Adamko, MD, FRCPC
Department of Pediatric Pulmonary Medicine
University of Saskatchewan
The ultimate goal of my research program is to help doctors prevent hospitalizations in people with chronic airways diseases like asthma, COPD, or cystic fibrosis. Currently, my main focus is to develop a clinic-based urine test that would drastically improve the diagnosis and management of asthma and other similar respiratory diseases. There is no single test that can accurately diagnose and help manage asthma and other inflammatory respiratory diseases non-invasively in a typical doctor’s office setting. This is a serious problem since inaccuracy of diagnosis and mismanagement of these diseases leads to exacerbations, or flare-ups, resulting in increased hospital visits and decreased quality of life for these patients. Asthma specifically, is the most common chronic illness in children, and it is a leading medical expense.
We propose that urine is an ideal fluid for such medical diagnostics due to its ease of collection in patients of all ages and richness of underlying chemical composition. Metabolomics is the study of small molecules generated from cellular metabolic activity. Tissue damage, resulting from inflammatory airway diseases generates markers in the body that are excreted and can be measured in the urine. The overall working hypothesis of my research program has been that the urine-specific metabolic signature will be different between people with respiratory inflammatory airway disease and those without.
We began this work using a technology called Nuclear Magnetic Resonance (NMR) because at that time we were interested in screening urine samples for potential biomarkers. NMR can identify and quantify metabolites in urine with little pretreatment in about a day or so. In contrast, another technology call Mass Spectrometry (MS) measures thousands more metabolites than NMR, but its methodology makes screening more difficult. As discussed below, for absolute quantification of metabolites, MS requires specific methods designed for each structurally-related metabolite. Once a set of candidate biomarkers is proposed, it is necessary to obtain isotope labeled standards to confirm metabolite identity, and then develop calibration curves in order to quantify concentrations. This requires an immense of amount of work and likely one could be creating methods to measure irrelevant metabolites.
Overall, while the decision to use NMR has allowed us to rapidly discover potential biomarkers associated with asthma, we have come to recognize two significant problems with NMR when considering its use clinically. First, NMR can require a day or more to get results, when results within hours are desirable. Secondly, the data generated from NMR analysis, while accurate, are less reproducible than that from targeted MS. MS is more sensitive and results can be obtained within an hour. Now that we have the NMR data in place, we are developing targeted MS-based quantitative methods for diagnostic purposes. We also propose to study chronic obstructive pulmonary disease (COPD) and CF as a relevant lung diseases. To do this, we continue to recruit and monitor subjects with asthma, COPD and CF. We believe this work will ultimately benefit all people affected by respiratory disease in this province, will lead to a marketable diagnostic product.
In addition, most asthma exacerbations are caused by infection with common airway viruses. Unlike healthy individuals, infection with common airway viruses in patients with allergic asthma leads to the development of inflammation characterized by an increased number of eosinophils. My other objective has been to investigate the way eosinophils and allergic inflammation can respond to airway virus infection with a view to understanding the way viruses induce asthma attacks. Based on past papers we believe the virus stimulates allergic inflammation to induce damage of the lung nervous system. In the lungs, dominant control of airway smooth muscle is through the parasympathetic nervous system. M2 muscarinic receptors act to control the stimulation of smooth muscle by inhibiting acetylcholine release.
Similar to the pathology seen in humans, we have demonstrated that virus-induced M2 receptor dysfunction and subsequent AHR in allergen-sensitized guinea pigs are mediated via eosinophil major basic protein. We have shown that in vitro eosinophils response to virus infection through memory CD4+ T cells. We have data showing that memory response to virus in vivo also induces airway hyperresponsiveness.
Finally as a pediatrician, I am involved with several clinical studies in infants and children.
- Darryl J. Adamko, B.L. Yost, A.D. Fryer, and D.B. Jacoby. Ovalbumin sensitization changes the inflammatory response to subsequent viral infection. Eosinophils mediate airway responsiveness, M2 muscarinic receptor dysfunction, and antiviral effects. 1999, J Exp Med; 190 (10): 1465-78.
- Darryl J. Adamko, A.D. Fryer, B.S. Bochner and D.B. Jacoby. CD8+ T-Lymphocytes in Viral Hyperreactivity and M2 Muscarinic Receptor Dysfunction. 2003. Am J Respir Crit Care Med 167(4):550-6.
- Francis Davoine, Min Cao, Yingqi Wu, Farnam Ajamian, Ramses Ilarraza, Andy Kokaji, Redwan Moqbel, and Darryl J. Adamko. Virus-induced eosinophil mediator release requires antigen presenting and CD4+ T-cells. J Allergy Clin Immunol. 2008 Jul;122(1):69-77, 77.e1-2. Epub 2008 May 9.
- Ramses Ilarraza, Yingqi Wu, Christopher D. Skappak, Farnam Ajamian, David Proud, and Darryl J. Adamko. Rhinovirus Has The Unique Ability To Directly Activate Human T Cells In Vitro. J Allergy Clin Immunol, 2013, Feb;131(2):395-404. (doi: 10.1016/j.jaci.2012.11.041). PMID: 23374267
These papers showed how viruses could be involved in asthma exacerbation. We demonstrated that the mechanism of virus-induced airway hyperreactivity (AHR) in an allergic inflammatory environment was secondary to an eosinophil-mediated process of M2 muscarinic receptor (M2R) dysfunction. This occurred through CD8+ T cells in animals with no previous exposure to virus. We then showed that viral antigen presentation is another important mechanism that could activate eosinophils, in vitro. We have a paper submitted that confirms this can also occur in vivo. In addition, Rhino virus appears to have a unique ability to activate T cells and eosinophils in vitro, which was not seen by other airway viruses. This might explain the importance of RV in the association with asthma attacks.
- Darryl Adamko, Ying Wu, Gerald Gleich, Paige Lacy and Redwan Moqbel. The Induction of Eosinophil Peroxidase Release: Improved Methods of Measurement and Stimulation. J Immunological Methods. 2004 Aug;291(1-2):101-8.
This paper described a novel method of measuring EPO activity. Further, from this in vitro work, I have isolated an important mechanism for EPO release, including the intracellular signaling pathway.
- Darryl J. Adamko, Yingqi Wu, Farnam Ajamian, Ramses Ilarraza, Redwan Moqbel, and Gerald J. Gleich. The Effect of Cationic Charge on Release of Eosinophil Mediators. 2008, J. Allergy Clin Immunol. 2008 Aug;122(2):383-90, 390.e1-4. Epub 2008 May 2. PMID: 18455220.
This publication showed that cationic charges induce eosinophil mediator release by the polarization of surface receptors. Hence, cationic proteins deposited on atopic tissues could enhance in situ autocrine activation.
- Ramses Ilarraza, Yingqi Wu, Francis Davoine, Cory Ebeling, and Darryl J. Adamko. Human dendritic cells promote an antiviral immune response when stimulated by CVT-E002. J Pharm Pharmacol. 2011 May;63(5):670-678. doi: 10.1111/j.2042-7158.2011.01257.x. Epub 2011 Apr 4. PMID: 21492169
- Cory Ebeling, Yinqi Wu , John R. Gordon, and Darryl J. Adamko. Compound CVT-E002 significantly reduces allergen induced airway inflammation and airway hyperresponsiveness in vivo. Molecular Nutrition and Food Research, 2011, Dec;55(12):1905-8. doi: 10.1002/mnfr.201100216. Epub 2011 Oct 31.
- Ramses Ilarraza, Yingqi Wu, and Darryl J. Adamko. Montelukast inhibits leukotriene stimulation of human dendritic cells in vitro. Int Arch Allergy Immunol 2012;159:422-427 (DOI: 10.1159/000338818).
These papers demonstrated the importance of human dendritic cells in the response to virus infection, and showed some potential modulating compounds.
- Farnam Ajamian, Yingqi Wu, Cory Ebeling, Ramses Ilarraza, Solomon O. Odemuyiwa, Redwan Moqbel, Darryl J. Adamko. Infection of Human Dendritic Cells by Respiratory Syncytial Virus Induces Th2 Imbalance Through Indoleamine 2,3 Dioxygenase. Clin Exp Allergy, 2015, Mar;45(3):644-59. doi: 10.1111/cea.12498.
This paper demonstrated a novel potential mechanism for the induction of Th2 responses after RSV infection.
- Erik J. Saude, Idongesit P. Obiefuna, Ray L. Somorjai, Farnam Ajamian, Christopher Skappak, Taisir Ahmad, Brion K. Dolenko, Brian D. Sykes, Redwan Moqbel, and Darryl J. Adamko. Metabolomic Biomarkers in an Animal Model of Asthma: Urine Nuclear Magnetic Resonance. Am J Respir Crit Care Med. 2009 Jan 1;179(1):25-34. Epub 2008 Oct 17. PMID: 18931331
- Erik J. Saude, Christopher Skappak, Kim Cook, Shana Regush, Brian H. Rowe, Redwan Moqbel, Brian D. Sykes, and Darryl J. Adamko. Metabolomic Profiling Pediatric Asthma: the Diagnostic Utility of Urine NMR. J Allergy Clin Immunol. 2011 Mar;127(3):757-64.e1-6. PMID: 21377043.
- Christopher Skappak, Shana Regush, Po-Yin Chung, and Darryl J Adamko, Identifying hypoxia in a fetal piglet model using urinary NMR metabolomic profiling. PLoS One. 2013, May 31;8(5):e65035. doi: 10.1371/journal.pone.0065035. PMID: 23741447
- Darryl J. Adamko, Parameswaran Nair, Irvin Mayers, Ross T. Tsuyuki, Shana Regush, and Brian H. Rowe. Metabolomic Profiling of Asthma and COPD: A Pilot Study Differentiating Diseases. J Allergy Clin Immunol. 2015 Jul 4. pii: S0091-6749(15)00728-9. doi: 10.1016/j.jaci.2015.05.022. [Epub ahead of print] PMID: 26152317.
These papers demonstrate show our work in the area of metabolomic analysis of airways diseases. We started in an animal model of asthma and found that urine metabolomic analysis could differentiate animal groups. This method was extended to children with asthma. It worked in an animal model of neonatal asphyxia. We then showed that different respiratory diseases (asthma vs. COPD) can also be differentiated.
Current Sources of Funding
The Western Diversification Program: Improved diagnosis of respiratory disease: Metabolomic analysis of urine. Lead Investigator of team grant ($235,000).
Children’s Hospital Foundation of Saskatchewan: Improved diagnosis of pediatric asthma: Metabolomic analysis of urine. Lead Investigator of team grant ($67,000/year).
AllerGen, NCE - Noninvasive assessment of atopic diseases: Metabolomic profiling of urine using Nuclear Magnetic Resonance (NMR); Lead Investigator of team grant ($50,000/year)
Saskatchewan Health Research Foundation (SHRF) Establishment Grant: Metabolomic analysis of urine: Improving the diagnosis of asthma and COPD; Lead Investigator of team grant ($60,000/year).
Current Research Personnel
- Yingqi (Chad) Wu, technician since 2001.
- Shana Regush, technician since 2010.
- Hanan Awad, research assistant since 2011.
- Mona Hamada, Grad Student (PhD) since 2011.