Journal: bioRxiv

Article Title: Metastasis-associated wound repair promotes reciprocal activation of the lung epithelium and breast cancer metastases during outgrowth

doi: 10.1101/2025.07.23.666437

Figure Lengend Snippet: A, Model indicating the cellular interactions being interrogated: targeting AT2-BC interactions during metastatic outgrowth. B, The proportion of AT2 secreted factor genes enriched following BC interactions that are CREB-regulated for all three AT2 RNAseq datasets, percentage indicated. C, t-SNE visualizations of cells from metastatic mouse lungs clustered by gene expression and colored by Pde4 (phosphodiesterase 4) isoform gene expression. D, Percent cell viability was measured by crystal violet assay after a 3 day treatment with vehicle DMSO (0) or increasing concentrations of the PDE4 inhibitor roflumilast. Data was normalized to the mean absorbance of DMSO-treated cells; mean ± SEM. E, AT2 cell viability was measured by crystal violet assay following co-culture with TNBC cells and treatment with 2nM roflumilast (ROF): 5 days for A549 cells and 7 days for iAT2 cells. The percent difference in cell viability, between DMSO and 2nM ROF treated cells, was calculated for AT2 cells cultured alone (-) or co-culture with TNBC cells. Mean ± SEM (one-way ANOVA with Tukey’s multiple comparison test); * p ≤0.05, *** p <0.001. F, TNBC cell viability was measured by crystal violet assay following co-culture with AT2 cells and treatment with 2nM ROF: 5 days for A549 cells and 7 days for iAT2 cells. The percent difference in cell viability, between DMSO and 2nM ROF treated cells, was calculated for TNBC cells cultured alone (-) or co-culture with AT2 cells. Mean ± SEM (unpaired t -tests); * p ≤0.05. G, Schematic of metastatic outgrowth experimental design using the late-stage Met-1 metastasis model ( n =5-8 mice per group). Met-1 cells were IV injected into female mice and oral treatment with 5mg/kg ROF began 3 days later. Mice were treated daily for 3 weeks and metastatic lung tissue was collected. H, Lungs were stained for cell turnover markers Ki67 and cleaved-caspase 3 (CC3) by IHC. Shown are representative images of lung metastases; scale bar = 100µm. The percentage of positively stained cells was scored per metastasis and averaged per mouse; mean (unpaired t -tests with Welch’s correction when appropriate). I, Metastatic lungs were stained for the Met-1 mammary-specific marker PyMT. Metastatic burden was quantified in serial sections as the area of PyMT-positive metastases, means per group are indicated as dotted lines (unpaired t -test). Representative data is shown from a single section indicating the number and size of metastases per mouse; each notch on the x-axis represents an individual mouse.

Article Snippet: Roflumilast (ROF) was purchased from Selleck Chemical LLC. (No. S2131), and cilomilast (CILO) was purchased from MedChemExpress (No. HY-10790).

Techniques: Gene Expression, Crystal Violet Assay, Co-Culture Assay, Cell Culture, Comparison, Injection, Staining, Marker

Journal: Molecular Cell

Article Title: Actionable Cytopathogenic Host Responses of Human Alveolar Type 2 Cells to SARS-CoV-2

doi: 10.1016/j.molcel.2020.11.028

Figure Lengend Snippet:

Article Snippet: Roflumilast , Selleck , S2131.

Techniques: Virus, Recombinant, Imaging, Software, Mass Spectrometry, Microscopy

Journal: Physiological Reports

Article Title: Roflumilast and aquaporin‐2 regulation in rat renal inner medullary collecting duct

doi: 10.14814/phy2.13121

Figure Lengend Snippet: Effect of roflumilast on AQP 2 phosphorylation changes in rat IMCD suspensions. IMCD suspensions were incubated with vehicle or roflumilast (30 nmol/L) in the absence or presence of 0.1 nmol/L dDAVP for 30 min. (A–E) immunoblots for the four vasopressin‐regulated serine sites (S256, S261, S264, and S269) of AQP 2 and total AQP 2. (F–O) band density analysis depicted as log 2 value of the effect of vehicle or roflumilast normalized by vehicle in the absence (F–J) or presence (K–O) of dDAVP . *Statistically significant ( P < 0.05). IMCD, inner medullary collecting duct.

Article Snippet: Samples were incubated with roflumilast (product no. S2131, Selleck Chemicals), roflumilast N ‐oxide (RNO; product no. R639710, Toronto Research Chemicals), or arginine vasopressin (product no. H1780, Bachem) as described in the figure legends.

Techniques: Phospho-proteomics, Incubation, Western Blot

Journal: Physiological Reports

Article Title: Roflumilast and aquaporin‐2 regulation in rat renal inner medullary collecting duct

doi: 10.14814/phy2.13121

Figure Lengend Snippet: Effect of roflumilast N ‐oxide on AQP 2 phosphorylation changes in rat IMCD suspensions. IMCD suspensions were incubated with vehicle or RNO (30 nmol/L) in the absence or presence of 0.1 nmol/L dDAVP for 30 min. (A–E) immunoblots for the four vasopressin‐regulated serine sites (S256, S261, S264, and S269) of AQP 2 and total AQP 2. (F–O) band density analysis depicted as log 2 value of the effect of vehicle or RNO normalized by vehicle in the absence (F–J) or presence (K–O) of dDAVP . *Statistically significant ( P < 0.05). IMCD, inner medullary collecting duct; RNO, roflumilast N ‐oxide.

Article Snippet: Samples were incubated with roflumilast (product no. S2131, Selleck Chemicals), roflumilast N ‐oxide (RNO; product no. R639710, Toronto Research Chemicals), or arginine vasopressin (product no. H1780, Bachem) as described in the figure legends.

Techniques: Phospho-proteomics, Incubation, Western Blot

Journal: Physiological Reports

Article Title: Roflumilast and aquaporin‐2 regulation in rat renal inner medullary collecting duct

doi: 10.14814/phy2.13121

Figure Lengend Snippet: Dose‐dependent effect of roflumilast and IBMX on AQP 2 phosphorylation changes in IMCD suspensions in the presence of 0.1 nmol/L AVP . The dose range (3 nmol/L, 30 nmol/L, 300 nmol/L, and 3000 nmol/L ) and the incubation time (30 min) were the same for both agents. (A&B) immunoblots for total AQP 2 and the four vasopressin‐regulated serine sites (S256, S261, S264, and S269) of AQP 2. (C&D) line graphs summarizing the changes in normalized band densities of the total and four phosphorylation sites of AQP 2 elicited by roflumilast (C) or IBMX (D). n = 6 for both groups. *Statistically significant ( P < 0.05). IBMX, isobutyl‐1‐methylxanthine; IMCD, inner medullary collecting duct.

Article Snippet: Samples were incubated with roflumilast (product no. S2131, Selleck Chemicals), roflumilast N ‐oxide (RNO; product no. R639710, Toronto Research Chemicals), or arginine vasopressin (product no. H1780, Bachem) as described in the figure legends.

Techniques: Phospho-proteomics, Incubation, Western Blot

Journal: Physiological Reports

Article Title: Roflumilast and aquaporin‐2 regulation in rat renal inner medullary collecting duct

doi: 10.14814/phy2.13121

Figure Lengend Snippet: Effect of roflumilast on AQP 2 phosphorylation changes in rat IMCD suspensions in the presence of a selective EP 4 agonist ONO ‐ AE 1‐329 (30 min incubation period). (A) immunoblots for the four vasopressin‐regulated serine sites (S256, S261, S264, and S269) of AQP 2 and total AQP 2. (B–F) bar graphs of the normalized band density analysis. IMCD, inner medullary collecting duct.

Article Snippet: Samples were incubated with roflumilast (product no. S2131, Selleck Chemicals), roflumilast N ‐oxide (RNO; product no. R639710, Toronto Research Chemicals), or arginine vasopressin (product no. H1780, Bachem) as described in the figure legends.

Techniques: Phospho-proteomics, Incubation, Western Blot

Journal: Physiological Reports

Article Title: Roflumilast and aquaporin‐2 regulation in rat renal inner medullary collecting duct

doi: 10.14814/phy2.13121

Figure Lengend Snippet: Immunofluorescence labeling of AQP 2 in microdissected IMCD segments. Freshly microdissected IMCD s were incubated with vehicle (A) 30 nmol/L roflumilast (C) or 0.1 nmol/L dDAVP (D) for 30 min, followed by fixation and immunofluorescence staining for AQP 2. B, using an AQP 1 antibody at a concentration similar to that used for AQP 2 antibody in A, C and D, AQP 1 identified only the thin structures attached to an IMCD segment (pretreated with 0.1 nmol/L dDAVP ) but not IMCD . Green: AQP 2 or AQP 1, blue: DAPI. ( n = 3). IMCD, inner medullary collecting duct.

Article Snippet: Samples were incubated with roflumilast (product no. S2131, Selleck Chemicals), roflumilast N ‐oxide (RNO; product no. R639710, Toronto Research Chemicals), or arginine vasopressin (product no. H1780, Bachem) as described in the figure legends.

Techniques: Immunofluorescence, Labeling, Incubation, Staining, Concentration Assay

Journal: Physiological Reports

Article Title: Roflumilast and aquaporin‐2 regulation in rat renal inner medullary collecting duct

doi: 10.14814/phy2.13121

Figure Lengend Snippet: Osmotic water permeability ( P f ) measurements in microdissected IMCD segments. IMCD tubules were isolated and perfused for osmotic water permeability measurements in three 30‐min experimental periods: basal, roflumilast and roflumilast+ dDAVP (upper panel) or basal, RNO and RNO + dDAVP (lower panel). Data from the same experimental period were pooled for statistical analysis. Water permeability measured at 30 nmol/L roflumilast or RNO alone was not statistically different from the basal condition, whereas in the presence of 0.1 nmol/L dDAVP , water permeability was significantly increased ( P < 0.001, n = 6). IMCD, inner medullary collecting duct; RNO, roflumilast N ‐oxide.

Article Snippet: Samples were incubated with roflumilast (product no. S2131, Selleck Chemicals), roflumilast N ‐oxide (RNO; product no. R639710, Toronto Research Chemicals), or arginine vasopressin (product no. H1780, Bachem) as described in the figure legends.

Techniques: Permeability, Isolation

Journal: International Journal of Chronic Obstructive Pulmonary Disease

Article Title: ABCD of the phosphodiesterase family: interaction and differential activity in COPD

doi:

Figure Lengend Snippet: PDE4 inhibitors in current and discontinued clinical development

Article Snippet: Roflumilast , Altana , Phase III , ( ) .

Techniques:

Journal: Drug Design, Development and Therapy

Article Title: Roflumilast: first phosphodiesterase 4 inhibitor approved for treatment of COPD

doi:

Figure Lengend Snippet: Structure of roflumilast and its metabolic inactivation.

Article Snippet: In contrast, nonxanthine-based compounds that selectively inhibit PDE4 do not share these limitations of theophylline, and have undergone extensive preclinical and clinical evaluation., , The most advanced compound within this class is the benzamide, roflumilast (see ), which is being developed jointly by Nycomed (Zurich, Switzerland, formerly Altana) in Europe and the Forest Research Institute in the US (ownership transferred from Nycomed in December 2009) for the treatment of COPD.

Techniques:

Journal: Drug Design, Development and Therapy

Article Title: Roflumilast: first phosphodiesterase 4 inhibitor approved for treatment of COPD

doi:

Figure Lengend Snippet: Patient demographics in the large, randomized roflumilast treatment trials

Article Snippet: In contrast, nonxanthine-based compounds that selectively inhibit PDE4 do not share these limitations of theophylline, and have undergone extensive preclinical and clinical evaluation., , The most advanced compound within this class is the benzamide, roflumilast (see ), which is being developed jointly by Nycomed (Zurich, Switzerland, formerly Altana) in Europe and the Forest Research Institute in the US (ownership transferred from Nycomed in December 2009) for the treatment of COPD.

Techniques:

Journal: Drug Design, Development and Therapy

Article Title: Roflumilast: first phosphodiesterase 4 inhibitor approved for treatment of COPD

doi:

Figure Lengend Snippet: Patient outcomes in the roflumilast treatment trials

Article Snippet: In contrast, nonxanthine-based compounds that selectively inhibit PDE4 do not share these limitations of theophylline, and have undergone extensive preclinical and clinical evaluation., , The most advanced compound within this class is the benzamide, roflumilast (see ), which is being developed jointly by Nycomed (Zurich, Switzerland, formerly Altana) in Europe and the Forest Research Institute in the US (ownership transferred from Nycomed in December 2009) for the treatment of COPD.

Techniques: