Antitumor activity of chLpMab-2, a human-mouse chimeric cancer-specific antihuman podoplanin antibody, via antibody-dependent cellular cytotoxicity

Our plain-English summary of Antitumor activity of chLpMab-2, a human-mouse chimeric cancer-specific antihuman podoplanin antibody, via antibody-dependent cellular cytotoxicity, by M.K. Kaneko, S. Yamada, T. Nakamura, et al (Cancer Medicine, 2017).

The study

• Authors: M.K. Kaneko, S. Yamada, T. Nakamura, S. Abe, Y. Nishioka, A. Kunita, M. Fukayama, Y. Fujii, S. Ogasawara, Y. Kato
• Journal: Cancer Medicine
• Published: 2017;6(4):768-777
• Cited by: 36 articles, as counted by NIH iCite
• Read it: PubMed · Publisher · Free full text

Abstract

Human podoplanin (hPDPN), a platelet aggregation-inducing transmembrane glycoprotein, is expressed in different types of tumors, and it binds to C-type lectin-like receptor 2 (CLEC-2). The overexpression of hPDPN is involved in invasion and metastasis. Anti-hPDPN monoclonal antibodies (mAbs) such as NZ-1 have shown antitumor and antimetastatic activities by binding to the platelet aggregation-stimulating (PLAG) domain of hPDPN. Recently, we developed a novel mouse anti-hPDPN mAb, LpMab-2, using the cancer-specific mAb (CasMab) technology. In this study we developed chLpMab-2, a human-mouse chimeric anti-hPDPN antibody, derived from LpMab-2. chLpMab-2 was produced using fucosyltransferase 8-knockout (KO) Chinese hamster ovary (CHO)-S cell lines. By flow cytometry, chLpMab-2 reacted with hPDPN-expressing cancer cell lines including glioblastomas, mesotheliomas, and lung cancers. However, it showed low reaction with normal cell lines such as lymphatic endothelial and renal epithelial cells. Moreover, chLpMab-2 exhibited high antibody-dependent cellular cytotoxicity (ADCC) against PDPN-expressing cells, despite its low complement-dependent cytotoxicity. Furthermore, treatment with chLpMab-2 abolished tumor growth in xenograft models of CHO/hPDPN, indicating that chLpMab-2 suppressed tumor development via ADCC. In conclusion, chLpMab-2 could be useful as a novel antibody-based therapy against hPDPN-expressing tumors.

© 2017 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.

In plain English

This laboratory study reported the development and testing of chLpMab-2, a human–mouse chimeric antibody against human podoplanin (hPDPN) derived from the mouse antibody LpMab-2 and produced in fucosyltransferase 8-knockout CHO-S cells. By flow cytometry, chLpMab-2 bound LN229/hPDPN cells and endogenous hPDPN in the glioblastoma line LN319 but not in LN319/hPDPN-knockout cells, and it detected PDPN in the lung squamous cell carcinoma line PC-10 and the mesothelioma line NCI-H226, while failing to detect hPDPN in HEK-293T renal epithelial cells and the Met-5A mesothelial line, which the comparison antibody chLpMab-7 did bind. In 51Cr release assays using human peripheral blood mononuclear cells as effector cells, chLpMab-2 produced antibody-dependent cellular cytotoxicity against LN319, PC-10, NCI-H226 and CHO/hPDPN cells, but not against hPDPN-negative CHO-K1 cells; in separate 51Cr release assays using baby rabbit complement, chLpMab-2 showed weak complement-dependent cytotoxicity. In BALB/c nude mice bearing CHO/hPDPN tumours (six per group) given human NK cells, tumour volume was significantly reduced by chLpMab-2 on days 15, 18 and 21 versus control human IgG. The authors concluded chLpMab-2 may be useful as antibody-based therapy against hPDPN-expressing tumours.

We wrote this plain-English summary from the full article with the help of AI, and a second AI model checked each sentence against the article. It is not the authors' wording; the abstract and the article are their own account.

What the study did and found

• Design: Laboratory study: antibody development with in vitro cell-line assays (flow cytometry, 51Cr release ADCC and CDC) and a mouse xenograft experiment
• Who was studied: Human and animal cell lines (LN229, LN229/hPDPN, LN319, LN319/hPDPN-KO (PDIS-6), PC-10, NCI-H226, Met-5A, HEK-293T, CHO-K1, CHO/hPDPN, CHO-S and glycan-deficient PDIS-9 and PDIS-14), human peripheral blood mononuclear cells from healthy donors, and BALB/c nude mice (6 per group) implanted with CHO/hPDPN cells; work done in Japan
• Exposure: Treatment with the chimeric anti-hPDPN antibody chLpMab-2 (afucosylated, produced in FUT8-KO CHO-S PDIS-5 cells), compared with chLpMab-7 or control human IgG; in vitro 0.01-10 µg/mL, in vivo 100 µg weekly for 4 weeks
• Main results:
  • chLpMab-2 bound LN229/hPDPN cells and endogenous hPDPN in LN319 but not in LN319/hPDPN-knockout cells (PDIS-6)
  • chLpMab-2 detected PDPN in the lung squamous cell carcinoma line PC-10 and the mesothelioma line NCI-H226; reaction with NCI-H226 was much lower than that of chLpMab-7
  • chLpMab-2 did not detect hPDPN in the normal cell lines HEK-293T (renal epithelial) and Met-5A (mesothelial), whereas chLpMab-7 did
  • In the glycan-deficient lines, chLpMab-2 reacted with CHO-S/hPDPN and PDIS-9/hPDPN but not with the sialic acid transporter-knockout PDIS-14/hPDPN cells
  • In 6-h 51Cr release assays using human mononuclear cells as effectors (E/T ratio 100), chLpMab-2 showed ADCC against LN319, PC-10 and NCI-H226, and against CHO/hPDPN but not hPDPN-negative CHO-K1 cells, in a dose-dependent manner
  • In 51Cr release assays using baby rabbit complement at a 1:32 dilution, chLpMab-2 showed weak complement-dependent cytotoxicity against CHO/hPDPN cells, and against LN319, PC-10 and NCI-H226 (data not shown)
  • In the CHO/hPDPN xenograft model, tumours formed in both groups but tumour volume was significantly reduced by chLpMab-2 on days 15, 18 and 21 compared with control human IgG (6 mice per group)
  • Reported as data not shown: ADCC of afucosylated chLpMab-2 was 3.4 times higher than that of fucosylated chLpMab-2, while afucosylated antibody showed lower CDC
• The authors' conclusion: The authors concluded that chLpMab-2 may be useful as a novel antibody-based therapy against hPDPN-expressing tumours, and that ADCC appeared important for its antitumour activity in the xenograft model
• Limitations the authors state:
  • The authors stated that the LN319, PC-10 and NCI-H226 cell lines were not tested in vivo because they were not suitable for xenograft models, so the antitumour test used only CHO/hPDPN cells

Topics

Medical terms in this study, explained in our Lexicon: Podoplanin (D2-40), Glycoprotein, Metastasis, Monoclonal, Mesothelioma, Lung Cancer.

Related studies

• Antitumor effect of novel anti-podoplanin antibody NZ-12 against malignant pleural mesothelioma in an orthotopic xenograft model — S. Abe, M.K. Kaneko, et al. (2016)
• Establishment of Mouse Monoclonal Antibody LpMab-13 Against Human Podoplanin — S. Ogasawara, M.K. Kaneko, et al. (2016)
• Therapeutic efficacy evaluation of radioimmunotherapy with 90 Y-labeled anti-podoplanin antibody NZ-12 for mesothelioma — H. Sudo, A.B. Tsuji, et al. (2019)
• A human single-domain antibody elicits potent antitumor activity by targeting an epitope in mesothelin close to the cancer cell surface — Z. Tang, M. Feng, et al. (2013)
• Podoplanin: An emerging cancer biomarker and therapeutic target — H. Krishnan, J. Rayes, et al. (2018)
• Lapatinib enhances trastuzumab-mediated antibody-dependent cellular cytotoxicity via upregulation of HER2 in malignant mesothelioma cells — R. Okita, K. Shimizu, et al. (2015)

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