Exposure in Epigenetic Regulation of Immune Response in Chronic Beryllium Disease (CBD)

This study will provide important results for each aim, while also providing an integrative transcriptional and epigenomic profile of CBD.

In Aim 1 the Investigator will define genome-wide epigenetic alterations of CBD, by determining genes that are DM in pivotal immune cells, in the target organ (CD4+ BAL cells) in CBD compared to BeS and healthy controls. In addition, the Investigator will determine the impact of Be exposure on the methylation profile of CBD and BeS cells compared to each other and normal controls. This information will be used to define DM regions, genes and their networks.

Using the cases and controls from Aim 1, we will evaluate the gene-expression from these same subjects in Aim 2 to define functional epigenetic loci based on DE in CD4+ BAL cells with and without Be exposure. The Investigator will also integrate ENCODE/RE methylation, histone modification, and chromatin accessibility data as well as our genome-wide association study (GWAS) data to prioritize epigenetic marks and networks for confirmation and validation in Aim 3. In Aim 3, the Investigator will test the generalizability of their findings, explore the potential of methylation marks as biomarkers of disease in PBMCs and determine if change in methylation of these targets with AZA or folic acid affects key immune and regulatory pathways in a second set of CBD and BeS subjects. Throughout the Aims, the Investigator will use both fresh CD4+ T cells to directly assess disease relevance and Be-stimulated cultured CD4+ T cells (compared to unstimulated cultured T cells) to assess the impact of environmental exposure .


Eligibility Criteria:
Inclusion Criteria:

Chronic Beryllium Disease (CBD):

1. History of Beryllium exposure
2. Positive blood and/or bronchoalveolar lavage (BAL) Beryllium Lymphocyte Proliferation Tests (BeLPT)
3. Biopsy-proven pathologic changes consistent with CBD, specifically non-caseating granulomas and/or mononuclear cell interstitial infiltrates.

Beryllium Sensitization:

1. History of Beryllium exposure
2. Two or more positive blood beryllium lymphocyte proliferation tests (BeLPT) or positive bronchoalveolar lavage (BAL) BeLPT
3. Normal lung tissue (no histology suggestive of CBD).

Normal Controls:

1. No history of beryllium exposure
2. Former smokers or never smokers -

Exclusion Criteria:

Chronic Beryllium Disease:

1. Immunosuppressive therapy within the last three months
2. Current cigarette smoking or smoking within six months prior to the study
3. Positive lung washing or biopsy cultures for fungi, mycobacteria or other respiratory pathogen consistent with an acute or chronic infection
4. Weight less than 110 lbs. (for venipuncture)
5. Pregnancy
6. Severe room air hypoxemia and or hypercapnia (precluding BAL), e.g., resting PaO2 \< 45, PaCO2 \> 45 mm Hg; (Denver altitude 5,280 feet)
7. Presence of another disease that may be expected to significantly affect patient mortality and or the immune response (e.g., HIV, HCV, cancer, uncorrected bleeding diathesis, acute hypercapnia with a resting PaCO2 above 45 mm Hg; serious cardiac arrhythmia, recent myocardial infarction within 6 weeks)
8. Patient inability to participate in the study, such as inability to undergo venipuncture and BAL procedures that form part of the inclusion/exclusion criteria or part of the outcome measure

Beryllium Sensitization:

1. Known underlying systemic or lung disease;
2. Current cigarette smoking or smoking within six months prior to the study
3. Positive lung pathology consistent with CBD
4. Pregnancy
5. Weight less than 110 lbs. (for venipuncture)
6. Presence of another disease that may be expected to significantly affect patient mortality and or the immune response (e.g., HIV, HCV, cancer, uncorrected bleeding diathesis, serious cardiac arrhythmia; recent myocardial infarction within 6 weeks)
7. Patient inability to participate in the study, such as inability to undergo venipuncture and BAL procedures that form part of the inclusion/exclusion criteria or part of the outcome measure

Normal Controls:

1. History of beryllium exposure
2. Known underlying systemic or lung disease;
3. Immunosuppressive therapy or other medication for as systemic disease process in the last 3 months;
4. Current smokers or smoking within 6 months of study
5. Pregnancy
6. Weight less than 110 lbs. (for venipuncture)
7. Inability to undergo BAL or venipuncture procedures -