AntiCD3Fv Fused to Human Interleukin-3 Deletion Variant Redirected T Cells Against Human Acute Myeloid Leukemic Stem Cells

Overview

Journal J Hematol Oncol

Publisher Biomed Central

Specialties Hematology
Oncology

Date 2015 Apr 17

PMID 25879549

Citations 16

Authors

Dongmei Fan

Zhenzhen Li

Xiaolong Zhang

Yuqi Yang

Xiangfei Yuan

Xiuli Zhang

Ming Yang

Yizhi Zhang

Dongsheng Xiong

Affiliations

Soon will be listed here.

Abstract

Background: Leukemic stem cells (LSCs) are frequently seen as a cause of treatment failure and relapse in patients with acute myeloid leukemia (AML). Thus, successful new therapeutic strategies for the treatment of AML should aim at eradicating LSCs. The identification of targets on the cell surface of LSCs is getting more and more attention. Among these, CD123, also known as the interleukin-3 (IL3)-receptor α chain, has been identified as a potential immunotherapeutic target due to its overexpression on LSCs in AML as well as on AML blasts, rather than normal hematopoietic stem cells.

Methods: We constructed a CD123-targeted fusion protein antiCD3Fv-⊿IL3, with one binding site for T cell antigen receptor (TCRCD3) and the other for CD123, by recombinant gene-engineering technology. Cysteine residues were introduced into the V domains of the antiCD3Fv segment to enhance its stability by locking the two chains of Fv together with disulfide covalent bonds. The stability and cytotoxicity of the two fusion proteins were detected in vitro and in vivo.

Results: Both fusion proteins were produced and purified from Escherichia coli 16C9 cells with excellent yields in fully active forms. High-binding capability was observed between these two fusion proteins and human IL3R, leading to the specific lysis of CD123-expressing cell lines KG1a; also, mononuclear cells from primary AML patients were inhibited in a colony forming assay in vitro, presumably by redirecting T lymphocytes in vitro. In addition, they displayed an antileukemic activity against KG1a xenografts in non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice, especially disulfide-stabilized (ds)-antiCD3Fv-⊿IL3 for its improved stability.

Conclusions: These results suggest that both fusion proteins display the antileukemic activity against CD123-expressing cell lines as well as leukemic progenitors in vitro and in vivo, especially ds-antiCD3Fv-⊿IL3. They could be the promising candidates for future immunotherapy of AML.

Citing Articles

Gene Therapy and Cardiovascular Diseases.

Lu D, Cushman S, Thum T, Bar C Adv Exp Med Biol. 2022; 1396:235-254.

PMID: 36454471 DOI: 10.1007/978-981-19-5642-3_16.

Pre-clinical development of a novel CD3-CD123 bispecific T-cell engager using cross-over dual-variable domain (CODV) format for acute myeloid leukemia (AML) treatment.

Bonnevaux H, Guerif S, Albrecht J, Jouannot E, De Gallier T, Beil C Oncoimmunology. 2021; 10(1):1945803.

PMID: 34484869 PMC: 8409758. DOI: 10.1080/2162402X.2021.1945803.

CD123 as a Biomarker in Hematolymphoid Malignancies: Principles of Detection and Targeted Therapies.

El Achi H, Dupont E, Paul S, Khoury J Cancers (Basel). 2020; 12(11).

PMID: 33113953 PMC: 7690688. DOI: 10.3390/cancers12113087.

Cancer Stem Cells-Origins and Biomarkers: Perspectives for Targeted Personalized Therapies.

Walcher L, Kistenmacher A, Suo H, Kitte R, Dluczek S, Strauss A Front Immunol. 2020; 11:1280.

PMID: 32849491 PMC: 7426526. DOI: 10.3389/fimmu.2020.01280.

Immune targets in the tumor microenvironment treated by radiotherapy.

Ozpiskin O, Zhang L, Li J Theranostics. 2019; 9(5):1215-1231.

PMID: 30867826 PMC: 6401500. DOI: 10.7150/thno.32648.

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