Abstract 187: Absence of S100A9 confers survival advantage in an aggressive de novo mouse model of glioblastoma multiforme
Authors: Calinescu Alexandra , Assi Hikmat , Kolb Bradley , Koschmann Carl , Lowenstein Pedro R. , Ohlfest John , Castro Maria G. 
Published: May 2015

Glioblastoma multiforme (GBM) is the most common (60%) primary brain tumor in adults and constitutes a major challenge for both patients and clinicians, with a median survival of only 15-21 months when treated with surgery, radiation therapy and temozolomide. To improve patient outcome, a promising and safe avenue of adjuvant treatment is immune therapy with dendritic cell vaccines or adoptive T cell transfer. These treatments are however hindered by the immune suppressive environment induced by GBMs. Understanding the mechanisms by which GBM suppress the anti-tumor immune response is paramount in order to develop successful immune therapies.

The Sleeping Beauty transposase system was used to modify the genetic makeup of stem cells in the sub-ventricular zone of neonatal mice and thus induce de novo glioblastomas with several combinations of genes. The most aggressive tumors where induced when combining NRAS and SV-40 Large T antigen (NLgT) generating invasive tumors which show the histological hallmarks human GBM (WHO grade IV) with pseudo-pallisading necrosis, neovascularization and hemorrhages, rendering the mice moribund with a median survival of 30 days.

Myeloid derived suppressive cells, a heterogeneous population of immature bone marrow derived cells, are induced by cancers and other consumptive diseases and strongly inhibit adaptive and innate immune responses. We show that MDSCs isolated from de novo GBMs, inhibit antigen-specific and antigen non-specific T cell proliferation. Tumor infiltrating MDSCs highly express the pro-inflammatory secreted calcium binding protein S100A9 and its cognate receptors RAGE and TLR4. In bone marrow cultures, conditioned media from primary cell lines derived from NLgT tumors induce a marked expansion (60-70%) of myeloid derived suppressor cells (MDSCs) and enhance the expression of S100A9. Mice deficient for S100A9 with de novo NLgT tumors show an increased survival compared to wild-type animals (median survival= 47days, a 56% improvement). Analysis of tumor-infiltrating mononuclear cells in moribund NLgT wild-type and S100A9KO mice shows no significant difference in percent infiltrating MDSCs, macrophages and dendritic cells (% of CD45+), however the percent of CD8+ cytotoxic T lymphocytes is increased in S100A9 KO (16% vs. 6% p=0.0228). In addition, expression of MHCII is increased in macrophages and dendritic cells from S100A9 KO mice when compared to wild type mice.

Taken together these data suggest that the absence of S100A9 confers a survival advantage by allowing a stronger anti-tumor immune response with:(1) increased number of cytotoxic T lymphocytes and (2) increased maturation of antigen presenting cells. Experiments are currently underway to identify the detailed cellular and molecular events, which give rise to this phenotype.

This study was supported by NIH/RO1 NS057711 and NS074387.

Citation Format: Alexandra Calinescu, Hikmat Assi, Bradley Kolb, Carl Koschmann, Pedro R. Lowenstein, John Ohlfest, Maria G. Castro. Absence of S100A9 confers survival advantage in an aggressive de novo mouse model of glioblastoma multiforme. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 187. doi:10.1158/1538-7445.AM2014-187