Michael Zhang1,2, Quan Zhou1,3, Chinghsin Huang2, Carmel T Chan2, Wei Wu2, Gordon Li1, Michael Lim1, Sanjiv S Gambhir2, Heike E Daldrup-Link4. 1. Department of Neurosurgery, Stanford University, Stanford, CA, USA. 2. Department of Radiology, Stanford University, Stanford, CA, USA. 3. Department of Otolaryngology-Head and Neck Surgery, Stanford University, Stanford, CA, USA. 4. Department of Radiology, Stanford University, Stanford, CA, USA. heiked@stanford.edu.
Abstract
PURPOSE: Current checkpoint inhibitor immunotherapy strategies in glioblastoma are challenged by mechanisms of resistance including an immunosuppressive tumor microenvironment. T cell immunoglobulin domain and mucin domain 3 (TIM3) is a late-phase checkpoint receptor traditionally associated with T cell exhaustion. We apply fluorescent imaging techniques to explore feasibility of in vivo visualization of the immune state in a glioblastoma mouse model. PROCEDURES: TIM3 monoclonal antibody was conjugated to a near-infrared fluorescent dye, IRDye-800CW (800CW). The TIM3 experimental conjugate and isotype control were assessed for specificity with immunofluorescent staining and flow cytometry in murine cell lines (GL261 glioma and RAW264.7 macrophages). C57BL/6 mice with orthotopically implanted GL261 cells were imaged in vivo over 4 days after intravenous TIM3-800CW injection to assess tumor-specific uptake. Cell-specific uptake was then assessed on histologic sections. RESULTS: The experimental TIM3-800CW, but not its isotype control, bound to RAW264.7 macrophages in vitro. Specificity to RAW264.7 macrophages and not GL261 tumor cells was quantitatively confirmed with the corresponding clone of TIM3 on flow cytometry. In vivo fluorescence imaging of the 800CW signal was localized to the intracranial tumor and significantly higher for the TIM3-800CW cohort, relative to non-targeting isotype control, immediately after tail vein injection and for up to 48 h after injection. Resected organs of tumor bearing mice showed significantly higher uptake in the liver and spleen. TIM3-800CW was seen to co-stain with CD3 (13%), CD11b (29%), and CD206 (26%). CONCLUSIONS: We propose fluorescent imaging of immune cell imaging as a potential strategy for monitoring and localizing immunologically relevant foci in the setting of brain tumors. Alternative markers and target validation will further clarify the temporal relationship of immunosuppressive effector cells throughout glioma resistance.
PURPOSE: Current checkpoint inhibitor immunotherapy strategies in glioblastoma are challenged by mechanisms of resistance including an immunosuppressive tumor microenvironment. T cell immunoglobulin domain and mucin domain 3 (TIM3) is a late-phase checkpoint receptor traditionally associated with T cell exhaustion. We apply fluorescent imaging techniques to explore feasibility of in vivo visualization of the immune state in a glioblastoma mouse model. PROCEDURES: TIM3 monoclonal antibody was conjugated to a near-infrared fluorescent dye, IRDye-800CW (800CW). The TIM3 experimental conjugate and isotype control were assessed for specificity with immunofluorescent staining and flow cytometry in murine cell lines (GL261 glioma and RAW264.7 macrophages). C57BL/6 mice with orthotopically implanted GL261 cells were imaged in vivo over 4 days after intravenous TIM3-800CW injection to assess tumor-specific uptake. Cell-specific uptake was then assessed on histologic sections. RESULTS: The experimental TIM3-800CW, but not its isotype control, bound to RAW264.7 macrophages in vitro. Specificity to RAW264.7 macrophages and not GL261 tumor cells was quantitatively confirmed with the corresponding clone of TIM3 on flow cytometry. In vivo fluorescence imaging of the 800CW signal was localized to the intracranial tumor and significantly higher for the TIM3-800CW cohort, relative to non-targeting isotype control, immediately after tail vein injection and for up to 48 h after injection. Resected organs of tumor bearing mice showed significantly higher uptake in the liver and spleen. TIM3-800CW was seen to co-stain with CD3 (13%), CD11b (29%), and CD206 (26%). CONCLUSIONS: We propose fluorescent imaging of immune cell imaging as a potential strategy for monitoring and localizing immunologically relevant foci in the setting of brain tumors. Alternative markers and target validation will further clarify the temporal relationship of immunosuppressive effector cells throughout glioma resistance.
Authors: Ana C Anderson; David E Anderson; Lisa Bregoli; William D Hastings; Nasim Kassam; Charles Lei; Rucha Chandwaskar; Jozsef Karman; Ee W Su; Mitsuomi Hirashima; Jeffrey N Bruce; Lawrence P Kane; Vijay K Kuchroo; David A Hafler Journal: Science Date: 2007-11-16 Impact factor: 47.728
Authors: Jennifer E Kim; Mira A Patel; Antonella Mangraviti; Eileen S Kim; Debebe Theodros; Esteban Velarde; Ann Liu; Eric W Sankey; Ada Tam; Haiying Xu; Dimitrios Mathios; Christopher M Jackson; Sarah Harris-Bookman; Tomas Garzon-Muvdi; Mary Sheu; Allison M Martin; Betty M Tyler; Phuoc T Tran; Xiaobu Ye; Alessandro Olivi; Janis M Taube; Peter C Burger; Charles G Drake; Henry Brem; Drew M Pardoll; Michael Lim Journal: Clin Cancer Res Date: 2016-06-29 Impact factor: 12.531
Authors: Haley L Wissler; Emily B Ehlerding; Zhigang Lyu; Yue Zhao; Si Zhang; Anisa Eshraghi; Zakey Yusuf Buuh; Jeffrey C McGuth; Yifu Guan; Jonathan W Engle; Sarah J Bartlett; Vincent A Voelz; Weibo Cai; Rongsheng E Wang Journal: Mol Pharm Date: 2019-03-25 Impact factor: 4.939
Authors: Naoki Nishio; Nynke S van den Berg; Stan van Keulen; Brock A Martin; Shayan Fakurnejad; Nutte Teraphongphom; Stefania U Chirita; Nicholas J Oberhelman; Guolan Lu; Crista E Horton; Michael J Kaplan; Vasu Divi; A Dimitrios Colevas; Eben L Rosenthal Journal: Nat Commun Date: 2019-11-06 Impact factor: 14.919
Authors: Quan Zhou; Johana C M Vega Leonel; Michelle Rai Santoso; Christy Wilson; Nynke S van den Berg; Carmel T Chan; Muna Aryal; Hannes Vogel; Romain Cayrol; Michael J Mandella; Frank Schonig; Guolan Lu; Sanjiv S Gambhir; Michael E Moseley; Eben L Rosenthal; Gerald A Grant Journal: Sci Rep Date: 2021-03-11 Impact factor: 4.379
Authors: David A Reardon; Alba A Brandes; Antonio Omuro; Paul Mulholland; Michael Lim; Antje Wick; Joachim Baehring; Manmeet S Ahluwalia; Patrick Roth; Oliver Bähr; Surasak Phuphanich; Juan Manuel Sepulveda; Paul De Souza; Solmaz Sahebjam; Michael Carleton; Kay Tatsuoka; Corina Taitt; Ricardo Zwirtes; John Sampson; Michael Weller Journal: JAMA Oncol Date: 2020-07-01 Impact factor: 31.777