cell line dohh2  (DSMZ)

Journal: Blood Advances

Article Title: Lymphoma B cells remodel bone marrow stromal cells into extracellular matrix–producing cancer-associated fibroblasts ∗ †

doi: 10.1182/bloodadvances.2024015616

Figure Lengend Snippet: Intrafemoral xenograft model reproduces lymphoma BM niche. (A) Tumor cell dissemination. Evolution over time of the proportion of viable human tumor cells (DAPI – hCD20 + ) compared with viable murine CD45 + cells in the blood (red), spleen (blue), and BM (grafted femur in green and contralateral femur in orange) of Rag −/− γc −/− mice that were transplanted with 0.5 × 10 6 DOHH2 cells by intrafemoral route (left). Luciferase imaging of representative mouse from 14 to 42 days after engraftment. Green star indicates grafted femur, and purple star indicates contralateral femur (right). (B) Experimental design of the B-cell and stromal cell transcriptomic characterization in Rag −/− γc −/− mice untreated, injected with PBS (sham), or grafted with DOHH2. (C) Using gene set enrichment analysis, enrichment for pathways upregulated in published data of primary FL B cells (FL) vs centrocytes were investigated in published data of DOHH2 cocultured with tonsil stromal cells and ECM in DT3D vs classical D2D and in DOHH2 recovered at the late time point (day 40) and/or at the early time point (day 19). Circle colors depict the NES and circle sizes the FDR (left). Spearman correlation plot of comparisons using NES values from previously selected pathways (right). ∗ P < .05. CC, centrocytes; D, day; D2D, 2-dimensional DOHH2 culture; DT3D, 3-dimensional spheroids with tonsil stromal cells; FDR, false discovery rate; hCD20, human CD20; mCD45, murine CD45; max, maximum; min, minimum; NES, normalized enrichment score; q.val, q value.

Article Snippet: The FL cell line DOHH2 was obtained from the DSMZ cell collection (Braunschweig, Germany) and the GCB-DLBCL cell line OCI-Ly-19 was a gift from Lou Staudt (National Cancer Institute, Bethesda, MD).

Techniques: Luciferase, Imaging, Injection

Journal: Blood Advances

Article Title: Lymphoma B cells remodel bone marrow stromal cells into extracellular matrix–producing cancer-associated fibroblasts ∗ †

doi: 10.1182/bloodadvances.2024015616

Figure Lengend Snippet: Lymphoma B cells drive LepR + MSC transcriptional reprogramming. (A) Uniform manifold approximation and projection (UMAP) plot of the scRNAseq data from endothelial and stromal cells of Rag −/− γc −/− mice untreated (Ctrl), injected with PBS (sham), or grafted with DOHH2 and euthanized at early (day 19) and late (day 40) time points (left). Heat map of the enrichment of mouse nonhematopoietic cell signature scores as previously defined by scRNA sequencing in steady-state C57BL/6 mice (right). (B) Stacked bar plot representing, for each cluster, the proportion of cells coming from the 4 conditions (control, sham, Gr.Early, and Gr.Late). The last bar of the plot represents the distribution of all cells between the different experimental conditions. The unbalance between experimental conditions in each cluster was evaluated using maximum residual values from a χ 2 test. (C) UMAP plot of the subclustering of the 3 most unbalanced clusters (4, 0, and 1) after analysis of differential abundance using the differentially abundant sequencing (DA-seq) algorithm. Four LepR + MSC subclusters were identified: MSC_Ctrl, MSC_Sh, MSC_Gr.Early, and MSC_Gr.Late. (D) Venn diagram of the differentially expressed genes between MSC_Gr.Late vs MSC_Gr.Early, MSC_Gr.Late vs MSC_Sh, and MSC_Gr.Early vs MSC_Sh (adjusted P value <.05; absolute value of log 2 fold change of >.25). (E) Heat map of scores from human FL LN stromal cell subcluster signatures (reprinted from Abe et al ; upper part), human FL-LSC signature (reprinted from Mourcin et al ; middle part), and BM-MSC clusters obtained from HDs and patients with MM (reprinted from de Jong et al ; lower part), relative to the LepR + MSC clusters identified by DA-seq.

Article Snippet: The FL cell line DOHH2 was obtained from the DSMZ cell collection (Braunschweig, Germany) and the GCB-DLBCL cell line OCI-Ly-19 was a gift from Lou Staudt (National Cancer Institute, Bethesda, MD).

Techniques: Injection, Sequencing, Control

Journal: Blood Advances

Article Title: Lymphoma B cells remodel bone marrow stromal cells into extracellular matrix–producing cancer-associated fibroblasts ∗ †

doi: 10.1182/bloodadvances.2024015616

Figure Lengend Snippet: Impact of malignant B cells on stromal cells in lymphoma-invaded BM. (A) Circos plot showing predicted interactions between ligands from DOHH2 cells and receptors from LepR + MSC at the late stage of graft. D40 DOHH2 were defined as senders, and MSC_Gr.Late as receivers. (B) Heat map representation of ligand activity of D40 DOHH2 vs MSC_Gr.Late cells. Colors correspond to Pearson coefficient (quantifying ligand activities) and text to P values assessed by random permutation. (C) Heat map of the top 50 differentially expressed regulons detected in the 4 LepR + MSC clusters using decoupleR algorithm and CollectTri database (left). Violin plots of Smad3 and Smad7 regulon activity for the LepR + MSC clusters (right). (D) Heat map generated with CellPhoneDB repository, representing the number of significative bidirectional interactions between LepR + clusters (MSC_Sh, MSC_Gr.Early, MSC_Gr.Late, and MSC_Ctrl) from this study and human primary FL B cells (left). Diagram of bidirectional interactions detected, by CellPhoneDB, between ligands from human FL B cells and receptors from MSC_Gr.Late, and observed in >8 of 20 patients with FL (right). Interactions also predicted in panel A are highlighted in red.

Article Snippet: The FL cell line DOHH2 was obtained from the DSMZ cell collection (Braunschweig, Germany) and the GCB-DLBCL cell line OCI-Ly-19 was a gift from Lou Staudt (National Cancer Institute, Bethesda, MD).

Techniques: Activity Assay, Generated

Journal: Blood Advances

Article Title: Lymphoma B cells remodel bone marrow stromal cells into extracellular matrix–producing cancer-associated fibroblasts ∗ †

doi: 10.1182/bloodadvances.2024015616

Figure Lengend Snippet: Impact of stromal cells on malignant B cells in lymphoma-invaded BM. (A) Circos plot showing predicted interactions between ligands from LepR + MSCs and receptors from DOHH2 recovered from the grafted mouse at day 40. MSC_Gr.Late were defined as senders, and DOHH2 D40 as receivers. (B) Heat map representation of ligand activity of MSC_Gr.Late vs DOHH2 D40 cells. Colors correspond to Pearson coefficient quantifying ligand activities and text to P values assessed by random permutation. (C) Diagram of significative bidirectional interactions detected, by CellPhoneDB, between ligands from MSC_Gr.Late and receptors from human primary FL B cells, and observed in >8 of 20 patients with FL. Interactions predicted in panel A are highlighted in red. (D) Quantification by Luminex of matrix proteins and cytokines in the BM plasma from patients with FL (n = 11; except for TGFβ1, n = 13) collected at diagnosis and after 12 months of treatment. Blue dots represent patients with FL and green dotted line represent HD median level. Statistical significance was determined using the Mann-Whitney nonparametric U test. ∗ P < .05; ∗∗ P < .01. ns, nonsignificant.

Article Snippet: The FL cell line DOHH2 was obtained from the DSMZ cell collection (Braunschweig, Germany) and the GCB-DLBCL cell line OCI-Ly-19 was a gift from Lou Staudt (National Cancer Institute, Bethesda, MD).

Techniques: Activity Assay, Luminex, Clinical Proteomics, Biomarker Discovery, MANN-WHITNEY