42 results on '"Michael M. Morrissey"'
Search Results
2. Supplementary Figure 3 from Demonstration of a Genetic Therapeutic Index for Tumors Expressing Oncogenic BRAF by the Kinase Inhibitor SB-590885
- Author
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Pearl S. Huang, David A. Tuveson, Michael M. Morrissey, Meenhard Herlyn, Keiran S.M. Smalley, Rooja G. Contractor, Jerry L. Adams, Ami S. Lakdawala, Lusong Luo, Michael D. Schaber, Cynthia M. Rominger, Earl W. May, Thomas J. Stout, David Chau, Jae Lee, Julie C. Lougheed, Florian Karreth, Lifu Wang, Erin Hugger, David M. Wilson, Andrew K. Takle, David W. Rusnak, Rakesh Kumar, Shu Yun Zhang, Hieu T. Do, Maureen L. Ho, Roberta S. Batorsky, Denis R. Patrick, and Alastair J. King
- Abstract
Supplementary Figure 3 from Demonstration of a Genetic Therapeutic Index for Tumors Expressing Oncogenic BRAF by the Kinase Inhibitor SB-590885
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- 2023
3. Supplementary Figure 1 from Demonstration of a Genetic Therapeutic Index for Tumors Expressing Oncogenic BRAF by the Kinase Inhibitor SB-590885
- Author
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Pearl S. Huang, David A. Tuveson, Michael M. Morrissey, Meenhard Herlyn, Keiran S.M. Smalley, Rooja G. Contractor, Jerry L. Adams, Ami S. Lakdawala, Lusong Luo, Michael D. Schaber, Cynthia M. Rominger, Earl W. May, Thomas J. Stout, David Chau, Jae Lee, Julie C. Lougheed, Florian Karreth, Lifu Wang, Erin Hugger, David M. Wilson, Andrew K. Takle, David W. Rusnak, Rakesh Kumar, Shu Yun Zhang, Hieu T. Do, Maureen L. Ho, Roberta S. Batorsky, Denis R. Patrick, and Alastair J. King
- Abstract
Supplementary Figure 1 from Demonstration of a Genetic Therapeutic Index for Tumors Expressing Oncogenic BRAF by the Kinase Inhibitor SB-590885
- Published
- 2023
4. Data from Demonstration of a Genetic Therapeutic Index for Tumors Expressing Oncogenic BRAF by the Kinase Inhibitor SB-590885
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Pearl S. Huang, David A. Tuveson, Michael M. Morrissey, Meenhard Herlyn, Keiran S.M. Smalley, Rooja G. Contractor, Jerry L. Adams, Ami S. Lakdawala, Lusong Luo, Michael D. Schaber, Cynthia M. Rominger, Earl W. May, Thomas J. Stout, David Chau, Jae Lee, Julie C. Lougheed, Florian Karreth, Lifu Wang, Erin Hugger, David M. Wilson, Andrew K. Takle, David W. Rusnak, Rakesh Kumar, Shu Yun Zhang, Hieu T. Do, Maureen L. Ho, Roberta S. Batorsky, Denis R. Patrick, and Alastair J. King
- Abstract
Oncogenic BRAF alleles are both necessary and sufficient for cellular transformation, suggesting that chemical inhibition of the activated mutant protein kinase may reverse the tumor phenotype. Here, we report the characterization of SB-590885, a novel triarylimidazole that selectively inhibits Raf kinases with more potency towards B-Raf than c-Raf. Crystallographic analysis revealed that SB-590885 stabilizes the oncogenic B-Raf kinase domain in an active configuration, which is distinct from the previously reported mechanism of action of the multi-kinase inhibitor, BAY43-9006. Malignant cells expressing oncogenic B-Raf show selective inhibition of mitogen-activated protein kinase activation, proliferation, transformation, and tumorigenicity when exposed to SB-590885, whereas other cancer cell lines and normal cells display variable sensitivities or resistance to similar treatment. These studies support the validation of oncogenic B-Raf as a target for cancer therapy and provide the first evidence of a correlation between the expression of oncogenic BRAF alleles and a positive response to a selective B-Raf inhibitor. (Cancer Res 2006; 66(23): 11100-5)
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- 2023
5. Reversible, orally available ADP receptor (P2Y12) antagonists Part I: Hit to lead process
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Shendong Yuan, Robert G. Wei, Andrea DiMella, Michael Snider, Dewan Zheng, Babu Subramanyam, Imadul Islam, Raju Mohan, Michael M. Morrissey, Brad O. Buckman, Laura Dunning, Wei Xu, Jih-Lie Tseng, and Judi Bryant
- Subjects
0301 basic medicine ,P2Y receptor ,Chemistry ,High-throughput screening ,Organic Chemistry ,Clinical Biochemistry ,Antagonist ,Pharmaceutical Science ,Hit to lead ,Plasma levels ,030204 cardiovascular system & hematology ,Pharmacology ,Metabolic stability ,Biochemistry ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,P2Y12 ,Drug Discovery ,Molecular Medicine ,Molecular Biology ,Liver microsomes - Abstract
A hit to lead process to identify reversible, orally available ADP receptor (P2Y12) antagonists lead compounds is described. High throughput screening afforded 1. Optimization of 1, using parallel synthesis methods, a methyl scan to identify promising regions for optimization, and exploratory SAR on these regions, provided 22 and 23. Compound 23 is an orally available, competitive reversible antagonist (KB = 94 nM for inhibition of ADP-induced platelet aggregation). It exhibits high metabolic stability in human, rat and dog liver microsomes and is orally absorbed. Although plasma level after oral dosing of 22 and 23 to rats is low, reasonable levels were achieved to merit extensive lead optimization of this structural class.
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- 2018
6. Design, Synthesis, and Activity of 2-Imidazol-1-ylpyrimidine Derived Inducible Nitric Oxide Synthase Dimerization Inhibitors
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Damian Arnaiz, Gary Phillips, Shawn David Erickson, Marc Whitlow, Margaret Kenrick, James J. Devlin, Gonghua Pan, Cecile Santos, William J. Guilford, Robert G. Wei, Marc Adler, Kurt W. Saionz, Bin Ye, Ron Vergona, John Parkinson, Zuchun Spring Zhao, David D. Davey, Michael Ohlmeyer, Babu Subramanyam, Vidyadhar M. Paradkar, Mark A. Polokoff, Keith A. Eagen, and Michael M. Morrissey
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Male ,Models, Molecular ,Gene isoform ,Nitric Oxide Synthase Type II ,Crystallography, X-Ray ,Chemical synthesis ,Cell Line ,Nitric oxide ,chemistry.chemical_compound ,Oxidoreductase ,Chlorocebus aethiops ,Drug Discovery ,Animals ,Benzodioxoles ,Binding site ,chemistry.chemical_classification ,biology ,Chemistry ,Anti-Inflammatory Agents, Non-Steroidal ,Imidazoles ,Biological activity ,Arthritis, Experimental ,Rats ,Nitric oxide synthase ,Pyrimidines ,Enzyme ,Biochemistry ,Rats, Inbred Lew ,biology.protein ,Molecular Medicine ,Dimerization - Abstract
By the screening of a combinatorial library for inhibitors of nitric oxide (NO) formation by the inducible isoform of nitric oxide synthase (iNOS) using a whole-cell assay, 2-(imidazol-1-yl)pyrimidines were identified. Compounds were found to inhibit the dimerization of iNOS monomers, thus preventing the formation of the dimeric, active form of the enzyme. Optimization led to the selection of the potent, selective, and orally available iNOS dimerization inhibitor, 21b, which significantly ameliorated adjuvant-induced arthritis in a rat model. Analysis of the crystal structure of the 21b--iNOS monomer complex provided a rationalization for both the SAR and the mechanism by which 21b blocks the formation of the protein--protein interaction present in the dimeric form of iNOS.
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- 2007
7. Demonstration of a Genetic Therapeutic Index for Tumors Expressing Oncogenic BRAF by the Kinase Inhibitor SB-590885
- Author
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Roberta S. Batorsky, Jerry L. Adams, Shu Yun Zhang, Michael D. Schaber, Rakesh Kumar, Julie Lougheed, Thomas J. Stout, Erin Hugger, Meenhard Herlyn, David Chau, Maureen L. Ho, Ami S. Lakdawala, Earl May, Rooja G. Contractor, Jae Lee, Pearl S. Huang, Andrew K. Takle, Hieu T. Do, Lusong Luo, David M. Wilson, Michael M. Morrissey, Lifu Wang, Cynthia M. Rominger, David A. Tuveson, Alastair J. King, Denis R. Patrick, David W. Rusnak, Keiran S.M. Smalley, and Florian A. Karreth
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Models, Molecular ,Proto-Oncogene Proteins B-raf ,Cancer Research ,Protein Conformation ,Blotting, Western ,Mice, Nude ,Biology ,Crystallography, X-Ray ,medicine.disease_cause ,Mice ,Cell Movement ,Mutant protein ,Cell Line, Tumor ,Neoplasms ,Gene expression ,medicine ,Animals ,Humans ,Phosphorylation ,Extracellular Signal-Regulated MAP Kinases ,Protein Kinase Inhibitors ,Alleles ,Cell Proliferation ,Molecular Structure ,Kinase ,Imidazoles ,Xenograft Model Antitumor Assays ,Molecular biology ,Blot ,Oncology ,Protein kinase domain ,Mechanism of action ,Cell culture ,Mutation ,Cancer research ,Female ,medicine.symptom ,Crystallization ,Carcinogenesis ,HT29 Cells - Abstract
Oncogenic BRAF alleles are both necessary and sufficient for cellular transformation, suggesting that chemical inhibition of the activated mutant protein kinase may reverse the tumor phenotype. Here, we report the characterization of SB-590885, a novel triarylimidazole that selectively inhibits Raf kinases with more potency towards B-Raf than c-Raf. Crystallographic analysis revealed that SB-590885 stabilizes the oncogenic B-Raf kinase domain in an active configuration, which is distinct from the previously reported mechanism of action of the multi-kinase inhibitor, BAY43-9006. Malignant cells expressing oncogenic B-Raf show selective inhibition of mitogen-activated protein kinase activation, proliferation, transformation, and tumorigenicity when exposed to SB-590885, whereas other cancer cell lines and normal cells display variable sensitivities or resistance to similar treatment. These studies support the validation of oncogenic B-Raf as a target for cancer therapy and provide the first evidence of a correlation between the expression of oncogenic BRAF alleles and a positive response to a selective B-Raf inhibitor. (Cancer Res 2006; 66(23): 11100-5)
- Published
- 2006
8. A Non-peptide Functional Antagonist of the CCR1 Chemokine Receptor Is Effective in Rat Heart Transplant Rejection
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Guo Ping Wei, Christian Weber, John G. Bauman, H. Daniel Perez, Ghannam Ameen F, Joseph Hesselgesser, Brad O. Buckman, Michael M. Morrissey, Ng Howard P, R. Michael Snider, Zhaohui Wang, Wei Xu, Richard Horuk, Peter J. Nelson, Imadul Islam, Mary Rosser, Kim S. C. Weber, Hermann Josef Gröne, Carol Clayberger, Alan M. Krensky, Monahan Sean D, Laura Dunning, Meina Liang, and Karen May
- Subjects
Graft Rejection ,Male ,Agonist ,CCR1 ,Chemokine ,medicine.drug_class ,medicine.medical_treatment ,Receptors, CCR1 ,Biology ,Biochemistry ,Cell Line ,Chemokine receptor ,Piperidines ,medicine ,Animals ,Humans ,Molecular Biology ,Heart transplantation ,Phenylurea Compounds ,Graft Survival ,Antagonist ,Cell Biology ,medicine.disease ,Receptor antagonist ,Rats ,Transplant rejection ,Rats, Inbred Lew ,Immunology ,Cyclosporine ,biology.protein ,Heart Transplantation ,Receptors, Chemokine - Abstract
Chemokines like RANTES appear to play a role in organ transplant rejection. Because RANTES is a potent agonist for the chemokine receptor CCR1, we examined whether the CCR1 receptor antagonist BX471 is efficacious in a rat heterotopic heart transplant rejection model. Treatment of animals with BX471 and a subtherapeutic dose of cyclosporin (2.5 mg/kg), which is by itself ineffective in prolonging transplant rejection, is much more efficacious in prolonging transplantation rejection than animals treated with either cyclosporin or BX471 alone. We have examined the mechanism of action of the CCR1 antagonist in in vitro flow assays over microvascular endothelium and have discovered that the antagonist blocks the firm adhesion of monocytes triggered by RANTES on inflamed endothelium. Together, these data demonstrate a significant role for CCR1 in allograft rejection.
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- 2001
9. Identification and Characterization of a Potent, Selective, and Orally Active Antagonist of the CC Chemokine Receptor-1
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Ghannam Ameen F, Monahan Sean D, H. Daniel Perez, Susan Harvey, Wei Xu, Laura Dunning, Joseph Hesselgesser, Richard Horuk, Jun Shen, Elena Ho, Ng Howard P, Ron Vergona, Mary Rosser, Karen May, Babu Subramanyam, Imadul Islam, Cornell Mallari, John G. Bauman, Zuchun Zhao, Huynh Oanh, R. Michael Snider, Guo-Ping Wei, Ken Shaw, Meina Liang, Brad O. Buckman, Dennis D. Taub, and Michael M. Morrissey
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Male ,CCR1 ,Leukocyte migration ,DNA, Complementary ,Receptors, CCR1 ,Administration, Oral ,Pharmacology ,Binding, Competitive ,Biochemistry ,Cell Line ,Chemokine Receptor Antagonist ,Dogs ,Piperidines ,medicine ,Animals ,Humans ,Receptor ,Molecular Biology ,Chemistry ,Phenylurea Compounds ,Monocyte ,Antagonist ,Chemotaxis ,Cell Biology ,Rats ,medicine.anatomical_structure ,Rats, Inbred Lew ,Receptors, Chemokine ,CC chemokine receptors - Abstract
The CC chemokine receptor-1 (CCR1) is a prime therapeutic target for treating autoimmune diseases. Through high capacity screening followed by chemical optimization, we identified a novel non-peptide CCR1 antagonist, R-N-[5-chloro-2-[2-[4-[(4-fluorophenyl)methyl]-2-methyl-1-piperazinyl ]-2-oxoethoxy]phenyl]urea hydrochloric acid salt (BX 471). Competition binding studies revealed that BX 471 was able to displace the CCR1 ligands macrophage inflammatory protein-1alpha (MIP-1alpha), RANTES, and monocyte chemotactic protein-3 (MCP-3) with high affinity (K(i) ranged from 1 nm to 5.5 nm). BX 471 was a potent functional antagonist based on its ability to inhibit a number of CCR1-mediated effects including Ca(2+) mobilization, increase in extracellular acidification rate, CD11b expression, and leukocyte migration. BX 471 demonstrated a greater than 10,000-fold selectivity for CCR1 compared with 28 G-protein-coupled receptors. Pharmacokinetic studies demonstrated that BX 471 was orally active with a bioavailability of 60% in dogs. Furthermore, BX 471 effectively reduces disease in a rat experimental allergic encephalomyelitis model of multiple sclerosis. This study is the first to demonstrate that a non-peptide chemokine receptor antagonist is efficacious in an animal model of an autoimmune disease. In summary, we have identified a potent, selective, and orally available CCR1 antagonist that may be useful in the treatment of chronic inflammatory diseases.
- Published
- 2000
10. Species selectivity of a small molecule antagonist for the CCR1 chemokine receptor
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Joseph Hesselgesser, Mary Rosser, M. Liang, Richard Horuk, R. M. Snider, Ghannam Ameen F, John G. Bauman, P. J. Kretschmer, Ng Howard P, Michael M. Morrissey, H. D. Perez, Imadul Islam, Laura Dunning, Karen May, and Haifeng Pu
- Subjects
CCR1 ,Chemokine ,DNA, Complementary ,Molecular Sequence Data ,Receptors, CCR1 ,Biology ,Piperazines ,Mice ,Chemokine receptor ,Piperidines ,Species Specificity ,Nitriles ,Functional selectivity ,medicine ,Animals ,Humans ,Amino Acid Sequence ,Cloning, Molecular ,Chemokine CCL4 ,Receptor ,Chemokine CCL5 ,Chemokine CCL3 ,Pharmacology ,Binding Sites ,Antagonist ,Callithrix ,Macrophage Inflammatory Proteins ,Molecular biology ,Mechanism of action ,Biochemistry ,Competitive antagonist ,biology.protein ,Calcium ,Receptors, Chemokine ,Rabbits ,medicine.symptom - Abstract
The species specificity of a small molecule antagonist for the human CCR1 chemokine receptor, 2-2-diphenyl-5-(4-chlorophenyl)piperidin-1-yl)valeronitrile (CCR1 antagonist 1), has been examined using cloned CCR1 receptors from various species. The compound was able to bind to rabbit, marmoset, and human CCR1, and was able to block the functional activation of these receptors. However, it failed to significantly displace radiolabeled macrophage inflammatory protein-1alpha (MIP-1alpha) binding to mouse CCR1 at concentrations up to 10 microM. These data suggested that the antagonist binding site is well-conserved in rabbit, marmoset and human CCR1, but not in mouse CCR1. The functional selectivity and mechanism of action for CCR1 antagonist 1 were further characterized. CCR1 antagonist 1 blocked the increase in intracellular Ca(2+) stimulated by CCR1 agonists, but had no effect on N-formyl-Met-Leu-Phe (FMLP), monocyte chemotactic protein-1 (MCP-1) and stromal-derived factor 1alpha (SDF1alpha)-induced Ca(2+) mobilization, demonstrating functional selectivity for CCR1. Since CCR1 antagonist 1 is a functional antagonist of marmoset and rabbit CCR1 receptors, it should be possible to test its efficacy in animal models of disease.
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- 2000
11. Discovery of Novel Non-Peptide CCR1 Receptor Antagonists
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Karen May, John G. Bauman, Joseph Hesselgesser, Michael M. Morrissey, Ng Howard P, Richard Horuk, M. Liang, Imadul Islam, R. M. Snider, H. D. Perez, and Ghannam Ameen F
- Subjects
CCR1 ,Chemokine ,medicine.drug_class ,Anti-Inflammatory Agents ,Drug Evaluation, Preclinical ,Receptors, CCR1 ,Pharmacology ,Cell Line ,Structure-Activity Relationship ,Chemokine receptor ,Piperidines ,Nitriles ,Drug Discovery ,medicine ,Enzyme-linked receptor ,Humans ,Structure–activity relationship ,Chemokine CCL4 ,Receptor ,Chemokine CCL5 ,Protease-activated receptor 2 ,Chemokine CCL3 ,biology ,Chemistry ,Macrophage Inflammatory Proteins ,Receptor antagonist ,Recombinant Proteins ,Biochemistry ,biology.protein ,Molecular Medicine ,Calcium ,Receptors, Chemokine - Abstract
Ligands for the CCR1 receptor (MIP-1alpha and RANTES) have been implicated in a number of chronic inflammatory diseases, most notably multiple sclerosis and rheumatoid arthritis. Because these ligands share a common receptor, CCR1, we sought to discover antagonists for this receptor as an approach to treating these disorders. A novel series of 4-hydroxypiperidines has been discovered by high throughput screening (HTS) which potently inhibits the binding of MIP-1alpha and RANTES to the recombinant human CCR1 chemokine receptor. The structure-activity relationships of various segments of this template are described as the initial HTS lead 1 was optimized synthetically to the highly potent receptor antagonist 6s. This compound has been shown to have at least 200-fold selectivity for inhibition of CCR1 over other human 7-TM receptors, including other chemokine receptors. In addition, data obtained from in vitro functional assays demonstrate the functional antagonism of compound 6s and structurally related analogues against the CCR1 receptor in a concentration dependent manner. The discovery and optimization of potent and selective CCR1 receptor antagonists represented by compound 6s potentially represent a novel approach to the treatment of chronic inflammatory diseases.
- Published
- 1999
12. Identification and Characterization of Small Molecule Functional Antagonists of the CCR1 Chemokine Receptor
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Ghannam Ameen F, Wei Zheng, Dennis D. Taub, John G. Bauman, M. Liang, H. D. Perez, Monahan Sean D, Michael M. Morrissey, Mary Rosser, Ng Howard P, Richard Horuk, Joseph Hesselgesser, Karen May, Imadul Islam, R. M. Snider, and Guo Ping Wei
- Subjects
CCR1 ,Multiple Sclerosis ,Chemokine receptor CCR5 ,Receptors, CCR1 ,C-C chemokine receptor type 6 ,Pharmacology ,Hydroxylation ,Ligands ,Biochemistry ,Cell Line ,Arthritis, Rheumatoid ,Chemokine receptor ,Piperidines ,Humans ,CCL17 ,Chemokine CCL4 ,CCL13 ,Receptor ,Chemokine CCL5 ,Molecular Biology ,Chemokine CCL3 ,biology ,Cell Biology ,Macrophage Inflammatory Proteins ,Chemotaxis, Leukocyte ,Kinetics ,biology.protein ,Receptors, Chemokine ,CC chemokine receptors - Abstract
The CC chemokines macrophage inflammatory protein-1alpha (MIP-1alpha) and RANTES (regulated on activation normal T cell expressed) have been implicated in rheumatoid arthritis and multiple sclerosis. Since their effects are mediated through the CCR1 chemokine receptor, we set up a small molecule CCR1 antagonist program to search for inhibitors. Through high capacity screening we discovered a number of 4-hydroxypiperidine compounds with CCR1 antagonist activity and report their synthesis and in vitro pharmacology here. Scatchard analysis of the competition binding data revealed that the compounds had Ki values ranging from 40 to 4000 nM. The pharmacological profile of the most potent member of this series, compound 1 (2-2-diphenyl-5-(4-chlorophenyl)piperidin-lyl)valeronitri te), was further evaluated. Compound 1 showed concentration-dependent inhibition of MIP-1alpha-induced extracellular acidification and Ca2+ mobilization demonstrating functional antagonism. When given alone, the compound did not elicit any responses, indicating the absence of intrinsic agonist activity. Compound 1 inhibited MIP-1alpha- and RANTES-induced migration in peripheral blood mononuclear cells in a dose-responsive manner. Selectivity testing against a panel of seven transmembrane domain receptors indicated that compound 1 is inactive on a number of receptors at concentrations up to 10 microM. This is the first description of CCR1 receptor antagonists that may be useful in the treatment of chronic inflammatory diseases involving MIP-1alpha, RANTES, and CCR1.
- Published
- 1998
13. Synthesis and evaluation of a library of peptidomimetics based upon the β-turn
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Andrew A. Bray, Jonathan A. Ellman, Lon Trinh, Michael Snyder, Wei Zhang, Michael M. Morrissey, and Alex A. Virgilio
- Subjects
Scaffold ,Chemistry ,Peptidomimetic ,Organic Chemistry ,Drug Discovery ,Receptor ,Biochemistry ,Combinatorial chemistry - Abstract
A diverse library of 1152 compounds based on a β-turn mimetic scaffold was synthesized. A subset of the library was characterized by ES-MS. Several compounds from the library with modest affinity to a cloned N -formyl-Met-Leu-Phe (tMLF) receptor were identified.
- Published
- 1997
14. High capacity screening of pooled compounds: Identification of the active compound without re-assay of pool members
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Amy Liang, Kathryn V. Dunn, Peter J. Kretschmer, Lan Trinh, Richard Spann, David Senator, Jason Kondracki, John Morser, Michael M. Morrissey, Samuel E. Lipson, Mark A. Polokoff, Wei Zheng, John Loughlin, and James J. Devlin
- Subjects
chemistry.chemical_classification ,biology ,Drug discovery ,High capacity ,Nitric oxide ,Matrix (chemical analysis) ,chemistry.chemical_compound ,Enzyme ,chemistry ,Biochemistry ,Active compound ,Enzyme inhibitor ,Drug Discovery ,biology.protein ,Bioassay - Abstract
A matrix-based compound pooling and deconvolution method has been developed that significantly increased the efficiency of a high-capacity screening program. This method is based on screening pools of 10 compounds. The matrix used to assemble the pools resulted in each compound being assayed twice—each time with a completely different set of pooled compounds. The active compound in an active pool was accurately predicted by determining which compound was present in an active pool both times that a pool containing that compound was tested. This has eliminated the need to re-assay each individual member of active pools. This approach has been tested with a set of 6,680 compounds in three different assays: inhibition of Factor Xa, inhibition of gastrin-releasing peptide receptor binding, and inhibition of nitric oxide synthase (isoform II). Thus significant time is saved not only by pooling the compounds for the initial assay but also by avoiding the need to re-assay all compounds in active pools. Moreover, no false negatives occurred in these assays—an important consideration from a drug discovery perspective. © 1996 Wiley-Liss, Inc.
- Published
- 1996
15. Optical Nanofibers for Probing Cold Atoms
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Laura Russell, Kieran Deasy, Michael M. Morrissey, Amy Watkins, and Síle Nic Chormaic
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Condensed Matter::Quantum Gases ,Novel technique ,Materials science ,business.industry ,Nanophotonics ,Physics::Optics ,Nanofiber ,Laser cooling ,Fibre optic sensors ,Atom optics ,Optoelectronics ,Spontaneous emission ,Physics::Atomic Physics ,Fiber ,Atomic physics ,business ,Astrophysics::Galaxy Astrophysics - Abstract
We present a novel technique for measuring the characteristics of a cloud of cold atoms by monitoring the spontaneous emission coupled into the guided mode of a nanofiber. We show that the fiber is very sensitive to the atoms close to its surface.
- Published
- 2009
16. Thiophene-anthranilamides as highly potent and orally available factor Xa inhibitors
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Yuo-Ling Chou, John Morser, Marc Adler, Wheeseong Lee, Joseph Post, David R. Light, Kochanny Monica, Karna Lyn Sacchi, Dao Lentz, Arnaiz Damian O, Marc Whitlow, Bin Ye, Amy Liang, Ron Vergona, Richard W. Fitch, Mark E. Sullivan, Sarah Cheeseman, Janice Ewing, Zuchun Zhao, Babu Subramanyam, Kathy White, Kenneth J. Shaw, Karanjawala Rushad E, Janette Walters, Brian D. Griedel, Yi-Xin Wang, William P. Dole, Michael M. Morrissey, Steven T. Sakata, Galina Rumennik, and Shung C. Wu
- Subjects
Male ,Models, Molecular ,medicine.drug_mechanism_of_action ,medicine.drug_class ,Factor Xa Inhibitor ,Aminopyridines ,Thiophenes ,Pharmacology ,In Vitro Techniques ,Crystallography, X-Ray ,Structure-Activity Relationship ,Dogs ,Oral administration ,In vivo ,Drug Discovery ,medicine ,Potency ,Animals ,Humans ,ortho-Aminobenzoates ,Rats, Wistar ,Serine protease ,Venous Thrombosis ,biology ,Chemistry ,Drug discovery ,Anticoagulant ,Anticoagulants ,Amides ,Rats ,Biochemistry ,Enzyme inhibitor ,biology.protein ,Prothrombin Time ,Molecular Medicine ,Factor Xa Inhibitors - Abstract
There remains a high unmet medical need for a safe oral therapy for thrombotic disorders. The serine protease factor Xa (fXa), with its central role in the coagulation cascade, is among the more promising targets for anticoagulant therapy and has been the subject of intensive drug discovery efforts. Investigation of a hit from high-throughput screening identified a series of thiophene-substituted anthranilamides as potent nonamidine fXa inhibitors. Lead optimization by incorporation of hydrophilic groups led to the discovery of compounds with picomolar inhibitory potency and micromolar in vitro anticoagulant activity. Based on their high potency, selectivity, oral pharmacokinetics, and efficacy in a rat venous stasis model of thrombosis, compounds ZK 814048 (10b), ZK 810388 (13a), and ZK 813039 (17m) were advanced into development.
- Published
- 2007
17. Design, synthesis, and biological activity of novel factor Xa inhibitors: 4-aryloxy substituents of 2,6-diphenoxypyridines
- Author
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Howard P, Ng, Brad O, Buckman, Keith A, Eagen, William J, Guilford, Monica J, Kochanny, Raju, Mohan, Kenneth J, Shaw, Shung C, Wu, Dao, Lentz, Amy, Liang, Lan, Trinh, Elena, Ho, David, Smith, Babu, Subramanyam, Ron, Vergona, Janette, Walters, Kathy A, White, Mark E, Sullivan, Michael M, Morrissey, and Gary B, Phillips
- Subjects
Structure-Activity Relationship ,Dogs ,Serine Proteinase Inhibitors ,Fibrinolytic Agents ,Pyridines ,Drug Design ,Administration, Oral ,Animals ,Biological Availability ,Cattle ,Heterocyclic Compounds, 4 or More Rings ,Factor Xa Inhibitors ,Rats - Abstract
A novel series of triaryloxypyridines have been designed to inhibit factor Xa, a serine protease strategically located in the coagulation cascade. Inhibitor 5e has a K(I) against factor Xa of 0.12nM and is greater than 8000- and 2000-fold selective over two related serine proteases, thrombin and trypsin, respectively. The 4-position of the central pyridine has been identified as a site that tolerates various substitutions without deleterious effects on potency and selectivity. This suggests that the 4-position of the pyridine ring is an ideal site for chemical modifications to identify inhibitors with improved pharmacokinetic characteristics. This investigation has resulted in inhibitor 5d, which has an oral availability of 6% in dogs. The synthesis, in vitro activity, and in vivo profile of this class of inhibitors is outlined.
- Published
- 2002
18. CCR1-specific non-peptide antagonist: efficacy in a rabbit allograft rejection model
- Author
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Ng Howard P, Imadul Islam, Joseph Hesselgesser, Colin Green, Guo Ping Wei, Wei Xu, Meina Liang, H. Daniel Perez, Laura Dunning, John G. Bauman, Mary Rosser, Michael M. Morrissey, Ghannam Ameen F, Karen May, Brad O. Buckman, Richard Horuk, Sandra Shurey, and R. Michael Snider
- Subjects
Agonist ,Graft Rejection ,Chemokine ,Pathology ,medicine.medical_specialty ,medicine.drug_class ,Immunology ,Receptors, CCR1 ,Pharmacology ,Cell Line ,chemistry.chemical_compound ,Jurkat Cells ,Piperidines ,medicine ,Immunology and Allergy ,Animals ,Humans ,Transplantation, Homologous ,Urea ,Chemokine CCL4 ,Chemokine CCL3 ,Creatinine ,biology ,business.industry ,Phenylurea Compounds ,Graft Survival ,Antagonist ,Macrophage Inflammatory Proteins ,medicine.disease ,Kidney Transplantation ,Transplant rejection ,Transplantation ,Disease Models, Animal ,chemistry ,Competitive antagonist ,biology.protein ,Receptors, Chemokine ,Rabbits ,business ,CC chemokine receptors - Abstract
The classic signs of acute cellular rejection during organ transplantation include the infiltration of mononuclear cells into the interstitium. This recruitment of leukocytes into the transplanted tissue is promoted by chemokines like RANTES. Since RANTES is a potent agonist for the CC chemokine receptor CCR1, we examined whether the CCR1 antagonist BX 471 was efficacious in a rabbit kidney transplant rejection model. BX 471 was able to compete with high affinity with the CCR1 ligands MIP-1α and RANTES for binding to HEK 293 cells expressing rabbit CCR1. BX 471 was a competitive antagonist of rabbit CCR1 in Ca 2+ flux studies. Two separate studies in which animals were subcutaneously implanted with slow release pellets of BX 471 demonstrated that animals implanted with BX 471 had increased survival compared with untreated controls or animals implanted with placebo. The mean survival time for the placebo group was 12.33±1.7 days. The animals in the BX 471 treated group had mean survival times of 16.9±2.1 and 16.0±1.7 days, respectively, for the two studies. Analysis of the combined data by Student t -test gave a P value of 0.03 that is significant at the 0.05 level. In addition, there was a marked reduction in the urea and creatinine levels in the BX 471 treated animals compared with the control and placebo groups in both studies. Finally, pathologic analysis of the kidneys in the rabbit renal transplantation model from animals in the different groups showed that BX 471 was similar to cyclosporin in its ability to prevent extensive infarction of transplanted kidneys. Based on the data from these studies, BX 471 shows clear efficacy at the single dose tested compared with animals treated with placebo.
- Published
- 2001
19. Allosteric inhibitors of inducible nitric oxide synthase dimerization discovered via combinatorial chemistry
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Kirk Mcmillan, Gonghua Pan, Michael M. Morrissey, Leslie J. Browne, Mark A. Polokoff, Baldwin John J, Ronald Vergona, Gary Phillips, Marc Adler, Michael Ohlmeyer, David D. Davey, John Parkinson, Cornell Mallari, Charles B. Glaser, Nolan H. Sigal, Shawn David Erickson, Keith A. Eagen, James J. Devlin, Tish A. Young, Ronald E. Dolle, Eric Blasko, Daniel Chelsky, Richard I. Feldman, Douglas S. Auld, and Marc Whitlow
- Subjects
Models, Molecular ,Oxygenase ,Allosteric regulation ,Nitric Oxide Synthase Type II ,Plasma protein binding ,Biology ,Endothelial NOS ,Nitric Oxide ,Nitric oxide ,Cell Line ,chemistry.chemical_compound ,Mice ,Allosteric Regulation ,Animals ,Combinatorial Chemistry Techniques ,Humans ,Binding site ,Enzyme Inhibitors ,Multidisciplinary ,Binding Sites ,Molecular Structure ,Biological Sciences ,Molecular biology ,Small molecule ,Rats ,Nitric oxide synthase ,Biochemistry ,chemistry ,biology.protein ,Nitric Oxide Synthase ,Dimerization ,Protein Binding - Abstract
Potent and selective inhibitors of inducible nitric oxide synthase (iNOS) (EC 1.14.13.39 ) were identified in an encoded combinatorial chemical library that blocked human iNOS dimerization, and thereby NO production. In a cell-based iNOS assay (A-172 astrocytoma cells) the inhibitors had low-nanomolar IC 50 values and thus were >1,000-fold more potent than the substrate-based direct iNOS inhibitors 1400W and N- methyl- l -arginine. Biochemical studies confirmed that inhibitors caused accumulation of iNOS monomers in mouse macrophage RAW 264.7 cells. High affinity ( K d ≈ 3 nM) of inhibitors for isolated iNOS monomers was confirmed by using a radioligand binding assay. Inhibitors were >1,000-fold selective for iNOS versus endothelial NOS dimerization in a cell-based assay. The crystal structure of inhibitor bound to the monomeric iNOS oxygenase domain revealed inhibitor–heme coordination and substantial perturbation of the substrate binding site and the dimerization interface, indicating that this small molecule acts by allosterically disrupting protein–protein interactions at the dimer interface. These results provide a mechanism-based approach to highly selective iNOS inhibition. Inhibitors were active in vivo , with ED 50 values of
- Published
- 2000
20. Erratum
- Author
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Michael M. Morrissey, Erik Postma, Alastair J. Wilson, Loeske E. B. Kruuk, Craig A. Walling, Denis Réale, Daniel H. Nussey, and Michelle N. Clements
- Subjects
Animal model ,Animal Science and Zoology ,Environmental ethics ,Biology ,Ecology, Evolution, Behavior and Systematics - Published
- 2011
21. Thiophene-Anthranilamides as Highly Potent and Orally Available Factor Xa Inhibitors1.
- Author
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Bin Ye, Damian O. Arnaiz, Yuo-Ling Chou, Brian D. Griedel, Rushad Karanjawala, Wheeseong Lee, Michael M. Morrissey, Karna L. Sacchi, Steven T. Sakata, Kenneth J. Shaw, Shung C. Wu, Zuchun Zhao, Marc Adler, Sarah Cheeseman, William P. Dole, Janice Ewing, Richard Fitch, Dao Lentz, Amy Liang, and David Light
- Published
- 2007
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- View/download PDF
22. Design, Synthesis, and Activity of 2-Imidazol-1-ylpyrimidine Derived Inducible Nitric Oxide Synthase Dimerization Inhibitors.
- Author
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David D. Davey, Marc Adler, Damian Arnaiz, Keith Eagen, Shawn Erickson, William Guilford, Margaret Kenrick, Michael M. Morrissey, Mike Ohlmeyer, Gonghua Pan, Vidyadhar M. Paradkar, John Parkinson, Mark Polokoff, Kurt Saionz, Cecile Santos, Babu Subramanyam, Ron Vergona, Robert G. Wei, Marc Whitlow, and Bin Ye
- Published
- 2007
- Full Text
- View/download PDF
23. Fine scale diversity in the lava: genetic and phenotypic diversity in small populations of Arctic charr Salvelinus alpinus.
- Author
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Leblanc CA, Räsänen K, Morrissey M, Skúlason S, Ferguson M, and Kristjánsson BK
- Subjects
- Animals, Trout genetics, Genetic Drift, Ecosystem
- Abstract
Background: A major goal in evolutionary biology is to understand the processes underlying phenotypic variation in nature. Commonly, studies have focused on large interconnected populations or populations found along strong environmental gradients. However, studies on small fragmented populations can give strong insight into evolutionary processes in relation to discrete ecological factors. Evolution in small populations is believed to be dominated by stochastic processes, but recent work shows that small populations can also display adaptive phenotypic variation, through for example plasticity and rapid adaptive evolution. Such evolution takes place even though there are strong signs of historical bottlenecks and genetic drift. Here we studied 24 small populations of the freshwater fish Arctic charr (Salvelinus alpinus) found in groundwater filled lava caves. Those populations were found within a few km2-area with no apparent water connections between them. We studied the relative contribution of neutral versus non-neutral evolutionary processes in shaping phenotypic divergence, by contrasting patterns of phenotypic and neutral genetic divergence across populations in relation to environmental measurements. This allowed us to model the proportion of phenotypic variance explained by the environment, taking in to account the observed neutral genetic structure., Results: These populations originated from the nearby Lake Mývatn, and showed small population sizes with low genetic diversity. Phenotypic variation was mostly correlated with neutral genetic diversity with only a small environmental effect., Conclusions: Phenotypic diversity in these cave populations appears to be largely the product of neutral processes, fitting the classical evolutionary expectations. However, the fact that neutral processes did not explain fully the phenotypic patterns suggests that further studies can increase our understanding on how neutral evolutionary processes can interact with other forces of selection at early stages of divergence. The accessibility of these populations has provided the opportunity for long-term monitoring of individual fish, allowing tracking how the environment can influence phenotypic and genetic divergence for shaping and maintaining diversity in small populations. Such studies are important, especially in freshwater, as habitat alteration is commonly breaking populations into smaller units, which may or may not be viable., (© 2024. The Author(s).)
- Published
- 2024
- Full Text
- View/download PDF
24. Transcriptomic Determinants of Response to Pembrolizumab Monotherapy across Solid Tumor Types.
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Cristescu R, Nebozhyn M, Zhang C, Albright A, Kobie J, Huang L, Zhao Q, Wang A, Ma H, Alexander Cao Z, Morrissey M, Ribas A, Grivas P, Cescon DW, McClanahan TK, Snyder A, Ayers M, Lunceford J, and Loboda A
- Subjects
- Antibodies, Monoclonal, Humanized, Humans, RNA, Transcriptome, Transforming Growth Factor beta genetics, Antineoplastic Agents, Immunological adverse effects, Neoplasms drug therapy, Neoplasms genetics, Neoplasms pathology
- Abstract
Purpose: To explore relationships between biological gene expression signatures and pembrolizumab response., Experimental Design: RNA-sequencing data on baseline tumor tissue from 1,188 patients across seven tumor types treated with pembrolizumab monotherapy in nine clinical trials were used. A total of 11 prespecified gene expression signatures [18-gene T-cell-inflamed gene expression profile (TcellinfGEP), angiogenesis, hypoxia, glycolysis, proliferation, MYC, RAS, granulocytic myeloid-derived suppressor cell (gMDSC), monocytic myeloid-derived suppressor cell (mMDSC), stroma/epithelial-to-mesenchymal transition (EMT)/TGFβ, and WNT] were evaluated for their relationship to objective response rate (per RECIST, version 1.1). Logistic regression analysis of response for consensus signatures was adjusted for tumor type, Eastern Cooperative Oncology Group performance status, and TcellinfGEP, an approach equivalent to evaluating the association between response and the residuals of consensus signatures after detrending them for their relationship with the TcellinfGEP (previously identified as a determinant of pembrolizumab response) and tumor type. Testing of the 10 prespecified non-TcellinfGEP consensus signatures for negative association [except proliferation (hypothesized positive association)] with response was adjusted for multiplicity., Results: Covariance patterns of the 11 signatures (including TcellinfGEP) identified in Merck-Moffitt and The Cancer Genome Atlas datasets showed highly concordant coexpression patterns in the RNA-sequencing data from pembrolizumab trials. TcellinfGEP was positively associated with response; signatures for angiogenesis, mMDSC, and stroma/EMT/TGFβ were negatively associated with response to pembrolizumab monotherapy., Conclusions: These findings suggest that features beyond IFNγ-related T-cell inflammation may be relevant to anti-programmed death 1 monotherapy response and may define other axes of tumor biology as candidates for pembrolizumab combinations. See related commentary by Cho et al., p. 1479., (©2021 The Authors; Published by the American Association for Cancer Research.)
- Published
- 2022
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25. Genomic profiling of NETs: a comprehensive analysis of the RADIANT trials.
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Yao J, Garg A, Chen D, Capdevila J, Engstrom P, Pommier R, Van Cutsem E, Singh S, Fazio N, He W, Riester M, Patel P, Voi M, Morrissey M, Pavel M, and Kulke MH
- Subjects
- Aged, Antineoplastic Agents therapeutic use, Everolimus therapeutic use, Female, Genomics, High-Throughput Nucleotide Sequencing, Humans, Male, Mutation, Neuroendocrine Tumors drug therapy, Exome Sequencing, Neuroendocrine Tumors genetics
- Abstract
Neuroendocrine tumors (NETs) have historically been subcategorized according to histologic features and the site of anatomic origin. Here, we characterize the genomic alterations in patients enrolled in 3 phase 3 clinical trials of NET of different anatomic origins and assessed the potential correlation with clinical outcomes. Whole-exome and targeted panel sequencing was used to characterize 225 NET samples collected in the RADIANT series of clinical trials. Genomic profiling of NET was analyzed along with nongenomic biomarker data on tumor grade and circulating chromogranin A (CgA) and neuron specific enolase (NSE) levels from these patients enrolled in clinical trials. Our results highlight recurrent large-scale chromosomal alterations as a common theme among NET. Although the specific pattern of chromosomal alterations differed between tumor subtypes, the evidence for generalized chromosomal instability (CIN) was observed across all primary sites of NET. In pancreatic NET, although the P-value was not significant, higher CIN suggests a trend towards longer survival (HR, 0.55, P=0.077); whereas in the gastrointestinal NET, lower CIN was associated with longer survival (HR, 0.44, P=0.0006). Our multivariate analyses demonstrated that when combined with other clinical data among patients with progressive advanced NETs, chromosomal level alteration adds important prognostic information. Large-scale CIN is a common feature of NET, and specific patterns of chromosomal gain and loss appeared to have independent prognostic value in NET subtypes. However, whether CIN in general has clinical significance in NET requires validation in larger patient cohort and warrants further mechanistic studies.
- Published
- 2019
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26. Genetic mechanisms of target antigen loss in CAR19 therapy of acute lymphoblastic leukemia.
- Author
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Orlando EJ, Han X, Tribouley C, Wood PA, Leary RJ, Riester M, Levine JE, Qayed M, Grupp SA, Boyer M, De Moerloose B, Nemecek ER, Bittencourt H, Hiramatsu H, Buechner J, Davies SM, Verneris MR, Nguyen K, Brogdon JL, Bitter H, Morrissey M, Pierog P, Pantano S, Engelman JA, and Winckler W
- Subjects
- Antigens, CD19 genetics, Antigens, CD19 immunology, Humans, Immunotherapy, Adoptive, Loss of Heterozygosity genetics, Mutation, Precursor Cell Lymphoblastic Leukemia-Lymphoma genetics, Precursor Cell Lymphoblastic Leukemia-Lymphoma immunology, Precursor Cell Lymphoblastic Leukemia-Lymphoma pathology, Receptors, Chimeric Antigen immunology, Receptors, Chimeric Antigen therapeutic use, T-Lymphocytes immunology, Drug Resistance, Neoplasm genetics, Precursor Cell Lymphoblastic Leukemia-Lymphoma drug therapy, Receptors, Antigen, T-Cell genetics, Receptors, Chimeric Antigen genetics
- Abstract
We identified genetic mutations in CD19 and loss of heterozygosity at the time of CD19
- relapse to chimeric antigen receptor (CAR) therapy. The mutations are present in the vast majority of resistant tumor cells and are predicted to lead to a truncated protein with a nonfunctional or absent transmembrane domain and consequently to a loss of surface antigen. This irreversible loss of CD19 advocates for an alternative targeting or combination CAR approach.- Published
- 2018
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- View/download PDF
27. Reversible, orally available ADP receptor (P2Y 12 ) antagonists Part I: Hit to lead process.
- Author
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Islam I, Yuan S, Wei RG, Xu W, Morrissey M, Mohan R, Zheng D, DiMella A, Dunning L, Snider M, Subramanyam B, Tseng JL, Bryant JA, and Buckman BO
- Subjects
- Administration, Oral, Animals, Dogs, Dose-Response Relationship, Drug, Fluorenes administration & dosage, Fluorenes chemistry, High-Throughput Screening Assays, Humans, Microsomes, Liver metabolism, Molecular Structure, Platelet Aggregation drug effects, Rats, Structure-Activity Relationship, Fluorenes pharmacology, Receptors, Purinergic P2Y12 metabolism
- Abstract
A hit to lead process to identify reversible, orally available ADP receptor (P2Y
12 ) antagonists lead compounds is described. High throughput screening afforded 1. Optimization of 1, using parallel synthesis methods, a methyl scan to identify promising regions for optimization, and exploratory SAR on these regions, provided 22 and 23. Compound 23 is an orally available, competitive reversible antagonist (KB = 94 nM for inhibition of ADP-induced platelet aggregation). It exhibits high metabolic stability in human, rat and dog liver microsomes and is orally absorbed. Although plasma level after oral dosing of 22 and 23 to rats is low, reasonable levels were achieved to merit extensive lead optimization of this structural class., (Copyright © 2018 Elsevier Ltd. All rights reserved.)- Published
- 2018
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28. General Methods for Evolutionary Quantitative Genetic Inference from Generalized Mixed Models.
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de Villemereuil P, Schielzeth H, Nakagawa S, and Morrissey M
- Subjects
- Genetics, Population methods, Quantitative Trait, Heritable, Evolution, Molecular, Models, Genetic, Software
- Abstract
Methods for inference and interpretation of evolutionary quantitative genetic parameters, and for prediction of the response to selection, are best developed for traits with normal distributions. Many traits of evolutionary interest, including many life history and behavioral traits, have inherently nonnormal distributions. The generalized linear mixed model (GLMM) framework has become a widely used tool for estimating quantitative genetic parameters for nonnormal traits. However, whereas GLMMs provide inference on a statistically convenient latent scale, it is often desirable to express quantitative genetic parameters on the scale upon which traits are measured. The parameters of fitted GLMMs, despite being on a latent scale, fully determine all quantities of potential interest on the scale on which traits are expressed. We provide expressions for deriving each of such quantities, including population means, phenotypic (co)variances, variance components including additive genetic (co)variances, and parameters such as heritability. We demonstrate that fixed effects have a strong impact on those parameters and show how to deal with this by averaging or integrating over fixed effects. The expressions require integration of quantities determined by the link function, over distributions of latent values. In general cases, the required integrals must be solved numerically, but efficient methods are available and we provide an implementation in an R package, QGglmm. We show that known formulas for quantities such as heritability of traits with binomial and Poisson distributions are special cases of our expressions. Additionally, we show how fitted GLMM can be incorporated into existing methods for predicting evolutionary trajectories. We demonstrate the accuracy of the resulting method for evolutionary prediction by simulation and apply our approach to data from a wild pedigreed vertebrate population., (Copyright © 2016 de Villemereuil et al.)
- Published
- 2016
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29. Association of MTOR Mutations With Developmental Brain Disorders, Including Megalencephaly, Focal Cortical Dysplasia, and Pigmentary Mosaicism.
- Author
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Mirzaa GM, Campbell CD, Solovieff N, Goold C, Jansen LA, Menon S, Timms AE, Conti V, Biag JD, Adams C, Boyle EA, Collins S, Ishak G, Poliachik S, Girisha KM, Yeung KS, Chung BHY, Rahikkala E, Gunter SA, McDaniel SS, Macmurdo CF, Bernstein JA, Martin B, Leary R, Mahan S, Liu S, Weaver M, Doerschner M, Jhangiani S, Muzny DM, Boerwinkle E, Gibbs RA, Lupski JR, Shendure J, Saneto RP, Novotny EJ, Wilson CJ, Sellers WR, Morrissey M, Hevner RF, Ojemann JG, Guerrini R, Murphy LO, Winckler W, and Dobyns WB
- Subjects
- Adolescent, Adult, Amino Acids pharmacology, Animals, Cells, Cultured, Cerebral Cortex cytology, Child, Child, Preschool, Developmental Disabilities diagnostic imaging, Developmental Disabilities genetics, Embryo, Mammalian, Female, Gene Expression Regulation drug effects, Gene Expression Regulation genetics, Genetic Association Studies, Humans, Intercellular Signaling Peptides and Proteins deficiency, Male, Malformations of Cortical Development diagnostic imaging, Mechanistic Target of Rapamycin Complex 1, Megalencephaly diagnostic imaging, Multiprotein Complexes pharmacology, Nerve Tissue Proteins genetics, Nerve Tissue Proteins metabolism, Neurons drug effects, Rats, Retrospective Studies, TOR Serine-Threonine Kinases metabolism, TOR Serine-Threonine Kinases pharmacology, Young Adult, Malformations of Cortical Development genetics, Megalencephaly genetics, Mosaicism, Mutation genetics, TOR Serine-Threonine Kinases genetics
- Abstract
Importance: Focal cortical dysplasia (FCD), hemimegalencephaly, and megalencephaly constitute a spectrum of malformations of cortical development with shared neuropathologic features. These disorders are associated with significant childhood morbidity and mortality., Objective: To identify the underlying molecular cause of FCD, hemimegalencephaly, and diffuse megalencephaly., Design, Setting, and Participants: Patients with FCD, hemimegalencephaly, or megalencephaly (mean age, 11.7 years; range, 2-32 years) were recruited from Pediatric Hospital A. Meyer, the University of Hong Kong, and Seattle Children's Research Institute from June 2012 to June 2014. Whole-exome sequencing (WES) was performed on 8 children with FCD or hemimegalencephaly using standard-depth (50-60X) sequencing in peripheral samples (blood, saliva, or skin) from the affected child and their parents and deep (150-180X) sequencing in affected brain tissue. Targeted sequencing and WES were used to screen 93 children with molecularly unexplained diffuse or focal brain overgrowth. Histopathologic and functional assays of phosphatidylinositol 3-kinase-AKT (serine/threonine kinase)-mammalian target of rapamycin (mTOR) pathway activity in resected brain tissue and cultured neurons were performed to validate mutations., Main Outcomes and Measures: Whole-exome sequencing and targeted sequencing identified variants associated with this spectrum of developmental brain disorders., Results: Low-level mosaic mutations of MTOR were identified in brain tissue in 4 children with FCD type 2a with alternative allele fractions ranging from 0.012 to 0.086. Intermediate-level mosaic mutation of MTOR (p.Thr1977Ile) was also identified in 3 unrelated children with diffuse megalencephaly and pigmentary mosaicism in skin. Finally, a constitutional de novo mutation of MTOR (p.Glu1799Lys) was identified in 3 unrelated children with diffuse megalencephaly and intellectual disability. Molecular and functional analysis in 2 children with FCD2a from whom multiple affected brain tissue samples were available revealed a mutation gradient with an epicenter in the most epileptogenic area. When expressed in cultured neurons, all MTOR mutations identified here drive constitutive activation of mTOR complex 1 and enlarged neuronal size., Conclusions and Relevance: In this study, mutations of MTOR were associated with a spectrum of brain overgrowth phenotypes extending from FCD type 2a to diffuse megalencephaly, distinguished by different mutations and levels of mosaicism. These mutations may be sufficient to cause cellular hypertrophy in cultured neurons and may provide a demonstration of the pattern of mosaicism in brain and substantiate the link between mosaic mutations of MTOR and pigmentary mosaicism in skin.
- Published
- 2016
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30. A 17-Year-Old Girl With Acute Onset of Hemiparesis.
- Author
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Morrissey M, Ciorciari AJ, and Cunningham SJ
- Subjects
- Acute Disease, Adolescent, Diagnosis, Differential, Diffuse Cerebral Sclerosis of Schilder rehabilitation, Female, Humans, Paresis rehabilitation, Diffuse Cerebral Sclerosis of Schilder complications, Diffuse Cerebral Sclerosis of Schilder diagnosis, Paresis diagnosis, Paresis etiology
- Abstract
The presentation of acute-onset hemiparesis in a teenager can be challenging and offers a wide differential diagnosis. We discuss the approach to the patient (which should begin with thorough history taking and physical examination) and advanced imaging as directed by the patient's signs and symptoms. We report the case of an otherwise well 17-year-old girl who presented to the pediatric emergency department with a 2-day history of left-sided weakness and difficulty ambulating. Her eventual diagnosis of Balo concentric sclerosis, a rare form of multiple sclerosis, is discussed.
- Published
- 2016
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31. Molecular analysis of a male breast cancer patient with prolonged stable disease under mTOR/PI3K inhibitors BEZ235/everolimus.
- Author
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Brannon AR, Frizziero M, Chen D, Hummel J, Gallo J, Riester M, Patel P, Cheung W, Morrissey M, Carbone C, Cottini S, Tortora G, and Melisi D
- Abstract
The mTORC1 inhibitor everolimus (Afinitor/RAD001) has been approved for multiple cancer indications, including ER(+)/HER2(-) metastatic breast cancer. However, the combination of everolimus with the dual PI3K/mTOR inhibitor BEZ235 was shown to be more efficacious than either everolimus or BEZ235 alone in preclinical models. Herein, we describe a male breast cancer (MBC) patient who was diagnosed with hormone receptor-positive (HR(+))/HER2(-) stage IIIA invasive ductal carcinoma and sequentially treated with chemoradiotherapy and hormonal therapy. Upon the development of metastases, the patient began a 200 mg twice-daily BEZ235 and 2.5 mg weekly everolimus combination regimen. The patient sustained a prolonged stable disease of 18 mo while undergoing the therapy, before his tumor progressed again. Therefore, we sought to both better understand MBC and investigate the underlying molecular mechanisms of the patient's sensitivity and subsequent resistance to the BEZ235/everolimus combination therapy. Genomic and immunohistochemical analyses were performed on samples collected from the initial invasive ductal carcinoma pretreatment and a metastasis postprogression on the BEZ235/everolimus combination treatment. Both tumors were relatively quiet genomically with no overlap to recurrent MBC alterations in the literature. Markers of PI3K/mTOR pathway hyperactivation were not identified in the pretreatment sample, which complements previous reports of HR(+) female breast cancers being responsive to mTOR inhibition without this activation. The postprogression sample, however, demonstrated greater than fivefold increased estrogen receptor and pathogenesis-related protein expression, which could have constrained the PI3K/mTOR pathway inhibition by BEZ235/everolimus. Overall, these analyses have augmented the limited episteme on MBC genetics and treatment.
- Published
- 2016
- Full Text
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32. High Affinity Antibodies against Influenza Characterize the Plasmablast Response in SLE Patients After Vaccination.
- Author
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Kaur K, Zheng NY, Smith K, Huang M, Li L, Pauli NT, Henry Dunand CJ, Lee JH, Morrissey M, Wu Y, Joachims ML, Munroe ME, Lau D, Qu X, Krammer F, Wrammert J, Palese P, Ahmed R, James JA, and Wilson PC
- Subjects
- Antibody Affinity, Antibody Formation, Case-Control Studies, Humans, Influenza Vaccines administration & dosage, Antibodies, Monoclonal immunology, Lupus Erythematosus, Systemic immunology, Orthomyxoviridae immunology
- Abstract
Breakdown of B cell tolerance is a cardinal feature of systemic lupus erythematosus (SLE). Increased numbers of autoreactive mature naïve B cells have been described in SLE patients and autoantibodies have been shown to arise from autoreactive and non-autoreactive precursors. How these defects, in the regulation of B cell tolerance and selection, influence germinal center (GC) reactions that are directed towards foreign antigens has yet to be investigated. Here, we examined the characteristics of post-GC foreign antigen-specific B cells from SLE patients and healthy controls by analyzing monoclonal antibodies generated from plasmablasts induced specifically by influenza vaccination. We report that many of the SLE patients had anti-influenza antibodies with higher binding affinity and neutralization capacity than those from controls. Although overall frequencies of autoreactivity in the influenza-specific plasmablasts were similar for SLE patients and controls, the variable gene repertoire of influenza-specific plasmablasts from SLE patients was altered, with increased usage of JH6 and long heavy chain CDR3 segments. We found that high affinity anti-influenza antibodies generally characterize the plasmablast responses of SLE patients with low levels of autoreactivity; however, certain exceptions were noted. The high-avidity antibody responses in SLE patients may also be correlated with cytokines that are abnormally expressed in lupus. These findings provide insights into the effects of dysregulated immunity on the quality of antibody responses following influenza vaccination and further our understanding of the underlying abnormalities of lupus.
- Published
- 2015
- Full Text
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33. Robust estimates of environmental effects on population vital rates: an integrated capture-recapture model of seasonal brook trout growth, survival and movement in a stream network.
- Author
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Letcher BH, Schueller P, Bassar RD, Nislow KH, Coombs JA, Sakrejda K, Morrissey M, Sigourney DB, Whiteley AR, O'Donnell MJ, and Dubreuil TL
- Subjects
- Age Factors, Animals, Demography, Ecosystem, Models, Theoretical, Population Dynamics, Rivers, Seasons, Trout growth & development, Temperature, Trout physiology, Water Movements
- Abstract
Modelling the effects of environmental change on populations is a key challenge for ecologists, particularly as the pace of change increases. Currently, modelling efforts are limited by difficulties in establishing robust relationships between environmental drivers and population responses. We developed an integrated capture-recapture state-space model to estimate the effects of two key environmental drivers (stream flow and temperature) on demographic rates (body growth, movement and survival) using a long-term (11 years), high-resolution (individually tagged, sampled seasonally) data set of brook trout (Salvelinus fontinalis) from four sites in a stream network. Our integrated model provides an effective context within which to estimate environmental driver effects because it takes full advantage of data by estimating (latent) state values for missing observations, because it propagates uncertainty among model components and because it accounts for the major demographic rates and interactions that contribute to annual survival. We found that stream flow and temperature had strong effects on brook trout demography. Some effects, such as reduction in survival associated with low stream flow and high temperature during the summer season, were consistent across sites and age classes, suggesting that they may serve as robust indicators of vulnerability to environmental change. Other survival effects varied across ages, sites and seasons, indicating that flow and temperature may not be the primary drivers of survival in those cases. Flow and temperature also affected body growth rates; these responses were consistent across sites but differed dramatically between age classes and seasons. Finally, we found that tributary and mainstem sites responded differently to variation in flow and temperature. Annual survival (combination of survival and body growth across seasons) was insensitive to body growth and was most sensitive to flow (positive) and temperature (negative) in the summer and fall. These observations, combined with our ability to estimate the occurrence, magnitude and direction of fish movement between these habitat types, indicated that heterogeneity in response may provide a mechanism providing potential resilience to environmental change. Given that the challenges we faced in our study are likely to be common to many intensive data sets, the integrated modelling approach could be generally applicable and useful., (Published 2014. This article is a U.S. Government work and is in the public domain in the USA.)
- Published
- 2015
- Full Text
- View/download PDF
34. The identification and characterization of a STAT5 gene signature in hematologic malignancies.
- Author
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Sonkin D, Palmer M, Rong X, Horrigan K, Regnier CH, Fanton C, Holash J, Pinzon-Ortiz M, Squires M, Sirulnik A, Radimerski T, Schlegel R, Morrissey M, and Cao ZA
- Subjects
- Animals, Cell Line, Tumor, Female, Hematologic Neoplasms drug therapy, Heterografts, Humans, Janus Kinases metabolism, Mice, Mice, Inbred NOD, Mice, SCID, Nitriles, Pyrazoles pharmacology, Pyrimidines, Signal Transduction drug effects, Hematologic Neoplasms genetics, Hematologic Neoplasms metabolism, STAT5 Transcription Factor genetics, STAT5 Transcription Factor metabolism
- Abstract
Background: The JAK-STAT pathway is an important signaling pathway downstream of multiple cytokine and growth factor receptors. Dysregulated JAK-STAT signaling has been implicated in the pathogenesis of multiple human malignancies., Objective: Given this pivotal role of JAK-STAT dysregulation, it is important to identify patients with an overactive JAK-STAT pathway for possible treatment with JAK inhibitors., Methods: We developed a gene signature assay to detect overactive JAK-STAT signaling. The cancer cell line encyclopedia and associated gene-expression data were used to correlate the activation status of STAT5 with the induction of a set of STAT5 target genes., Results: Four target genes were identified (PIM1, CISH, SOCS2, and ID1), the expression of which correlated significantly with pSTAT5 status in 40 hematologic tumor cell lines. In pSTAT5-positive models, the expression of the gene signature genes decreased following ruxolitinib treatment, which corresponded to pSTAT5 downmodulation. In pSTAT5-negative cell lines, neither pSTAT5 modulation nor a change in signature gene expression was observed following ruxolitinib treatment., Conclusions: The gene signature can potentially be used to stratify or enrich for patient populations with activated JAK-STAT5 signaling that might benefit from treatments targeting JAK-STAT signaling. Furthermore, the 4-gene signature is a predictor of the pharmacodynamic effects of ruxolitinib.
- Published
- 2015
- Full Text
- View/download PDF
35. High abundance of plasma cells secreting transglutaminase 2-specific IgA autoantibodies with limited somatic hypermutation in celiac disease intestinal lesions.
- Author
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Di Niro R, Mesin L, Zheng NY, Stamnaes J, Morrissey M, Lee JH, Huang M, Iversen R, du Pré MF, Qiao SW, Lundin KE, Wilson PC, and Sollid LM
- Subjects
- Autoantibodies blood, B-Lymphocytes immunology, Celiac Disease blood, Escherichia coli genetics, Escherichia coli metabolism, GTP-Binding Proteins blood, Glutens immunology, HLA-DQ Antigens immunology, Humans, Immunoglobulin A blood, Immunoglobulin D blood, Immunoglobulin D immunology, Immunoglobulin G blood, Immunoglobulin G immunology, Immunoglobulin M blood, Immunoglobulin M immunology, Intestinal Mucosa enzymology, Intestinal Mucosa immunology, Mutation, Plasma Cells immunology, Plasma Cells metabolism, Protein Glutamine gamma Glutamyltransferase 2, Single-Cell Analysis, Somatic Hypermutation, Immunoglobulin, Transglutaminases blood, Antibody-Producing Cells immunology, Autoantibodies immunology, Celiac Disease immunology, GTP-Binding Proteins immunology, GTP-Binding Proteins metabolism, Immunoglobulin A immunology, Transglutaminases immunology, Transglutaminases metabolism
- Abstract
Celiac disease is an immune-mediated disorder in which mucosal autoantibodies to the enzyme transglutaminase 2 (TG2) are generated in response to the exogenous antigen gluten in individuals who express human leukocyte antigen HLA-DQ2 or HLA-DQ8 (ref. 3). We assessed in a comprehensive and nonbiased manner the IgA anti-TG2 response by expression cloning of the antibody repertoire of ex vivo-isolated intestinal antibody-secreting cells (ASCs). We found that TG2-specific plasma cells are markedly expanded within the duodenal mucosa in individuals with active celiac disease. TG2-specific antibodies were of high affinity yet showed little adaptation by somatic mutations. Unlike infection-induced peripheral blood plasmablasts, the TG2-specific ASCs had not recently proliferated and were not short-lived ex vivo. Altogether, these observations demonstrate that there is a germline repertoire with high affinity for TG2 that may favor massive generation of autoreactive B cells. TG2-specific antibodies did not block enzymatic activity and served as substrates for TG2-mediated crosslinking when expressed as IgD or IgM but not as IgA1 or IgG1. This could result in preferential recruitment of plasma cells from naive IgD- and IgM-expressing B cells, thus possibly explaining why the antibody response to TG2 bears signs of a primary immune response despite the disease chronicity.
- Published
- 2012
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36. Broadly cross-reactive antibodies dominate the human B cell response against 2009 pandemic H1N1 influenza virus infection.
- Author
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Wrammert J, Koutsonanos D, Li GM, Edupuganti S, Sui J, Morrissey M, McCausland M, Skountzou I, Hornig M, Lipkin WI, Mehta A, Razavi B, Del Rio C, Zheng NY, Lee JH, Huang M, Ali Z, Kaur K, Andrews S, Amara RR, Wang Y, Das SR, O'Donnell CD, Yewdell JW, Subbarao K, Marasco WA, Mulligan MJ, Compans R, Ahmed R, and Wilson PC
- Subjects
- Adult, Animals, Cross Reactions, Epitopes, B-Lymphocyte immunology, Female, Humans, Immunologic Memory, Lymphocyte Activation, Male, Mice, Mice, Inbred BALB C, Middle Aged, Young Adult, Antibodies, Viral immunology, B-Lymphocytes immunology, Influenza A Virus, H1N1 Subtype immunology, Influenza, Human immunology
- Abstract
The 2009 pandemic H1N1 influenza pandemic demonstrated the global health threat of reassortant influenza strains. Herein, we report a detailed analysis of plasmablast and monoclonal antibody responses induced by pandemic H1N1 infection in humans. Unlike antibodies elicited by annual influenza vaccinations, most neutralizing antibodies induced by pandemic H1N1 infection were broadly cross-reactive against epitopes in the hemagglutinin (HA) stalk and head domain of multiple influenza strains. The antibodies were from cells that had undergone extensive affinity maturation. Based on these observations, we postulate that the plasmablasts producing these broadly neutralizing antibodies were predominantly derived from activated memory B cells specific for epitopes conserved in several influenza strains. Consequently, most neutralizing antibodies were broadly reactive against divergent H1N1 and H5N1 influenza strains. This suggests that a pan-influenza vaccine may be possible, given the right immunogen. Antibodies generated potently protected and rescued mice from lethal challenge with pandemic H1N1 or antigenically distinct influenza strains, making them excellent therapeutic candidates.
- Published
- 2011
- Full Text
- View/download PDF
37. Interfering with resistance to smoothened antagonists by inhibition of the PI3K pathway in medulloblastoma.
- Author
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Buonamici S, Williams J, Morrissey M, Wang A, Guo R, Vattay A, Hsiao K, Yuan J, Green J, Ospina B, Yu Q, Ostrom L, Fordjour P, Anderson DL, Monahan JE, Kelleher JF, Peukert S, Pan S, Wu X, Maira SM, García-Echeverría C, Briggs KJ, Watkins DN, Yao YM, Lengauer C, Warmuth M, Sellers WR, and Dorsch M
- Subjects
- Aminopyridines therapeutic use, Animals, Cell Proliferation drug effects, Gene Amplification drug effects, Hedgehog Proteins metabolism, Insulin-Like Growth Factor I metabolism, Kruppel-Like Transcription Factors metabolism, Medulloblastoma drug therapy, Medulloblastoma genetics, Medulloblastoma pathology, Mice, Morpholines therapeutic use, Mutation genetics, Phosphatidylinositol 3-Kinases metabolism, Protein Kinase Inhibitors therapeutic use, Receptors, G-Protein-Coupled metabolism, Smoothened Receptor, Tumor Suppressor Protein p53 metabolism, Up-Regulation drug effects, Zinc Finger Protein Gli2, Aminopyridines pharmacology, Drug Resistance, Neoplasm drug effects, Medulloblastoma enzymology, Morpholines pharmacology, Phosphoinositide-3 Kinase Inhibitors, Protein Kinase Inhibitors pharmacology, Receptors, G-Protein-Coupled antagonists & inhibitors, Signal Transduction drug effects
- Abstract
The malignant brain cancer medulloblastoma is characterized by mutations in Hedgehog (Hh) signaling pathway genes, which lead to constitutive activation of the G protein (heterotrimeric guanosine triphosphate-binding protein)-coupled receptor Smoothened (Smo). The Smo antagonist NVP-LDE225 inhibits Hh signaling and induces tumor regression in animal models of medulloblastoma. However, evidence of resistance was observed during the course of treatment. Molecular analysis of resistant tumors revealed several resistance mechanisms. We noted chromosomal amplification of Gli2, a downstream effector of Hh signaling, and, more rarely, point mutations in Smo that led to reactivated Hh signaling and restored tumor growth. Analysis of pathway gene expression signatures also, unexpectedly, identified up-regulation of phosphatidylinositol 3-kinase (PI3K) signaling in resistant tumors as another potential mechanism of resistance. Probing the relevance of increased PI3K signaling, we demonstrated that addition of the PI3K inhibitor NVP-BKM120 or the dual PI3K-mTOR (mammalian target of rapamycin) inhibitor NVP-BEZ235 to the initial treatment with the Smo antagonist markedly delayed the development of resistance. Our findings may be useful in informing treatment strategies for medulloblastoma.
- Published
- 2010
- Full Text
- View/download PDF
38. The Polycomb group protein Bmi-1 is essential for the growth of multiple myeloma cells.
- Author
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Jagani Z, Wiederschain D, Loo A, He D, Mosher R, Fordjour P, Monahan J, Morrissey M, Yao YM, Lengauer C, Warmuth M, Sellers WR, and Dorsch M
- Subjects
- Animals, Apoptosis genetics, Apoptosis Regulatory Proteins genetics, Apoptosis Regulatory Proteins metabolism, Bcl-2-Like Protein 11, Cell Growth Processes genetics, Cell Line, Tumor, Female, Gene Expression Regulation, Neoplastic, Humans, Membrane Proteins genetics, Membrane Proteins metabolism, Mice, Mice, Inbred NOD, Mice, SCID, Multiple Myeloma genetics, Multiple Myeloma metabolism, Nuclear Proteins genetics, Polycomb Repressive Complex 1, Proto-Oncogene Proteins genetics, Repressor Proteins genetics, Transcription, Genetic, Multiple Myeloma pathology, Nuclear Proteins metabolism, Proto-Oncogene Proteins metabolism, Repressor Proteins metabolism
- Abstract
Bmi-1 is a member of the Polycomb group family of proteins that function in the epigenetic silencing of genes governing self-renewal, differentiation, and proliferation. Bmi-1 was first identified through its ability to accelerate c-Myc-induced lymphomagenesis. Subsequent studies have further supported an oncogenic role for Bmi-1 in several cancers including those of the breast, lung, prostate, and brain. Using a stable and inducible shRNA system to silence Bmi-1 gene expression, we show a novel role for Bmi-1 in regulating the growth and clonogenic capacity of multiple myeloma cells both in vitro and in vivo. Moreover, to elucidate novel gene targets controlled by Bmi-1, global transcriptional profiling studies were performed in the setting of induced loss of Bmi-1 function. We found that the expression of the proapoptotic gene Bim is negatively regulated by Bmi-1 and that Bim knockdown functionally rescues the apoptotic phenotype induced upon loss of Bmi-1. Therefore, these studies not only highlight Bmi-1 as a cancer-dependent factor in multiple myeloma, but also elucidate a novel antiapoptotic mechanism for Bmi-1 function involving the suppression of Bim., (Copyright 2010 AACR.)
- Published
- 2010
- Full Text
- View/download PDF
39. Fourier transform infrared spectroscopy investigation of native tissue matrix modifications using a gamma irradiation process.
- Author
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Shah NB, Wolkers WF, Morrissey M, Sun WQ, and Bischof JC
- Subjects
- Amides chemistry, Humans, Protein Denaturation radiation effects, Protein Structure, Secondary, Solvents, Spectroscopy, Fourier Transform Infrared, Temperature, Water, Collagen metabolism, Collagen radiation effects, Extracellular Matrix metabolism, Extracellular Matrix radiation effects, Gamma Rays
- Abstract
In this study, Fourier transform infrared spectroscopy (FTIR) was used to evaluate the effects of a gamma irradiation process on AlloDerm, an implantable tissue device derived from human cadaver skin. Inspection of protein amide bands of dried tissue matrix revealed little difference in overall protein secondary structure between gamma-treated and control tissue matrices, although effects of gamma irradiation became apparent after rehydration. The temperature at which protein denaturation began decreased significantly, from 62.4 degrees C +/- 1.0 degrees C in the nonsterilized control tissue matrix to 53.4 degrees C +/- 1.2 degrees C, 48.7 degrees C +/- 0.3 degrees C, and 46.7 degrees C +/- 0.1 degrees C for 5-, 14- and 20-kGy gamma-treated AlloDerm samples, respectively (p < 0.05). Differences were particularly significant in the details of the tissue matrix denaturation profile, which consisted of multiple transitions. The solvent accessibility of tissue matrix was studied by following the rate of protein proton exchange in heavy water (D(2)O), which was evident from the appearance of the amide-II' band as a function of time to D(2)O exposure. In control tissue matrix, a rapid hydrogen/deuterium exchange was observed, with 50% exchange in 30 min, whereas the gamma-treated tissue matrix exhibited a much slower exchange, reaching the 50% exchange level after more than 2 h. The data indicate significant alterations of extracellular milieu in the tissue matrix after the gamma irradiation process. This study shows that FTIR is a valuable tool for studying protein stability and interactions in complex biological scaffolds such as extracellular tissue matrix.
- Published
- 2009
- Full Text
- View/download PDF
40. Contribution of polycomb homologues Bmi-1 and Mel-18 to medulloblastoma pathogenesis.
- Author
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Wiederschain D, Chen L, Johnson B, Bettano K, Jackson D, Taraszka J, Wang YK, Jones MD, Morrissey M, Deeds J, Mosher R, Fordjour P, Lengauer C, and Benson JD
- Subjects
- Animals, Cell Death, Cell Proliferation, Cell Survival, DNA-Binding Proteins genetics, Down-Regulation genetics, Fibroblasts cytology, Gene Expression, Gene Expression Profiling, Gene Expression Regulation, Neoplastic, HeLa Cells, Humans, Medulloblastoma genetics, Mice, Nuclear Proteins genetics, Polycomb Repressive Complex 1, Polycomb-Group Proteins, Proto-Oncogene Proteins genetics, RNA, Small Interfering metabolism, Rats, Repressor Proteins genetics, Transplantation, Heterologous, DNA-Binding Proteins metabolism, Medulloblastoma pathology, Nuclear Proteins metabolism, Proto-Oncogene Proteins metabolism, Repressor Proteins chemistry, Repressor Proteins metabolism, Sequence Homology, Amino Acid
- Abstract
Bmi-1 and Mel-18 are structural homologues that belong to the Polycomb group of transcriptional regulators and are believed to stably maintain repression of gene expression by altering the state of chromatin at specific promoters. While a number of clinical and experimental observations have implicated Bmi-1 in human tumorigenesis, the role of Mel-18 in cancer cell growth has not been investigated. We report here that short hairpin RNA-mediated knockdown of either Bmi-1 or Mel-18 in human medulloblastoma DAOY cells results in the inhibition of proliferation, loss of clonogenic survival, anchorage-independent growth, and suppression of tumor formation in nude mice. Furthermore, overexpression of both Bmi-1 and Mel-18 significantly increases the clonogenic survival of Rat1 fibroblasts. In contrast, stable downregulation of Bmi-1 or Mel-18 alone does not affect the growth of normal human WI38 fibroblasts. Proteomics-based characterization of Bmi-1 and Mel-18 protein complexes isolated from cancer cells revealed substantial similarities in their respective compositions. Finally, gene expression analysis identified a number of cancer-relevant pathways that may be controlled by Bmi-1 and Mel-18 and also showed that these Polycomb proteins regulate a set of common gene targets. Taken together, these results suggest that Bmi-1 and Mel-18 may have overlapping functions in cancer cell growth.
- Published
- 2007
- Full Text
- View/download PDF
41. GP perceptions of workforce shortage in a rural setting.
- Author
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May J, Jones PD, Cooper RJ, Morrissey M, and Kershaw G
- Subjects
- Cohort Studies, Follow-Up Studies, Health Care Surveys, Humans, New South Wales, Social Perception, Workforce, Attitude of Health Personnel, Family Practice, Physicians, Family supply & distribution, Rural Health Services
- Abstract
Introduction: Currently Australia is experiencing a rural medical workforce shortage, especially among GPs. Strategies aimed at improving this shortage have generally been directed at small and remote rural communities (RRMA 4-7); however, longstanding GP shortages also continue in large (RRMA 3) rural communities. The key to the understanding the rural workforce is the perceptions of GPs themselves. This article compared GP perceptions of workforce shortages in Tamworth, New South Wales, Australia (an RRMA 3 town) with actual levels of workforce participation., Methods: A survey of 31/33 GPs working in the New South Wales town of Tamworth was conducted in 2005. Participating GPs were individually interviewed and were asked to estimate local GP workforce needs, calculate their weekly consulting time sessions and advise if they were accepting new patients. The survey was repeated 12 months later with the same cohort to track workforce change., Results: In May 2005 there were 27.8 full time equivalent (FTE) GPs working in Tamworth (population 42 000). In May 2006 this had risen to 31.5 FTE practitioners. Initially, all practitioners surveyed believed there was a workforce shortage, with no practice accepting new patients. This shortage was perceived to be >10 FTE GPs (6.5%), between 5-9 GPs (64.5%) and between 1-4 GPs by 29% of surveyed GPs. In June 2006 there were 31.5 FTE GPs working in Tamworth. The follow-up survey of 29 GPs revealed a significant shift in their perceptions with only 41.4% of GPs perceiving the shortage as 1-4 FTE GPs (p = 0.2), 17.2% between 5-9 GPs and 41.4% nil. No GPs in the follow-up survey perceived the shortage as >10. At the end of the 12 month study period, 8 of 17 practices were accepting new patients., Conclusion: GP perceptions of shortage largely reflected concurrent workforce changes that occurred during the study period where there was a 12% improvement after a prolonged period of workforce stagnation. This change drove improvements in patient access and in many GPs' minds ameliorated much of the perceived shortage. Many factors may be involved, including the increased use of practice nurses, private billing and start-up capacity. General practitioner perceptions appear to be sensitive to workforce changes, with sampled GPs working with higher patient ratios than those seen as acceptable in metropolitan areas.
- Published
- 2007
42. Community, social capital and Indigenous health in the Northern Territory.
- Author
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Morrissey M
- Subjects
- Delivery of Health Care, Health Status, Humans, Northern Territory, Terminology as Topic, Native Hawaiian or Other Pacific Islander psychology, Residence Characteristics, Social Support
- Abstract
Objective: The objective of this paper is to explore the discourse of 'community' and its offshoots, 'social capital' and 'community capacity building', in the contexts of health service delivery to, and the health status of, Indigenous people in the Northern Territory of Australia, and to link this discourse to the wider context of social control and the management of diversity in a multicultural society., Design: The discourse is subjected to critical theoretical and historical analysis and comparisons are drawn between this and similar discourses in the immigration and settlement area., Results/conclusions: The constitution of Indigenous society as a series of 'communities' and the orientation of primary health care policy towards 'capacity building' has the effect, if not the intention, of depoliticising Indigenous health, whilst reproducing, legitimising and mystifying relations of white dominance and permitting the maintenance of a health service delivery system for Indigenous people which, in relation to need, is grotesquely underfunded and incapable of making serious inroads into the appalling health problems of the Indigenous population.
- Published
- 2006
- Full Text
- View/download PDF
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