43 results on '"Jessen, Bart"'
Search Results
2. Correction to: An IQ consortium analysis of starting dose selection for oncology small molecule first‑in‑patient trials suggests an alternative NOAEL‑based method can be safe while reducing time to the recommended phase 2 dose
- Author
-
Jessen, Bart A., Cornwell, Paul, Redmond, Sean, Visalli, Thomas, Lemper, Marie, Bunch, Todd, and Hart, Timothy
- Published
- 2023
- Full Text
- View/download PDF
3. A distal region of the human TGM1 promoter is required for expression in transgenic mice and cultured keratinocytes
- Author
-
Phillips, Marjorie A, Jessen, Bart A, Lu, Ying, Qin, Qin, Stevens, Mary E, and Rice, Robert H
- Subjects
Biomedical and Clinical Sciences ,Clinical Sciences ,Biotechnology ,Genetics ,Animals ,Cell Line ,Gene Expression ,Genes ,Reporter ,Humans ,Keratinocytes ,Mice ,Mice ,Transgenic ,Mutation ,Promoter Regions ,Genetic ,Transcription Factors ,Transcription ,Genetic ,Transglutaminases ,Public Health and Health Services ,Dermatology & Venereal Diseases ,Clinical sciences - Abstract
BackgroundTGM1(transglutaminase 1) is an enzyme that crosslinks the cornified envelope of mature keratinocytes. Appropriate expression of the TGM1 gene is crucial for proper keratinocyte function as inactivating mutations lead to the debilitating skin disease, lamellar ichthyosis. TGM1 is also expressed in squamous metaplasia, a consequence in some epithelia of vitamin A deficiency or toxic insult that can lead to neoplasia. An understanding of the regulation of this gene in normal and abnormal differentiation states may contribute to better disease diagnosis and treatment.MethodsIn vivo requirements for expression of the TGM1 gene were studied by fusing various lengths of promoter DNA to a reporter and injecting the DNA into mouse embryos to generate transgenic animals. Expression of the reporter was ascertained by Western blotting and immunohistochemistry. Further delineation of a transcriptionally important distal region was determined by transfections of progressively shortened or mutated promoter DNA into cultured keratinocytes.ResultsIn vivo analysis of a reporter transgene driven by the TGM1 promoter revealed that 1.6 kilobases, but not 1.1 kilobases, of DNA was sufficient to confer tissue-specific and cell layer-specific expression. This same region was responsible for reporter expression in tissues undergoing squamous metaplasia as a response to vitamin A deprivation. Mutation of a distal promoter AP1 site or proximal promoter CRE site, both identified as important transcriptional elements in transfection assays, did not prevent appropriate expression. Further searching for transcriptional elements using electrophoretic mobility shift (EMSA) and transfection assays in cultured keratinocytes identified two Sp1 elements in a transcriptionally active region between -1.6 and -1.4 kilobases. While mutation of either Sp1 site or the AP1 site singly had only a small effect, mutation of all three sites eliminated nearly all the transcriptional activity.ConclusionsA distal region of the TGM1 gene promoter, containing AP1 and Sp1 binding sites, is evolutionarily conserved and responsible for high level expression in transgenic mice and in transfected keratinocyte cultures.
- Published
- 2004
4. The contribution of physicochemical properties to multiple in vitro cytotoxicity endpoints
- Author
-
Lu, Shuyan, Jessen, Bart, Strock, Christopher, and Will, Yvonne
- Published
- 2012
- Full Text
- View/download PDF
5. Transient knock down of checkpoint kinase 1 in hematopoietic progenitors is linked to bone marrow toxicity
- Author
-
Hu, Wenyue, Zong, Qing, John-Baptiste, Annette, and Jessen, Bart
- Published
- 2011
- Full Text
- View/download PDF
6. A high content screening assay for identifying lysosomotropic compounds
- Author
-
Nadanaciva, Sashi, Lu, Shuyan, Gebhard, David F., Jessen, Bart A., Pennie, William D., and Will, Yvonne
- Published
- 2011
- Full Text
- View/download PDF
7. Cardiovascular differentiation of imatinib and bosutinib in the rat
- Author
-
Heyen, Jonathan R., Hu, Wenyue, Jamieson, Joseph, Thibault, Stephane, Batugo, Minerva, Loi, Cho-Ming, Burns-Naas, Leigh Ann, McHarg, Aileen D., and Jessen, Bart
- Published
- 2013
- Full Text
- View/download PDF
8. Comparison of preservative-induced toxicity on monolayer and stratified Chang conjunctival cells
- Author
-
Yanochko, Gina M., Khoh-Reiter, Su, Evans, Mark G., and Jessen, Bart A.
- Published
- 2010
- Full Text
- View/download PDF
9. Toxicity and toxicokinetics of the cyclin-dependent kinase inhibitor AG-024322 in cynomolgus monkeys following intravenous infusion
- Author
-
Brown, Alan P., Courtney, Cynthia L., Criswell, Kay A., Holliman, Christopher L., Evering, Winston, and Jessen, Bart A.
- Published
- 2008
- Full Text
- View/download PDF
10. Mechanistic Investigation of Imatinib-Induced Cardiac Toxicity and the Involvement of c-Abl Kinase
- Author
-
Hu, Wenyue, Lu, Shuyan, McAlpine, Indrawan, Jamieson, Joseph D., Lee, Dong U., D. Marroquin, Lisa, Heyen, Jonathan R., and Jessen, Bart A.
- Published
- 2012
- Full Text
- View/download PDF
11. Lack of evidence for a link between latanoprost use and malignant melanoma: an analysis of safety databases and a review of the literature
- Author
-
Tressler, Charles S, Wiseman, Robert L, Dombi, Theresa M, Jessen, Bart, Huang, Kui, Kwok, Kenneth K, and Wirostko, Barbara M
- Published
- 2011
- Full Text
- View/download PDF
12. Corneal Neovascularization and Ocular Irritancy Responses in Dogs Following Topical Ocular Administration of an EP4-Prostaglandin E2 Agonist
- Author
-
Aguirre, Shirley A., Huang, Wenhu, Prasanna, Ganesh, and Jessen, Bart
- Published
- 2009
- Full Text
- View/download PDF
13. Effect of the Multitargeted Tyrosine Kinase Inhibitors Imatinib, Dasatinib, Sunitinib, and Sorafenib on Mitochondrial Function in Isolated Rat Heart Mitochondria and H9c2 Cells
- Author
-
Will, Yvonne, Dykens, James A., Nadanaciva, Sashi, Hirakawa, Brad, Jamieson, Joseph, Marroquin, Lisa D., Hynes, James, Patyna, Shem, and Jessen, Bart A.
- Published
- 2008
14. The Relevance of a 3D Corneal Model in Assessing the Safety of the Topical Ocular Formulation Preservative Benzalkonium Chloride in Xalatan and Its Vehicle: Poster No.: GC-D2-147 Panel No.: 147
- Author
-
Khoh-Reiter, Su and Jessen, Bart
- Published
- 2008
15. Retinal and Peripheral Nerve Toxicity Induced by the Administration of a Pan-Cyclin Dependent Kinase (cdk) Inhibitor in Mice
- Author
-
Illanes, Oscar, Anderson, Scott, Niesman, Michael, Zwick, Laura, and Jessen, Bart A.
- Published
- 2006
- Full Text
- View/download PDF
16. Gastric and Pancreatic Lesions in Rats Treated with a Pan-CDK Inhibitor
- Author
-
Ramiro-Ibáñez, Fernando, Trajkovic, Dusko, and Jessen, Bart
- Published
- 2005
- Full Text
- View/download PDF
17. Assessment of Hepatocytes and Liver Slices as in Vitro Test Systems to Predict in Vivo Gene Expression
- Author
-
Jessen, Bart A., Mullins, Jennifer S., de Peyster, Ann, and Stevens, Gregory J.
- Published
- 2003
18. Role of Sp1 Response Element in Transcription of the Human Transglutaminase 1 Gene
- Author
-
Jessen, Bart A., Phillips, Marjorie A., Hovnanian, Alain, and Rice, Robert H.
- Published
- 2000
19. Evaluation of the cytotoxic effects of ophthalmic solutions containing benzalkonium chloride on corneal epithelium using an organotypic 3-D model
- Author
-
Jessen Bart A and Khoh-Reiter Su
- Subjects
Ophthalmology ,RE1-994 - Abstract
Abstract Background Benzalkonium chloride (BAC) is a common preservative used in ophthalmic solutions. The aim of this study was to compare the cytotoxic effects of BAC-containing ophthalmic solutions with a BAC-free ophthalmic solution using an organotypic 3-dimensional (3-D) corneal epithelial model and to determine the effects of latanoprost ophthalmic solution and its BAC-containing vehicle on corneal thickness in a monkey model. Methods The cytotoxicity of commercially available BAC-containing ophthalmic formulations of latanoprost (0.02% BAC) and olopatadine (0.01% BAC) was compared to that of BAC-free travoprost and saline in a corneal organotypic 3-D model using incubation times of 10 and 25 minutes. To compare the extent of differentiation of 3-D corneal cultures to monolayer transformed human corneal epithelial (HCE-T) cell cultures, expression levels (mRNA and protein) of the corneal markers epidermal growth factor receptor, transglutaminase 1 and involucrin were quantified. Finally, latanoprost ophthalmic solution or its vehicle was administered at suprapharmacologic doses (two 30 μL drops twice daily in 1 eye for 1 year) in monkey eyes, and corneal pachymetry was performed at baseline and at weeks 4, 13, 26 and 52. Results In the 3-D corneal epithelial culture assays, there were no significant differences in cytotoxicity between the BAC-containing latanoprost and olopatadine ophthalmic solutions and BAC-free travoprost ophthalmic solution at either the 10- or 25-minute time points. The 3-D cultures expressed higher levels of corneal epithelial markers than the HCE-T monolayers, indicating a greater degree of differentiation. There were no significant differences between the corneal thickness of monkey eyes treated with latanoprost ophthalmic solution or its vehicle (both containing 0.02% BAC) and untreated eyes. Conclusion The lack of cytotoxicity demonstrated in 3-D corneal cultures and in monkey studies suggests that the levels of BAC contained in ophthalmic solutions are not likely to cause significant direct toxicity to epithelium of otherwise normal corneas.
- Published
- 2009
- Full Text
- View/download PDF
20. Keratinocyte Differentiation Marker Suppression by Arsenic: Mediation by AP1 Response Elements and Antagonism by Tetradecanoylphorbol Acetate
- Author
-
Jessen, Bart A., Qin, Qin, Phillips, Marjorie A., Phillips, Donald L., and Rice, Robert H.
- Published
- 2001
- Full Text
- View/download PDF
21. Phenotypic Characterization of Targeted Knockdown of Cyclin-Dependent Kinases in the Intestinal Epithelial Cells.
- Author
-
Lu, Shuyan, Sung, Tae, Amaro, Marina, Hirakawa, Brad, Jessen, Bart, and Hu, Wenyue
- Subjects
CYCLIN-dependent kinases ,EPITHELIAL cells ,SMALL molecules ,CELL death ,INHIBITION of cellular proliferation - Abstract
Cyclin-dependent kinases (CDKs) are serine/threonine kinases that regulate cell cycle and have been vigorously pursued as druggable targets for cancer. There are over 20 members of the CDK family. Given their structural similarity, selective inhibition by small molecules has been elusive. In addition, collateral damage to highly proliferative normal cells by CDK inhibitors remains a safety concern. Intestinal epithelial cells are highly proliferative and the impact of individual CDK inhibition on intestinal cell proliferation has not been well studied. Using the rat intestinal epithelial (IEC6) cells as an in vitro model, we found that the selective CDK4/6 inhibitor palbociclib lacked potent anti-proliferative activity in IEC6 relative to the breast cancer cell line MCF7, indicating the absence of intestinal cell reliance on CDK4/6 for cell cycle progression. To further illustrate the role of CDKs in intestinal cells, we chose common targets of CDK inhibitors (CDK 1, 2, 4, 6, and 9) for targeted gene knockdown to evaluate phenotypes. Surprisingly, only CDK1 and CDK9 knockdown demonstrated profound cell death or had moderate growth effects, respectively. CDK2, 4, or 6 knockdowns, whether single, double, or triple combinations, did not have substantial impact. Studies evaluating CDK1 knockdown under various cell seeding densities indicate direct effects on viability independent of proliferation state and imply a potential noncanonical role for CDK1 in intestinal epithelial biology. This research supports the concept that CDK1 and CDK9, but not CDKs 2, 4, or 6, are essential for intestinal cell cycle progression and provides safety confidence for interphase CDK inhibition. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
22. Lysosomal adaptation: How cells respond to lysosomotropic compounds.
- Author
-
Lu, Shuyan, Sung, Tae, Lin, Nianwei, Abraham, Robert T., and Jessen, Bart A.
- Subjects
CELLULAR signal transduction ,LYSOSOMES ,RHODOPSIN ,PHYSIOLOGICAL stress ,PHYSIOLOGICAL adaptation ,GLYCERALDEHYDEPHOSPHATE dehydrogenase - Abstract
Lysosomes are acidic organelles essential for degradation and cellular homoeostasis and recently lysosomes have been shown as signaling hub to respond to the intra and extracellular changes (e.g. amino acid availability). Compounds including pharmaceutical drugs that are basic and lipophilic will become sequestered inside lysosomes (lysosomotropic). How cells respond to the lysosomal stress associated with lysosomotropism is not well characterized. Our goal is to assess the lysosomal changes and identify the signaling pathways that involve in the lysosomal changes. Eight chemically diverse lysosomotropic drugs from different therapeutic areas were subjected to the evaluation using the human adult retinal pigmented epithelium cell line, ARPE-19. All lysosomotropic drugs tested triggered lysosomal activation demonstrated by increased lysosotracker red (LTR) and lysosensor green staining, increased cathepsin activity, and increased LAMP2 staining. However, tested lysosomotropic drugs also prompted lysosomal dysfunction exemplified by intracellular and extracellular substrate accumulation including phospholipid, SQSTM1/p62, GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) and opsin. Lysosomal activation observed was likely attributed to lysosomal dysfunction, leading to compensatory responses including nuclear translocation of transcriptional factors TFEB, TFE3 and MITF. The adaptive changes are protective to the cells under lysosomal stress. Mechanistic studies implicate calcium and mTORC1 modulation involvement in the adaptive changes. These results indicate that lysosomotropic compounds could evoke a compensatory lysosomal biogenic response but with the ultimate consequence of lysosomal functional impairment. This work also highlights a pathway of response to lysosomal stress and evidences the role of TFEB, TFE3 and MITF in the stress response. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
- View/download PDF
23. Validation of Cross-Species Reactivity of the VEGF-A/PDGFRβ Bifunctional Antibody PF-06653157.
- Author
-
Ashoor, Roshan, Lee, Dong, Cheng, Alvan, Jessen, Bart, and Huang, Wenhu
- Subjects
MONOCLONAL antibodies ,NEOVASCULARIZATION inhibitors ,REVERSE transcriptase polymerase chain reaction ,VASCULAR endothelial growth factors ,QUALITATIVE research ,VASCULAR endothelial growth factor antagonists ,ANIMAL experimentation ,ANTIGEN-antibody reactions ,BIOCHEMISTRY ,CELL receptors ,DOGS ,DOSE-effect relationship in pharmacology ,PHENOMENOLOGY ,MICE ,PRIMATES ,RABBITS ,RATS ,SWINE ,PATHOLOGIC neovascularization - Abstract
Purpose: PF-06653157 is a bifunctional antagonist monoclonal antibody (mAb) that targets human VEGF-A ligand and PDGF-Rβ. With the advent of PF-06653157 as an angiogenesis inhibitor and potential treatment for angiogenesis deregulation diseases, a relevant toxicology species is needed for toxicity and efficacy studies. Investigative studies were conducted to validate the mAb dual antagonist properties in a human system and determine its cross-reactive pharmacology in nonhuman cells.Methods: Sequence alignment was used to determine percent sequence identity of VEGF and PDGF receptors and ligands; qualitative reverse transcription polymerase chain reaction (qRT-PCR) was used to determine the presence of PDGF-Rβ on cells of interest. The functional activity of PF-06653157 antibody was assessed in human, dog, porcine, rabbit, rat, mouse, and cynomolgus monkey cells treated with VEGF and PDGF ligands through cell proliferation assays and western blot analysis of AKT and p44/p42 (ERK1/2) protein phosphorylation and enzyme-linked immunosorbent assay.Results: PF-06653157 attenuated phosphorylation of AKT and p44/p42 proteins in human and cynomolgus monkey cells. The antibody did not attenuate AKT nor p44/p42 phosphorylation in any other species tested. PDGFR signaling could not be activated with human PDGF ligand in the porcine cells, so PF-06653157 activity in porcine remains inconclusive.Conclusion: The PF-06653157 mAb cross-reacts with cynomolgus monkey cells in a similar manner to human cells. Therefore, cynomolgus monkeys are considered the appropriate species for efficacy and regulatory toxicology studies in PF-06653157 development. [ABSTRACT FROM AUTHOR]- Published
- 2016
- Full Text
- View/download PDF
24. Contribution of Membrane Trafficking Perturbation to Retinal Toxicity.
- Author
-
Khoh-Reiter, Su, Sokolowski, Sharon A., Jessen, Bart, Evans, Mark, Dalvie, Deepak, and Shuyan Lu
- Subjects
PHOTORECEPTORS ,ELECTROPHYSIOLOGY ,EPITHELIAL cells ,PHAGOCYTOSIS ,ANTIGEN-antibody reactions - Abstract
The retina is a highly structured tissue that is formed by layers containing 7 different cell types. The photoreceptor cell is a specialized type of neuron in the retina that is capable of absorbing and converting light into electrophysiological signals. There is a constant renewal process for photoreceptors consisting of intermittent shedding of the distal tips of the photosensitive outer segment and subsequent phagocytosis (uptake, degradation and recycling) by retinal pigmented epithelial (RPE) cells. This rebuilding process is essential for vision and the survival of photoreceptors and RPE cells. Drugs with a basic moiety have the potential to accumulate in the lysosome and impair its functions including the phagocytosis process, which could hinder clearance of outer segments and ultimately induce retinopathy. To determine the prevalence of this cellular mechanism in retinal toxicity, a collection of proprietary compounds associated with retinal toxicity were subjected to a battery of in vitro tests using the human adult retinal pigmented epithelium cell line, ARPE-19. The tests included a phagocytosis assay, and lysosomal and autophagosomal staining. The compounds that induced retinopathy clustered in the basic and lipophilic region, which drives lysosomal sequestration. This accumulation coincided with phagocytosis inhibition and an increase in autophagosome staining, suggesting a blockage of the membrane trafficking process. A correlation between the physicochemical properties and in vitro lysosomal pathway effects was established. These data reveal the importance of physicochemical properties of compounds and lysosome accumulation as a potential mechanism for drug-induced retinopathy and demonstrate the usefulness of in vitro screening in predicting this liability. [ABSTRACT FROM AUTHOR]
- Published
- 2015
- Full Text
- View/download PDF
25. Investigation of Ocular Events Associated with Taprenepag Isopropyl, a Topical EP2 Agonist in Development for Treatment of Glaucoma.
- Author
-
Yanochko, Gina M., Affolter, Timothy, Eighmy, Johnnie J., Evans, Mark G., Khoh-Reiter, Su, Lee, Dong, Miller, Paul E., Shiue, Michael H. I., Trajkovic, Dusko, and Jessen, Bart A.
- Subjects
HEALTH outcome assessment ,OCULAR pharmacology ,GLAUCOMA treatment ,EYE diseases ,INTRAOCULAR pressure ,LABORATORY monkeys ,PROSTAGLANDINS ,PROSTAGLANDIN receptors ,THERAPEUTICS - Abstract
Purpose: Taprenepag isopropyl is an EP2 receptor agonist that is in development for the treatment of glaucoma. Iritis, photophobia, and increased corneal thickness observed in a Phase 2 clinical trial with taprenepag isopropyl were not previously observed in topical ocular toxicity studies in rabbits and dogs. In vivo studies using cynomolgus monkeys and in vitro models were used to elucidate the mechanisms underlying these ocular events. Methods: Monkeys were dosed daily for 28 days in 1 eye with taprenepag and in the other with vehicle control. Complete ophthalmic examinations were performed at baseline and weekly thereafter. Serial sections of eyes were examined histopathologically at the end of the study. Recovery after the discontinuation of taprenepag was assessed for 28 days in the monkeys in the high-dose group. In vitro studies evaluated cell viability, paracellular permeability, and cytokine induction with human corneal epithelial or endothelial cell cultures. Results: Monkeys demonstrated a dose-related incidence of iritis and increased corneal thickness that resolved within 28 days of discontinuing taprenepag. There was no evidence in vivo of taprenepag toxicity to the corneal endothelium or epithelium. Cell viability of stratified epithelial cells was primarily affected by excipients and was similar to Xalatan
® . The viability of HCEC-12 cells was not affected by taprenepag at concentrations up to 100 μM. Conclusions: The lack of in vivo or in vitro endothelial cytotoxicity and the reversibility of the increase in corneal thickness and iritis in the monkey provide confidence to permit further clinical development of taprenepag. [ABSTRACT FROM AUTHOR]- Published
- 2014
- Full Text
- View/download PDF
26. Safety Assessment of Subconjunctivally Implanted Devices Containing Latanoprost in Dutch-Belted Rabbits.
- Author
-
Jessen, Bart A., Shiue, Michael H.I., Kaur, Harjeet, Miller, Paul, Leedle, Robert, Guo, Hong, and Evans, Mark
- Subjects
- *
GLAUCOMA treatment , *INTRAOCULAR pressure , *DISEASE progression , *DRUG efficacy , *DRUG tolerance , *ELECTRORETINOGRAPHY , *LABORATORY rabbits - Abstract
Purpose: Latanoprost is used for the treatment of an increased intraocular pressure (IOP) to prevent the progression of glaucoma. Since the lack of compliance with topical ocular dosing may compromise efficacy, alternate methods of delivery are being sought. A 9-month study was conducted to assess the safety and tolerability of latanoprost-containing subconjunctivally implanted devices. Methods: Dutch-belted rabbits were implanted subconjunctivally with up to 5 placebo or drug-loaded devices containing from 50 to 180 μg of latanoprost per device. Study assessment consisted of irritation scoring, clinical signs, ophthalmic exams, IOP, electroretinography (ERG), ocular histology of cohorts at 3 and 9 months postimplantation, and systemic exposure to latanoprost acid. Results: The implants were well tolerated, with minimal-to-mild clinical and microscopic ocular findings attributable to either the placebo or drug-loaded devices. Mild conjunctival congestion persisted through week 13 of the study and tended to correlate with the number of devices and presence of drug. Ophthalmic examinations revealed no effects beyond conjunctival surface hyperemia. No effects on the IOP, corneal thickness, or ERG parameters were observed. The lack of changes in the IOP was expected due to the known lack of the IOP-lowering effects in rabbits from latanoprost. Microscopically, implants at the 3-month necropsy were associated with subconjunctival tissue cavities (containing the implants), fibrous encapsulation, and an infiltrate of lymphocytes and macrophages, sometimes as multinucleate cells, into the subconjunctival implant cavity. The drug-containing implants were often associated with inflammatory cell infiltrates, including heterophils (neutrophils), within the implant subconjunctival cavities and adjacent to the implant sites. At the 9-month necropsy, heterophils were no longer common among the inflammatory cell infiltrates; macrophages and lymphocytes persisted; most of the biodegradable implants were fragmented and disintegrating; and fibrovascular proliferation was present within implant luminal remnants. None of the findings were considered adverse. Systemic exposures were above the limit of quantification (0.1 ng/mL) for up to 96 h in the higher-dose groups, consistent with the initial burst phase of compound release. Conclusion: Overall, the study supports the safety of the latanoprost-containing subconjunctival device as a means of extended delivery of the antiglaucoma medication. Latanoprost-containing subconjunctival implants were well tolerated by Dutch-belted rabbits for up to 9 months. Such devices may improve patient compliance and serve as a means of extended delivery of antiglaucoma medications. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
27. A tyrosine kinase inhibitor-induced myocardial degeneration in rats through off-target phosphodiesterase inhibition.
- Author
-
Hu, Wenyue, Hirakawa, Brad, Jessen, Bart, Lee, Michelle, and Aguirre, Shirley
- Subjects
PROTEIN-tyrosine kinase inhibitors ,MYOCARDIAL infarction ,LABORATORY rats ,PHOSPHODIESTERASE inhibitors ,HEART beat ,CONTRACTILITY (Biology) - Abstract
ABSTRACT PF-04254644 is a selective kinase inhibitor of mesenchymal epithelial transition factor/hepatocyte growth factor receptor with known off-target inhibitory activity against the phosphodiesterase (PDE) family. Rats given repeated oral doses of PF-04254644 developed a mild to moderate myocardial degeneration accompanied by sustained increase in heart rate and contractility. Investigative studies were conducted to delineate the mechanisms of toxicity. Microarray analysis of Sprague-Dawley rat hearts in a 6 day repeat dose study with PF-04254644 or milrinone, a selective PDE3 inhibitor, revealed similar perturbation of the cyclic adenosine monophosphate (c-AMP) pathway. PDE inhibition and activation of c-AMP were further substantiated using PDE3B immunofluorescence staining and through a c-AMP response element reporter gene assay. The intracellular calcium and oxidative stress signaling pathways were more perturbed by treatment with PF-04254644 than milrinone. The rat cardiomyocytes calcium assay found a dose-dependent increase in intracellular calcium with PF-04254644 treatment. These data suggest that cardiotoxicity of PF-04254644 was probably due to activation of c-AMP signaling, and possibly subsequent disruption of intracellular calcium and oxidative stress signaling pathways. The greater response with PF-04254644 as compared with milrinone in gene expression and micro- and ultrastructural changes is probably due to the broader panel of PDEs inhibition. Copyright © 2012 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]
- Published
- 2012
- Full Text
- View/download PDF
28. Retina Expression and Cross-Species Validation of Gene Silencing by PF-655, a Small Interfering RNA Against RTP801 for the Treatment of Ocular Disease.
- Author
-
Lee, Dong U., Huang, Wenhu, Rittenhouse, Kay D., and Jessen, Bart
- Subjects
GENE expression ,GENE silencing ,SMALL interfering RNA ,TREATMENT of eye diseases ,OPHTHALMIC drugs ,LABORATORY rats - Abstract
Purpose: PF-655, a synthetic 19-mer siRNA, targeting the RTP801 gene is currently in clinical trials for the treatment of wet age-related macular degeneration and diabetic macular edema. Preclinical studies have shown a dose-related suppression of RTP801 expression in rat disease models. Investigative studies were conducted with PF-655 to validate the Dutch-Belted rabbit as a biologically relevant species for gene silencing to support nonclinical ocular toxicity and continual dosing studies. Methods: Cross-species comparison and DNA sequencing was done to determine the level of homology between PF-655 and rabbit RTP801. Human (HEK 293) and rabbit (SIRC cornea) cell lines were stimulated with CoCl
2 to mimic hypoxic stress (an inducer of RTP801 expression) and treated with PF-655. Taqman-polymerase chain reaction and immunoblot analysis were performed to gauge RTP801 expression in cell culture and rabbit retinas. Results: Sequence analysis showed a 1-base mismatch in the PF-655 targeting site from genomic DNA of Dutch-Belted rabbit and the SIRC cell line, a cornea cell derived from the New Zealand White rabbit. HEK and SIRC CoCl2 -stressed cells induced RTP801 expression 10-20-fold above control conditions. Treatment with 20 or 100 nM PF-655 showed a decrease in gene expression, 40%-50% relative to appropriate controls. RTP801 mRNA was detectable in primary rabbit retina tissues, with cycle threshold values showing a large linear range for the assay. Conclusion: These results support our investigation into cross-species validation of gene suppression by a therapeutic siRNA designed to a human gene. The SIRC cell line was utilized as a surrogate to test the degree of RTP801 gene silencing induced by PF-655 in vitro. With a 1-base mismatch, the level of silencing in a rabbit ocular cell line was comparable to that of a human cell line. Sequence analysis and expression data confirmed the relevance of the RTP801 target gene in rabbits and the utility of this species as a relevant animal model. Additionally, our work outlines a tractable method that validates relevant larger non-rodent species for ophthalmic drug testing. [ABSTRACT FROM AUTHOR]- Published
- 2012
- Full Text
- View/download PDF
29. Off-target immune cell toxicity caused by AG-012986, a pan-CDK inhibitor, is associated with inhibition of p38 MAPK phosphorylation.
- Author
-
Lee, Dong U. and Jessen, Bart
- Abstract
AG-012986 is a pan-CDK (cyclin-dependent kinase) inhibitor that has in vitro and in vivo antitumor properties but was stopped in development due in part to rapid bone-marrow-independent white blood cell toxicity in preclinical studies and the potential for acute and delayed immunosuppression in humans. Because peripheral lymphocytes are largely nonproliferating, it was hypothesized the toxicity of AG-012986 was due to an off-target mechanism and not driven by the intended pharmacology. We show the toxicity mechanism in primary human immune cells is caspase driven. T-cells treated with AG-012986 and acutely stimulated through the T-cell receptor exhibited decreased toxicity while still maintaining cell division inhibition. This indicated that the pharmacology of AG-012986 functioned as expected but the toxicity had now been decoupled through activation. Induced phosphorylation of p38 and IL-2 production was impaired with AG-012986. Thus, AG-012986 could cause apoptosis of T-cells by targeting upstream kinases in the p38 Mitogen-activated protein kinase (MAPK) pathway and impairing cellular survival. © 2012 Wiley Periodicals, Inc. J Biochem Mol Toxicol 26:101-108 2012; View this article online at . DOI 10.1002/jbt.20415 [ABSTRACT FROM AUTHOR]
- Published
- 2012
- Full Text
- View/download PDF
30. Peripheral white blood cell toxicity induced by broad spectrum cyclin-dependent kinase inhibitors.
- Author
-
Jessen, Bart A., Leo Lee, Koudriakova, Tatiana, Haines, Morgan, Lundgren, Karen, Price, Sharon, Nonomiya, Jim, Lewis, Cristina, and Stevens, Gregory J.
- Subjects
LEUCOCYTES ,CYCLIN-dependent kinases ,PROTEIN kinases ,IMMUNOSUPPRESSION ,IMMUNOREGULATION ,BONE marrow ,BLOOD cells ,TOXICOLOGY ,TOXICITY testing - Abstract
The article relates a study which characterizes an unusual bone marrow-independent acute toxicity toward leukocytes from broad spectrum cyclin-dependent kinase (CDK) inhibitors in monkeys and rodents. It was noted that CDK inhibitors are expected to slow the rate of cell division and potentially increase the apoptotic fraction of rapidly dividing cells. In the study, the potential combination of both acute and delayed immunosuppression would restrict the application of the compounds. It was assumed in the study that the toxicity was not driven by the intended pharmacological mechanism to facilitate the development of a testing strategy to identify compounds with a reduced potential for leukocyte toxicity.
- Published
- 2007
- Full Text
- View/download PDF
31. Expression profiling during adipocyte differentiation of 3T3-L1 fibroblasts
- Author
-
Jessen, Bart A. and Stevens, Greg J.
- Subjects
- *
FAT cells , *GENE expression , *DNA microarrays - Abstract
The 3T3-L1 cell line is a well-established and commonly used in vitro model to assess adipocyte differentiation. Over the course of several days confluent 3T3-L1 cells can be converted to adipocytes in the presence of an adipogenic cocktail. Changes in gene expression were measured by DNA microarrays at three time points (24 h, 4 days, and 1 week) during the course of differentiation from preadipocytes to mature adipocytes. Several functional categories of genes were affected by adipocyte conversion. In addition, seven genes were found to be commonly altered by 5-fold or more by adipocyte conversion at all three time points. Lipocalin 2, haptoglobin, serum amyloid A3, stearoyl-CoA desaturase, and 11β-hydroxysteroid dehydrogenase 1 were induced while actin α2 and procollagen VIII α1 were suppressed by adipocyte differentiation. Further study of the regulation of these genes and pathways will lead to an increased understanding of the biochemical pathways involved in adipocyte differentiation and possibly to the identification of new therapeutic targets for treatment of obesity and other metabolic diseases. [Copyright &y& Elsevier]
- Published
- 2002
- Full Text
- View/download PDF
32. The Contribution of Lysosomotropism to Autophagy Perturbation.
- Author
-
Ashoor, Roshan, Yafawi, Rolla, Jessen, Bart, and Lu, Shuyan
- Subjects
CELL metabolism ,LYSOSOMES ,AUTOPHAGY ,EUKARYOTIC cells ,CYTOPLASM ,DRUG delivery devices ,HOMEOSTASIS - Abstract
Autophagy refers to the catabolic process in eukaryotic cells that delivers cytoplasmic material to lysosomes for degradation. This highly conserved process is involved in the clearance of long-lived proteins and damaged organelles. Consequently, autophagy is important in providing nutrients to maintain cellular function under starvation, maintaining cellular homeostasis, and promoting cell survival under certain conditions. Several pathways, including mTOR, have been shown to regulate autophagy. However, the impact of lysosomal function impairment on the autophagy process has not been fully explored. Basic lipophilic compounds can accumulate in lysosomes via pH partitioning leading to perturbation of lysosomal function. Our hypothesis is that these types of compounds can disturb the autophagy process. Eleven drugs previously shown to accumulate in lysosomes were selected and evaluated for their effects on cytotoxicity and autophagy using ATP depletion and LC3 assessment, respectively. All eleven drugs induced increased staining of endogenous LC3 and exogenous GFP-LC3, even at non toxic dose levels. In addition, an increase in the abundance of SQSTM1/p62 by all tested compounds denotes that the increase in LC3 is due to autophagy perturbation rather than enhancement. Furthermore, the gene expression profile resulting from in vitro treatment with these drugs revealed the suppression of plentiful long-lived proteins, including structural cytoskeletal and associated proteins, and extracellular matrix proteins. This finding indicates a retardation of protein turnover which further supports the notion of autophagy inhibition. Interestingly, upregulation of genes containing antioxidant response elements, e.g. glutathione S transferase and NAD(P)H dehydrogenase quinone 1 was observed, suggesting activation of Nrf2 transcription factor. These gene expression changes could be related to an increase in SQSTM1/p62 resulting from autophagy deficiency. In summary, our data indicate that lysosomal accumulation due to the basic lipophilic nature of xenobiotics could be a general mechanism contributing to the perturbation of the autophagy process. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
33. Lysosome Impairment, Phagocytosis Inhibition and Retinopathy.
- Author
-
Shuyan Lu, Sokolowski, Sharon A., Blasi, Eileen, and Jessen, Bart
- Abstract
Retina is a highly compact neural structure composed of distinct layers. The retinal pigmented epithelium (RPE) is the pigmented cell layer just outside the neurosensory and serves a number of crucial metabolic functions including the phagocytosis and removal of shed photoreceptor cell outer segments. Drugs with a basic moiety can accumulate in the lysososme and disturb lysosome function including phagocytosis, which will retard the clearance of outer segments and ultimately induce retinopathy. To test this hypothesis we chose a compound that contains a basic moiety and induced retinopathy in long term in vivo studies in rats In vitro assays including viability, LC-3 staining, and phagocytosis were conducted using the human retinal pigmented epithelium cell line ARPE-19. These studies showed that this compound caused cellular vacuolation and dose dependent cytotoxicity that was related to the initial cell number and dosing volume, indicating an accumulation effect. This compound also decreased phagocytosis function of ARPE-19, and induced accumulation of LC-3, suggesting a blockage of the autophagy process. EM analysis of eyes taken from an in vivo study showed the presence of large phagolysosomes, supporting the hypothesis of impairment of lysosome function as a mechanism for retinal toxicity. Twenty three proprietary compounds for which the in vivo effects on the retina were known were tested in a phagocytosis assay to examine the prevalence of this cellular mechanism in retinal toxicity. A correlation amongst the physiochemical properties, in vitro lysosomal pathway effects, and in vivo retinal effects was established. This data reveals the importance of physiochemical properties and lysosome accumulation as a common mechanism for drug induced retinopathy and demonstrates the potential usefulness of in vitro screening in predicting this liability. [ABSTRACT FROM AUTHOR]
- Published
- 2010
- Full Text
- View/download PDF
34. Microphysiological systems in early stage drug development: Perspectives on current applications and future impact.
- Author
-
Kopec, Anna K., Ryuji Yokokawa, Khan, Nasir, Ikuo Horii, Finley, James E., Bono, Christine P., Donovan, Carol, Roy, Jessica, Harney, Julie, Burdick, Andrew D., Jessen, Bart, Shuyan Lu, Collinge, Mark, Sadeghian, Ramin Banan, Derzi, Mazin, Tomlinson, Lindsay, and Burkhardt, John E.
- Subjects
- *
DRUG development , *CARDIOVASCULAR system , *IN vivo studies , *LABORATORY animals , *PHYSIOLOGICAL models , *ORGANS (Anatomy) - Abstract
Microphysiological systems (MPS) are making advances to provide more standardized and predictive physiologically relevant responses to test articles in living tissues and organ systems. The excitement surrounding the potential of MPS to better predict human responses to medicines and improving clinical translation is overshadowed by their relatively slow adoption by the pharmaceutical industry and regulators. Collaboration between multiorganizational consortia and regulators is necessary to build an understanding of the strengths and limitations of MPS models and closing the current gaps. Here, we review some of the advances in MPS research, focusing on liver, intestine, vascular system, kidney and lung and present examples highlighting the context of use for these systems. For MPS to gain a foothold in drug development, they must have added value over existing approaches. Ideally, the application of MPS will augment in vivo studies and reduce the use of animals via tiered screening with less reliance on exploratory toxicology studies to screen compounds. Because MPS support multiple cell types (e.g. primary or stem-cell derived cells) and organ systems, identifying when MPS are more appropriate than simple 2D in vitro models for understanding physiological responses to test articles is necessary. Once identified, MPS models require qualification for that specific context of use and must be reproducible to allow future validation. Ultimately, the challenges of balancing complexity with reproducibility will inform the promise of advancing the MPS field and are critical for realization of the goal to reduce, refine and replace (3Rs) the use of animals in nonclinical research. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
35. Antisense inhibition of 11βhydroxysteroid dehydrogenase type 1 improves diabetes in a novel cortisone-induced diabetic KK mouse model
- Author
-
Bhat, B. Ganesh, Younis, Husam, Herrera, Jocelyn, Palacio, Kimbie, Pascual, Bernadette, Hur, George, Jessen, Bart, Ogilvie, Kathleen M., and Rejto, Paul A.
- Subjects
- *
DIABETES , *ENDOCRINE diseases , *CARBOHYDRATE intolerance , *HYDROCORTISONE - Abstract
Abstract: The inhibition of 11βhydroxysteroid dehydrogenase 1 (11βHSD1), an enzyme that catalyzes the conversion of inactive cortisone to active cortisol, is an attractive target to treat diabetes by suppressing hepatic gluconeogenesis. To test this hypothesis, we developed a novel glucocorticoid-induced diabetic KK mouse model and used 11βHSD1 antisense oligonucleotide (ASO) as an inhibitory tool. KK mice were treated with 25 or 50mg/kg/day of 11βHSD1 ASO for 28 days. On day 25, cortisone pellets were surgically implanted to induce diabetes. In the ASO-treated mice, plasma blood glucose levels were significantly reduced by up to 54%. In parallel, cortisol and other diabetes endpoints were also significantly reduced. Hepatic 11βHSD1 mRNA was suppressed by up to 84% with a concomitant respective decrease of up to 49% in the expression of PEPCK. The results suggest that inhibition of 11βHSD1 activity reduces the availability of cortisol to activate the glucocorticoid receptor, down regulates gluconeogenesis and thus reduces plasma glucose levels in cortisone-induced diabetic KK mice. [Copyright &y& Elsevier]
- Published
- 2008
- Full Text
- View/download PDF
36. Pharmacologic Properties and Preclinical Activity of Sasanlimab, A High-affinity Engineered Anti-Human PD-1 Antibody.
- Author
-
Al-Khami AA, Youssef S, Abdiche Y, Nguyen H, Chou J, Kimberlin CR, Chin SM, Kamperschroer C, Jessen B, Kern B, Budimir N, Dillon CP, Xu A, Clark JD, Chou J, Kraynov E, Rajpal A, Lin JC, and Salek-Ardakani S
- Subjects
- Animals, Cell Line, Tumor, Humans, Immune Checkpoint Inhibitors pharmacology, Mice, Immune Checkpoint Inhibitors therapeutic use
- Abstract
Development of antagonistic mAbs that specifically target the immune checkpoint receptor, programmed cell death protein-1 (PD-1), is of great interest for cancer immunotherapy. Here, we report the biophysical characteristics and nonclinical antagonistic activities of sasanlimab (PF-06801591), a humanized anti-PD-1 antibody of IgG4 isotype. We show that sasanlimab binds selectively and with similar high potency to human and cynomolgus monkey PD-1 receptor and blocks its interaction with PD-L1 and PD-L2, with no detectable Fc-dependent effector function. The binding of sasanlimab to human and cynomolgus PD-1 is associated with the formation of a stable complex, which is likely to be the main driver of this high-affinity interaction. In vitro , sasanlimab significantly augmented T-cell proliferation and cytokine production in mixed lymphocyte reaction and superantigen stimulation assays. In vivo , sasanlimab accelerated the incidence of GvHD by enhancing T-cell proliferation and cytokine secretion in a xenogeneic model of acute GvHD and halted the growth of MC-38 colon adenocarcinoma tumors in human PD-1 knock-in mice. Pharmacokinetic and toxicokinetic findings from cynomolgus monkey showed that sasanlimab was active and well-tolerated. Taken together, the data presented here support the clinical development of sasanlimab for the treatment of patients with advanced cancers as a single agent or in combination with other immunotherapies., (©2020 American Association for Cancer Research.)
- Published
- 2020
- Full Text
- View/download PDF
37. Intestinal Toxicity in Rats Following Administration of CDK4/6 Inhibitors Is Independent of Primary Pharmacology.
- Author
-
Thibault S, Hu W, Hirakawa B, Kalabat D, Franks T, Sung T, Khoh-Reiter S, Lu S, Finkelstein M, Jessen B, and Sacaan A
- Subjects
- Aminopyridines adverse effects, Animals, Benzimidazoles adverse effects, Cyclin-Dependent Kinase 4 antagonists & inhibitors, Cyclin-Dependent Kinase 6 antagonists & inhibitors, Diarrhea genetics, Diarrhea metabolism, Disease Models, Animal, Gene Expression Regulation drug effects, Male, Piperazines adverse effects, Protein Kinase Inhibitors adverse effects, Purines adverse effects, Pyridines adverse effects, Rats, Rats, Sprague-Dawley, Aminopyridines administration & dosage, Benzimidazoles administration & dosage, Diarrhea chemically induced, Piperazines administration & dosage, Protein Kinase Inhibitors administration & dosage, Purines administration & dosage, Pyridines administration & dosage
- Abstract
Recently three different cyclin-dependent kinase 4 and 6 (CDK4/6) dual inhibitors were approved for the treatment of breast cancer (palbociclib, ribociclib, and abemaciclib), all of which offer comparable therapeutic benefits. Their safety profiles, however, are different. For example, neutropenia is observed at varying incidences in patients treated with these drugs; however, it is the most common adverse event for palbociclib and ribociclib, whereas diarrhea is the most common adverse event observed in patients treated with abemaciclib. To understand the mechanism of diarrhea observed with these drugs and in an effort to guide the development of safer drugs, we compared the effects of oral administration of palbociclib, ribociclib, and abemaciclib on the gastrointestinal tract of rats using doses intended to produce comparable CDK4/6 inhibition. Rats administered abemaciclib, but not palbociclib or ribociclib, had fecal alterations, unique histopathologic findings, and distinctive changes in intestinal gene expression. Morphologic changes in the intestine were characterized by proliferation of crypt cells, loss of goblet cells, poorly differentiated and degenerating enterocytes with loss of microvilli, and mucosal inflammation. In the jejunum of abemaciclib-treated rats, downregulation of enterocyte membrane transporters and upregulation of genes associated with cell proliferation were observed, consistent with activation of the Wnt pathway and downstream transcriptional regulation. Among these CDK4/6 inhibitors, intestinal toxicity was unique to rats treated with abemaciclib, suggesting a mechanism of toxicity not due to primary pharmacology (CDK4/6 inhibition), but to activity at secondary pharmacologic targets., (©2018 American Association for Cancer Research.)
- Published
- 2019
- Full Text
- View/download PDF
38. Mechanistic Investigation of Bone Marrow Suppression Associated with Palbociclib and its Differentiation from Cytotoxic Chemotherapies.
- Author
-
Hu W, Sung T, Jessen BA, Thibault S, Finkelstein MB, Khan NK, and Sacaan AI
- Subjects
- Animals, Apoptosis drug effects, Blood Cell Count, Bone Marrow Cells drug effects, Bone Marrow Cells metabolism, Cell Differentiation drug effects, Cell Proliferation, Cell Survival drug effects, Cellular Senescence drug effects, Dogs, Drug Synergism, Estradiol analogs & derivatives, Estradiol pharmacology, Female, Fulvestrant, Hematopoietic Stem Cells cytology, Hematopoietic Stem Cells drug effects, Hematopoietic Stem Cells metabolism, Humans, Male, Models, Animal, Antineoplastic Agents pharmacology, Bone Marrow drug effects, Bone Marrow pathology, Hematopoiesis drug effects, Piperazines pharmacology, Protein Kinase Inhibitors pharmacology, Pyridines pharmacology
- Abstract
Purpose: Palbociclib (PD-0332991) is the first selective cyclin-dependent kinase (CDK) 4/6 inhibitor approved for metastatic breast cancer. Hematologic effects, especially neutropenia, are dose-limiting adverse events for palbociclib in humans., Experimental Design: Reversible hematologic effects and bone marrow hypocellularity have been identified in toxicology studies in rats and dogs after palbociclib treatment. To understand the mechanism by which the hematologic toxicity occurs, and to further differentiate it from the myelotoxicity caused by cytotoxic chemotherapeutic agents, anin vitroassay using human bone marrow mononuclear cells (hBMNC) was utilized., Results: This work demonstrated that palbociclib-induced bone marrow suppression occurred through cell-cycle arrest, with no apoptosis at clinically relevant concentrations, was not lineage-specific, and was reversible upon palbociclib withdrawal. In contrast, treatment with chemotherapeutic agents (paclitaxel and doxorubicin) resulted in DNA damage and apoptotic cell death in hBMNCs. In the presence or absence of the antiestrogen, palbociclib-treated hBMNCs did not become senescent and resumed proliferation following palbociclib withdrawal, consistent with pharmacologic quiescence. The breast cancer cells, MCF-7, conversely, became senescent following palbociclib or antiestrogen treatment with additive effects in combination and remained arrested in the presence of antiestrogen., Conclusions: Palbociclib causes reversible bone marrow suppression, clearly differentiating it from apoptotic cell death caused by cytotoxic chemotherapeutic agents. This study also distinguished the cell-cycle arresting action of palbociclib on normal bone marrow cells from the senescent effects observed in breast cancer cells. These results shed light on the mechanism and support risk management of palbociclib-induced bone marrow toxicity in the clinic., (©2015 American Association for Cancer Research.)
- Published
- 2016
- Full Text
- View/download PDF
39. Combination of 4-1BB agonist and PD-1 antagonist promotes antitumor effector/memory CD8 T cells in a poorly immunogenic tumor model.
- Author
-
Chen S, Lee LF, Fisher TS, Jessen B, Elliott M, Evering W, Logronio K, Tu GH, Tsaparikos K, Li X, Wang H, Ying C, Xiong M, VanArsdale T, and Lin JC
- Subjects
- Animals, Antibodies, Monoclonal administration & dosage, Antibodies, Monoclonal immunology, Antigens, CD immunology, CD8-Positive T-Lymphocytes immunology, Cell Differentiation immunology, Cytotoxicity, Immunologic immunology, Female, Immunologic Memory immunology, Immunophenotyping methods, Immunotherapy methods, Interferon-gamma immunology, Liver enzymology, Melanoma, Experimental pathology, Melanoma, Experimental therapy, Mice, Inbred C57BL, Tumor Microenvironment immunology, Lymphocyte Activation Gene 3 Protein, Antineoplastic Combined Chemotherapy Protocols immunology, CD8-Positive T-Lymphocytes drug effects, Melanoma, Experimental immunology, Programmed Cell Death 1 Receptor antagonists & inhibitors, Tumor Necrosis Factor Receptor Superfamily, Member 9 agonists
- Abstract
Immunotherapies targeting the programmed death 1 (PD-1) coinhibitory receptor have shown great promise for a subset of patients with cancer. However, robust and safe combination therapies are still needed to bring the benefit of cancer immunotherapy to broader patient populations. To search for an optimal strategy of combinatorial immunotherapy, we have compared the antitumor activity of the anti-4-1BB/anti-PD-1 combination with that of the anti-PD-1/anti-LAG-3 combination in the poorly immunogenic B16F10 melanoma model. Pronounced tumor inhibition occurred only in animals receiving anti-PD-1 and anti-4-1BB concomitantly, while combining anti-PD-1 with anti-LAG-3 led to a modest degree of tumor suppression. The activity of the anti-4-1BB/anti-PD-1 combination was dependent on IFNγ and CD8(+) T cells. Both 4-1BB and PD-1 proteins were elevated on the surface of CD8(+) T cells by anti-4-1BB/anti-PD-1 cotreatment. In the tumor microenvironment, an effective antitumor immune response was induced as indicated by the increased CD8(+)/Treg ratio and the enrichment of genes such as Cd3e, Cd8a, Ifng, and Eomes. In the spleen, the combination treatment shaped the immune system to an effector/memory phenotype and increased the overall activity of tumor-specific CD8(+) CTLs, reflecting a long-lasting systemic antitumor response. Furthermore, combination treatment in C57BL/6 mice showed no additional safety signals, and only minimally increased severity of the known toxicity relative to 4-1BB agonist alone. Therefore, in the absence of any cancer vaccine, anti-4-1BB/anti-PD-1 combination therapy is sufficient to elicit a robust antitumor effector/memory T-cell response in an aggressive tumor model and is therefore a candidate for combination trials in patients., (©2014 American Association for Cancer Research.)
- Published
- 2015
- Full Text
- View/download PDF
40. Evaluation of the effects of a VEGFR-2 inhibitor compound on alanine aminotransferase gene expression and enzymatic activity in the rat liver.
- Author
-
Fuentealba C, Bera M, Jessen B, Sace F, Stevens GJ, Trajkovic D, Yang AH, and Evering W
- Abstract
Background: Traditional assessment of drug-induced hepatotoxicity includes morphological examination of the liver and evaluation of liver enzyme activity in serum. The objective of the study was to determine the origin of drug-related elevation in serum alanine aminotransferase (ALT) activity in the absence of morphologic changes in the liver by utilizing molecular and immunohistochemical techniques., Methods: Sixteen female Sprague-Dawley rats were divided into 2 groups (control and treated, n = 4 per group) and treated rats were dosed orally twice daily (400 mg/kg/day) for 7 days with a VEGFR-2 compound (AG28262), which in a previous study caused ALT elevation without morphological changes. Serum of both treated and control animals were evaluated on day 3 of treatment and at day 8. Three separate liver lobes (caudate, right medial, and left lateral) were examined for determination of ALT tissue activity, ALT gene expression and morphological changes., Results: ALT activity was significantly (p < 0.01) elevated on day 3 and further increased on day 8. Histologic changes or increase in TUNEL and caspase3 positive cells were not observed in the liver lobes examined. ALT gene expression in the caudate lobe was significantly up-regulated by 63%. ALT expression in the left lateral lobe was not significantly affected. Statistically significant increased liver ALT enzymatic activity occurred in the caudate (96%) and right medial (41%) lobes but not in the left lateral lobe., Conclusions: AG28262, a VEFG-r2 inhibitor, causes an increase in serum ALT, due in part to both gene up-regulation. Differences between liver lobes may be attributable to differential distribution of blood from portal circulation. Incorporation of molecular data, such as gene and protein expression, and sampling multiple liver lobes may shed mechanistic insight to the evaluation of hepatotoxicity.
- Published
- 2011
- Full Text
- View/download PDF
41. Evaluation of the cytotoxic effects of ophthalmic solutions containing benzalkonium chloride on corneal epithelium using an organotypic 3-D model.
- Author
-
Khoh-Reiter S and Jessen BA
- Subjects
- Animals, Anti-Allergic Agents adverse effects, Antihypertensive Agents adverse effects, Benzalkonium Compounds administration & dosage, Cell Line, Cloprostenol adverse effects, Cloprostenol analogs & derivatives, Dibenzoxepins adverse effects, Drug Evaluation, Epithelium, Corneal pathology, Humans, Latanoprost, Macaca fascicularis, Models, Biological, Olopatadine Hydrochloride, Ophthalmic Solutions administration & dosage, Preservatives, Pharmaceutical administration & dosage, Prostaglandins F, Synthetic adverse effects, Time Factors, Travoprost, Benzalkonium Compounds adverse effects, Epithelium, Corneal drug effects, Ophthalmic Solutions adverse effects, Preservatives, Pharmaceutical adverse effects
- Abstract
Background: Benzalkonium chloride (BAC) is a common preservative used in ophthalmic solutions. The aim of this study was to compare the cytotoxic effects of BAC-containing ophthalmic solutions with a BAC-free ophthalmic solution using an organotypic 3-dimensional (3-D) corneal epithelial model and to determine the effects of latanoprost ophthalmic solution and its BAC-containing vehicle on corneal thickness in a monkey model., Methods: The cytotoxicity of commercially available BAC-containing ophthalmic formulations of latanoprost (0.02% BAC) and olopatadine (0.01% BAC) was compared to that of BAC-free travoprost and saline in a corneal organotypic 3-D model using incubation times of 10 and 25 minutes. To compare the extent of differentiation of 3-D corneal cultures to monolayer transformed human corneal epithelial (HCE-T) cell cultures, expression levels (mRNA and protein) of the corneal markers epidermal growth factor receptor, transglutaminase 1 and involucrin were quantified. Finally, latanoprost ophthalmic solution or its vehicle was administered at suprapharmacologic doses (two 30 microL drops twice daily in 1 eye for 1 year) in monkey eyes, and corneal pachymetry was performed at baseline and at weeks 4, 13, 26 and 52., Results: In the 3-D corneal epithelial culture assays, there were no significant differences in cytotoxicity between the BAC-containing latanoprost and olopatadine ophthalmic solutions and BAC-free travoprost ophthalmic solution at either the 10- or 25-minute time points. The 3-D cultures expressed higher levels of corneal epithelial markers than the HCE-T monolayers, indicating a greater degree of differentiation. There were no significant differences between the corneal thickness of monkey eyes treated with latanoprost ophthalmic solution or its vehicle (both containing 0.02% BAC) and untreated eyes., Conclusion: The lack of cytotoxicity demonstrated in 3-D corneal cultures and in monkey studies suggests that the levels of BAC contained in ophthalmic solutions are not likely to cause significant direct toxicity to epithelium of otherwise normal corneas.
- Published
- 2009
- Full Text
- View/download PDF
42. Pharmacologic properties of AG-012986, a pan-cyclin-dependent kinase inhibitor with antitumor efficacy.
- Author
-
Zhang C, Lundgren K, Yan Z, Arango ME, Price S, Huber A, Higgins J, Troche G, Skaptason J, Koudriakova T, Nonomiya J, Yang M, O'Connor P, Bender S, Los G, Lewis C, and Jessen B
- Subjects
- Animals, Benzamides pharmacokinetics, Blotting, Western, Cell Cycle drug effects, Colonic Neoplasms metabolism, Colonic Neoplasms pathology, Colony-Forming Units Assay, Humans, Immunoenzyme Techniques, In Situ Nick-End Labeling, Mice, Mice, Nude, Mice, SCID, Phosphorylation drug effects, Retinoblastoma Protein metabolism, Thiazoles pharmacokinetics, Tissue Distribution, Tumor Cells, Cultured, Xenograft Model Antitumor Assays, Antineoplastic Agents pharmacology, Apoptosis drug effects, Benzamides pharmacology, Cell Proliferation drug effects, Colonic Neoplasms drug therapy, Cyclin-Dependent Kinases antagonists & inhibitors, Thiazoles pharmacology
- Abstract
AG-012986 is a multitargeted cyclin-dependent kinase (CDK) inhibitor active against CDK1, CDK2, CDK4/6, CDK5, and CDK9, with selectivity over a diverse panel of non-CDK kinases. Here, we report the potent antitumor efficacies of AG-012986 against multiple tumor lines in vitro and in vivo. AG-012986 showed antiproliferative activities in vitro with IC(50)s of <100 nmol/L in 14 of 18 tumor cell lines. In vivo, significant antitumor efficacy induced by AG-012986 was seen (tumor growth inhibition, >83.1%) in 10 of 11 human xenograft tumor models when administered at or near the maximum tolerated dose for 8 or 12 days. AG-012986 caused dose-dependent hypophosphorylation at Ser(795) of the retinoblastoma protein, cell cycle arrest, and apoptosis in vitro. Colony-forming assays indicated that the potency of AG-012986 substantially decreased with treatment time of <24 h. In vivo, AG-012986 also showed dose-dependent retinoblastoma Ser(795) hypophosphorylation, cell cycle arrest, decreased Ki-67 tumor staining, and apoptosis in conjunction with antitumor activity. Studies comparing i.p. bolus with s.c. implanted minipump dosing regimens revealed that in vivo efficacy correlated with the duration of minimally effective plasma levels rather than maximal drug plasma levels. Dosing optimization of AG-012986 provided guidance for selecting a treatment schedule to achieve the best antitumor efficacy while minimizing the risk of adverse side effects.
- Published
- 2008
- Full Text
- View/download PDF
43. Inhibiting matrix metalloproteinases with prinomastat produces abnormalities in fetal growth and development in rats.
- Author
-
Younis HS, Jessen BA, Wu EY, and Stevens GJ
- Subjects
- Animals, Body Weight drug effects, Bone and Bones abnormalities, Eating drug effects, Embryo Culture Techniques, Embryo Loss chemically induced, Embryo, Mammalian abnormalities, Embryo, Mammalian drug effects, Enzyme Inhibitors pharmacokinetics, Female, Fetus drug effects, Fetus enzymology, Organ Size drug effects, Organic Chemicals pharmacokinetics, Pregnancy, Rats, Rats, Sprague-Dawley, Uterus anatomy & histology, Uterus drug effects, Enzyme Inhibitors toxicity, Fetus abnormalities, Matrix Metalloproteinase Inhibitors, Organic Chemicals toxicity
- Abstract
Background: Matrix metalloproteinases (MMPs) play key roles in remodeling of the extracellular matrix during embryogenesis and fetal development. The objective of this study was to determine the effects of prinomastat, a potent selective MMP inhibitor, on fetal growth and development., Methods: Prinomastat (25, 100, 250 mg/kg/day, p.o.) was administered to pregnant female Sprague-Dawley rats on gestational days (GD) 6-17. A Cesarian section was carried out on GD 20 and the fetuses were evaluated for viability and skeletal and soft tissue abnormalities., Results: Prinomastat treatment at the 250 mg/kg/day dose produced a decrease in body weight and food consumption in the dams. A dose-dependent increase in post-implantation loss was observed in the 100 and 250 mg/kg/day-dose groups, resulting in only 22% of the dams having viable litters for evaluation at the 250 mg/kg/day dose. Fetal skeletal tissue variations and malformations were present in all prinomastat treated groups and their frequency increased with dose. Variations and malformation in fetal soft tissue were also increased at the 100 and 250 mg/kg/day doses. Prinomastat also interfered with fetal growth of rat embryo cultures in vitro., Conclusions: These data confirm that MMP inhibition has a profound effect on fetal growth and development in vivo and in vitro., (Birth Defects Research (Part B) 77:95-103, 2006. (c) 2006 Wiley-Liss, Inc.)
- Published
- 2006
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.