Brad Gouker
A5031597263- Peptidase Inhibition and Analysis
- Renal cell carcinoma treatment
- Mechanisms of cancer metastasis
- Protease and Inhibitor Mechanisms
- Hormonal Regulation and Hypertension
- Ubiquitin and proteasome pathways
- Renal and related cancers
- Cancer, Lipids, and Metabolism
- Cancer Research and Treatments
- Cancer Cells and Metastasis
- Cancer Genomics and Diagnostics
- Endoplasmic Reticulum Stress and Disease
- Cancer Mechanisms and Therapy
- Statistical Methods in Clinical Trials
- Histiocytic Disorders and Treatments
- Lymphoma Diagnosis and Treatment
- Autophagy in Disease and Therapy
- Phagocytosis and Immune Regulation
- Cancer, Hypoxia, and Metabolism
- Cell Adhesion Molecules Research
- Oral and Maxillofacial Pathology
- Molecular Biology Techniques and Applications
- Sarcoma Diagnosis and Treatment
- Biosimilars and Bioanalytical Methods
Frederick National Laboratory for Cancer Research
2015-2024
Leidos (United States)
2015-2021
Leidos Biomedical Research Inc. (United States)
2015-2017
Lung squamous cell carcinoma (LSCC) is the second most prevalent type of lung cancer. Despite extensive genomic characterization, no targeted therapies are approved for treatment LSCC. Distal amplification 3q chromosome frequent alteration in LSCC, and there an urgent need to identify efficacious druggable targets within this amplicon. We protein kinase TNIK as a therapeutic target amplified approximately 50% LSCC cases. genetic depletion or pharmacologic inhibition reduces growth cells...
<p>NAD<sup>+</sup> and NADH analysis of UOK365 cells. Analysis the relative levels NAD<sup>+</sup> their ratio after 24 hour treatment cell line model with a range concentrations (0.5-100 nM) either GNE-618 (A-B) or OT-82 (C-D). All were compared to cells treated amount DMSO present in highest concentration utilized drugs.</p>
<p>Real-time invasion analysis of UOK262 and UOK365 cells treated with OT-82. Real-time demonstrating cell line over 5 days (120 hrs) was performed (A) to show that DMSO (used as the vehicle for OT-82) did not affect in comparison untreated (B) demonstrate effects OT-82 treatment at either 1 nM or 10 alone (DMSO) non-invasive control (no serum lower chamber). In each case, a representative graph from one three separate experiments is shown. (C) The inhibition repeats dose shown line....
<p>In vitro analysis of NAMPT inhibition on glycolysis in FH-deficient tumor cells. (A) Effects the inhibitor OT-82 (100 nM) extracellular acidification rate (ECAR) UOK262 and UOK365 HLRCC cell lines RPTEC normal line. (B) Representation different effects versus DMSO at time point 5 (designated by a star) after injection glucose UOK262, UOK365, RPTEC. 2-DG, 2-deoxyglucose.</p>
<p>Structural figures of NAMPT inhibitors.</p>
<p>In vivo effects of NAMPT inhibitors in HLRCC xenograft models. Mean body weights are constant throughout the drug study for animals harboring UOK262 xenografts (A) and UOK365 (B; shown Figure 3A-B, <i>n</i> = 10 mice per arm, 95mg/kg OT-82 8 week duration). (C-D) H&E stained sections retinas obtained from NSG treated with vehicle or 5 days. (E-F) Caspase 3 95 mg/kg Histological were evaluated signs retinal toxicity by a trained veterinary pathologist. No overt...
<div>Abstract<p>Hereditary leiomyomatosis and renal cell cancer (HLRCC) is an inherited syndrome caused by germline pathogenic variants in the fumarate hydratase (<i>FH</i>) gene. Affected individuals are at risk for developing cutaneous uterine leiomyomas aggressive FH-deficient carcinoma (RCC) with a papillary histology. Due to disrupted tricarboxylic acid cycle, kidney cancers rely on aerobic glycolysis energy production, potentially creating compensatory metabolic...
<p>Expression levels of NAD<sup>+</sup> biosynthetic and salvage pathway genes in HLRCC tumors, normal kidney samples pan-kidney data from TCGA. (A) Overview pathways. (B-E) Box plots for selected NAD+ derived RNA-seq analysis tumors specimens. (F) TCGA data. NAMPT (nicotinamide phosphoribosyltransferase), NAPRT (nicotinate phosphoribosyltransferase ), QPRT (quinolinate NMNAT mononucleotide adenylyl transferase), KIRP (kidney renal papillary cancer), CIMP (CpG island...
<p>Validation studies to evaluate inhibition of additional HLRCC cell lines by NAMPT inhibitors. A) Effects the inhibitor GNE-618 on viability in FH <sup>-/</sup>- cells UOK348, UOK271, UOK350, UOK268, UOK365, restored UOK268WT and non-transformed kidney epithelial RPTEC. Cell was assessed Titer-Glo assay at 96 h. B) OT-82 -/- RPTEC.</p>
<p>Proliferation analysis of UOK268 and RPTEC in response to OT-82 nicotinic acid rescue NAD<sup>+</sup>/NADH depletion. (A-B) or cells were grown 96 well plates from an initial plating 2,000 cellular confluency was monitored real-time by Incucyte S3 Live-Cell Imaging System. After 24 hours measurements treated with either DMSO, 1 nM OT-82, 5 10 plus mM nicotinamide mononucleotide (NMN) evaluated for additional days. (C-D) The effect NMN on depletion is shown. (E) Effects...
<p>Cell proliferation rates of HLRCC cell lines treated with low and high concentrations NAMPT inhibitors. A) Effect OT-82 at 0.8nM or 100nM (left) GNE-618 8nM 200nM (right) on UOK262 after 96 hours. B) UOK268 C) UOK365 hours.</p>
<p>HLRCC spheroid characterization and OT-82 effect on 3D culture. A) UOK262 (B) UOK365 spheroids plated at different cell densities (8000, 4000, 2000 cells/well in triplicate) after 24h, showing degrees of aggregation. Cell viability (C) (D) treated with GNE-618 (1 nM-500 nM) or (0.2 nM-100 assessed Cell-Titer Glo assay decreases a dose dependent manner.</p>
<p>Time course study design for imaging and OT-82 treatment hyperpolarized pyruvate 13C-MRSI. Post MRSI was performed 5 hours post dose #2.</p>
<p>Pharmacokinetic profile of OT-82 in nude and NSG mice. (A) Bioanalysis plasma concentrations were made using a validated LC-MS/MS assay (<i>n</i>=3 mice, 50 mg/kg, 7 intervals; <i>n</i>=3 75mg/kg, intervals). (B) Pharmacokinetic parameters calculated after single dose OT-82. The data from all mice each group pooled together to assume one “average” mouse per group.</p>
<p>NAMPT immunohistochemical analysis in HLRCC tumors and normal kidney. (A) In patient #1, both a primary kidney tumor (upper panel) an associated metastatic mass (lower demonstrated strong NAMPT staining. Surrounding non-tumor tissues show little (B) #4, shows staining comparison to tissue present on the same slide panel). (C) #5, Material from this was used derive UOK262 cell line, which similar positive when grown as xenograft (D) #6, Matching H&E is included for all samples...
Abstract Hereditary leiomyomatosis and renal cell cancer (HLRCC) is an inherited syndrome caused by germline pathogenic variants in the fumarate hydratase (FH) gene. Affected individuals are at risk for developing cutaneous uterine leiomyomas aggressive FH-deficient carcinoma (RCC) with a papillary histology. Due to disrupted tricarboxylic acid cycle, kidney cancers rely on aerobic glycolysis energy production, potentially creating compensatory metabolic vulnerabilities. This study conducted...
Rhabdomyosarcoma (RMS) is the most common pediatric soft tissue sarcoma. Despite decades of clinical trials, overall survival rate for patients with relapsed and metastatic disease remains below 30%, underscoring need novel treatments. FGFR4, a receptor tyrosine kinase that overexpressed in RMS mutationally activated 10% cases, promising target treatment. Here, we show futibatinib, an irreversible pan-FGFR inhibitor, inhibits growth cell lines vitro by inhibiting phosphorylation FGFR4 its...
Abstract We previously reported the generation of rabbit monoclonal antibodies to twelve EMT (epithelial-to-mesenchymal transition) transcription factors and cancer stem cell (CSC) markers for development pharmacodynamic assays inform clinical trials new anticancer therapies (Pfister et al., AACR 2013). Here we demonstrate functional utility some these reagents in detecting HGF-induced changes CSC biology a xenograft tumor model. Initial antibody characterization was performed vitro subset...
Abstract Epithelial-mesenchymal transition (EMT) is a dynamic process whereby epithelial cells acquire mesenchymal properties. Despite the clinical significance of acquired properties for metastasis and drug resistance, histopathological evidence transitional in patient samples lacking EMT remains an unproven hypothesis. We previously developed validated multiplex immunofluorescence assay (IFA) that quantifies levels biomarkers (E-Cadherin, Vimentin, β-catenin) snap-frozen, formalin-fixed,...
Abstract Robust PD assay results are valuable for informing decisions about continued preclinical and clinical development of new agents identifying effective combinations targeted agents. The National Cancer Institute's Division Treatment Diagnosis (DCTD) develops validates assays to obtain accurate information drug effect on intended molecular targets in first-in-human trials. Our group utilizes quantitative immunofluorescence (qIFA) biomarkers FFPE slides prepared from pre- post-dose...
<p>Uncropped immunoblots reported in this study.</p>