TY - CHAP
T1 - High-dimensional multiplexed immunohistochemical characterization of immune contexture in human cancers
AU - Banik, Grace
AU - Betts, Courtney B.
AU - Liudahl, Shannon M.
AU - Sivagnanam, Shamilene
AU - Kawashima, Rie
AU - Cotechini, Tiziana
AU - Larson, William
AU - Goecks, Jeremy
AU - Pai, Sara I.
AU - Clayburgh, Daniel R.
AU - Tsujikawa, Takahiro
AU - Coussens, Lisa M.
N1 - Publisher Copyright:
© 2020 Elsevier Inc.
PY - 2020
Y1 - 2020
N2 - Biomarker assessments of tumor specimens is widely used in cancer research to audit tumor cell intrinsic as well as tumor cell extrinsic features including the diversity of immune, stromal, and mesenchymal cells. To comprehensively and quantitatively audit the tumor-immune microenvironment (TiME), we developed a novel multiplex immunohistochemistry (mIHC) platform and computational image processing workflow using a single formalin-fixed paraffin-embedded (FFPE) tissue section. Herein, we validated this platform using nine matched primary newly diagnosed and recurrent head and neck squamous cell carcinoma (HNSCC) sections sequentially subjected to immunodetection with a panel of 29 antibodies identifying malignant tumor cells, and 17 distinct leukocyte lineages and their functional states. Image cytometric analysis was applied to interpret chromogenic signals from digitally scanned and coregistered light microscopy-based images enabling identification and quantification of individual tumor cells, structural features, immune cell phenotypes and their functional state. In agreement with our previous study via a 12-plex imaging mIHC platform, myeloid-inflamed status in newly diagnosed primary tumors associated with significantly short progression free survival, independent of lymphoid-inflamed status. Spatial distribution of tumor and immune cell lineages in TiME was also examined and revealed statistically significant CD8+ T cell exclusion from tumor nests, whereas regulatory T cells and myeloid cells, when present in close proximity to tumor cells, highly associated with rapid cancer recurrence. These findings indicate presence of differential immune-spatial profiles in newly diagnosed and recurrent HNSCC, and establish the robustness of the 29-plex mIHC platform and associated analytics for quantitative analysis of single tissue sections revealing longitudinal TiME changes.
AB - Biomarker assessments of tumor specimens is widely used in cancer research to audit tumor cell intrinsic as well as tumor cell extrinsic features including the diversity of immune, stromal, and mesenchymal cells. To comprehensively and quantitatively audit the tumor-immune microenvironment (TiME), we developed a novel multiplex immunohistochemistry (mIHC) platform and computational image processing workflow using a single formalin-fixed paraffin-embedded (FFPE) tissue section. Herein, we validated this platform using nine matched primary newly diagnosed and recurrent head and neck squamous cell carcinoma (HNSCC) sections sequentially subjected to immunodetection with a panel of 29 antibodies identifying malignant tumor cells, and 17 distinct leukocyte lineages and their functional states. Image cytometric analysis was applied to interpret chromogenic signals from digitally scanned and coregistered light microscopy-based images enabling identification and quantification of individual tumor cells, structural features, immune cell phenotypes and their functional state. In agreement with our previous study via a 12-plex imaging mIHC platform, myeloid-inflamed status in newly diagnosed primary tumors associated with significantly short progression free survival, independent of lymphoid-inflamed status. Spatial distribution of tumor and immune cell lineages in TiME was also examined and revealed statistically significant CD8+ T cell exclusion from tumor nests, whereas regulatory T cells and myeloid cells, when present in close proximity to tumor cells, highly associated with rapid cancer recurrence. These findings indicate presence of differential immune-spatial profiles in newly diagnosed and recurrent HNSCC, and establish the robustness of the 29-plex mIHC platform and associated analytics for quantitative analysis of single tissue sections revealing longitudinal TiME changes.
KW - Cancer immunology
KW - Histopathology
KW - Immunohistochemistry
KW - Multiplex imaging
UR - http://www.scopus.com/inward/record.url?scp=85068177219&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85068177219&partnerID=8YFLogxK
U2 - 10.1016/bs.mie.2019.05.039
DO - 10.1016/bs.mie.2019.05.039
M3 - Chapter
C2 - 32122539
AN - SCOPUS:85068177219
SN - 9780128186770
T3 - Methods in Enzymology
SP - 1
EP - 20
BT - Tumor Immunology and Immunotherapy - Integrated Methods Part A
A2 - Galluzzi, Lorenzo
A2 - Rudqvist, Nils-Petter
PB - Academic Press Inc.
ER -