Journal: Blood Advances

Article Title: Dual role of signaling pathways in myeloma requires cell type–specific targeting of ligand-receptor interactions ∗

doi: 10.1182/bloodadvances.2023011463

Figure Lengend Snippet: Differential enrichment of B-cell subsets associated with prognosis. (A) Analysis of the cell types composing the flow-sorted CD3 + , CD14 + , CD19 + , and CD45 + TN immune compartments according to gene signatures from azimuth. (B) Experimental workflow. Low-input RNA-seq was performed of individual cell populations sorted based on indicated markers from the bone marrow of 25 patients with NDMM. Cibersort was used to deconvolute cell type composition. (C-F) Bar graphs showing the composition of individual immune cell compartments based on deconvolution in patients with NDMM. Shown are percentages of cell types inferred by Cibersort for CD19 + cells (C), CD3 + cells (D), CD45 + TN cells (E), and CD14 + cells (F). (G) Patients with early (TTPlow) and late (TTPhigh) relapse were divided by median TTP. (H) Heat map showing relative differences in cell type abundance between patients with poor vs good prognosis. (I-L) Dot plots showing the distribution of values for the indicated cell populations comparing TTPlow with TTPhigh. Significance was assessed using an unpaired t test; ∗ P ≤ .05. (K) Log2 ratio of naive/memory B cells in TTPlow vs TTPhigh patients. Significance was assessed using an unpaired t test; ∗ P ≤ .05. (M) Relative scores for a gene set for naive B cells in TTPlow vs TTPhigh patients. ∗ P ≤ .05, by unpaired t test. Correlation of relative scores (bottom) for naive B-cell gene set vs TTP value ( R = 0.56; P = .036). (N) Expression of canonical marker genes for naive and memory B cells in TTPlow vs TTPhigh patients. (O) Expression of marker genes distinguishing B cells in TTPlow and TTPhigh patients. (P) Heat map comparing activity of signaling pathways between TTPlow vs TTPhigh patients. ASDC, AXL + dendritic cell; BaEoMa, basophil eosinophil mast progenitor; cDC1, CD141 + myeloid dendritic cell; cDC2, CD1c + myeloid dendritic cell; CD8 effector.1, memory-like CD8 + GZMK + alpha-beta T cell; CD8 effector.2, late differentiated CD8 + GZMB + cytotoxic alpha-beta T cell; CD8 effector.3, NKT-like CD8 + alpha-beta T cell; ILC, innate lymphoid cell; MAIT, mucosal associated invariant T; Mono, monocyte; NKdim, CD56dim NK cell; NKbright, CD56bright NK cell.

Article Snippet: Cells were stained with antibodies against CD45 fluorescein isothiocyanate (CD45 FITC) (HI30, eBioscience; 1:100), CD3 peridinin-chlorophyll-protein complex-cyanine 5.5 (CD3 PerCP-Cy5.5) (OKT3, eBioscience; 1:100), CD14 allophycocyanin-cyanine 7 (CD14 APC-Cy7) (MoP9, BD Biosciences; 1:100), and CD19 phycoerythrin (CD19 PE) (HIB19, BioLegend; 1:100).

Techniques: RNA Sequencing Assay, Expressing, Marker, Activity Assay

Journal: Blood Advances

Article Title: Dual role of signaling pathways in myeloma requires cell type–specific targeting of ligand-receptor interactions ∗

doi: 10.1182/bloodadvances.2023011463

Figure Lengend Snippet: Dual interactions in B cells. (A) Model of dual interactions. Model of “Cellular-partner selectivity” (left). Given molecule A and molecule B (undefined ligand-receptor interacting pair), B cells (illustrated as CD19, gray) express exclusively molecule A on the cell surface and the other cells in their vicinity present molecule B. Prognosis is based on the engaged cell type. Model based on “directionality” of the signaling (right). Given molecule A and molecule B (undefined ligand-receptor interacting pair), the B cell (illustrated as CD19, gray) presents both ligand and receptor molecules. Prognosis is determined by the directionality of the signal upon binding of the complementary molecule, which is expressed by other cells in the surroundings. (B) Proportion of dual interactions established between each possible immune pair under study and their distribution in each prognostic condition. (C) Heat map showing dual interactions with potential targetability in MM. Displayed are the percentage of cells expressing the molecule and the median expression of both molecules by the pair of engaged cell types. Highlighted are those interactions which are significant ( P < .05) in 1 condition but not the other. In the shown interactions, the first molecule is expressed in all cases by CD19 + cells (B cells); the second molecule is expressed by the 4 cell types (CD3 + , CD14 + , CD19 + , or CD45 + TN). On the left side of the list of interactions, the column represents TTPlow patients; on the right side, the column represents TTPhigh patients. (D) Model illustrating IL-15–IL-15RA interactions on diverse cell types as example.

Article Snippet: Cells were stained with antibodies against CD45 fluorescein isothiocyanate (CD45 FITC) (HI30, eBioscience; 1:100), CD3 peridinin-chlorophyll-protein complex-cyanine 5.5 (CD3 PerCP-Cy5.5) (OKT3, eBioscience; 1:100), CD14 allophycocyanin-cyanine 7 (CD14 APC-Cy7) (MoP9, BD Biosciences; 1:100), and CD19 phycoerythrin (CD19 PE) (HIB19, BioLegend; 1:100).

Techniques: Binding Assay, Expressing