Th2-biased anti-tumor immunity promotes glioma growth in a mouse model Sami Sauma April 12,2016

Background – Astrocytoma Grade II low grade astrocytoma Grade III anaplastic astrocytoma Grade IV glioblastoma multiforme (GBM) Most common primary CNS tumor Aggressive, short clinical history, less than 3 months Aggressive; median survival 15 months

Background – Astrocytoma AIII CD163low CD163high AII AIII GBM Prosniak et al., 2013; Clin Cancer Res 19:3776-3786

Background – The Immune System Helper T Cell Zhu, 2008; Blood: 112 (5)

Background – The Immune System Alternatively Activated Monoctye Zhu, 2008; Blood: 112 (5)

Background – The Immune System Tumor Associated Macrophages

Background – GL261 Glioma Cell Line Commercialization NCI Repository Methylcholanthrene Seligman and Shear, 1939 Ex vivo transplantation Serial passaging Orthotopic Model Subcutaneous Model

Background – Antisense Oligodeoxynucleotides and the Tbet-/- Mouse (AS ODN) C57BL/6J Widely used inbred strain of mice Natural bias towards Th1 immunity Refractory to tumor development Antisense technology Tbet-/- C57BL/6 background A targeted mutation of the T-box transcription factor Tbx21 CD4 T cells do not produce the Th1-type cytokine interferon-γ Elevated levels of Th2-type cytokines

Background – Published Vaccine Data GL261 GL261 + AS ODN Gl261 (left flank) + AS ODN (right flank) Harshyne et al., 2015; Cancer Immunol Immunother 64(3):299–309 Morin-Brureau et al., 2015; Cancer Immunol Immunother 64(4):447-57

Results – Subcutaneous Model (Vaccine) GL261 GL261 + AS ODN GL261 GL261 + AS ODN

Results – Intracranial Titration of GL261

Results – Orthotopic Model Vaccine Intervention PBS GL261 + AS ODN PBS GL261 + AS ODN

Results – Orthotopic Model Vaccine Intervention PBS GL261 chambered + AS ODN PBS GL261 ch.+ AS ODN

Results – Orthotopic Model Vaccine Intervention PBS GL261 exosomes + AS ODN PBS GL261 exo + AS ODN

Results – Orthotopic Model PBS GL261 exo + AS ODN GL261 + AS ODN qPCR analysis of left and right cortices, and tumor tissue reveals no significant difference between two vaccination groups and mock. Elevated CD204 seen throughout.

Results – Subcutaneous Model PBS AS ODN Results – Subcutaneous Model AS ODN Intervention Day 0 flank administration of untreated GL261 cells in C57 and Tbet-/- mice Day 19/20 (dashed line) intraperitoneal injection of antisense or vehicle (PBS)

Results – Subcutaneous Model AS ODN Intervention PBS AS ODN

Results – Orthotopic Model AS ODN Intervention PBS AS ODN PBS AS ODN

Results – Orthotopic Model AS ODN Intervention PBS AS ODN PBS AS ODN

Results – Orthotopic Model AS ODN Intervention PBS AS ODN PBS AS ODN PBS AS ODN PBS AS ODN PBS AS ODN PBS AS ODN PBS AS ODN