Journal Article
. 2017 Sep; 114(38):10184-10189.
doi: 10.1073/pnas.1710776114.

Localized CD47 blockade enhances immunotherapy for murine melanoma

Jessica R Ingram 1 Olga S Blomberg 1 Jonathan T Sockolosky 2 Lestat Ali 3 Florian I Schmidt 1 Novalia Pishesha 1 Camilo Espinosa 1 Stephanie K Dougan 3 K Christopher Garcia 2 Hidde L Ploegh 4 Michael Dougan 4 
Affiliations
  • PMID: 28874561
  •     24 References
  •     46 citations

Abstract

CD47 is an antiphagocytic ligand broadly expressed on normal and malignant tissues that delivers an inhibitory signal through the receptor signal regulatory protein alpha (SIRPα). Inhibitors of the CD47-SIRPα interaction improve antitumor antibody responses by enhancing antibody-dependent cellular phagocytosis (ADCP) in xenograft models. Endogenous expression of CD47 on a variety of cell types, including erythrocytes, creates a formidable antigen sink that may limit the efficacy of CD47-targeting therapies. We generated a nanobody, A4, that blocks the CD47-SIRPα interaction. A4 synergizes with anti-PD-L1, but not anti-CTLA4, therapy in the syngeneic B16F10 melanoma model. Neither increased dosing nor half-life extension by fusion of A4 to IgG2a Fc (A4Fc) overcame the issue of an antigen sink or, in the case of A4Fc, systemic toxicity. Generation of a B16F10 cell line that secretes the A4 nanobody showed that an enhanced response to several immune therapies requires near-complete blockade of CD47 in the tumor microenvironment. Thus, strategies to localize CD47 blockade to tumors may be particularly valuable for immune therapy.

Keywords: T cell; cancer; immunotherapy; macrophage; protein engineering.

Nanobodies: natural single-domain antibodies.
Serge Muyldermans.
Annu Rev Biochem, 2013 Mar 19; 82. PMID: 23495938
Highly Cited. Review.
Anti-CTLA-4 antibodies of IgG2a isotype enhance antitumor activity through reduction of intratumoral regulatory T cells.
Mark J Selby, John J Engelhardt, +4 authors, Alan J Korman.
Cancer Immunol Res, 2014 Apr 30; 1(1). PMID: 24777248
Highly Cited.
Cancer immunotherapy targeting the CD47/SIRPα axis.
Kipp Weiskopf.
Eur J Cancer, 2017 Mar 14; 76. PMID: 28286286
Highly Cited. Review.
Disruption of SIRPα signaling in macrophages eliminates human acute myeloid leukemia stem cells in xenografts.
Alexandre P A Theocharides, Liqing Jin, +9 authors, Jean C Y Wang.
J Exp Med, 2012 Sep 05; 209(10). PMID: 22945919    Free PMC article.
Targeting Immunotherapy to the Tumor Microenvironment.
Michael Dougan, Stephanie K Dougan.
J Cell Biochem, 2017 Mar 24; 118(10). PMID: 28332219
Review.
Nanobodies as Versatile Tools to Understand, Diagnose, Visualize and Treat Cancer.
Isabel Van Audenhove, Jan Gettemans.
EBioMedicine, 2016 Jul 20; 8. PMID: 27428417    Free PMC article.
Review.
CD47 is upregulated on circulating hematopoietic stem cells and leukemia cells to avoid phagocytosis.
Siddhartha Jaiswal, Catriona H M Jamieson, +6 authors, Irving L Weissman.
Cell, 2009 Jul 28; 138(2). PMID: 19632178    Free PMC article.
Highly Cited.
Engineered erythrocytes covalently linked to antigenic peptides can protect against autoimmune disease.
Novalia Pishesha, Angelina M Bilate, +10 authors, Hidde L Ploegh.
Proc Natl Acad Sci U S A, 2017 Mar 09; 114(12). PMID: 28270614    Free PMC article.
The CD47-signal regulatory protein alpha (SIRPa) interaction is a therapeutic target for human solid tumors.
Stephen B Willingham, Jens-Peter Volkmer, +41 authors, Irving L Weissman.
Proc Natl Acad Sci U S A, 2012 Mar 28; 109(17). PMID: 22451913    Free PMC article.
Highly Cited.
CD47 blockade triggers T cell-mediated destruction of immunogenic tumors.
Xiaojuan Liu, Yang Pu, +7 authors, Meng Michelle Xu.
Nat Med, 2015 Sep 01; 21(10). PMID: 26322579    Free PMC article.
Highly Cited.
Engineered SIRPα variants as immunotherapeutic adjuvants to anticancer antibodies.
Kipp Weiskopf, Aaron M Ring, +8 authors, K Christopher Garcia.
Science, 2013 Jun 01; 341(6141). PMID: 23722425    Free PMC article.
Highly Cited.
CD47 is an adverse prognostic factor and therapeutic antibody target on human acute myeloid leukemia stem cells.
Ravindra Majeti, Mark P Chao, +5 authors, Irving L Weissman.
Cell, 2009 Jul 28; 138(2). PMID: 19632179    Free PMC article.
Highly Cited.
Durable antitumor responses to CD47 blockade require adaptive immune stimulation.
Jonathan T Sockolosky, Michael Dougan, +5 authors, K Christopher Garcia.
Proc Natl Acad Sci U S A, 2016 Apr 20; 113(19). PMID: 27091975    Free PMC article.
Highly Cited.
Coinhibitory Pathways in Immunotherapy for Cancer.
Susanne H Baumeister, Gordon J Freeman, Glenn Dranoff, Arlene H Sharpe.
Annu Rev Immunol, 2016 Mar 02; 34. PMID: 26927206
Highly Cited. Review.
Noninvasive imaging of immune responses.
Mohammad Rashidian, Edmund J Keliher, +8 authors, Hidde L Ploegh.
Proc Natl Acad Sci U S A, 2015 Apr 23; 112(19). PMID: 25902531    Free PMC article.
Highly Cited.
IAP inhibitors enhance co-stimulation to promote tumor immunity.
Michael Dougan, Stephanie Dougan, +11 authors, Glenn Dranoff.
J Exp Med, 2010 Sep 15; 207(10). PMID: 20837698    Free PMC article.
Pre-Clinical Development of a Humanized Anti-CD47 Antibody with Anti-Cancer Therapeutic Potential.
Jie Liu, Lijuan Wang, +10 authors, Ravindra Majeti.
PLoS One, 2015 Sep 22; 10(9). PMID: 26390038    Free PMC article.
Highly Cited.
"Velcro" engineering of high affinity CD47 ectodomain as signal regulatory protein α (SIRPα) antagonists that enhance antibody-dependent cellular phagocytosis.
Chia Chi M Ho, Nan Guo, +6 authors, K Christopher Garcia.
J Biol Chem, 2015 Apr 04; 290(20). PMID: 25837251    Free PMC article.
PD-1 and its ligands in tolerance and immunity.
Mary E Keir, Manish J Butte, Gordon J Freeman, Arlene H Sharpe.
Annu Rev Immunol, 2008 Jan 05; 26. PMID: 18173375
Highly Cited. Review.
The neonatal Fc receptor, FcRn, as a target for drug delivery and therapy.
Jonathan T Sockolosky, Francis C Szoka.
Adv Drug Deliv Rev, 2015 Feb 24; 91. PMID: 25703189    Free PMC article.
Review.
Combination immunotherapy of B16 melanoma using anti-cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) and granulocyte/macrophage colony-stimulating factor (GM-CSF)-producing vaccines induces rejection of subcutaneous and metastatic tumors accompanied by autoimmune depigmentation.
A van Elsas, A A Hurwitz, J P Allison.
J Exp Med, 1999 Aug 03; 190(3). PMID: 10430624    Free PMC article.
Highly Cited.
The interaction between signal regulatory protein alpha (SIRPα) and CD47: structure, function, and therapeutic target.
A Neil Barclay, Timo K Van den Berg.
Annu Rev Immunol, 2013 Nov 13; 32. PMID: 24215318
Highly Cited. Review.
Synergistic innate and adaptive immune response to combination immunotherapy with anti-tumor antigen antibodies and extended serum half-life IL-2.
Eric F Zhu, Shuning A Gai, +14 authors, K Dane Wittrup.
Cancer Cell, 2015 Apr 16; 27(4). PMID: 25873172    Free PMC article.
Highly Cited.
Fc-dependent depletion of tumor-infiltrating regulatory T cells co-defines the efficacy of anti-CTLA-4 therapy against melanoma.
Tyler R Simpson, Fubin Li, +11 authors, Sergio A Quezada.
J Exp Med, 2013 Jul 31; 210(9). PMID: 23897981    Free PMC article.
Highly Cited.
Targeting Cytokine Therapy to the Pancreatic Tumor Microenvironment Using PD-L1-Specific VHHs.
Michael Dougan, Jessica R Ingram, +12 authors, Stephanie K Dougan.
Cancer Immunol Res, 2018 Feb 21; 6(4). PMID: 29459478    Free PMC article.
Single-Domain Antibodies and the Promise of Modular Targeting in Cancer Imaging and Treatment.
María Elena Iezzi, Lucía Policastro, +2 authors, Gabriela Alicia Canziani.
Front Immunol, 2018 Mar 10; 9. PMID: 29520274    Free PMC article.
Review.
Anti-CTLA-4 therapy requires an Fc domain for efficacy.
Jessica R Ingram, Olga S Blomberg, +15 authors, Michael Dougan.
Proc Natl Acad Sci U S A, 2018 Mar 28; 115(15). PMID: 29581255    Free PMC article.
Highly Cited.
Preclinical Development of a Bispecific Antibody that Safely and Effectively Targets CD19 and CD47 for the Treatment of B-Cell Lymphoma and Leukemia.
Vanessa Buatois, Zoë Johnson, +28 authors, Walter G Ferlin.
Mol Cancer Ther, 2018 May 11; 17(8). PMID: 29743205    Free PMC article.
CD47-signal regulatory protein α signaling system and its application to cancer immunotherapy.
Yoji Murata, Yasuyuki Saito, Takenori Kotani, Takashi Matozaki.
Cancer Sci, 2018 Jun 07; 109(8). PMID: 29873856    Free PMC article.
Highly Cited. Review.
PD-L1 and CD47 co-expression in pulmonary sarcomatoid carcinoma: a predictor of poor prognosis and potential targets of future combined immunotherapy.
Zhenlin Yang, Jiachen Xu, +2 authors, Jie He.
J Cancer Res Clin Oncol, 2019 Sep 16; 145(12). PMID: 31522278
Macrophages induce CD47 upregulation via IL-6 and correlate with poor survival in hepatocellular carcinoma patients.
Jing Chen, Dan-Xue Zheng, +4 authors, Jing Xu.
Oncoimmunology, 2019 Oct 28; 8(11). PMID: 31646099    Free PMC article.
Theranostics in immuno-oncology using nanobody derivatives.
Quentin Lecocq, Yannick De Vlaeminck, +6 authors, Karine Breckpot.
Theranostics, 2019 Nov 07; 9(25). PMID: 31695800    Free PMC article.
Review.
Single-cell RNA sequencing reveals compartmental remodeling of tumor-infiltrating immune cells induced by anti-CD47 targeting in pancreatic cancer.
Yu Pan, Fengchun Lu, +10 authors, Heguang Huang.
J Hematol Oncol, 2019 Nov 28; 12(1). PMID: 31771616    Free PMC article.
Inducing Tumor Suppressive Microenvironments through Genome Edited CD47-/- Syngeneic Cell Vaccination.
Subhadra Jayaraman Rukmini, Huanjing Bi, +3 authors, Kaiming Ye.
Sci Rep, 2019 Dec 29; 9(1). PMID: 31882679    Free PMC article.
WNT Signaling in Tumors: The Way to Evade Drugs and Immunity.
Elena Martin-Orozco, Ana Sanchez-Fernandez, Irene Ortiz-Parra, Maria Ayala-San Nicolas.
Front Immunol, 2020 Jan 11; 10. PMID: 31921125    Free PMC article.
Review.
Glutaminyl cyclase is an enzymatic modifier of the CD47- SIRPα axis and a target for cancer immunotherapy.
Meike E W Logtenberg, J H Marco Jansen, +18 authors, Ton N Schumacher.
Nat Med, 2019 Mar 06; 25(4). PMID: 30833751    Free PMC article.
Highly Cited.
Nanobody-based CAR T cells that target the tumor microenvironment inhibit the growth of solid tumors in immunocompetent mice.
Yushu Joy Xie, Michael Dougan, +8 authors, Hidde Ploegh.
Proc Natl Acad Sci U S A, 2019 Apr 03; 116(16). PMID: 30936321    Free PMC article.
Highly Cited.
m6A mRNA demethylase FTO regulates melanoma tumorigenicity and response to anti-PD-1 blockade.
Seungwon Yang, Jiangbo Wei, +8 authors, Yu-Ying He.
Nat Commun, 2019 Jun 27; 10(1). PMID: 31239444    Free PMC article.
Highly Cited.
Programmable bacteria induce durable tumor regression and systemic antitumor immunity.
Sreyan Chowdhury, Samuel Castro, +3 authors, Tal Danino.
Nat Med, 2019 Jul 05; 25(7). PMID: 31270504    Free PMC article.
Highly Cited.
Macrophages show higher levels of engulfment after disruption of cis interactions between CD47 and the checkpoint receptor SIRPα.
Brandon H Hayes, Richard K Tsai, +7 authors, Dennis E Discher.
J Cell Sci, 2020 Jan 23; 133(5). PMID: 31964705    Free PMC article.
Engineering macrophages to phagocytose cancer cells by blocking the CD47/SIRPɑ axis.
Hongcheng Yang, Ruoyang Shao, +3 authors, Ying Lin.
Cancer Med, 2019 Jun 12; 8(9). PMID: 31183992    Free PMC article.
Review.
Intratumoral accumulation of gut microbiota facilitates CD47-based immunotherapy via STING signaling.
Yaoyao Shi, Wenxin Zheng, +7 authors, Yang-Xin Fu.
J Exp Med, 2020 Mar 07; 217(5). PMID: 32142585    Free PMC article.
The Role of Circulating Tumor Cells in the Metastatic Cascade: Biology, Technical Challenges, and Clinical Relevance.
Hassan Dianat-Moghadam, Mehdi Azizi, +4 authors, Catherine Alix-Panabières.
Cancers (Basel), 2020 Apr 09; 12(4). PMID: 32260071    Free PMC article.
Review.
ImmunoPET: Concept, Design, and Applications.
Weijun Wei, Zachary T Rosenkrans, +3 authors, Weibo Cai.
Chem Rev, 2020 Mar 24; 120(8). PMID: 32202104    Free PMC article.
Review.
The CD47-SIRPα Immune Checkpoint.
Meike E W Logtenberg, Ferenc A Scheeren, Ton N Schumacher.
Immunity, 2020 May 21; 52(5). PMID: 32433947    Free PMC article.
Review.
Antisense targeting of CD47 enhances human cytotoxic T-cell activity and increases survival of mice bearing B16 melanoma when combined with anti-CTLA4 and tumor irradiation.
Anthony L Schwartz, Pulak R Nath, +7 authors, David D Roberts.
Cancer Immunol Immunother, 2019 Oct 20; 68(11). PMID: 31628526    Free PMC article.
Nanobodies: Next Generation of Cancer Diagnostics and Therapeutics.
Emily Y Yang, Khalid Shah.
Front Oncol, 2020 Aug 15; 10. PMID: 32793488    Free PMC article.
Review.
Improved Antitumor Efficacy of Chimeric Antigen Receptor T Cells that Secrete Single-Domain Antibody Fragments.
Yushu Joy Xie, Michael Dougan, +4 authors, Hidde L Ploegh.
Cancer Immunol Res, 2020 Feb 06; 8(4). PMID: 32019780    Free PMC article.
Nanobody Engineering: Toward Next Generation Immunotherapies and Immunoimaging of Cancer.
Timothée Chanier, Patrick Chames.
Antibodies (Basel), 2019 Sep 24; 8(1). PMID: 31544819    Free PMC article.
Review.
Tumor-Directed Blockade of CD47 with Bispecific Antibodies Induces Adaptive Antitumor Immunity.
Elie Dheilly, Stefano Majocchi, +9 authors, Krzysztof Masternak.
Antibodies (Basel), 2018 Jan 03; 7(1). PMID: 31544856    Free PMC article.
Potential New Cancer Immunotherapy: Anti-CD47-SIRPα Antibodies.
Quansheng Lu, Xi Chen, +3 authors, Guan Jiang.
Onco Targets Ther, 2020 Oct 17; 13. PMID: 33061420    Free PMC article.
Review.
Emerging technologies for local cancer treatment.
Corrine Ying Xuan Chua, Jeremy Ho, +2 authors, Alessandro Grattoni.
Adv Ther (Weinh), 2020 Oct 20; 3(9). PMID: 33072860    Free PMC article.
Preclinical and Clinical Development of Therapeutic Antibodies Targeting Functions of CD47 in the Tumor Microenvironment.
Sukhbir Kaur, Kyle V Cicalese, Rajdeep Bannerjee, David D Roberts.
Antib Ther, 2020 Nov 28; 3(3). PMID: 33244513    Free PMC article.
Discovery of high affinity, pan-allelic, and pan-mammalian reactive antibodies against the myeloid checkpoint receptor SIRPα.
Janet Sim, Jonathan T Sockolosky, +13 authors, Jaume Pons.
MAbs, 2019 Jul 02; 11(6). PMID: 31257988    Free PMC article.
Current Progresses and Challenges of Immunotherapy in Triple-Negative Breast Cancer.
Karan Mediratta, Sara El-Sahli, Vanessa D'Costa, Lisheng Wang.
Cancers (Basel), 2020 Dec 02; 12(12). PMID: 33256070    Free PMC article.
Review.
Engineered CAR-T cells targeting TAG-72 and CD47 in ovarian cancer.
Runzhe Shu, Vera J Evtimov, +7 authors, Richard L Boyd.
Mol Ther Oncolytics, 2021 Feb 23; 20. PMID: 33614914    Free PMC article.
Exploiting Phagocytic Checkpoints in Nanomedicine: Applications in Imaging and Combination Therapies.
Madeleine R Landry, Joshua M Walker, Conroy Sun.
Front Chem, 2021 Mar 23; 9. PMID: 33748077    Free PMC article.
Review.
Effect of cabazitaxel on macrophages improves CD47-targeted immunotherapy for triple-negative breast cancer.
Xu Cao, Bolei Li, +12 authors, Mingye Feng.
J Immunother Cancer, 2021 Mar 24; 9(3). PMID: 33753567    Free PMC article.
Intratumoral SIRPα-deficient macrophages activate tumor antigen-specific cytotoxic T cells under radiotherapy.
Zhen Bian, Lei Shi, +3 authors, Yuan Liu.
Nat Commun, 2021 May 30; 12(1). PMID: 34050181    Free PMC article.
Nanobodies in cancer.
Elisha R Verhaar, Andrew W Woodham, Hidde L Ploegh.
Semin Immunol, 2020 Dec 05; 52. PMID: 33272897    Free PMC article.
Review.
Bacteria-Inspired Nanomedicine.
Maya Holay, Zhongyuan Guo, +4 authors, Liangfang Zhang.
ACS Appl Bio Mater, 2021 Aug 10; 4(5). PMID: 34368643    Free PMC article.
Review.
Blockade of immune checkpoints in lymph nodes through locoregional delivery augments cancer immunotherapy.
David M Francis, Margaret P Manspeaker, +6 authors, Susan N Thomas.
Sci Transl Med, 2020 Oct 02; 12(563). PMID: 32998971    Free PMC article.
cIAP1/2 antagonism eliminates MHC class I-negative tumors through T cell-dependent reprogramming of mononuclear phagocytes.
Kevin Roehle, Li Qiang, +26 authors, Stephanie K Dougan.
Sci Transl Med, 2021 May 21; 13(594). PMID: 34011631    Free PMC article.
Progress of CD47 immune checkpoint blockade agents in anticancer therapy: a hematotoxic perspective.
Yu-Chi Chen, Wei Shi, Jia-Jie Shi, Jin-Jian Lu.
J Cancer Res Clin Oncol, 2021 Oct 06; 148(1). PMID: 34609596
Review.
An oncolytic virus expressing a full-length antibody enhances antitumor innate immune response to glioblastoma.
Bo Xu, Lei Tian, +10 authors, Jianhua Yu.
Nat Commun, 2021 Oct 10; 12(1). PMID: 34625564    Free PMC article.
Emerging new therapeutic antibody derivatives for cancer treatment.
Shijie Jin, Yanping Sun, +5 authors, Liqiang Pan.
Signal Transduct Target Ther, 2022 Feb 09; 7(1). PMID: 35132063    Free PMC article.
Review.
Promoting antibody-dependent cellular phagocytosis for effective macrophage-based cancer immunotherapy.
Xu Cao, Jing Chen, +9 authors, Mingye Feng.
Sci Adv, 2022 Mar 19; 8(11). PMID: 35302839    Free PMC article.
Nanobody-based CAR-T cells for cancer immunotherapy.
Pouya Safarzadeh Kozani, Abdolhossein Naseri, +4 authors, Pooria Safarzadeh Kozani.
Biomark Res, 2022 Apr 27; 10(1). PMID: 35468841    Free PMC article.
Review.
CD47-SIRPα Checkpoint Disruption in Metastases Requires Tumor-Targeting Antibody for Molecular and Engineered Macrophage Therapies.
Jason C Andrechak, Lawrence J Dooling, +6 authors, Dennis E Discher.
Cancers (Basel), 2022 Apr 24; 14(8). PMID: 35454837    Free PMC article.
Role of CD47-SIRPα Checkpoint in Nanomedicine-Based Anti-Cancer Treatment.
Haiqin Liao, Chengcheng Niu.
Front Bioeng Biotechnol, 2022 May 14; 10. PMID: 35557862    Free PMC article.
Review.