This proposal outlines a comprehensive three year training plan for the candidate to become an independent scientist in academic pulmonary and critical care medicine. The candidate earned her Ph.D. from the Tongji Medical College, Huazhong University of Science &Technology (HUST) and M.D. from University of Ulm, Germany. Her research focused on the development of novel immunotherapeutic strategies for patients with acute myeloid leukemia (AML). She subsequently received postdoctoral training in the Department of Dermatology, University of Colorado Denver (UCD). Her research was highly focused on the cell-cell immunological synapse interactions between activated memory T lymphocytes and antigen presenting cells. In addition, she received his first academic appointment at National Jewish Health (NJH) in the Division of Environmental &Occupational Health Sciences (DEOHS) in the Department of Medicine studying the granulomatous lung disease chronic beryllium disease (CBD), which results from exposure to beryllium. Soon after her appointment, she was successful in obtaining a career development award from an institutional K12 through the Pulmonary Division, UCD. This grant provided her with a faculty appointment at an Instructor level at UCD. There, she developed proficiency in genomics, and the immunopathogenic mechanisms of CBD. With Dr. Maier, an expert in environmental sciences, genetics, genomics and bioinformatics, head of DEOHS, serving as the primary mentor, this program provides the candidate additional training in three essential areas: 1) education in genetics, genomics, biostatistics and updates in essential laboratory skills;2) supervised preparation of research publications;3) mentored development of her independent research proposal. Additional mentoring is provided by co-mentor Dr. Day, Ph.D., an expert in pulmonary innate immunity and inflammation with extensive prior mentoring experience, and an expert panel of scientists. This program will take place at UCD and NJH, both of which provide outstanding environment for training scientists. It offers state-of-the-art research and clinical facilities, outstanding opportunities for education and the exchange of scientific ideas, and strong commitment to the candidate's development as an independent investigator. The scientific proposal explores the novel hypothesis that patients with CBD have differential regulation of immune-related genes that ultimately leads to abnormalities in immune function and heightened susceptibility to be. Specifically, the central hypothesis is that Be-exposure results in the inappropriate activation of the JAK/STAT pathway and MAPK pathway and contributes to CBD pathogenesis. Therefore, the goal of this study is to gain insight into the mechanisms and pathogenic pathways important in CBD by conducting an in- depth analysis of the differentially expressed candidate genes in CBD.
Its specific aims are: 1) To demonstrate that the JAK/STAT pathway and MAPK pathway related genes are inappropriately activated in CBD compared to BeS and Be-exposed controls with beryllium-exposure, validating these potentially important CBD candidate gene identified in the microarray analysis using qRT-PCR. 2) To functionally assess the role of the candidate genes in the JAK/STAT pathway and MAPK pathway in CBD pathogenesis by assessing either the over- expression of the gene product using the Flp-in expression analysis system in vitro, or the reduction of the gene product using siRNA "knock down" analysis. 3) To investigate whether the differentially genes are associated with CBD (n=100) and BeS (n=100) in a larger separate population and thus reveal potential novel biomarkers for CBD. Insights garnered from these data will significantly add to the understanding of molecular mechanisms of CDB and may lead to development of potential novel biomarker and new targets for study as potential therapeutic approaches for CBD. Public Health Relevance: Exciting preliminary studies show that the peripheral blood cells from CBD and BeS demonstrate differential gene expression profiles relevant to the immune function and pathogenesis of these processes, compared to Be-exposed non-diseased controls. This proposal is to gain insight into the mechanisms and pathogenic pathways important in CBD by conducting an in-depth analysis of the differentially expressed candidate genes in CBD focusing on a pathway that is to a Th1 immune response. The results may improve our understanding of factors involved in the development of CBD, aspects which determine prognosis, and targets for therapy, serves as a model of exposure-related immunologic disease with potential application to other environmentally induced diseases.

Public Health Relevance

CBD is a granulomatous lung disorder caused by beryllium exposure in the workplace and is characterized by the accumulation of Be-specific CD4-T cells in the lung. Depending on the genetic susceptibility and the nature of the exposure, CBD can occur in up to 20% of exposed workers. Another health effect from exposure to beryllium, beryllium sensitization (BeS), was recently recognized with the use of a diagnostic blood test to determine the immune response to Be, the Be lymphocyte proliferation test (BeLPT). These workers have developed a peripheral immune response to Be, yet have no evidence of chronic lung disease and/or granuloma formation on lung biopsy. The prevalence of BeS ranges from 2-5% of all exposed workers and BeS progresses to CBD at a rate of approximately 8% per year. The molecular mechanisms involved in the development of BeS and CBD still remain poorly understood, including factors important in the cytokine network, immunological synapses (the interface between an antigen-presenting cell and a lymphocyte), and regulatory mechanisms, to name a few. In this study we plan to explore the novel hypothesis that patients with CBD have differential regulation of immune-related genes that ultimately leads to abnormalities in immune function and heightened susceptibility to beryllium. Specifically, we hypothesize that beryllium-exposure results in the inappropriate activation of the JAK/STAT (The Janus kinase/signal transducers and activators of transcription) pathway and MAPK (mitogen-activated protein kinases) pathway and contributes to CBD pathogenesis. In this project we seek to improve our understanding of factors involved in the development of CBD, aspects which determine prognosis, and targets for therapy. While this study will help us understand CBD better, it may also help us understand other similar immune-mediated lung and other diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Research Scientist Development Award - Research & Training (K01)
Project #
5K01ES020857-04
Application #
8520310
Study Section
Special Emphasis Panel (ZES1-LKB-J (K9))
Program Officer
Shreffler, Carol K
Project Start
2011-09-21
Project End
2014-07-31
Budget Start
2013-08-01
Budget End
2014-07-31
Support Year
4
Fiscal Year
2013
Total Cost
$125,172
Indirect Cost
$9,272
Name
National Jewish Health
Department
Type
DUNS #
076443019
City
Denver
State
CO
Country
United States
Zip Code
80206
Li, L; Huang, Z; Gillespie, M et al. (2014) p38 Mitogen-Activated Protein Kinase in beryllium-induced dendritic cell activation. Hum Immunol 75:1155-62