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A novel CD47-blocking peptide fused to pro-apoptotic KLA repeat inhibits lung cancer growth in mice

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Abstract

CD47 is highly expressed in many tumor tissues and induces immune evasion by interaction with SIRP-alpha (signal regulatory protein-alpha) expressed on tumor-associated macrophages. In this study, we identified a novel CD47-blocking peptide VK17 by phage display technique. A pro-apoptotic VK30 peptide was obtained after VK17 was fused to KLA amino acid repeat at C-termini. The VK30 was specifically bound to CD47 on lung cancer cells, and subsequently inducing lung cancer cell apoptosis. Meanwhile, the expression of Bax was increased, whereas the expression of Bcl-2 and Ki-67 were reduced in the VK30-treated lung cancer cells. In addition, VK30 effectively improved the phagocytic activity of macrophages against VK30-pretreated lung cancer cells. Combinational treatment of lung cancer cells with blocking antibody anti-CD47 and VK30 additively enhanced VK30 binding to CD47, subsequently increasing lung cancer cell apoptosis and macrophage phagocytosis. Intraperitoneal administration of 2 mg/kg VK30 induced effective trafficking of VK30 into tumor tissues, and suppressing lung cancer cell growth in mice, associated with increased tumor cell apoptosis, macrophage activation and phagocytosis in vivo. The expression of CD47 was reduced in the VK30-treated tumor tissues and the expression level was positively correlated to tumor size. In addition, VK30 reduced the infiltration of CD11b+Ly6G+ neutrophils and CD11b+Ly6C+Ly6G+ granulocytic myeloid-derived suppressor cells (Gr-MDSCs) in tumor tissues, associated with suppressed expression of tumorigenic IL-6 and TNF-alpha from these cell types. Thereby, VK30 exerted anti-tumor effects in mice through inducing tumor cell apoptosis and macrophage phagocytosis. VK30 would be a novel therapeutic peptide in lung cancer immunotherapy.

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Funding

This study was supported by grants from the National Natural Science Foundation of China to ZLJ (82370082) and ZHC (81970023); Shanghai Municipal Key Clinical Specialty and Science and Technology Commission of Shanghai Municipality to YLS (shslczdzk02201, 20DZ2261200).

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  1. Linyue Pan and Lu Hu have contributed equally to this work.

Authors and Affiliations

  1. Department of Pulmonary Medicine, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai, 200032, China

    Linyue Pan, Mengjie Chen, Yuanlin Song, Zhihong Chen, Chun Li & Zhilong Jiang

  2. Department of Pulmonary and Critical Care Medicine, The Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China

    Lu Hu

  3. Shanghai Key Laboratory of Lung Inflammation and Injury, Shanghai, China

    Yuanlin Song

  4. Department of Orthopedic Surgery, Zhongshan Hospital, Fudan University, Shanghai, China

    Yutong Gu

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  1. Linyue Pan
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Contributions

LP, LH, MC and YG participated in collection and analysis of lung cancer tissues, cell culture, Western blot and animal experiments. YS, ZC and CL participated in generation of hypothesis, supervision and discussion. ZJ was responsible for generation of hypothesis, performing flow cytometry analysis, data analysis and assembly, manuscript writing and revision. All authors read and approved the final manuscript.

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Correspondence to Chun Li or Zhilong Jiang.

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The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

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Human tissues were collected following written informed consent of all participants, according to the Declaration of Helsinki and granted by the Ethics Committee of Zhongshan Hospital. All animal protocols were approved by Institutional Animal Care and Use Committee at Zhongshan Hospital.

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262_2023_3554_MOESM1_ESM.jpg

Supplemental Figure S1. Identification of bone marrow-derived macrophages (BMDMs) by flow cytometry analysis. Bone marrow cells were differentiated for 7 days and F4/80+ BMDMs were gated on CD11b+ cells. Representative histogram. (JPG 147 kb)

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Pan, L., Hu, L., Chen, M. et al. A novel CD47-blocking peptide fused to pro-apoptotic KLA repeat inhibits lung cancer growth in mice. Cancer Immunol Immunother 72, 4179–4194 (2023). https://doi.org/10.1007/s00262-023-03554-9

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