ImmunoPET imaging of CD38 in murine lymphoma models using 89Zr-labeled daratumumab
CD38 is considered a potential biomarker for multiple myeloma (MM) and has shown a strong link with chronic lymphocytic leukemia due to high and uniform expression on plasma cells. In vivo evaluation of CD38 expression may provide useful information about lesion detection and prognosis of treatment in MM. In this study, immunoPET imaging with 89Zr-labeled daratumumab was used for differentiation of CD38 expression in murine lymphoma models to provide a potential non-invasive method for monitoring CD38 in the clinic.
Daratumumab was radiolabeled with 89Zr (t1/2 = 78.4 h) via conjugation with desferrioxamine (Df). After Western blot (WB) was used to screen CD38 expression in five lymphoma cell lines, flow cytometry and cellular binding assays were performed to test the binding ability of labeled or conjugated daratumumab with CD38 in vitro. PET imaging and biodistribution studies were performed to evaluate CD38 expression after injection of 89Zr-Df-daratumumab. 89Zr-Df-IgG was also evaluated as a non-specific control group in the Ramos model. Finally, CD38 expression in tumor tissues was verified by histological analysis.
Using WB screening, the Ramos cell line was found to express the highest level of CD38 while the HBL-1 cell line had the lowest expression. Df-conjugated and 89Zr-labeled daratumumab displayed similar high binding affinities with Ramos cells. PET imaging of 89Zr-Df-daratumumab showed a high tumor uptake of up to 26.6 ± 8.0 %ID/g for Ramos at 120 h post-injection, and only up to 6.6 ± 2.9 %ID/g for HBL-1 (n = 4). Additionally, 89Zr-Df-IgG demonstrated a low tumor uptake in the Ramos model (only 4.3 ± 0.8 %ID/g at 120 h post-injection). Ex vivo biodistribution studies showed similar trends with imaging results. Immunofluorescence staining of tumor tissues verified higher CD38 expression of Ramos than that of HBL-1.
The role of 89Zr-Df-daratumumab was investigated for evaluating CD38 expression in lymphoma models non-invasively and was found to be to a promising imaging agent of CD38-positive hematological diseases such as MM in future clinical applications.
KeywordsDaratumumab Positron emission tomography (PET) CD38 B-cell lymphoma ImmunoPET Cancer 89Zr
This work was supported by the University of Wisconsin - Madison, the National Institutes of Health (NIBIB/NCI 1R01CA169365, 1R01EB021336, P30CA014520, T32CA009206), the American Cancer Society (125246-RSG-13-099-01-CCE), the National Natural Science Foundation of China (81672602, 81472589, 31771036, 51703132, 51573096), the Beijing Nova Program (Z141102001814055, Z171100001117024), the Beijing Capital Special Development Application Program (Z141107002514159), PKU medicine-engineer collaborative seed program (BMU20160574), Logistics Scientific Research Project (BWS16J010) and the Basic Research Program of Shenzhen (JCYJ20170412111100742, JCYJ20160422091238319)
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflicts of interest.
All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.
- 2.Regelink JC, Minnema MC, Terpos E, Kamphuis MH, Raijmakers PG, Pieters-van den Bos IC, et al. Comparison of modern and conventional imaging techniques in establishing multiple myeloma-related bone disease: a systematic review. Br J Haematol. 2013;162:50–61. https://doi.org/10.1111/bjh.12346.CrossRefPubMedGoogle Scholar
- 11.de Weers M, Tai YT, van der Veer MS, Bakker JM, Vink T, Jacobs DC, et al. Daratumumab, a novel therapeutic human CD38 monoclonal antibody, induces killing of multiple myeloma and other hematological tumors. J Immunol. 2011;186:1840–8. https://doi.org/10.4049/jimmunol.1003032.CrossRefPubMedGoogle Scholar
- 12.Overdijk MB, Verploegen S, Bogels M, van Egmond M, Lammerts van Bueren JJ, Mutis T, et al. Antibody-mediated phagocytosis contributes to the anti-tumor activity of the therapeutic antibody daratumumab in lymphoma and multiple myeloma. MAbs. 2015;7:311–21. https://doi.org/10.1080/19420862.2015.1007813.CrossRefPubMedPubMedCentralGoogle Scholar
- 13.Overdijk MB, Jansen JH, Nederend M, Lammerts van Bueren JJ, Groen RW, Parren PW, et al. The therapeutic CD38 monoclonal antibody Daratumumab induces programmed cell death via Fcgamma receptor-mediated cross-linking. J Immunol. 2016;197:807–13. https://doi.org/10.4049/jimmunol.1501351.CrossRefPubMedGoogle Scholar
- 18.Bailly C, Clery PF, Faivre-Chauvet A, Bourgeois M, Guerard F, Haddad F, et al. Immuno-PET for clinical Theranostic approaches. Int J Mol Sci. 2016;18 https://doi.org/10.3390/ijms18010057.
- 21.Dimopoulos M, Terpos E, Comenzo RL, Tosi P, Beksac M, Sezer O, et al. International myeloma working group consensus statement and guidelines regarding the current role of imaging techniques in the diagnosis and monitoring of multiple myeloma. Leukemia. 2009;23:1545–56. https://doi.org/10.1038/leu.2009.89.CrossRefPubMedGoogle Scholar
- 23.Bartel TB, Haessler J, Brown TL, Shaughnessy JD Jr, van Rhee F, Anaissie E, et al. F18-fluorodeoxyglucose positron emission tomography in the context of other imaging techniques and prognostic factors in multiple myeloma. Blood. 2009;114:2068–76. https://doi.org/10.1182/blood-2009-03-213280.CrossRefPubMedPubMedCentralGoogle Scholar
- 25.Rajkumar SV, Harousseau JL, Durie B, Anderson KC, Dimopoulos M, Kyle R, et al. Consensus recommendations for the uniform reporting of clinical trials: report of the international myeloma workshop consensus panel 1. Blood. 2011;117:4691–5. https://doi.org/10.1182/blood-2010-10-299487.CrossRefPubMedPubMedCentralGoogle Scholar
- 27.Ak I, Aslan V, Vardareli E, Gulbas Z. Tc-99m methoxyisobutylisonitrile bone marrow imaging for predicting the levels of myeloma cells in bone marrow in multiple myeloma: correlation with CD38/CD138 expressing myeloma cells. Ann Hematol. 2003;82:88–92. https://doi.org/10.1007/s00277-002-0600-2.CrossRefPubMedGoogle Scholar
- 29.Pavon EJ, Zumaquero E, Rosal-Vela A, Khoo KM, Cerezo-Wallis D, Garcia-Rodriguez S, et al. Increased CD38 expression in T cells and circulating anti-CD38 IgG autoantibodies differentially correlate with distinct cytokine profiles and disease activity in systemic lupus erythematosus patients. Cytokine. 2013;62:232–43. https://doi.org/10.1016/j.cyto.2013.02.023.CrossRefPubMedGoogle Scholar
- 32.Boix F, Millan O, San Segundo D, Mancebo E, Rimola A, Fabrega E, et al. High expression of CD38, CD69, CD95 and CD154 biomarkers in cultured peripheral T lymphocytes correlates with an increased risk of acute rejection in liver allograft recipients. Immunobiology. 2016;221:595–603. https://doi.org/10.1016/j.imbio.2016.01.008.CrossRefPubMedGoogle Scholar
- 40.Witzig TE, Gordon LI, Cabanillas F, Czuczman MS, Emmanouilides C, Joyce R, et al. Randomized controlled trial of yttrium-90-labeled ibritumomab tiuxetan radioimmunotherapy versus rituximab immunotherapy for patients with relapsed or refractory low-grade, follicular, or transformed B-cell non-Hodgkin’s lymphoma. J Clin Oncol. 2002;20:2453–63. https://doi.org/10.1200/JCO.2002.11.076.CrossRefPubMedGoogle Scholar
- 42.Teiluf K, Seidl C, Blechert B, Gaertner FC, Gilbertz KP, Fernandez V, et al. Alpha-Radioimmunotherapy with (2)(1)(3)bi-anti-CD38 immunoconjugates is effective in a mouse model of human multiple myeloma. Oncotarget. 2015;6:4692–703. https://doi.org/10.18632/oncotarget.2986.CrossRefPubMedGoogle Scholar