguish normal hematopoietic stem cells (HSC) from CML LSCs. Because T...

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Autores principales: Eldesouki, Raghda E, Wu, Chengxiang, Saleh, Fayez M, Mohammed, Eman Abdel-Moemen, Younes, Soha, Hassan, Naglaa Elsayed, Brown, Theresa C, Alt, Eckhard U, Robinson, James E, Badr, Fouad Mohamed, Braun, Stephen E
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Dove 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7837560/
https://www.ncbi.nlm.nih.gov/pubmed/33519209
http://dx.doi.org/10.2147/OTT.S255299
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author Eldesouki, Raghda E
Wu, Chengxiang
Saleh, Fayez M
Mohammed, Eman Abdel-Moemen
Younes, Soha
Hassan, Naglaa Elsayed
Brown, Theresa C
Alt, Eckhard U
Robinson, James E
Badr, Fouad Mohamed
Braun, Stephen E
author_facet Eldesouki, Raghda E
Wu, Chengxiang
Saleh, Fayez M
Mohammed, Eman Abdel-Moemen
Younes, Soha
Hassan, Naglaa Elsayed
Brown, Theresa C
Alt, Eckhard U
Robinson, James E
Badr, Fouad Mohamed
Braun, Stephen E
author_sort Eldesouki, Raghda E
collection PubMed
description INTRODUCTION: Quiescent leukemia stem cells (LSCs) play a major role in therapeutic resistance and disease progression of chronic myeloid leukemia (CML). LSCs belong to the primitive population; CD34+CD38-Lin-, which does not distinguish normal hematopoietic stem cells (HSC) from CML LSCs. Because Thomsen–Friedenreich/CD176 antigen is expressed on CD34+ HSC and IL1RAP is tightly correlated to BCR-ABL expression, we sought to increase the specificity towards LSC by using additional biomarkers. METHODS: We evaluated the co-expression of both antigens on CD34+ peripheral blood mononuclear cells (PBMCs) from both healthy volunteers and CML patients, using flow cytometry. Then, we used site-directed mutagenesis to induce knob-in-hole mutations in the human IgG heavy chain and the human lambda light chain to generate the bi-specific antibody (Bis-Ab) TF/RAP that binds both antigens simultaneously. We measured complement-directed cytotoxicity (CDC) in CML samples with the Bis-Ab by flow cytometry. RESULTS: In contrast to healthy volunteers, CML samples displayed a highly significant co-expression of CD176 and IL1RAP. When either a double-positive cell line or CML samples were treated with increasing doses of Bis-Ab, increased binding and CDC was observed indicating co-operative binding of the Bis-Ab as compared to monoclonal antibodies. DISCUSSION: These results show that the bi-specific antibody is capable of targeting IL1RAP+ and CD176+ cell population among CML PBMCs, but not corresponding normal cells in CDC assay. We hereby offer a novel strategy for the depletion of CML stem cells from the bulk population in clinical hematopoietic stem cell transplantation.
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spelling pubmed-78375602021-01-28 Identification and Targeting of Thomsen–Friedenreich and IL1RAP Antigens on Chronic Myeloid Leukemia Stem Cells Using Bi-Specific Antibodies Eldesouki, Raghda E Wu, Chengxiang Saleh, Fayez M Mohammed, Eman Abdel-Moemen Younes, Soha Hassan, Naglaa Elsayed Brown, Theresa C Alt, Eckhard U Robinson, James E Badr, Fouad Mohamed Braun, Stephen E Onco Targets Ther Original Research INTRODUCTION: Quiescent leukemia stem cells (LSCs) play a major role in therapeutic resistance and disease progression of chronic myeloid leukemia (CML). LSCs belong to the primitive population; CD34+CD38-Lin-, which does not distinguish normal hematopoietic stem cells (HSC) from CML LSCs. Because Thomsen–Friedenreich/CD176 antigen is expressed on CD34+ HSC and IL1RAP is tightly correlated to BCR-ABL expression, we sought to increase the specificity towards LSC by using additional biomarkers. METHODS: We evaluated the co-expression of both antigens on CD34+ peripheral blood mononuclear cells (PBMCs) from both healthy volunteers and CML patients, using flow cytometry. Then, we used site-directed mutagenesis to induce knob-in-hole mutations in the human IgG heavy chain and the human lambda light chain to generate the bi-specific antibody (Bis-Ab) TF/RAP that binds both antigens simultaneously. We measured complement-directed cytotoxicity (CDC) in CML samples with the Bis-Ab by flow cytometry. RESULTS: In contrast to healthy volunteers, CML samples displayed a highly significant co-expression of CD176 and IL1RAP. When either a double-positive cell line or CML samples were treated with increasing doses of Bis-Ab, increased binding and CDC was observed indicating co-operative binding of the Bis-Ab as compared to monoclonal antibodies. DISCUSSION: These results show that the bi-specific antibody is capable of targeting IL1RAP+ and CD176+ cell population among CML PBMCs, but not corresponding normal cells in CDC assay. We hereby offer a novel strategy for the depletion of CML stem cells from the bulk population in clinical hematopoietic stem cell transplantation. Dove 2021-01-22 /pmc/articles/PMC7837560/ /pubmed/33519209 http://dx.doi.org/10.2147/OTT.S255299 Text en © 2021 Eldesouki et al. http://creativecommons.org/licenses/by-nc/3.0/ This work is published and licensed by Dove Medical Press Limited. The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License (http://creativecommons.org/licenses/by-nc/3.0/). By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. For permission for commercial use of this work, please see paragraphs 4.2 and 5 of our Terms (https://www.dovepress.com/terms.php).
spellingShingle Original Research
Eldesouki, Raghda E
Wu, Chengxiang
Saleh, Fayez M
Mohammed, Eman Abdel-Moemen
Younes, Soha
Hassan, Naglaa Elsayed
Brown, Theresa C
Alt, Eckhard U
Robinson, James E
Badr, Fouad Mohamed
Braun, Stephen E
Identification and Targeting of Thomsen–Friedenreich and IL1RAP Antigens on Chronic Myeloid Leukemia Stem Cells Using Bi-Specific Antibodies
title Identification and Targeting of Thomsen–Friedenreich and IL1RAP Antigens on Chronic Myeloid Leukemia Stem Cells Using Bi-Specific Antibodies
title_full Identification and Targeting of Thomsen–Friedenreich and IL1RAP Antigens on Chronic Myeloid Leukemia Stem Cells Using Bi-Specific Antibodies
title_fullStr Identification and Targeting of Thomsen–Friedenreich and IL1RAP Antigens on Chronic Myeloid Leukemia Stem Cells Using Bi-Specific Antibodies
title_full_unstemmed Identification and Targeting of Thomsen–Friedenreich and IL1RAP Antigens on Chronic Myeloid Leukemia Stem Cells Using Bi-Specific Antibodies
title_short Identification and Targeting of Thomsen–Friedenreich and IL1RAP Antigens on Chronic Myeloid Leukemia Stem Cells Using Bi-Specific Antibodies
title_sort identification and targeting of thomsen–friedenreich and il1rap antigens on chronic myeloid leukemia stem cells using bi-specific antibodies
topic Original Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7837560/
https://www.ncbi.nlm.nih.gov/pubmed/33519209
http://dx.doi.org/10.2147/OTT.S255299
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