Abstract B18: Presence of tumor-associated macrophages in SHH medulloblastoma

Abstract

The concept of tumor-promoting inflammation is a recognized enabling characteristic of cancer. Tumor associated macrophages (TAMs), one of the main contributors to the tumor microenvironment, have been identified in many adult malignancies and in the MYCN non-amplified high risk neuroblastomas. TAMs have been shown to promote cancer via multiple mechanisms including tumor cell growth and survival, invasion, metastasis, angiogenesis, inflammation, and immune regulation. Recent studies in adult gliomas have demonstrated increased expression of macrophage/microglia associated genes in subtypes of gliomas, however little is known about the role of inflammation in medulloblastomas, the most common malignant childhood brain tumor. In this study we examined the expression of inflammation-related genes and presence of TAMs in molecular subgroups of pediatric medulloblastoma. The molecular subgroups of medulloblastomas were initially identified using Human Exon Array (HuEx) microarray data from 168 samples (65 patients profiled at Children's Hospital Los Angeles and 103 from previously published cohort) using a modified algorithm based on non-negative matrix factorization. We then examined the inflammation and immunology related genes that were differentially expressed among the molecular subgroups and discovered greater expression of inflammation-related genes in tumors of the SHH molecular subgroup compared with those of the Group 3 and Group 4 subgroups. Gene expression analysis was then performed on 83 medulloblastoma samples using a custom-built TaqMan Low Density Array (TLDA) card containing 41 tumor-related and 4 inflammation-related genes that were significantly deregulated among medulloblastoma subgroups in the HuEx microarray analyses. Thirty-one of these genes were used to develop a signature predictive of molecular subgroup. The internal cross-validted error rate of the TLDA 31-gene signature was 8%, with predominant misclassifications occurring between Groups 3 and 4. The expression levels of CD163 and CSF1R, two markers associated with TAMs, were higher in tumors of the SHH subgroup compared to those in the WNT, Group 3, and Group 4 subgroups (CD163, t-test p=.02 for WNT and p<0.001 for Groups 3 and 4; CSF1R, t-test p<0.01 for all groups). To validate these findings, we performed immunohistochemical analysis of 51 primary and 3 recurrent medulloblastoma tumors using antibodies directed against CD163, which were then blindly graded by two neuropathologists. There were significantly greater numbers of macrophages observed in tumors of the SHH subgroup compared to those in Group 3 and Group 4 (t test p<0.001 for both groups). Interestingly, macrophages in the desmoplastic or SHH tumors were concentrated in the poorly differentiated and highly proliferative internodular areas suggesting a tumor-proliferating role for these TAMs. Our study reports the first evidence of the presence of TAMs in medulloblastomas and provides a novel 31-gene TLDA signature that accurately determines molecular subgroups of medulloblastomas. The increase in expression of macrophage related genes and intratumoral macrophages was strongly associated with the SHH subgroup of patients. The identification of TAMs in medulloblastomas provides opportunity for testing new therapies directed against TAMs such as CSF1R inhibitors, and the recognition of the importance of tumor microenvironment in childhood brain tumors. Citation Format: Ashley Margol, Janahan Gnanachandran, Nathan Robison, Long Hung, Rebekah Kennedy, Marzieh Vali, Girish Dhall, Jonathan Finlay, Anat Epstein, Mark Krieger, Kathleen Dorris, Maryam Fouladi, Floyd Gilles, Alexander Judkins, Richard Sposto, Shahab Asgharzadeh. Presence of tumor-associated macrophages in SHH medulloblastoma. [abstract]. In: Proceedings of the AACR Special Conference on Pediatric Cancer at the Crossroads: Translating Discovery into Improved Outcomes; Nov 3-6, 2013; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2013;74(20 Suppl):Abstract nr B18.

Complement C3a Activates Astrocytes to Promote Medulloblastoma Progression through TNF -α

“…For example, C3a-C3aR signaling in microglia contributes to β-amyloid pathology and neuroinflammation in Alzheimer's disease [55]; and this signaling in microglia was recently reported to drive the pathogenesis of neuromyelitis optica [56]. Meanwhile, microglia/macrophages were demonstrated to participate in MB TME [57], although the conclusions are paradoxical that Maxmov et al found tumor-associated macrophages impair MB growth [58], whereas Yao et al demonstrated microglia promotes MB progression through IGF1 production [59]. In our study, the ideal approach to exclude C3a's effect on microglia/macrophages during MB development is to conditionally knockout C3aR in astrocytes of MB mice, that means to generate transgenic mice by crossmating GFAP-cre and C3aR loxp/loxp mice, then to crossmate these mice with MB mice.…”