Detroit R & D, Inc.

Other

Oxford Biomedical Research, Inc.

Vanderbilt University

Hyesook Kim

Deanna Kroetz

Jorge H Capdevila

Raymond R Novak

Raymond F Novak

Denis Marc Callewaert

L Jackson Roberts

Brian Brummeler Haab

Jason D Morrow

Sohee Kim

Alan A Dombkowski

Edward Y Kim

David A Putt

Elizabeth Starr Roberts-Kirchhoff

Jonathan Charnecki

This invention discloses diagnosis of risk of chemotherapy-induced cardiotoxicity by measurement of increased expression of soluble epoxide hydrolase in vitro and in vivo in cells, tissues or animals including measurement of increased levels of soluble epoxide hydrolase metabolites, e.g., 14,15-DHET and 11,12-DHET, in biological fluids. This invention also includes diagnosis of risk of chemotherapy-induced cardiotoxicity by measuring increased levels of oxidative stress in cells, tissues or animals including measurement of increased levels of oxidative stress biomarkers, e.g., 8-isoprostane, in biological fluids. Fatty acid and protein biomarkers to diagnose the risk of chemotherapy-induced cardiotoxicity are detected using various detection methods including mass spectrometry and immunoassay such as ELISA, Western blot analysis or label-free microwell and nanowell technologies. This invention discloses targeted medical intervention for a subject who is at risk or with chemotherapy-induced cardiotoxicity by treating with soluble epoxide hydrolase inhibitor(s) with or without antioxidants to prevent or ameliorate the chemotherapy-induced cardiotoxicity.

Methods for identifying risk of chemotherapy-induced cardiotoxicity and targeted medical intervention

A method of characterizing the protein O-GlcNAcylation site-specificity of an antibody. A method of detecting or quantitating the expression of site-specific O-GlcNAcylated proteins expressed in cells and biological samples. A method of diagnosing cancer in a host based on the cellular expression of site-specific O-GlcNAcylated proteins. A method of screening anti-cancer compounds according to their ability to increase a level O-GlcNAcylation of oncogene or tumor suppressor proteins. Methods of treating cancer in an animal host by administering compounds that increase a level of O-GlcNAcylated c-myc or p53 in cancer cells. A method of distinguishing subclasses of pancreatic cancer according to the sensitivity of pancreatic cancer cells to an imidazole derivative, and a method of personalized pancreatic cancer treatment delivered according to the sensitivity subclasses.

Glycosylation site-specific antibodies and anti-cancer compounds

A method of producing form-specific anti-peptide antibodies for a wild type protein and its one amino acid mutated protein using a peptide antigen, by obtaining a protein sequence of the wild type protein and its one amino acid mutated protein, selecting a continuous amino acid sequence without any internal cysteine residues that includes the one amino acid mutated sequence and wild type sequence corresponding to the mutated site at the end of the sequence to obtain a synthetic mutation peptide and a synthetic wild type peptide, conjugating the synthetic peptides to a carrier protein, and immunizing an animal to produce antibodies. Methods of detecting cancer and methods of treating cancer.

Form-specific antibodies for NAG-1 (MIC-1, GDF-15), H6D and other TGF-&#x3b2; subfamily and heart disease and cancer diagnoses

An antibody microarray screen including a substrate, monoclonal and polyclonal antibodies that are purified immunoglobins, wherein the antibodies are spotted on predetermined positions on the substrate, and fluids unprocessed for immunoglobulin isolation (e.g., anti-sera, ascites fluids, or hybridoma culture media), wherein the unprocessed fluids are spotted on the predetermined positions on the substrate. Production of drug-metabolizing enzyme antibody microarrays containing closely related cytochromes P450 is disclosed. Methods of manufacturing an antibody microarray, an internal control molecule for use in an antibody microarray, a method of determining optimal spotting concentrations of IgG and a method to increase a detectable signal with microarray analysis are disclosed.

Chip production, hybridization and data interpretation for antibody and protein microarrays

A method of assessing arachidonic acid (AA) metabolites-dependent hypertension by measuring glucuronidated dihydroxyeicosatrienoic acids (DHETs) and DHET metabolites in a biological sample which contains the epitopes unique to DHET (using any methods including GC/MS, LC/MS or ELISA). An example of the glucuronidated DHET metabolite is DHET-alcohols such as omega or omega-1 oxidated DHET and DHET esterified glycerol. The method further includes determining the amount of glucuronidated molecules containing a DHET-specific epitope which is immunoreactive with antibodies produced against DHETs.

Detection of hypertension using glucuronidated metabolic products

A method to assess arachidonic acid (AA) metabolites-dependent hypertension by measuring glucuronidated dihydroxyeicosatrienoic acids (DHETs) and DHET metabolites in a biological sample which contains the epitopes unique to DHET (using any methods including GC/MS, LC/MS or ELISA) is disclosed. An example of the glucuronidated DHET metabolite is DHET-alcohols such as omega or omega-1 oxidated DHET and DHET esterified glycerol. The method further includes determining the amount of glucuronidated molecules containing a DHET-specific epitope which is immunoreactive with antibodies produced against DHETs. The present invention measuring glucuronidated DHET levels in a biological sample is useful for drug development and monitoring efficiency of drug treatment of a mammal who has AA epoxygenase-, epoxide hydrolase-and/or UDP-glucuronosyl transferase-dependent hypertension.

There is provided a method of assessing oxidant stress by measuring polymerization of proteins. Also provided is a marker for oxidant stress which includes a polymerized protein. A kit for use in assessing oxidant stress, the kit including an assay for detecting polymerized proteins is also provided. A method of lowering oxidant stress by administering to a patient an effective amount of at least one reducing agent is also provided. A pharmaceutical composition for lowering oxidant stress, the pharmaceutical having an effective amount of reducing agent and a pharmaceutically acceptable carrier is also provided.

Assessment of oxidant stress in vitro and in vivo

A method to assess hypertension by measuring the amount of free and conjugated hydroxyeicosatrienoic acids (DHETs) and metabolites of DHETs, which are metabolites of arachidonic acid (AA) epoxygenases and epoxide hydrolases, in a biological sample which contains the DHETs (using any methods including GC/MS or ELISA) is disclosed. The method further included determining the amount of molecules containing a DHET-specific epitope immunoreactive with antibodies produced against DHETs present in the sample. This amount is compared with a control sample(s). Hypertension is determined through the comparison wherein the amount of increase of free and conjugated DHETs and metabolites of DHETs in the sample isolated from an organism. The present invention also provides a method to assess catalytic activity of AA epoxygenases using immunoassays by subtracting the amounts of NADPH-independent epoxyeicosatriencic acids (EETs) from total (NADPH-dependent&plus;independent) EETs. The present invention also provides a method to decrease hepatic M epoxygenase expression including 2C23 by treatment of rats with a glucocorticoid including dexamethasone.

Detection of hypertension using immunoreactive metabolic products

A method to assess hypertension by measuring the amount of free and conjugated hydroxyeicosatrienoic acids (DHETS) and metabolites of DHETs, which are metabolites of arachidonic acid (AA) epoxygenases and epoxide hydrolases, in a biological sample which contains the DHETs (using any methods including GC/MS or ELISA) is disclosed. The method further included determining the amount of molecules containing a DHET-specific epitope immunoreactive with antibodies produced against DHETs present in the sample. This amount is compared with a control sample(s). Hypertension is determined through the comparison wherein the amount of increase of free and conjugated DHETs and metabolites of DHETs in the sample isolated from an organism. The present invention also provides a method to assess catalytic activity of AA epoxygenases using immunoassays by measuring the amounts of NADPH-independent epoxyeicosatrienoic acids (EETs) and total (NADPH-dependent&plus;independent) EETs. The present invention also provides a method to decrease hepatic AA epoxygenase expression including 2C23 by treatment of rats with a glucocorticoid including dexamethasone.

A method to assess oxidative stress in vivo by measuring the amount of free, esterified and glucuronidated forms of isoprostanes (8EPGF2) in a biological sample which contains the isoprostanes is disclosed. The method further includes determining the amount of total isoprostanes present in the sample. This amount is compared with a control sample. The oxidative stress is determined through the comparison wherein the amount of isoprostanes increase in the sample isolated from an organism undergoing oxidative stress compared to the control. Alternatively the method of the present invention provides for only the measurement of the glucuronidated form wherein the amount of glucuronidated isoprostanes increase in the sample isolated from an organism undergoing oxidative stress compared to the control.

Assessment of oxidative stress in vivo

A method for production of a form-specific and/or inhibitory antibody against a cytochrome P450 protein is disclosed. The method includes the steps of selecting a cytochrome P450 protein for which a form-specific and/or inhibitory antibody is needed. The amino acid sequence of the selected cytochrome P450 protein is determined and aligned with a comparison amino acid sequence using an alignment algorithm. A peptide sequence corresponding to a region of a substrate recognition site is identified and a peptide of the selected sequence prepared. Using the peptide as an immunogen, an inhibitory and/or form-specific antibody is produced.

Growing community of inventors

Bloomfield Hills, MI, United States of America

Hyesook Kim