Spatially Resolved Tumor Microenvironment Predicts Treatment Outcomes in Relapsed/Refractory Hodgkin Lymphoma.

PURPOSE: About a third of patients with relapsed or refractory classic Hodgkin lymphoma (r/r CHL) succumb to their disease after high-dose chemotherapy followed by autologous stem-cell transplantation (HDC/ASCT). Here, we aimed to describe spatially resolved tumor microenvironment (TME) ecosystems to establish novel biomarkers associated with treatment failure in r/r CHL. PATIENTS AND METHODS: We performed imaging mass cytometry (IMC) on 71 paired primary diagnostic and relapse biopsies using a marker panel specific to CHL biology. For each cell type in the TME, we calculated a spatial score measuring the distance of nearest neighbor cells to the malignant Hodgkin Reed Sternberg cells within the close interaction range. Spatial scores were used as features in prognostic model development for post-ASCT outcomes. RESULTS: Highly multiplexed IMC data revealed shared TME patterns in paired diagnostic and early r/r CHL samples, whereas TME patterns were more divergent in pairs of diagnostic and late relapse samples. Integrated analysis of IMC and single-cell RNA sequencing data identified unique architecture defined by CXCR5+ Hodgkin and Reed Sternberg (HRS) cells and their strong spatial relationship with CXCL13+ macrophages in the TME. We developed a prognostic assay (RHL4S) using four spatially resolved parameters, CXCR5+ HRS cells, PD1+CD4+ T cells, CD68+ tumor-associated macrophages, and CXCR5+ B cells, which effectively separated patients into high-risk versus low-risk groups with significantly different post-ASCT outcomes. The RHL4S assay was validated in an independent r/r CHL cohort using a multicolor immunofluorescence assay. CONCLUSION: We identified the interaction of CXCR5+ HRS cells with ligand-expressing CXCL13+ macrophages as a prominent crosstalk axis in relapsed CHL. Harnessing this TME biology, we developed a novel prognostic model applicable to r/r CHL biopsies, RHL4S, opening new avenues for spatial biomarker development.

Three-Plane-assembled Deep Learning Segmentation of Gliomas.

PURPOSE: To design a computational method for automatic brain glioma segmentation of multimodal MRI scans with high efficiency and accuracy. MATERIALS AND METHODS: The 2018 Multimodal Brain Tumor Segmentation Challenge (BraTS) dataset was used in this study, consisting of routine clinically acquired preoperative multimodal MRI scans. Three subregions of glioma-the necrotic and nonenhancing tumor core, the peritumoral edema, and the contrast-enhancing tumor-were manually labeled by experienced radiologists. Two-dimensional U-Net models were built using a three-plane-assembled approach to segment three subregions individually (three-region model) or to segment only the whole tumor (WT) region (WT-only model). The term three-plane-assembled means that coronal and sagittal images were generated by reformatting the original axial images. The model performance for each case was evaluated in three classes: enhancing tumor (ET), tumor core (TC), and WT. RESULTS: On the internal unseen testing dataset split from the 2018 BraTS training dataset, the proposed models achieved mean Sørensen-Dice scores of 0.80, 0.84, and 0.91, respectively, for ET, TC, and WT. On the BraTS validation dataset, the proposed models achieved mean 95% Hausdorff distances of 3.1 mm, 7.0 mm, and 5.0 mm, respectively, for ET, TC, and WT and mean Sørensen-Dice scores of 0.80, 0.83, and 0.91, respectively, for ET, TC, and WT. On the BraTS testing dataset, the proposed models ranked fourth out of 61 teams. The source code is available at https://github.com/GuanLab/Brain_Glioma. CONCLUSION: This deep learning method consistently segmented subregions of brain glioma with high accuracy, efficiency, reliability, and generalization ability on screening images from a large population, and it can be efficiently implemented in clinical practice to assist neuro-oncologists or radiologists. Supplemental material is available for this article. © RSNA, 2020.

Cellular, transcriptomic and isoform heterogeneity of breast cancer cell line revealed by full-length single-cell RNA sequencing.

Tumor heterogeneity is generated through a combination of genetic and epigenetic mechanisms, the latter of which plays an important role in the generation of stem like cells responsible for tumor formation and metastasis. Although the development of single cell transcriptomic technologies holds promise to deconvolute this complexity, a number of these techniques have limitations including drop-out and uneven coverage, which challenge the further delineation of tumor heterogeneity. We adopted deep and full-length single-cell RNA sequencing on Fluidigm's Polaris platform to reveal the cellular, transcriptomic, and isoform heterogeneity of SUM149, a triple negative breast cancer (TNBC) cell line. We first validate the quality of the TNBC sequencing data with the sequencing data from erythroleukemia K562 cell line as control. We next scrutinized well-defined marker genes for cancer stem-like cell to identify different cell populations. We then profile the isoform expression data to investigate the heterogeneity of alternative splicing patterns. Though classified as triple-negative breast cancer, the SUM149 stem cells show heterogeneous expression of marker receptors (ER, PR, and HER2) across the cells. We identified three cell populations that express patterns of stemness: epithelial-mesenchymal transition (EMT) cancer stem cells (CSCs), mesenchymal-epithelial transition (MET) CSCs and Dual-EMT-MET CSCs. These cells also manifested a high level of heterogeneity in alternative splicing patterns. For example, CSCs have shown different expression patterns of the CD44v6 exon, as well as different levels of truncated EGFR transcripts, which may suggest different potentials for proliferation and invasion among cancer stem cells. Our study identified features of the landscape of previously underestimated cellular, transcriptomic, and isoform heterogeneity of cancer stem cells in triple-negative breast cancers.