Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Mechanisms of Acquired Drug Resistance to the HDAC6 Selective Inhibitor Ricolinostat Reveals Rational Drug : Drug Combination with Ibrutinib

doi: 10.1158/1078-0432.CCR-16-2022

Figure Lengend Snippet: (A) Resistant (R) cells and parental cells (P) were evaluated by RNA Seq for gene expression. The two cells lines were distinct as demonstrated by principle component analysis. The heat map represents the top 100 genes with significant overall 2-log fold change between the resistant and parental lines. (B) Gene set enrichment analysis (GSEA) was performed to compare enrichment of pathways in resistant verses parental lines. (C) Differentially expressed genes of interest were confirmed via PCR for the resistant line as compared to the parental line. (D) Protein expression of genes of interest was confirmed with immunoblot analysis. (E) Concentration : effect relationships of ibrutinib in resistant and parental lines was evaluated at 48 and 72 hours.

Article Snippet: Deuterated internal standard ibrutinib-d5 was purchased from TLC Pharmaceutical Standards Ltd (Ontario, Canada).

Techniques: RNA Sequencing Assay, Expressing, Western Blot, Concentration Assay

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Mechanisms of Acquired Drug Resistance to the HDAC6 Selective Inhibitor Ricolinostat Reveals Rational Drug : Drug Combination with Ibrutinib

doi: 10.1158/1078-0432.CCR-16-2022

Figure Lengend Snippet: (A). Heat map represents the viability of a panel of cell lines following treatment with ACY-1215, ibrutinib or the combination at 24, 48, and 72 hours. Red boxes indicate lower viability. Synergy was calculated by the relative risk ration (RRR). RRR < 1 connotes synergy and is represented by red boxes. (B) Primary human lymphoma samples, chronic lymphocytic leukemia (CLL), lymphoplasmacytic lymphoma (LPL), and 17p deleted marginal zone lymphoma (MZL), were treated with ACY-1215, ibrutinib or the combination over 24 to 96 hours. Viability was measured and synergy was calculated by RRR and represented in the heat maps. (C) The IRE-1 and BTK pathways were evaluated following treatment with the combination of ibrutinib and ACY-1215 by immunoblot analysis.

Article Snippet: Deuterated internal standard ibrutinib-d5 was purchased from TLC Pharmaceutical Standards Ltd (Ontario, Canada).

Techniques: Western Blot

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Mechanisms of Acquired Drug Resistance to the HDAC6 Selective Inhibitor Ricolinostat Reveals Rational Drug : Drug Combination with Ibrutinib

doi: 10.1158/1078-0432.CCR-16-2022

Figure Lengend Snippet: (A) SCID-beige mice were injected with LY10 107 in their flanks and treated with ACY-1215 50 mg/kg days 1-5, 8-12, 15-19, ibrutinib 3 mg/kg days 1-20, or the combination via the intraperitoneal route. (B) Mice were weighed every 3-4 days as a measurement for toxicity. (C) Tumor volume was measure over time as a function of treatment group (D) Kaplan-Meier Curve was calculated for the resistant control mice as compared to the parental control and treatment cohorts.

Article Snippet: Deuterated internal standard ibrutinib-d5 was purchased from TLC Pharmaceutical Standards Ltd (Ontario, Canada).

Techniques: Injection

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Mechanisms of Acquired Drug Resistance to the HDAC6 Selective Inhibitor Ricolinostat Reveals Rational Drug : Drug Combination with Ibrutinib

doi: 10.1158/1078-0432.CCR-16-2022

Figure Lengend Snippet: Serum and tumor tissue was collected from mice at sequential time points and analyzed for concentration of ACY-1215 and ibrutinib by LC-MS/MS. Mice were treated with ACY-1215 at 50 mg/kg alone and ACY-1215 50 mg/kg with ibrutinib 3 mg/kg. Drug concentrations are represented as mean values. (A) Graphical representation of ACY-1215 concentration over time. Mice received one dose of ACY-1215 50 mg/kg or ACY-1215 50 mg/kg plus ibrutinib 3 mg/kg for analysis of serum concentration of ACY-1215. For analysis of drug concentration in tumor tissue, ACY-1215 was administered at 50 mg/kg with or without ibrutinib. (B) Graphical representation of ibrutinib concentration over time analyzed in serum and tumor tissue. Mice received one dose of ibrutinib 3 mg/kg via i.p. route. (C) Summary of pharmacokinetic data for ACY-1215 and ibrutinib. (D) Immunoblot analysis of the IRE1 pathway of the UPR and the BTK pathway from whole cell lysates of mouse tumor tissue treated with ACY-1215, ibrutinib or the combination. Mice were treated with a single i.p. injection and analyzed at 6 hours.

Article Snippet: Deuterated internal standard ibrutinib-d5 was purchased from TLC Pharmaceutical Standards Ltd (Ontario, Canada).

Techniques: Concentration Assay, Liquid Chromatography with Mass Spectroscopy, Western Blot, Injection

Journal: Scientific Reports

Article Title: Behavior of 1-Deoxy-, 3-Deoxy- and N -Methyl-Ceramides in Skin Barrier Lipid Models

doi: 10.1038/s41598-020-60754-4

Figure Lengend Snippet: Structure of natural N -lignoceroyl sphingosine (Cer NS in Motta nomenclature , referred to as Cer here for brevity), its unnatural analogs, i.e ., 1-deoxy-Cer (the chemical modification highlighted in light blue), 3-deoxy-Cer (dark blue) and N -Me-Cer (orange), and other SC lipids used to construct the lipid barrier models.

Article Snippet: 1-Deoxy-sphingosine (synthetic, over 99% stereochemically pure, i.e ., (2 S ,3 R ,4 E )), 3-deoxy-sphingosine (synthetic, over 99% stereochemically pure, i.e ., (2 R ,4 E )), N -methyl-sphingosine (synthetic, over 99% stereochemically pure, i.e ., (2 S ,3 R ,4 E )) and Cer (CerNS; synthetic, over 99% stereochemically pure, i.e ., (2 S ,3 R ,4 E )) were purchased from Avanti Polar Lipids (Alabaster, USA).

Techniques: Modification, Construct

Journal: Scientific Reports

Article Title: Behavior of 1-Deoxy-, 3-Deoxy- and N -Methyl-Ceramides in Skin Barrier Lipid Models

doi: 10.1038/s41598-020-60754-4

Figure Lengend Snippet: XRD diffractograms of SC lipid models containing either Cer (panel A), or 1-deoxy-Cer (B), or 3-deoxy-Cer (C), or N -Me-Cer (D), along with LIG, Chol, and CholS. The intensity is given in arbitrary units (a. u.). Full and dashed grid lines predict the positions of SLP (panel A and D), MLP (panel C), and other structure reflections. Asterisks mark the separated Chol. The determined repeat distances are shown in the respective panels. The peaks at 0.41 nm and 0.37 nm in the wide-angle region (Q = 14–18 nm −1 ) indicate the orthorhombic packing of polymethylene lipid chains.

Article Snippet: 1-Deoxy-sphingosine (synthetic, over 99% stereochemically pure, i.e ., (2 S ,3 R ,4 E )), 3-deoxy-sphingosine (synthetic, over 99% stereochemically pure, i.e ., (2 R ,4 E )), N -methyl-sphingosine (synthetic, over 99% stereochemically pure, i.e ., (2 S ,3 R ,4 E )) and Cer (CerNS; synthetic, over 99% stereochemically pure, i.e ., (2 S ,3 R ,4 E )) were purchased from Avanti Polar Lipids (Alabaster, USA).

Techniques:

Journal: Scientific Reports

Article Title: Behavior of 1-Deoxy-, 3-Deoxy- and N -Methyl-Ceramides in Skin Barrier Lipid Models

doi: 10.1038/s41598-020-60754-4

Figure Lengend Snippet: Lipid chain order in the SC lipid models containing Cer (black), 1-deoxy-Cer (light blue), 3-deoxy-Cer (dark blue), N -Me-Cer (orange), along with LIG, Chol, and CholS. Panels A and B show an example infrared spectrum and enlarged methylene symmetric stretching vibrations at 32 °C, respectively. Panel C shows the means ± SEM of the wavenumbers of the methylene symmetric vibrations; n = 2–3.

Article Snippet: 1-Deoxy-sphingosine (synthetic, over 99% stereochemically pure, i.e ., (2 S ,3 R ,4 E )), 3-deoxy-sphingosine (synthetic, over 99% stereochemically pure, i.e ., (2 R ,4 E )), N -methyl-sphingosine (synthetic, over 99% stereochemically pure, i.e ., (2 S ,3 R ,4 E )) and Cer (CerNS; synthetic, over 99% stereochemically pure, i.e ., (2 S ,3 R ,4 E )) were purchased from Avanti Polar Lipids (Alabaster, USA).

Techniques:

Journal: Scientific Reports

Article Title: Behavior of 1-Deoxy-, 3-Deoxy- and N -Methyl-Ceramides in Skin Barrier Lipid Models

doi: 10.1038/s41598-020-60754-4

Figure Lengend Snippet: Lipid chain order and phase transitions in the SC lipid models containing the studied Cer (black; A), 1-deoxy-Cer (light blue; B), 3-deoxy-Cer (dark blue; C), or N -Me-Cer (orange; D), along with ( d )-LIG, Chol, and CholS. First graphs in each panel show the unlabeled lipid mixtures, i.e ., the thermal evolution of infrared symmetric methylene stretching vibrations. Second graphs in each panel show the lipid mixtures with d -LIG, i.e ., the temperature dependence of the symmetric CH 2 (mainly from Cer, filled circles) and CD 2 stretching vibrations (from d -LIG, open circles). The temperatures of the main phase transitions (estimated from the first derivatives of the curves) are indicated. Data are shown as means ± SEM; n = 2–3.

Article Snippet: 1-Deoxy-sphingosine (synthetic, over 99% stereochemically pure, i.e ., (2 S ,3 R ,4 E )), 3-deoxy-sphingosine (synthetic, over 99% stereochemically pure, i.e ., (2 R ,4 E )), N -methyl-sphingosine (synthetic, over 99% stereochemically pure, i.e ., (2 S ,3 R ,4 E )) and Cer (CerNS; synthetic, over 99% stereochemically pure, i.e ., (2 S ,3 R ,4 E )) were purchased from Avanti Polar Lipids (Alabaster, USA).

Techniques:

Journal: Scientific Reports

Article Title: Behavior of 1-Deoxy-, 3-Deoxy- and N -Methyl-Ceramides in Skin Barrier Lipid Models

doi: 10.1038/s41598-020-60754-4

Figure Lengend Snippet: Lateral lipid packing in the SC lipid model containing either Cer (panel A), or 1-deoxy-Cer (panel B), or 3-deoxy-Cer (panel C), or N -Me-Cer (panel D), along with ( d )-LIG, Chol, and CholS. First graphs in each panel show the representative infrared spectra of the methylene scissoring vibrations of the unlabeled lipid mixtures at 32 °C. Second and third graphs in each panel show the CH 2 scissoring vibration (mainly from Cer) and CD 2 scissoring vibrations ( d -LIG; dashed line), respectively, of the lipid mixtures with d -LIG at 32 °C. The fourth graphs in each panel show the representative infrared spectra of the methylene rocking vibrations of the unlabeled mixtures at 32 °C. A presence of a CH 2 /CD 2 doublet in the spectrum is indicative of an orthorhombic packing. Intensity is given in arbitrary units (a. u.).

Article Snippet: 1-Deoxy-sphingosine (synthetic, over 99% stereochemically pure, i.e ., (2 S ,3 R ,4 E )), 3-deoxy-sphingosine (synthetic, over 99% stereochemically pure, i.e ., (2 R ,4 E )), N -methyl-sphingosine (synthetic, over 99% stereochemically pure, i.e ., (2 S ,3 R ,4 E )) and Cer (CerNS; synthetic, over 99% stereochemically pure, i.e ., (2 S ,3 R ,4 E )) were purchased from Avanti Polar Lipids (Alabaster, USA).

Techniques:

Journal: Scientific Reports

Article Title: Behavior of 1-Deoxy-, 3-Deoxy- and N -Methyl-Ceramides in Skin Barrier Lipid Models

doi: 10.1038/s41598-020-60754-4

Figure Lengend Snippet: The permeabilities of the studied SC lipid models composed either of Cer (black), or 1-deoxy-Cer (blue), or 3-deoxy-Cer (dark blue), or N -Me-Cer (orange), along with LIG, Chol, and CholS. ( A ) Water loss; ( B ) permeation profile for TH; ( C ) the flux of TH; ( D ) permeation profile for IND; ( E ) the flux of IND; ( F ) electrical impedance. Data are presented as the means ± standard error of the mean (SEM), n = 5–16. The asterisks show statistically significant differences against the Cer-containing sample ( p <0.05).

Article Snippet: 1-Deoxy-sphingosine (synthetic, over 99% stereochemically pure, i.e ., (2 S ,3 R ,4 E )), 3-deoxy-sphingosine (synthetic, over 99% stereochemically pure, i.e ., (2 R ,4 E )), N -methyl-sphingosine (synthetic, over 99% stereochemically pure, i.e ., (2 S ,3 R ,4 E )) and Cer (CerNS; synthetic, over 99% stereochemically pure, i.e ., (2 S ,3 R ,4 E )) were purchased from Avanti Polar Lipids (Alabaster, USA).

Techniques: