A5083716484- Extracellular vesicles in disease
- Ubiquitin and proteasome pathways
- Advanced Proteomics Techniques and Applications
- Monoclonal and Polyclonal Antibodies Research
- Circular RNAs in diseases
- Pulmonary Hypertension Research and Treatments
- Immune cells in cancer
- PI3K/AKT/mTOR signaling in cancer
- Cancer Research and Treatments
- MicroRNA in disease regulation
- Cancer Genomics and Diagnostics
Amsterdam University Medical Centers
2024
Vrije Universiteit Amsterdam
2024
The Netherlands Cancer Institute
2023
Abstract Purpose: Therapy resistance is a major clinical hurdle in bone cancer treatment and seems to be largely driven by poorly understood microenvironmental factors. Recent evidence suggests critical role for unique subpopulation of mesenchymal stem cells with inflammatory features (iMSC), though their origin function remained unexplored. We demonstrate that cancer-secreted extracellular vesicles (EV) trigger the development iMSCs, which hinder therapy response vivo, set out identify...
Functional interactions between cytotoxic T cells and tumor are central to anti-cancer immunity. However, our understanding of the proteins involved is limited. Here, we present HySic (hybrid quantification stable isotope labeling by amino acids in cell culture [SILAC]-labeled interacting cells) as a method quantify protein phosphorylation dynamics within physically cells. Using co-cultured cells, directly measure proteome phosphoproteome engaged without need for physical separation. We...
<p>Suppl. Figure 4. a-b) scRNA-seq mapping of human osteosarcoma tumors. MSC and other indicated clusters were identified through the UMAP (a) marker gene expression analysis (b).</p>
<p>Suppl. Figure 3. a-e) Relative expression levels of IL6, IL8, CXCL2, CXCL3 and CXCL6 in MSCs from an independent donor (MSC#2) exposed to plasma-derived EVs healthy #1 (H#1) MM patient (MM#1).</p>
<p>Suppl. Figure 6. a-b) Expression of the most upregulated TGFβ-dependent (a) and independent/partially dependent (b) genes in bone marrow MSCs based on RNA-seq normalized counts. c-f) Relative expression levels CXCL1, CXCL2, CXCL5, CXCL6 mRNAs (donor #1) exposed to 143B EVs presence or absence TGFBR1 inhibitor SB-431542 as assessed by RT-qPCR. g-m) IL6, IL8, CXCL3, #2) EW-7197. n-o) IL6 IL8 adipose-derived #3) EW-7197.</p>
<div>AbstractPurpose:<p>Therapy resistance is a major clinical hurdle in bone cancer treatment and seems to be largely driven by poorly understood microenvironmental factors. Recent evidence suggests critical role for unique subpopulation of mesenchymal stem cells with inflammatory features (iMSC), though their origin function remained unexplored. We demonstrate that cancer-secreted extracellular vesicles (EV) trigger the development iMSCs, which hinder therapy response...
<p>Suppl. Figure 2. a-b) scRNA-seq mapping of the non-hematopoietic mononuclear cell fraction from bone marrow aspirates MM patients and non-cancer control individuals. UMAP plot (a) violin plots (b) marker gene expression in identified clusters. c) split into datasets. d-i) Density differentially expressed cytokines chemokine samples.</p>
<p>Suppl. Figure 5. a-b) CD63 (a) and CD81 (b) protein quantification (western blot pixel density) in EVs released by engineered (shGFP, shSyntenin, shRab11b, shRab35) 143B cells. c) Growth curve of wt shRab35, shSyntenin) showing no differences cell growth over 72 hours). d-e) Relative mRNA expression levels IL6 (d) IL8 (e) MSCs exposed to indicated volumes wild type EVs.</p>
<p>Suppl. Figure 8. a-c) RNA class distribution of osteosarcoma (n=10) (a) and multiple myeloma (n=5) (b) patient plasma EVs, normalized counts (rpm) known (TLR-3 RIG-I activating) inflammatory RNAs in MM EVs 143B (c). d) Expression levels (endosomal cytosolic) pattern recognition receptors MSCs based on RNA-seq counts. e-h) Knockdown confirmation TLR3 (e), (f), MDA5 (g) LGP2 (h) primary as compared to control (shGFP-transduced) cells. i) IL8 protein production transduced with...
<p>Suppl. Figure 1. a-b) Tumor growth measured by caliper (a) and BLI (b) in osteosarcoma-bearing mice receiving the TGFBR1 inhibitor EW-7197 injected or not with 143B EV-educated MSCs. c) Western blot for CD63 CD81 EVs released human fibroblasts (hF) 143B, MDA-MB-231, PC3, HOS MCF7 cell lines. d) GSEA enrichment plot gene set ‘TNFα signaling via NFKB’ EV-exposed MSC compared to hF (ctrl) e) List of sets enriched MSCs MSCs.</p>
<p>Suppl. Figure 9. a) BLI images of tumor-bearing mice receiving Ladarixin and injected or not with human EV-induced iMSCs. b-c) Tumor growth measured by caliper (b) lung nodules count (c) in ladarixin combination EW-7197 (TGFBR1 inhibitor) iMSCs.</p>
<p>Suppl. Figure 7. a) Dynasore-mediated inhibition of PKH67-labeled EV uptake by MSCs as assessed FACS. b-e) Relative expression levels CXCL2, CXCL5, CXCL3, CXCL6 in treated with 143B EVs the presence or absence dynasore. Transcript are normalized to GAPDH and expressed fold increase relative experimental controls (untreated dynasore-treated MSCs). Graphs show average 3 (CXCL2 CXCL6) 2 (CXCL3 CXCL5) experiments. Statistics was calculated on data, *p < 0.05, two-tailed t test. f)...
Abstract Therapeutic interference of cell-cell communication mechanisms has allowed the clinical development multiple cancer treatments. For example, therapeutic intervention T cell-tumor cell interactions PD-1 and PD-L1, respectively, by immune checkpoint blockade (ICB) improved outcome patients. However, only a minority patients are currently benefiting from ICB. Thus, there is an urgent need to improve our understanding interplay. Recently developed algorithms serve deconvolute gene...
Abstract Functional interactions between cytotoxic T cells and tumor are central to anti-cancer immunity. Some of the proteins involved, particularly immune checkpoints expressed by cells, serve as promising clinical targets in immunotherapy. However, our understanding complexity dynamics is only rudimentary. Here we present HySic (for Hy brid quantification S ILAC (Stable Isotope Labelling Amino acids Cell culture)-labeled interacting c ells) an innovative method quantify protein...