Focusing diagnostics and therapeutics on those most likely to benefit is a key to success at both the public health and clinical levels. The goal of the Spivack laboratory is to identify individuals at particularly high risk for lung malignancy, and selected non-malignant lung diseases, upon whom to focus smoking/toxin exposure cessation (primary prevention), chemoprevention (secondary prevention), and early disease detection efforts.
The laboratory is currently exploring individual susceptibility markers by exploring quantitative gene (mRNA) expression phenotypes, and DNA sequence, methylation and other epigenetic features potentially underlying those expression phenotypes, in vitro and in human populations, in the setting of tobacco, air pollutant, as well as chemopreventive agent exposures. There are both mechanistic and translational components to the studies.
Mechanistically, the role of promoter sequence and epigenetic variation in promoter regions in the 5' regulatory region of carcinogenesis and oxidant pathway genes is being explored in vitro, using human genomic DNA reporter constructs, and native gene regulation models. Technologies include the realtime quantitation of native mRNA by the laboratory's RNA-specific strategy (patent pending), along with the laboratory's recently-developed tagged-bisulfite genomic sequencing strategy to determine CpG methylation status (tBGS, patent pending). Mechanisms of sequence-defined individual differences in chemopreventive responsiveness are being explored. Whole genome approaches to identifying selected early molecular events in lung cells upon various exposures are underway. Additionally, a recent endeavor is participation in the development of a comprehensive strategy for assaying microRNA and mRNA in parallel, initiated by collaborators, for determining the role of miRNA in candidate gene regulation and carcinogenesis.
Translationally, biomarkers are being established by pairing laser capture microdissected lung and several unique, non-invasively collected surrogate specimens developed in the laboratory, such as mRNA from brush-exfoliated buccal mucosa cells, and DNA from exhaled breath condensate. These specimens continue to accrue from a sampling (currently n>500) of a population assembled in a lung cancer case-control context. The specimens are being studied for quantitative gene expression in the carcinogen and oxidant metabolism, and cell cycle regulatory pathways, by both our RNA-specific RT-PCR approach and by the tBGS DNA methylation approach. These expression, genetic, and epigenetic data are being linked to precise plasma measurements of tobacco exposure, and downstream DNA adduction and mutation events, as an approach to putting a metric to gene-environment interaction. Testing of chemopreventive mixture and individual compounds for antimutagen and antioxidant activity is underway. The overall aim is to develop informative non-invasive risk profiling, preventive, and early disease detection strategies for the lung in human populations.
Selected Publications
- Han W, Reilly AA, Spivack SD. DNA methylation ascertained non-invasively from exhaled breath condensate. [In preparation].
- Hurteau GJ, Carlson AJ, Spivack, SD, Brock GJ. Restoration of E-Cadherin expression by over-expression of the microRNA hsa-miR-200c via reduced expression of the transcription factor TCF8. Cancer Res. 67:7972-76, 2007 [priority report].
- Hurteau, GJ, Spivack SD, Brock G. Parallel identification of miRNA and target mRNA by combined informatics and qRT-PCR approaches: application to has-miR-200c. Cell Cycle 5(17):1951-56, 2006.
- Han W, Cauchi S, Herman JG, Spivack SD. Methylation mapping of DNA by tag-modified bisulfite genomic DNA sequencing. Analytic Biochem. 355: 50-61, 2006.
- Cauchi S, Han W, Kumar SV, Spivack SD. Haplotype-environment interactions regulating the human GSTP1 promoter Cancer Res. 66(12): 6439-6448, 2006.
- Kumar SV, Hurteau GJ, Spivack SD. Validity of mRNA expression analyses of human saliva. Clin. Cancer Res. 12: 5033-39, 2006.
- Han W, Pentecost BT, Pietropaolo RL, Fasco MJ, Spivack SD. ERá increases basal and cigarette smoke-induced expression of CYP1A1 and CYP1B1, but not GSTP1 in normal human bronchial epithelial cells. Molec. Carcinogenesis, 44(3):202-211, 2005.
- Spivack SD, Hurteau GJ, Jain R, Kumar SV, Aldous KM, Gierthy JF, Kaminsky LS. Gene-environment interaction signatures by quantitative mRNA profiling in exfoliated buccal mucosal cells. Cancer Res, 64:6805-6813, 2004.
- Spivack SD, Hurteau GJ, Fasco MJ, Kaminsky LS. Phase I and II carcinogen metabolism gene expression in human lung tissue and tumors. Clinical Cancer Research, 9:6002-6011, 2003.
- Han W, Pentecost BT, Spivack SD. Functional evaluation of novel SNPs and haplotypes in the promoter region of CYP1B1 and CYP1A1 genes. Molec. Carcinogenesis 37:158-69, 2003.
- Fasco MJ*, Hurteau, GJ, SpivackSD. Gender-dependent expression of alpha and beta estrogen receptors in human nontumor and tumor lung tissue. Molecular Cellular Endocrin, 188(1-2):125-40. 2002.
- Hurteau GJ, Spivack SD. mRNA-Specific RT-PCR from human tissue extracts. Analytic Biochemistry 307:304-15. 2002.
- Spivack SD, EJ Mark*. Clinical-pathologic correlation (CPC), talcosilicosis and lung carcinoma. New England J of Medicine. 341(3):182. July 15, 1999.