A novel in situ multiplex immunofluorescence panel for the assessment of tumor immunopathology and response to virotherapy in pediatric glioblastoma reveals a role for checkpoint protein inhibition

Joshua D Bernstock; Nunzio Vicario; Li Rong; Pablo A Valdes; Bryan D Choi; Jason A Chen; Daniel DiToro; Diana S Osorio; Kara Kachurak; Florian Gessler; James M Johnston Jr; T Prescott Atkinson; Richard J Whitley; Asim K Bag; G Yancey Gillespie; James M Markert; Dragan Maric; Gregory K Friedman

doi:10.1080/2162402X.2019.1678921

A novel in situ multiplex immunofluorescence panel for the assessment of tumor immunopathology and response to virotherapy in pediatric glioblastoma reveals a role for checkpoint protein inhibition

Oncoimmunology. 2019 Oct 21;8(12):e1678921. doi: 10.1080/2162402X.2019.1678921. eCollection 2019.

Authors

Joshua D Bernstock^{1

2}, Nunzio Vicario³, Li Rong⁴, Pablo A Valdes², Bryan D Choi⁵, Jason A Chen⁶, Daniel DiToro^{1

7}, Diana S Osorio⁸, Kara Kachurak⁹, Florian Gessler¹⁰, James M Johnston Jr¹¹, T Prescott Atkinson¹², Richard J Whitley¹³, Asim K Bag¹⁴, G Yancey Gillespie¹¹, James M Markert¹¹, Dragan Maric¹⁵, Gregory K Friedman^{9

11}

Affiliations

¹ Medical Scientist Training Program, University of Alabama at Birmingham, Birmingham, AL, USA.
² Department of Neurosurgery, Brigham and Women's, Harvard Medical School, Boston, MA, USA.
³ Department of Biomedical and Biotechnological Sciences, Physiology Section, University of Catania, Catania, Italy.
⁴ Department of Pathology, Children's of Alabama, University of Alabama at Birmingham, Birmingham, AL, USA.
⁵ Department of Neurosurgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.
⁶ Medical Scientist Training Program, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
⁷ Department of Pathology, Brigham and Women's, Harvard Medical School, Boston, MA, USA.
⁸ Division of Pediatric Hematology/Oncology, Nationwide Children's Hospital, The Ohio State University, Columbus, OH, USA.
⁹ Division of Pediatric Hematology/Oncology, Department of Pediatrics, University of Alabama at Birmingham, Birmingham, AL, USA.
¹⁰ Department for Neurosurgery, Goethe University Frankfurt, Frankfurt am Main, Germany.
¹¹ Department of Neurosurgery, University of Alabama at Birmingham, Birmingham, AL, USA.
¹² Division of Pediatric Allergy, Asthma & Immunology, Department of Pediatrics and Diagnostic Mycoplasma Laboratory, Department of Pathology, University of Alabama at Birmingham, Birmingham, AL, USA.
¹³ Division of Infectious Diseases, Department of Pediatrics, University of Alabama at Birmingham, Birmingham, AL, USA.
¹⁴ Division of Neuroradiology, Department of Radiology, University of Alabama at Birmingham, Birmingham, AL, USA.
¹⁵ Flow and Imaging Cytometry Core Facility, National Institute of Neurological Disordersand Stroke, National Institutes of Health (NINDS/NIH), Bethesda, MD, USA.

Abstract

Immunotherapy with oncolytic herpes simplex virus-1 therapy offers an innovative, targeted, less-toxic approach for treating brain tumors. However, a major obstacle in maximizing oncolytic virotherapy is a lack of comprehensive understanding of the underlying mechanisms that unfold in CNS tumors/associated microenvironments after infusion of virus. We demonstrate that our multiplex biomarker screening platform comprehensively informs changes in both topographical location and functional states of resident/infiltrating immune cells that play a role in neuropathology after treatment with HSV G207 in a pediatric Phase 1 patient. Using this approach, we identified robust infiltration of CD8⁺ T cells suggesting activation of the immune response following virotherapy; however there was a corresponding upregulation of checkpoint proteins PD-1, PD-L1, CTLA-4, and IDO revealing a potential role for checkpoint inhibitors. Such work may ultimately lead to an understanding of the governing pathobiology of tumors, thereby fostering development of novel therapeutics tailored to produce optimal responses.

Keywords: HSV; checkpoint proteins; glioma; multiplex immunofluorescence; oncolytic virotherapy.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.

Grants and funding

R01 FD005379/FD/FDA HHS/United States