Isolation of human mAbs that directly modulate FMS-related tyrosine kinase 3 signaling

Yukiya Yamamoto, Sachiko Tsuzuki, Yasushi Akahori, Yoshinori Ukai, Mariko Sumitomo, Yuko Murayama, Kiyoko Yamamoto, Youko Inaguma, Masutaka Tokuda, Akihiro Abe, Yoshiki Akatsuka, Nobuhiko Emi, Yoshikazu Kurosawa

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

FMS-related tyrosine kinase 3 (FLT3) is a class III receptor tyrosine kinase that plays important roles in hematopoiesis, including early progenitors and dendritic cell development. FLT3 is expressed at high levels in 70-100% of cases of AML and in virtually all cases of B-lineage acute lymphoblastic leukemia. FLT3 is regarded as a molecular target in the development of novel therapies for acute leukemia patients. Currently, many small-molecule FLT3 inhibitors have been developed, but clinical trials have resulted in limited antileukemia effects because of off-target toxicities and drug resistance. The development of anti-FLT3 Abs might overcome these difficulties and enhance the antileukemia efficacy of FLT3 inhibitors. In the present study, we demonstrate the isolation of novel human mAbs against FLT3 with antagonistic or agonistic activities. An antagonistic Ab, designated A2, continuously inhibits FLT3 ligand (FL)-induced phosphorylation of FLT3 and MAPK. A2 cooperatively induces apoptosis with daunorubicin, even in the presence of FL. An agonistic Ab, designated 3E6, surprisingly induces the phosphorylation of FLT3 and MAPK, and supports the growth of a factor-dependent cell line independently of FL addition. In addition, A2 showed complement-dependent cytotoxicity activity, but was devoid of Ab-dependent cell mediated cytotoxicity. Finally, we evaluated Ab internalization in a cell line. Immunofluorescence and flow cytometry analyses revealed that A2 is efficiently internalized. Collectively, these data demonstrate that A2 is a potent human Ab that might be capable of delivering cytotoxic reagents and that has antagonistic effects on FLT3 signaling. In addition, 3E6 might be a potential scaffold for novel dendritic cell-based immunotherapies.

Original languageEnglish
Pages (from-to)350-359
Number of pages10
JournalCancer Science
Volume103
Issue number2
DOIs
Publication statusPublished - 01-02-2012

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varespladib methyl
Protein-Tyrosine Kinases
Ligands
Dendritic Cells
Phosphorylation
Cell Line
Daunorubicin
Hematopoiesis
Receptor Protein-Tyrosine Kinases
Precursor Cell Lymphoblastic Leukemia-Lymphoma
Drug Resistance
Immunotherapy
Fluorescent Antibody Technique
Intercellular Signaling Peptides and Proteins
Flow Cytometry
Leukemia
Stem Cells
Clinical Trials
Apoptosis
Therapeutics

All Science Journal Classification (ASJC) codes

  • Oncology
  • Cancer Research

Cite this

Yamamoto, Y., Tsuzuki, S., Akahori, Y., Ukai, Y., Sumitomo, M., Murayama, Y., ... Kurosawa, Y. (2012). Isolation of human mAbs that directly modulate FMS-related tyrosine kinase 3 signaling. Cancer Science, 103(2), 350-359. https://doi.org/10.1111/j.1349-7006.2011.02141.x
Yamamoto, Yukiya ; Tsuzuki, Sachiko ; Akahori, Yasushi ; Ukai, Yoshinori ; Sumitomo, Mariko ; Murayama, Yuko ; Yamamoto, Kiyoko ; Inaguma, Youko ; Tokuda, Masutaka ; Abe, Akihiro ; Akatsuka, Yoshiki ; Emi, Nobuhiko ; Kurosawa, Yoshikazu. / Isolation of human mAbs that directly modulate FMS-related tyrosine kinase 3 signaling. In: Cancer Science. 2012 ; Vol. 103, No. 2. pp. 350-359.
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abstract = "FMS-related tyrosine kinase 3 (FLT3) is a class III receptor tyrosine kinase that plays important roles in hematopoiesis, including early progenitors and dendritic cell development. FLT3 is expressed at high levels in 70-100{\%} of cases of AML and in virtually all cases of B-lineage acute lymphoblastic leukemia. FLT3 is regarded as a molecular target in the development of novel therapies for acute leukemia patients. Currently, many small-molecule FLT3 inhibitors have been developed, but clinical trials have resulted in limited antileukemia effects because of off-target toxicities and drug resistance. The development of anti-FLT3 Abs might overcome these difficulties and enhance the antileukemia efficacy of FLT3 inhibitors. In the present study, we demonstrate the isolation of novel human mAbs against FLT3 with antagonistic or agonistic activities. An antagonistic Ab, designated A2, continuously inhibits FLT3 ligand (FL)-induced phosphorylation of FLT3 and MAPK. A2 cooperatively induces apoptosis with daunorubicin, even in the presence of FL. An agonistic Ab, designated 3E6, surprisingly induces the phosphorylation of FLT3 and MAPK, and supports the growth of a factor-dependent cell line independently of FL addition. In addition, A2 showed complement-dependent cytotoxicity activity, but was devoid of Ab-dependent cell mediated cytotoxicity. Finally, we evaluated Ab internalization in a cell line. Immunofluorescence and flow cytometry analyses revealed that A2 is efficiently internalized. Collectively, these data demonstrate that A2 is a potent human Ab that might be capable of delivering cytotoxic reagents and that has antagonistic effects on FLT3 signaling. In addition, 3E6 might be a potential scaffold for novel dendritic cell-based immunotherapies.",
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Yamamoto, Y, Tsuzuki, S, Akahori, Y, Ukai, Y, Sumitomo, M, Murayama, Y, Yamamoto, K, Inaguma, Y, Tokuda, M, Abe, A, Akatsuka, Y, Emi, N & Kurosawa, Y 2012, 'Isolation of human mAbs that directly modulate FMS-related tyrosine kinase 3 signaling', Cancer Science, vol. 103, no. 2, pp. 350-359. https://doi.org/10.1111/j.1349-7006.2011.02141.x

Isolation of human mAbs that directly modulate FMS-related tyrosine kinase 3 signaling. / Yamamoto, Yukiya; Tsuzuki, Sachiko; Akahori, Yasushi; Ukai, Yoshinori; Sumitomo, Mariko; Murayama, Yuko; Yamamoto, Kiyoko; Inaguma, Youko; Tokuda, Masutaka; Abe, Akihiro; Akatsuka, Yoshiki; Emi, Nobuhiko; Kurosawa, Yoshikazu.

In: Cancer Science, Vol. 103, No. 2, 01.02.2012, p. 350-359.

Research output: Contribution to journalArticle

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AU - Yamamoto, Yukiya

AU - Tsuzuki, Sachiko

AU - Akahori, Yasushi

AU - Ukai, Yoshinori

AU - Sumitomo, Mariko

AU - Murayama, Yuko

AU - Yamamoto, Kiyoko

AU - Inaguma, Youko

AU - Tokuda, Masutaka

AU - Abe, Akihiro

AU - Akatsuka, Yoshiki

AU - Emi, Nobuhiko

AU - Kurosawa, Yoshikazu

PY - 2012/2/1

Y1 - 2012/2/1

N2 - FMS-related tyrosine kinase 3 (FLT3) is a class III receptor tyrosine kinase that plays important roles in hematopoiesis, including early progenitors and dendritic cell development. FLT3 is expressed at high levels in 70-100% of cases of AML and in virtually all cases of B-lineage acute lymphoblastic leukemia. FLT3 is regarded as a molecular target in the development of novel therapies for acute leukemia patients. Currently, many small-molecule FLT3 inhibitors have been developed, but clinical trials have resulted in limited antileukemia effects because of off-target toxicities and drug resistance. The development of anti-FLT3 Abs might overcome these difficulties and enhance the antileukemia efficacy of FLT3 inhibitors. In the present study, we demonstrate the isolation of novel human mAbs against FLT3 with antagonistic or agonistic activities. An antagonistic Ab, designated A2, continuously inhibits FLT3 ligand (FL)-induced phosphorylation of FLT3 and MAPK. A2 cooperatively induces apoptosis with daunorubicin, even in the presence of FL. An agonistic Ab, designated 3E6, surprisingly induces the phosphorylation of FLT3 and MAPK, and supports the growth of a factor-dependent cell line independently of FL addition. In addition, A2 showed complement-dependent cytotoxicity activity, but was devoid of Ab-dependent cell mediated cytotoxicity. Finally, we evaluated Ab internalization in a cell line. Immunofluorescence and flow cytometry analyses revealed that A2 is efficiently internalized. Collectively, these data demonstrate that A2 is a potent human Ab that might be capable of delivering cytotoxic reagents and that has antagonistic effects on FLT3 signaling. In addition, 3E6 might be a potential scaffold for novel dendritic cell-based immunotherapies.

AB - FMS-related tyrosine kinase 3 (FLT3) is a class III receptor tyrosine kinase that plays important roles in hematopoiesis, including early progenitors and dendritic cell development. FLT3 is expressed at high levels in 70-100% of cases of AML and in virtually all cases of B-lineage acute lymphoblastic leukemia. FLT3 is regarded as a molecular target in the development of novel therapies for acute leukemia patients. Currently, many small-molecule FLT3 inhibitors have been developed, but clinical trials have resulted in limited antileukemia effects because of off-target toxicities and drug resistance. The development of anti-FLT3 Abs might overcome these difficulties and enhance the antileukemia efficacy of FLT3 inhibitors. In the present study, we demonstrate the isolation of novel human mAbs against FLT3 with antagonistic or agonistic activities. An antagonistic Ab, designated A2, continuously inhibits FLT3 ligand (FL)-induced phosphorylation of FLT3 and MAPK. A2 cooperatively induces apoptosis with daunorubicin, even in the presence of FL. An agonistic Ab, designated 3E6, surprisingly induces the phosphorylation of FLT3 and MAPK, and supports the growth of a factor-dependent cell line independently of FL addition. In addition, A2 showed complement-dependent cytotoxicity activity, but was devoid of Ab-dependent cell mediated cytotoxicity. Finally, we evaluated Ab internalization in a cell line. Immunofluorescence and flow cytometry analyses revealed that A2 is efficiently internalized. Collectively, these data demonstrate that A2 is a potent human Ab that might be capable of delivering cytotoxic reagents and that has antagonistic effects on FLT3 signaling. In addition, 3E6 might be a potential scaffold for novel dendritic cell-based immunotherapies.

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Yamamoto Y, Tsuzuki S, Akahori Y, Ukai Y, Sumitomo M, Murayama Y et al. Isolation of human mAbs that directly modulate FMS-related tyrosine kinase 3 signaling. Cancer Science. 2012 Feb 1;103(2):350-359. https://doi.org/10.1111/j.1349-7006.2011.02141.x