mif inhibitor 4 ipp  (MedChemExpress)

Journal: bioRxiv

Article Title: Single-cell profiling of synchronous multi-organ metastasis reveals a systemic CD74 + lipid-associated macrophage niche driving polymetastatic breast cancer

doi: 10.64898/2026.01.31.701004

Figure Lengend Snippet: a. Quantification of metastatic burden using ex vivo BLI of FVB mice injected VO-PyMT cells transduced with control vectors (shControl) and Mif-knockdown vectors (shMif 1; shMif 2). Data was acquired at day 12 post i.c. injections. shControl, n = 11; shMif (1), n = 11; shMif (2), n = 5. Data represents results from 2 independent experiments. In each experiment, raw photon flux values (p/s) from each organ were normalized to the average photon flux (p/s) values of the shControl group. Boxes depict 25th and 75th percentiles. Median is shown as horizontal lines. Whiskers depict data range. All data points are shown as dots. P values were determined using two-tailed Mann-Whitney tests by comparing shControl ( n = 11) versus the two shMif groups combined ( n = 16). Images show representative ex vivo BLI in brain, lung, liver or lower limb bones from each experimental group. b. Quantification of multi-organ metastatic burden ex vivo using BLI in albino C57/BL6 mice injected intracardially with Py8119 breast cancer cells transduced with control (shControl) and Mif-knockdown vector (shMif 2). Representative images are shown for each organ analyzed. Ex vivo metastatic burden was quantified 10 days after cancer cell injection. shControl, n = 14; shMif (2), n = 14. Data is representative of 3 independent experiments. Data in each organ was normalized to the average photon flux (p/s) of the shControl group in each experiment. Boxes boundaries define the interquartile ranges. Horizontal lines depict median values in each group. The whiskers show data range. The dots depict data points. P values were calculated by one-tailed Mann-Whitney t tests. c. Schematic showing pre-clinical model of MIF-CD74 targeting using 4-IPP, a small molecule inhibitor of MIF binding capacity. Multi-metastatic VO-PyMT cells were injected intracardially at day 0. Seeding and micrometastatic growth was allowed for 3 days, followed by i.p. administration of either a vehicle solution or 4-IPP. At day 12 post i.c. injection of VO-PyMT cells, metastatic colonization was analyzed ex vivo in brain, lung, liver and lower limb bones. d. Box plots showing quantification of metastatic burden of experiment as determined by ex vivo BLI in brain, lung, liver and bones of FVB mice injected intracardially with VO-PyMT cells and treated with the MIF inhibitor 4-IPP as described in (c). Vehicle group, n = 14; 4-IPP group, n = 13. For bone metastatic burden, right and left lower limb bones were analyzed separately. Data represents results from 2 independent experiments. In each experiment, photon flux (p/s) in each organ was normalized to the average photon flux (p/s) in the Vehicle control group. Boxes boundaries indicate 25 th and 75 th percentiles. Median is indicated by a horizontal line in boxes. Whiskers show data range. Data points are indicated as dots. P values were calculated using one-tailed Mann-Whitney t tests.

Article Snippet: For MIF-CD74 axis disruption in vivo , 4-IPP (MedChemExpress, cat no HY-110063) was first prepared in a vehicle solution consisting of 5% DMSO, 45% polyethylene glycol 300 (PEG300, MedChemExpress, cat no HY-Y0873), and 45% normal saline (0.9% NaCl).

Techniques: Ex Vivo, Injection, Transduction, Control, Knockdown, Two Tailed Test, MANN-WHITNEY, Plasmid Preparation, One-tailed Test, Binding Assay

Journal: bioRxiv

Article Title: Single-cell profiling of synchronous multi-organ metastasis reveals a systemic CD74 + lipid-associated macrophage niche driving polymetastatic breast cancer

doi: 10.64898/2026.01.31.701004

Figure Lengend Snippet: a. Quantification of metastatic burden using ex vivo BLI of FVB mice injected VO-PyMT cells transduced with control vectors (shControl) and Mif-knockdown vectors (shMif 1; shMif 2). Data was acquired at day 12 post i.c. injections. shControl, n = 11; shMif (1), n = 11; shMif (2), n = 5. Data represents results from 2 independent experiments. In each experiment, raw photon flux values (p/s) from each organ were normalized to the average photon flux (p/s) values of the shControl group. Boxes depict 25th and 75th percentiles. Median is shown as horizontal lines. Whiskers depict data range. All data points are shown as dots. P values were determined using two-tailed Mann-Whitney tests by comparing shControl ( n = 11) versus the two shMif groups combined ( n = 16). Images show representative ex vivo BLI in brain, lung, liver or lower limb bones from each experimental group. b. Quantification of multi-organ metastatic burden ex vivo using BLI in albino C57/BL6 mice injected intracardially with Py8119 breast cancer cells transduced with control (shControl) and Mif-knockdown vector (shMif 2). Representative images are shown for each organ analyzed. Ex vivo metastatic burden was quantified 10 days after cancer cell injection. shControl, n = 14; shMif (2), n = 14. Data is representative of 3 independent experiments. Data in each organ was normalized to the average photon flux (p/s) of the shControl group in each experiment. Boxes boundaries define the interquartile ranges. Horizontal lines depict median values in each group. The whiskers show data range. The dots depict data points. P values were calculated by one-tailed Mann-Whitney t tests. c. Schematic showing pre-clinical model of MIF-CD74 targeting using 4-IPP, a small molecule inhibitor of MIF binding capacity. Multi-metastatic VO-PyMT cells were injected intracardially at day 0. Seeding and micrometastatic growth was allowed for 3 days, followed by i.p. administration of either a vehicle solution or 4-IPP. At day 12 post i.c. injection of VO-PyMT cells, metastatic colonization was analyzed ex vivo in brain, lung, liver and lower limb bones. d. Box plots showing quantification of metastatic burden of experiment as determined by ex vivo BLI in brain, lung, liver and bones of FVB mice injected intracardially with VO-PyMT cells and treated with the MIF inhibitor 4-IPP as described in (c). Vehicle group, n = 14; 4-IPP group, n = 13. For bone metastatic burden, right and left lower limb bones were analyzed separately. Data represents results from 2 independent experiments. In each experiment, photon flux (p/s) in each organ was normalized to the average photon flux (p/s) in the Vehicle control group. Boxes boundaries indicate 25 th and 75 th percentiles. Median is indicated by a horizontal line in boxes. Whiskers show data range. Data points are indicated as dots. P values were calculated using one-tailed Mann-Whitney t tests.

Article Snippet: For MIF-CD74 axis disruption in vivo , 4-IPP (MedChemExpress, cat no HY-110063) was first prepared in a vehicle solution consisting of 5% DMSO, 45% polyethylene glycol 300 (PEG300, MedChemExpress, cat no HY-Y0873), and 45% normal saline (0.9% NaCl).

Techniques: Ex Vivo, Injection, Transduction, Control, Knockdown, Two Tailed Test, MANN-WHITNEY, Plasmid Preparation, One-tailed Test, Binding Assay

Journal: bioRxiv

Article Title: Single-cell profiling of synchronous multi-organ metastasis reveals a systemic CD74 + lipid-associated macrophage niche driving polymetastatic breast cancer

doi: 10.64898/2026.01.31.701004

Figure Lengend Snippet: a. UMAP showing scRNA-seq dataset of human metastatic breast cancer , colored by main cell types. The number of cells per cell type are indicated in the legend. Data was collected from 6 different sites: liver, axilla, breast, bone, and lung. n = 30 donors. Data was accessed and plots were generated using Cellxgene , . b. Feature plots depicting MIF and CD74 relative RNA expression. c. In situ MIF and CD74 protein expression analyzed by immunohistochemistry in a cohort of 103 human breast cancer metastases in brain, lung, liver and bone. Micrographs show representative stainings. Black dashed lines delineate areas of cancer cells (CC) in metastatic lesions. Black arrows show tumor-adjacent or tumor-infiltrating CD74 + stromal cells. MIF and CD74 semi-quantitative IHC scores are indicated in the color legends and frequencies of MIF and CD74 scores in either cancer cells (CC) or stroma (STR) are shown in stacked bars for each organ. Scale bar: 100 µm. d. Heatmap showing CD74 IHC scores in the stroma of human breast-to-brain, -lung, -liver and -bone metastasis samples, grouped by corresponding MIF IHC scores from cancer cells. e. Relative percentages of cases of CD74 IHC score in stroma grouped by MIF IHC score in cancer cells in all metastases analyzed. n = 103 samples (Brain metastases, n = 39; Lung metastases, n = 13; Liver metastases, n = 20; Bone metastases, n = 31). Chi-squared value and P value as calculated by Chi-squared test are shown. Spearman correlation test was calculated, and the Rho and P value of Spearman test are indicated. f. Stratification of post-brain metastasis survival in breast cancer brain metastasis samples was done based on MIF expression in cancer cell areas. MIF-high samples were determined by MIF IHC score in cancer cells of 3, while MIF-low samples were defined by MIF IHC score in cancer cells of 0-2. Post-brain metastasis survival in each patient was determined by calculating the time in days elapsed from the date of collection of the brain metastasis sample analyzed, and the date of death. MIF-high group, n = 11; MIF-low group, n = 18. Hazard ratio was calculated by a log-rank test, and P value was determined by a Gehan-Breslow-Wilcoxon test. g. Kaplan-Meier plot showing probability of overall survival (OS) or distant metastasis-free survival (DMFS) in breast cancer patients with high or low expression of MIF RNA expression in primary tumor tissues. OS: MIF high, n = 770; MIF low, n = 1109. DMFS: MIF-high, n = 1119; MIF low, n = 1646. Plots were generated using the KM plotter compiled dataset of mRNA breast cancer gene chip and visualized using the KM plotter tool . h. Conceptual model depicting the role of MIF-CD74 paracrine interaction between cancer cells and CD74 + MAMs during multi-organ breast cancer metastasis. Overt, MIF-expressing breast cancer metastatic lesions are infiltrated by immunosuppressive, lipid-associated (LA) CD74 + MAMs, leading to immunosuppressive milieu and increased T cell exhaustion, which enables metastatic colonization at multiple distant sites, thus promoting progression to systemic, multi-metastatic disease. Targeting MIF blocks the accumulation of immunosuppressive MAMs, leading to dramatically reduced multi-organ metastasis, therefore representing a promising therapeutic avenue to prevent progression to polymetastatic disease.

Article Snippet: For MIF-CD74 axis disruption in vivo , 4-IPP (MedChemExpress, cat no HY-110063) was first prepared in a vehicle solution consisting of 5% DMSO, 45% polyethylene glycol 300 (PEG300, MedChemExpress, cat no HY-Y0873), and 45% normal saline (0.9% NaCl).

Techniques: Generated, RNA Expression, In Situ, Expressing, Immunohistochemistry