Engineering a CD123xCD3 bispecific scFv immunofusion for the treatment of leukemia and elimination of leukemia stem cells

Kuo, Shu-Ru; Wong, Lucas; Liu, Jen-Sing

doi:10.1093/protein/gzs040

Cited by 46 publications

(38 citation statements)

References 62 publications

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“…Single-chain Fv antibody fragments to CD123 have been generated and demonstrated to induce ADCC in CD123-expressing cell lines. 20 In addition, Kuo et al 21 demonstrated in in vitro studies the antileukemic activity of a CD33CD123 bispecific scFv immunofusion construct, but nonspecific activation of T cells and interferon g secretion were noted at low concentrations. Furthermore, the murine anti-CD3 sequence used raises a concern of an increased immunogenicity in humans.…”

Section: Discussionmentioning

confidence: 99%

“…Furthermore, the murine anti-CD3 sequence used raises a concern of an increased immunogenicity in humans. 21 Finally, both bispecific (CD1233CD16) and trispecific (CD1233CD163CD33 and CD1233CD163CD123) single-chain Fv reagents capable of redirecting NK cells (via CD16) to kill tumor targets have also been developed and tested. 20,22 Although NK cell-mediated ADCC has been demonstrated in vitro against AML cells, these reagents still require humanization to reduce immunogenicity, and their in vivo activity has yet to be reported.…”

Section: Discussionmentioning

confidence: 99%

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Targeting CD123 in acute myeloid leukemia using a T-cell–directed dual-affinity retargeting platform

Al-Hussaini¹,

Rettig²,

Ritchey³

et al. 2016

Blood

159

103

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Key Points• A novel CD33CD123DART agent induces T-cell-target-specific association, activation, and proliferation.• The CD33CD123 DART induces a dose-dependent killing of AML cell lines and primary AML blasts in vitro and in vivo.T-cell-directed killing of tumor cells using bispecific antibodies is a promising approach for the treatment of hematologic malignancies. Here we describe our preclinical work with a dual-affinity retargeting (DART) molecule generated from antibodies to CD3 and CD123, designed to redirect T cells against acute myeloid leukemia blasts. The CD33CD123 DART (also referred to as MGD006/S80880) consists of 2 independent polypeptides, each composed of the V H of 1 antibody in tandem with the V L of the other antibody. The target antigen CD123 (interleukin 3RA) is highly and differentially expressed in acute myeloid leukemia (AML) blasts compared with normal hematopoietic stem and progenitor cells.In this study we demonstrate that the CD33CD123 DART binds to both human CD3 and CD123 to mediate target-effector cell association, T-cell activation, proliferation, and receptor diversification. The CD33CD123 DART also induces a dose-dependent killing of AML cell lines and primary AML blasts in vitro and in vivo. These results provide the basis for testing the CD33CD123 DART in the treatment of patients with CD123 1 AML. (Blood.2016;127(1):122-131)

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Targeting CD123 in acute myeloid leukemia using a T-cell–directed dual-affinity retargeting platform

Al-Hussaini¹,

Rettig²,

Ritchey³

et al. 2016

Blood

159

103

View full text Add to dashboard Cite

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“…Patients with AML in remission Phase I clinical study [114] Fusion of anti-CD123 scFv and anti-CD3 scFv BiTE; human recombinant bispecific mAb construct, AML blasts and AML SCs Preclinical, in vitro [115] Fusion of anti-CD123 scFv, anti-CD33 scFv and anti-CD16 scFv…”

Section: Humanized Recombinant Chimeric Igg1 Mabmentioning

confidence: 99%

“…When expressed from CHO-S cells, CD123 scFv and CD3 scFv form a homodimer that closely resembles the structure of a natural antibody. This bispecific antibody construct is capable of redirecting and activating resting T cells (CD3 + ) via its CD3 scFv and simultaneously binding to CD123, thereby inducing T-cell mediated lysis and apoptosis of CD123 + AML blasts and AML SCs [115]. The trispecific constructs consist of scFvs binding to CD123, CD33 and CD16, and are able to target CD123/CD33 AML blasts and to mediate ADCC via binding of the Fcγ-receptor III (CD16) expressed on macrophages and NK cells.…”

Section: Anti-cd123mentioning

confidence: 99%

Targeting Human Cancer Stem Cells with Monoclonal Antibodies

Naujokat¹

2012

J Clin Cell Immunol

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Cancer stem cells (CSCs) constitute a distinct subpopulation of tumor cells that exhibit self-renewal and tumor initiation capacity and the ability to give rise to the heterogenous lineages of cancer cells that comprise the tumor. CSCs possess various intrinsic mechanisms of resistance to conventional chemotherapeutics, novel tumor-targeted drugs and radiation therapy, permitting them to survive current cancer therapies and to initiate tumor recurrence and metastasis. Different cell surface and transmembrane proteins expressed by CSCs, including CD44, CD47, EpCAM (CD326), CD123, CD133, GD2, Lgr5, insulin-like growth factor receptor I (IGF-IR), and members of the Notch and Wnt signaling pathways, have been identified and mainly used for the characterization of CSCs in experimental settings. Recently, monoclonal antibodies and antibody constructs such as Triomabs and BiTEs raised against these CSC proteins have shown efficacy against CSCs in human xenograft mice, and some of them have been demonstrated to induce tumor regression in clinical trials.Since current cancer therapies fail to eliminate CSCs, ultimately leading to cancer recurrence and progression, selective targeting of CSCs with mAbs and antibody constructs reviewed herein may represent a novel and promising therapeutic strategy to eradicate cancer.

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“…A number of therapeutic agents with similar molecular formats as those described for CD33 have been developed, including immunoglobulins, 47–49 a radio-immunoconjugate, 50 ADCs and a fusion protein between the cytokine IL-3 and a fragment of diphtheria toxin, 51–54 bi-specific T cell-recruiting agents (BiTEs) and Dual-Antigen Recruiting T-cell engagers (DARTs), 55–58 dual-targeting triplebodies, 59,60 and CAR-transfected T cells. 61–64 Expression of CD123 is largely restricted to hematopoietic cells, but expression on endothelial cells has been reported.…”

Section: Introductionmentioning

confidence: 99%

Dual-targeting triplebody 33-16-123 (SPM-2) mediates effective redirected lysis of primary blasts from patients with a broad range of AML subtypes in combination with natural killer cells

et al. 2018

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A number of agents designed for immunotherapy of Acute Myeloid Leukemia (AML) are in preclinical and early clinical development. Most of them target a single antigen on the surface of AML cells. Here we describe the development and key biological properties of a tri-specific agent, the dual-targeting triplebody SPM-2, with binding sites for target antigens CD33 and CD123, and for CD16 to engage NK cells as cytolytic effectors. Primary blasts of nearly all AML patients carry at least one of these target antigens and the pair is particularly promising for the elimination of blasts and leukemia stem cells (LSCs) from a majority of AML patients by dual-targeting agents. The cytolytic activity of NK cells mediated by SPM-2 was analyzed in vitro for primary leukemic cells from 29 patients with a broad range of AML-subtypes. Blasts from all 29 patients, including patients with genomic alterations associated with an unfavorable genetic subtype, were lysed at nanomolar concentrations of SPM-2. Maximum susceptibility was observed for cells with a combined density of CD33 and CD123 above 10,000 copies/cell. Cell populations enriched for AML-LSCs (CD34pos and CD34pos CD38neg cells) from 2 AML patients carried an increased combined antigen density and were lysed at correspondingly lower concentrations of SPM-2 than unsorted blasts. These initial findings raise the expectation that SPM-2 may also be capable of eliminating AML-LSCs and thus of prolonging survival. In the future, patients with a broad range of AML subtypes may benefit from treatment with SPM-2.

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Engineering a CD123xCD3 bispecific scFv immunofusion for the treatment of leukemia and elimination of leukemia stem cells

Cited by 46 publications

References 62 publications

Targeting CD123 in acute myeloid leukemia using a T-cell–directed dual-affinity retargeting platform

Targeting CD123 in acute myeloid leukemia using a T-cell–directed dual-affinity retargeting platform

Targeting Human Cancer Stem Cells with Monoclonal Antibodies

Dual-targeting triplebody 33-16-123 (SPM-2) mediates effective redirected lysis of primary blasts from patients with a broad range of AML subtypes in combination with natural killer cells

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