Central nervous system immune interactome is a function of cancer lineage, tumor microenvironment, and STAT3 expression
Hinda Najem, … , Jared K. Burks, Amy B. Heimberger
Hinda Najem, … , Jared K. Burks, Amy B. Heimberger
Published March 22, 2022
Citation Information: JCI Insight. 2022;7(9):e157612. https://doi.org/10.1172/jci.insight.157612.
View: Text | PDF
Research Article Immunology Oncology

Central nervous system immune interactome is a function of cancer lineage, tumor microenvironment, and STAT3 expression

Abstract

BACKGROUND Immune cell profiling of primary and metastatic CNS tumors has been focused on the tumor, not the tumor microenvironment (TME), or has been analyzed via biopsies.METHODS En bloc resections of gliomas (n = 10) and lung metastases (n = 10) were analyzed via tissue segmentation and high-dimension Opal 7-color multiplex imaging. Single-cell RNA analyses were used to infer immune cell functionality.RESULTS Within gliomas, T cells were localized in the infiltrating edge and perivascular space of tumors, while residing mostly in the stroma of metastatic tumors. CD163+ macrophages were evident throughout the TME of metastatic tumors, whereas in gliomas, CD68+, CD11c+CD68+, and CD11c+CD68+CD163+ cell subtypes were commonly observed. In lung metastases, T cells interacted with CD163+ macrophages as dyads and clusters at the brain-tumor interface and within the tumor itself and as clusters within the necrotic core. In contrast, gliomas typically lacked dyad and cluster interactions, except for T cell CD68+ cell dyads within the tumor. Analysis of transcriptomic data in glioblastomas revealed that innate immune cells expressed both proinflammatory and immunosuppressive gene signatures.CONCLUSION Our results show that immunosuppressive macrophages are abundant within the TME and that the immune cell interactome between cancer lineages is distinct. Further, these data provide information for evaluating the role of different immune cell populations in brain tumor growth and therapeutic responses.FUNDING This study was supported by the NIH (NS120547), a Developmental research project award (P50CA221747), ReMission Alliance, institutional funding from Northwestern University and the Lurie Comprehensive Cancer Center, and gifts from the Mosky family and Perry McKay. Performed in the Flow Cytometry & Cellular Imaging Core Facility at MD Anderson Cancer Center, this study received support in part from the NIH (CA016672) and the National Cancer Institute (NCI) Research Specialist award 1 (R50 CA243707). Additional support was provided by CCSG Bioinformatics Shared Resource 5 (P30 CA046592), a gift from Agilent Technologies, a Research Scholar Grant from the American Cancer Society (RSG-16-005-01), a Precision Health Investigator Award from University of Michigan (U-M) Precision Health, the NCI (R37-CA214955), startup institutional research funds from U-M, and a Biomedical Informatics & Data Science Training Grant (T32GM141746).

Authors

Hinda Najem, Martina Ott, Cynthia Kassab, Arvind Rao, Ganesh Rao, Anantha Marisetty, Adam M. Sonabend, Craig Horbinski, Roel Verhaak, Anand Shankar, Santhoshi N. Krishnan, Frederick S. Varn, Víctor A. Arrieta, Pravesh Gupta, Sherise D. Ferguson, Jason T. Huse, Gregory N. Fuller, James P. Long, Daniel E. Winkowski, Ben A. Freiberg, Charles David James, Leonidas C. Platanias, Maciej S. Lesniak, Jared K. Burks, Amy B. Heimberger

×

Figure 1

Study workflow and key observations of the TME.

Options: View larger image (or click on image) Download as PowerPoint
Study workflow and key observations of the TME. (A) Study workflow. Afte...
(A) Study workflow. After en bloc gross total resection of diagnosed brain tumor, the surgeon cuts a wedge that includes an infiltrating edge or normal brain (in the case of brain metastasis), tumor, and necrotic core. Formalin-fixed paraffin-embedded slides were prepared subsequently for analysis. The 7-color Opal multiplex-staining panel includes DAPI (dark blue nuclei), CD3 (Opal 480, cyan blue), p-STAT3 (Opal 520 [Akoya Biosciences], green), CD68 (Opal 570 [Akoya Biosciences], yellow), CD163 (Opal 690 [Akoya Biosciences], red), CD11c (Opal 780 [Akoya Biosciences], white), and GFAP or AE1/AE2 (Opal 620 [Akoya Biosciences], orange). (B) Representative multiplex imaging of a whole-mount section of a glioblastoma (GBM), spanning from infiltrating edge to necrotic core, visualized using Phenochart software (original magnification, ×0.2). (C) Representative images of 3 different regions of glioma TME (infiltrating edge, tumor, and necrotic core) (original magnification, ×20). There is a predominance of CD68+ cells within the tumor area in contrast to the edge and necrotic core where CD163+ macrophages predominate. (D) Gradient of CD163+ macrophages in which the density is highest near brain metastasis and decreases toward normal brain (original magnification, ×20). (E) p-STAT3+ reactive astrocytes at the infiltrating edge of glioma. The 2 white arrows point to CD11c+CD68+ microglia. The red arrow denotes red blood cells (RBCs) in which the signal from Opal 480 (cyan) was differentiated from the T cells by the absence of DAPI (original magnification, ×40). (F) CD3+ T cell infiltration of brain metastasis observed in the stroma of the tumor (original magnification, ×5). (G) CD3+ T cells, within the tumor, have extravasated from the vessel but are located adjacent to the vessel, and CD163+ macrophages as well as CD68+ monocytes line the vasculature wall in GBM. Some RBCs are also present inside the vessel (original magnification, ×20). (H) CD3+ T cell infiltration shown at the gliotic plane (infiltrating edge) in low-grade glioma (original magnification, ×20). (I) p-STAT3– nonreactive astrocyte (white arrow) in close proximity to a CD3+p-STAT3– T cell (blue arrow) and a CD163+p-STAT3– macrophage (red arrow) located in the normal brain/infiltrating edge in low-grade glioma (original magnification, ×40).