Using a combination of genetic linkage, microarray gene expression and genetic association studies, a group of Brigham and Women’s Hospital/ Harvard Medical School researchers have identified a serine protease inhibitor clade E, member 2, or SERPINE2, “as a novel candidate susceptibility gene for COPD,” according to Sorachai Srisuma, who is presenting the research at the 35th Congress of the International Union of Physiological Sciences in San Diego, March 31 - April 5, 2005.
The collaborative, multi-disciplinary team includes: Sorachai Srisuma, Dawn L. DeMeo, Brigham H. Mecham, Edwin K. Silverman, Scott T. Weiss, Kathleen J. Haley, John J. Reilly, Steven D. Shapiro, and Thomas J. Mariani. Mariani, head of the lab where Srisuma works, said the gene is “the most promising susceptibility candidate due to its biological relevance, its expression correlation with disease characteristics, and the allelic association in COPD families and replication in non-familial COPD patients.”
First major study to seek SERPINE2’s physiological role in lung
Srisuma said SERPINE2 “was of particular interest due to its pattern of expression and relationship to alpha-1-antitypsin, the only gene proven to modify risk to COPD (chronic obstructive pulmonary disease). Our team of human genetic epidemiologists, led by Dawn DeMeo and Edwin Silverman, previously identified a region on chromosome 2 they thought might contain a gene conferring susceptibility to COPD.
“We used DNA chips, or gene expression microarrays, to identify genes within this region that were expressed in the lung,” Srisuma noted. “Subsequently we showed that specific cells in the lungs express SERPINE2, and that its expression is altered in individuals with certain clinical characteristics of COPD. Furthermore, specific forms of the gene, termed polymorphisms or SNPs, were more common in people who developed COPD,” he said. Taken together, “these data strongly suggest SERPINE2 is a gene capable of modifying COPD risk, particularly in response to smoking,” he added.
SERPINE2 is a major tissue and cell-associated inhibitor of thrombin and plasmin, but not elastase, Srisuma noted. But no significant study of this protease inhibitor’s expression in normal or diseased lungs had been undertaken previously. “In an effort to begin to gain insight into the physiological role of SERPINE2 in the lung, we investigated the temporal and spatial expression pattern of the gene in mouse and human lungs,” he said.
Analysis of two independent microarray data sets describing normal mouse lung development revealed prominent SERPINE2 expression, which was maximal during formation of the airspaces, which is related to lung maturation. Immunostaining was performed to identify the location of SERPINE2 within the lung.
SERPINE2 shows analogous pattern in diseased human, normal mouse lungs
Prominent immunolocalization of SERPINE2 was observed in a cell-associated pattern within bronchiolar airway epithelial cells and in an extracellular matrix-associated pattern in the vascular adventitia. Immunohistochemistry in human lungs demonstrated an analogous staining pattern. “ Our studies revealed cell-specific and developmentally-regulated expression of SERPINE2 in the lung, which supports further investigation of this gene’s role in human lung diseases,” Srisuma said.
Next steps. Already the group has characterized where and when SERPINE2 is expressed in the lung at various developmental stages. “We are also investigating the specific role of SERPINE2 mutations in lung function and the risk of developing COPD in humans and genetically modified animals,” Srisuma noted. “We hope this line of investigation will show how SERPINE2 affects the risk of developing COPD and how smoking contributes to this process,” he added.
In the future, Srisuma will pursue a project identifying potential biomarkers in COPD patients from New England and also in his native Thailand. When he completes his current post-doctoral fellowship and returns home, a collaborative study will proceed related to lung development and pathogenesis of COPD and pediatric lung diseases.