权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Development of a carbon monoxide scavenging hemoprotein as a novel antidotal therapy to treat inhaled CO poisoning

开发一氧化碳清除血红蛋白作为治疗吸入性一氧化碳中毒的新型解毒疗法

基本信息

批准号：
10387161
负责人：
Matthew Ryan Dent
金额：
$ 7.68万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-01 至 2023-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10387161
关键词：
Accident and Emergency department Aerobic Affinity Ambulances Amino Acids Animal Model Antidotes Area Behavior monitoring Binding Biochemical Biology Biophysics Blood Pressure Carbon Monoxide Carbon Monoxide Poisoning Carboxyhemoglobin Cardiac Catheters Cessation of life Chemistry Data Development Diagnosis Disease Emergency department visit Erythrocytes Escherichia coli Exhibits Functional disorder Goals Health Heme Hemeproteins Hemoglobin Human Hyperbaric Oxygen In Vitro Inhalation Intravenous Investigation Knowledge Laboratories Leadership Ligands Mediating Medicine Mentors Metabolism Methods Microbe Modeling Molecular Cloning Morbidity - disease rate Mus Neurocognitive Organ Outcome Oxidation-Reduction Oxygen Patient-Focused Outcomes Patients Persons Physicians Physiological Poisoning Principal Investigator Property Protein Engineering Proteins Recombinant Proteins Recombinants Research Safety Scientist Signal Transduction Site Site-Directed Mutagenesis Soil Spectrum Analysis Survivors Technical Expertise Testing Therapeutic Toxic effect Training United States Variant Work autooxidation base career career development chemical stability clinically relevant conventional therapy drug development experience heart rate monitor hemodynamics human disease human model improved in vivo long-term sequelae microbial mortality mouse model nanomolar novel novel therapeutics point of care pre-clinical prevent protein metabolism skills transcription factor translational medicine

项目摘要

PROJECT SUMMARY/ABSTRACT Accidental carbon monoxide (CO) poisoning is the leading cause of human poisoning in the United States, resulting in approximately 50,000 cases and at least 1,500 deaths annually. No point-of-care antidotal therapy exists for CO poisoning to date, and conventional treatments are limited to inhalation of 100% normobaric oxygen or hyperbaric oxygen. While these therapies enhance CO clearance, delays in patient diagnosis and transport contribute to excess morbidity and mortality. Consequently, a fast-acting CO scavenger that can be deployed in the field, ambulance, or emergency room could significantly increase survival and long-term outcomes for patients. Given that CO binds tightly to ferrous heme, our lab seeks to develop a hemoprotein-based CO scavenger that can bind and eliminate CO as a novel therapy for CO poisoning. Based on preliminary studies of recombinant hemoproteins, we have identified four key criteria for a safe and efficacious hemoprotein-based CO scavenger: (1) high (nanomolar) CO affinity to maximize CO scavenging from physiological heme sites, (2) CO selectivity to minimize competitive inhibition by oxygen binding, (3) thermal and chemical stability to prevent heme release and adverse reactivity, and (4) redox stability of the Fe(II) heme to prevent autooxidation to the inactive, Fe(III) heme state. Early investigations of the regulator of CO metabolism (RcoM) protein, a CO- sensing transcription factor from soil microbes, suggest that this protein exhibits high CO affinity and unprecedented selectivity for CO over oxygen. The primary objective of this proposal is to develop RcoM into a safe and efficacious CO scavenger that will serve as an improved therapeutic treatment for CO poisoning. In Aim 1, we will utilize in vitro spectroscopic methods developed in our lab to identify 1) the minimum functional RcoM subunit, and 2) key amino acid residues that confer high CO affinity, selectivity, and heme stability. In addition to characterizing basic biochemical properties, we will assess the ability of recombinantly expressed RcoM variants to scavenge CO from hemoglobin in CO-saturated red blood cells in vitro. In Aim 2, we will evaluate the safety and efficacy of two recombinant RcoM truncates in vivo. We will assess systemic and organ- specific effects of intravenous RcoM delivery in healthy mice in vivo and quantify the ability of RcoM to reverse hemodynamic collapse and prevent death in a preclinical mouse model of CO poisoning previously developed in our laboratory. Completion of the proposed aims will advance our fundamental understanding of hemoprotein ligand selectivity while also advancing the translational development of a novel antidotal therapy to treat inhaled CO poisoning. These outcomes, in addition to career development, mentored training, and didactic coursework, will ultimately provide me with the technical expertise, background knowledge, and leadership skills necessary to accomplish my long-term academic career goal of directing a research team to study CO-dependent signaling mechanisms relevant to human health and disease.

项目总结/文摘