normal lung fibroblast imr90  (ATCC)

Journal: bioRxiv

Article Title: TRAIL pathway suppression of cancer cell growth and immune cell-mediated tumor cell-killing in a senescent fibroblast-constructed tumor microenvironment

doi: 10.1101/2023.11.30.569479

Figure Lengend Snippet: A. Schema of IMR90 senescence induced by etoposide. B. SA-β-gal staining of senescent IMR90 fibroblasts at 3 weeks post-treatment with etoposide (right panel). 200x magnification, scale bar represents 100 µm. C. Cytokine profiles of analytes that decreased post- etoposide treatment in IMR90 cells. D. Cytokine profiles of analytes that increased post-etoposide treatment in IMR90 cells. Blue circles represent pre-etoposide analyte concentrations in picograms per milliliter (pg/mL), red squares represent analytes concentrations 1-week post- etoposide, and purple inverted triangles represent analyte concentrations 3-weeks post- etoposide.

Article Snippet: Cell lines used in this study include human colorectal adenocarcinoma HT-29 cells (ATCC), normal lung fibroblast IMR90 cells (ATCC), human CD8+ cytotoxic TALL-104 cells (ATCC), and human natural killer NK-92 cells (previously provided by Dr. Kerry Campbell at Fox Chase Cancer Center).

Techniques: Staining

Journal: bioRxiv

Article Title: TRAIL pathway suppression of cancer cell growth and immune cell-mediated tumor cell-killing in a senescent fibroblast-constructed tumor microenvironment

doi: 10.1101/2023.11.30.569479

Figure Lengend Snippet: Senescent fibroblasts promote tumor cell proliferation in co-culture and tumor growth in vivo .. A. Schema of co-culture system with constitutive Luciferase reporter expression in HT29 cancer cells. B. Relative bioluminescence in a two-cell co-culture system using HT29 tumor cells and IMR90 fibroblasts. Relative bioluminescence was normalized to that of HT29-luc cocultured with senescent IMR90 at each day indicated. C. Imaging of tumor cells in co-culture system at day 6. Scale bar represents 100 microns. D. Bioluminescence in co-culture system at 3 and 6 days post-ABT263 treatment. E. Bioluminescence in co-culture system at 3 and 6 days post-3TC treatment. F. Heat map representing cytokine, chemokine, and growth factor profiles. Fold changes post-treatment with ABT-263 (5µM) or 3TC (10µM) are shown in both senescent and proliferating IMR90s. G. HT29 mouse xenograft average tumor growth volumes on day 5 after HT29 cells were subcutaneously implanted with senescent or proliferating IMR90 cells. H. Tumor growth curves of HT29 CRC xenografts in mice. Tumor growth is reported according to tumor size measurements. Tumor volumes (G and H) were measured by a caliper every three days. Relative bioluminescence was normalized to control in each cohort, respectively. Data are expressed as mean ± SD. *, p <0.05.

Article Snippet: Cell lines used in this study include human colorectal adenocarcinoma HT-29 cells (ATCC), normal lung fibroblast IMR90 cells (ATCC), human CD8+ cytotoxic TALL-104 cells (ATCC), and human natural killer NK-92 cells (previously provided by Dr. Kerry Campbell at Fox Chase Cancer Center).

Techniques: Co-Culture Assay, In Vivo, Luciferase, Expressing, Imaging, Control

Journal: bioRxiv

Article Title: TRAIL pathway suppression of cancer cell growth and immune cell-mediated tumor cell-killing in a senescent fibroblast-constructed tumor microenvironment

doi: 10.1101/2023.11.30.569479

Figure Lengend Snippet: HCT116-Luc cells were cocultured with the senescent IMR90 or proliferating MR90 cells. Relative bioluminescence was normalized to the control in each cohort, respectively. Data are expressed as mean ± SD. *, p <0.05.

Article Snippet: Cell lines used in this study include human colorectal adenocarcinoma HT-29 cells (ATCC), normal lung fibroblast IMR90 cells (ATCC), human CD8+ cytotoxic TALL-104 cells (ATCC), and human natural killer NK-92 cells (previously provided by Dr. Kerry Campbell at Fox Chase Cancer Center).

Techniques: Control

Journal: bioRxiv

Article Title: TRAIL pathway suppression of cancer cell growth and immune cell-mediated tumor cell-killing in a senescent fibroblast-constructed tumor microenvironment

doi: 10.1101/2023.11.30.569479

Figure Lengend Snippet: A. Cell viability graph representing senescent and proliferating IMR90 cell viability post treatment with ABT263 at indicated concentrations. B. Cell viability graph representing senescent and proliferating IMR90 cell viability post treatment with 3TC at indicated concentrations.

Article Snippet: Cell lines used in this study include human colorectal adenocarcinoma HT-29 cells (ATCC), normal lung fibroblast IMR90 cells (ATCC), human CD8+ cytotoxic TALL-104 cells (ATCC), and human natural killer NK-92 cells (previously provided by Dr. Kerry Campbell at Fox Chase Cancer Center).

Techniques:

Journal: bioRxiv

Article Title: TRAIL pathway suppression of cancer cell growth and immune cell-mediated tumor cell-killing in a senescent fibroblast-constructed tumor microenvironment

doi: 10.1101/2023.11.30.569479

Figure Lengend Snippet: A. Schema of culture of p21-knockdown senescent IMR90 with bystander cancer cells. B. Bioluminescence of bystander cells in two-cell co-culture system on day 3. C. SA-β-gal staining of p21-Knockdown senescent IMR90 (lower panel). p21 expression in p21-knockdown or p53-knockdown senescent IMR90 cells is shown (upper panel). D. Heat map representing cytokine, chemokine, and growth factor profiles. Fold changes are shown for p21-knockdown senescent IMR90 cells. Relative cytokine levels were normalized to those of si-Ctrl in senescent IMR90 cells. Relative cytokine levels from senescent IMR90 with si-Ctrl were normalized to those from proliferating IMR90 cells. E. Bioluminescence of bystander HT29 cancer cells in two-cell co- culture system with TNFα- knockdown senescent IMR90 cells. Data are expressed as mean ± SD. #, p≤0.1. F. Western blot assay for TNFα and p21 expression in senescent IMR90 cells (E). G. Schematic representation of senescent fibroblasts promote bystander cancer cell growth via p21-driven SASP in the TME. Relative bioluminescence was normalized to control for each cohort, respectively. Data are expressed as mean ± SD. *, p <0.05.

Article Snippet: Cell lines used in this study include human colorectal adenocarcinoma HT-29 cells (ATCC), normal lung fibroblast IMR90 cells (ATCC), human CD8+ cytotoxic TALL-104 cells (ATCC), and human natural killer NK-92 cells (previously provided by Dr. Kerry Campbell at Fox Chase Cancer Center).

Techniques: Knockdown, Co-Culture Assay, Staining, Expressing, Western Blot, Control

Journal: bioRxiv

Article Title: TRAIL pathway suppression of cancer cell growth and immune cell-mediated tumor cell-killing in a senescent fibroblast-constructed tumor microenvironment

doi: 10.1101/2023.11.30.569479

Figure Lengend Snippet: A. Bioluminescence of HT29-luc cells in two-cell co-culture system treated with TRAIL and TIC10/ONC201 as indicated. Relative bioluminescence was normalized to control for each cohort, respectively. B. Cell viability of IMR90 fibroblasts treated with TRAIL and ONC201. C. HT29 CRC colony formation using co-culture system treated with 3 µM of ONC201 or 100 ng/ml of TRAIL. D. HT29 colony formation in co-culture system treated with 5-FU (1 µg/ml). E. Imaging of HT29 CRC colony formation (quantified in panels C and D). F. Heat map representing cytokine, chemokine, and growth factor profiles. Fold-changes post-treatment with ONC201 (5 µM) or ONC212 (5 µM) are shown for both senescent and proliferating IMR90s. Data are expressed as mean ± SD. *, p <0.05. P+H, HT29 cells were cocultured with the proliferating IMR90 cells; S+H, HT29 cells were cocultured with senescent IMR90 cells.

Article Snippet: Cell lines used in this study include human colorectal adenocarcinoma HT-29 cells (ATCC), normal lung fibroblast IMR90 cells (ATCC), human CD8+ cytotoxic TALL-104 cells (ATCC), and human natural killer NK-92 cells (previously provided by Dr. Kerry Campbell at Fox Chase Cancer Center).

Techniques: Co-Culture Assay, Control, Imaging

Journal: bioRxiv

Article Title: TRAIL pathway suppression of cancer cell growth and immune cell-mediated tumor cell-killing in a senescent fibroblast-constructed tumor microenvironment

doi: 10.1101/2023.11.30.569479

Figure Lengend Snippet: A. Tri-culture of HT29 CRC cells (blue), senescent IMR90 fibroblasts (green), and NK-92 cells (unlabeled). B. Tri-culture of HT29 CRC cells (blue), proliferating IMR90 fibroblasts (green), and NK-92 cells (unlabeled) at 8-hour post- treatment timepoint. C. Quantification of senescent IMR90 + HT29 CRC cells + NK-92 tri-culture (panel A). D. Quantification of proliferating IMR90 + HT29 + NK-92 tri-culture (panel B). E. Quantification of senescent IMR90 + HT29 + TALL-104 tri-culture (panel G). F. Quantification of proliferating IMR90 + HT29 + TALL-104 tri-culture (panel H). G. Tri-culture of HT29 tumor cells (blue), senescent IMR90 fibroblasts (green), and TALL-104 T cells (unlabeled). H. Tri-culture of HT29 tumor cells (blue), proliferating IMR90 fibroblasts (green), and TALL-104 T cells (unlabeled). Ethidium homodimer (EthD-1) was used to visualize dead cells, 10x magnification, scale bar indicates 100 μm. One-way Anova followed by post-hoc Tukey’s multiple comparisons test was used to determine statistical significance at p < 0.05. The following symbols * and ** represent, p < 0.05 and p < 0.01, respectively. I. TALL-104 cell Granzyme B secretion post-ABT263 or ONC201 treatment was quantified using an IsoPlexis innate-immune chip single-cell cytokine profiling assay. J. A Uniform Manifold Approximation and Projection (UMAP) plot was used to cluster the single TALL-104 cells post-treatment as indicated using IsoPlexis technology. The cells were treated with ABT263 (2 µM) and ONC201 (2 µM), or the agents alone.

Article Snippet: Cell lines used in this study include human colorectal adenocarcinoma HT-29 cells (ATCC), normal lung fibroblast IMR90 cells (ATCC), human CD8+ cytotoxic TALL-104 cells (ATCC), and human natural killer NK-92 cells (previously provided by Dr. Kerry Campbell at Fox Chase Cancer Center).

Techniques: