SPORE in Lung Cancer
University of Pittsburgh
Principal Investigator:
James G. Herman, M.D.
Overall Abstract
The University of Pittsburgh Cancer Institute (UPCI) is conducting a Specialized Program of Research Excellence (SPORE) in Lung Cancer. The overall goals of the Lung Cancer SPORE are to improve detection and treatment of lung cancer and to understand the mechanisms of increased susceptibility of women to lung cancer. The SPORE program consists of four major translational research projects in lung cancer, three research cores, an administrative core, a developmental research program, and a career development program. The Lung Cancer SPORE uses interdisciplinary approaches to meet its objectives by carrying out projects with co-investigators in basic, applied, and clinical science. It is also organ-specific in its approach; all projects test hypotheses about lung cancer biology, susceptibility, detection, or treatment. The long-term goal of the UPCI SPORE is to conduct clinical trials, based on research results from its translational research projects that will serve as the basis for improving the outcome of patients diagnosed with lung cancer.
The main projects are:
- Selective Targeting of Estrogen Pathways in Lung Cancer Using Biomarkers
- Vitamin D Modulation of inflammation and Lung Cancer Risk
- Lung Cancer Risk Prediction in PLuSS
- Targeting Neutrophil Elastase in Lung Cancer
The research cores will assist the main research projects, developmental research projects, and career development investigators in carrying out lung cancer translational research. The research cores are Tissue and Blood Bank Core, Clinical Core, and Biostatistics Core. The Administrative Core will provide scientific and fiscal oversight for the program. The Clinical Core interacts with the laboratory investigators and other cores to initiate and implement the clinical activities. UPCI SPORE investigators work together as a team to meet the goals of the program and will also interact with investigators from Lung Cancer SPOREs at other institutions to improve outcome for lung cancer patients.
Project 1: Selective Targeting of Estrogen Pathways in Lung Cancer Using Biomarkers
Project Co-Leaders:
Jill M. Siegfried, Ph.D.
Sanja Dacic, M.D.
There is a growing body of literature showing that steroid hormone pathways are involved in the development and progression of lung cancer in both men and women. Primary non-small cell lung carcinomas (NSCLCs) have been found to express the estrogen receptors α and β (ERα and ERβ), the progesterone receptor (PR), and aromatase, the enzyme that synthesizes β-estradiol from steroid precursors. This project is a continuation of SPORE Project 1 from the previous grant period, where we found that non-genomic ERβ signaling to epidermal growth factor receptor (EGFR) is important in mediating NSCLC growth. Co-inhibition of ER and EGFR also showed enhanced anti-tumor effects. We recently discovered in a lung cancer tissue microarray (TMA) that ERβ is a negative prognostic indicator in NSCLC while PR is a positive prognostic indicator. We will translate these findings to optimally target steroid hormones for lung cancer treatment and to understand the mechanism of PR down-regulation in lung cancer. Specific Aim 1 will carry out an immunohistochemical (IHC) validation study of the combined association of ERβ and PR with patient survival in NSCLC tumors of known EGFR and K-ras mutation status. The hypothesis of Aim 1 is that elevated ERβ expression is associated with poor outcome in NSCLC, while elevated PR expression is associated with good outcome. Extent of tumor expression of EGFR and aromatase may modulate the associations of ERβ and PR with NSCLC outcome. Presence of EGFR mutations or increased EGFR copy number may also modify these survival associations. Specific Aim 2 will determine which form of PR, PRA and/or PRB, is important in NSCLC survival and determine the mechanism of loss of PR expression in lung tumor cells. The hypothesis of Aim 2 is that PR subtypes A and B may by differentially involved in biology or survival of NSCLC patients and that PR may be reduced in lung tumor cells as a result of heightened growth factor signaling. Specific Aim 3 will develop an mRNA expression-based molecular classification of NSCLC by comparing gene expression arrays in tumors characterized by ERβ/PR level. Specific Aim 4 will examine the relationship of pathway biomarkers that show survival effects in Aims 1-3 to clinical response in tissues from lung cancer patients enrolled in two SPORE phase II clinical trials utilizing anti-estrogens. Completion of these Aims will demonstrate which patients are most likely to benefit from treatment with anti-estrogens and provide additional survival biomarkers related to hormonal pathways for assessing aggressiveness of NSCLCs.
Project 2: Vitamin D Modulation of inflammation and Lung Cancer Risk
Project Co-Leaders:
Pamela Hershberger, Ph.D.
Brenda B. Diergaarde, Ph.D.
Frank Sciurba, M.D.
The studies proposed in this new SPORE project are designed to provide strong rationale for a vitamin D3-based approach to lung cancer prevention. We recently demonstrated that 1,25-dihydroxyvitamin D3 [1,25(OH)2D3], the active metabolite of vitamin D3, significantly inhibits the growth of lung cancer cells and antagonizes nuclear factor-κB (NF-κB) action. NF-κB signaling plays a key role in inflammation, and has been shown to underlie smoking-associated lung inflammation and carcinogenesis. Together, this suggests that 1,25(OH)2D3 may be useful for chemoprevention of lung cancer. Because systemic 1,25(OH)2D3 administration is complicated by its hypercalcemia-inducing properties, we propose to use oral supplementation with vitamin D3 to safely achieve chemopreventive1,25(OH)2D3 levels within lung tissues. Upon ingestion, vitamin D3 is readily converted to the non-toxic circulating precursor, 25(OH)D3, which is subsequently converted to 1,25(OH)2D3 within the lung by CYP27B1-expressing bronchial epithelial cells and alveolar macrophages. To determine the impact of vitamin D3 exposure on pulmonary inflammation and lung cancer risk we will: (Aim 1) utilize samples and data from 548 cases from our SPORE Lung Tumor Registry, and 180 cases and 993 controls from the Pittsburgh Lung Screening Study (PLuSS) to evaluate the relationship between variation in vitamin D3 and NF-κB pathway genes, 25(OH)D3 serum levels, and risk of lung cancer; (Aim 2) use banked samples from 150 PLuSS participants to establish the association between 25(OH)D3 serum levels and inflammation and lung cancer risk biomarkers in sputum; (Aim 3) use murine models to quantify the effects of vitamin D3 status on NNK-induced lung carcinogenesis and cigarette smoke-induced pulmonary inflammation, and examine the impact of cigarette smoke exposure on 25(OH)D3 levels and expression of vitamin D3-metabolizing enzymes; and, (Aim 4) conduct a bioeffectiveness study of vitamin D3 supplementation in individuals at increased risk for lung cancer. We will evaluate whether supplementation corrects vitamin D3 deficiency and also investigate effects on inflammation and lung cancer risk biomarkers in sputum, circulating inflammation markers, and pulmonary function in this aim.
Project 3: Lung Cancer Risk Prediction in PLuSS
Project Co-Leaders:
William L. Bigbee, Ph.D.
Joel L. Weissfeld, M.D., M.P.H.
This new Project 3 in the UPCI Lung Cancer SPORE builds on both clinical and population based resource development and substantive research findings from the prior SPORE funding periods. Project 3 will pursue three translational aims to refine and validate a lung cancer risk prediction model and a diagnostic serum biomarker panel. In Specific Aim 1, Project 3 will construct a genetic risk index from single nucleotide polymorphism (SNP) genotype information, measure the index’s association with lung cancer (Aim 1A), and evaluate the index’s contribution to our published lung cancer prediction model that incorporates demographic, cigarette smoking exposure, and clinical variables (Aim 1B). In Specific Aim 2, Project 3 will evaluate the lung cancer discrimination performance of a promising lung cancer diagnostic rule model constructed from the serum concentrations of 10 biomarker proteins (Aim 2A) and verify its performance in subjects with intermediate suspicion pulmonary nodules on screening computerized tomography (CT) (Aim 2B) and diagnostic performance independent of demographic, smoking, clinical, and genetic variables (Aim 2C). These aims use shared lung cancer cases (n=180) emerging from the Pittsburgh Lung Screening Study (PLuSS), a comparable lung cancer case series (n=548) derived from clinical sources, and controls [n=993 (Aim 2A) or n=482 (Aim 2B)] sampled from the PLuSS cohort. Collaboration with the Biostatistics and Bioinformatics Core will develop improved models based on these expansive datasets. A concluding translational aim uses independent comparison groups from the ACRIN/NLST and ACOSOG Z4031 cohorts to validate these prediction and diagnostic strategies (Aim 3). Completion of Aim 1 will produce a lung cancer risk prediction model that capably selects individuals for primary chemoprevention or enhanced screening. Aim 2 will yield a lung cancer diagnostic test to improve early lung cancer detection in the contexts of clinical trials of primary chemoprevention and management of high-risk subjects with CT-detected nodules. Lastly, Aim 3 will evaluate the performance of both the lung cancer prediction and diagnostic models in two large and similar case-control cohorts for independent validation and refinement of the final models.
Project 4: Targeting Neutrophil Elastase in Lung Cancer
Project Co-Leaders:
Steven D. Shapiro, M.D.
John J. Reilly, M.D.
This new SPORE project will test the overall hypothesis that NE is a critical mediator of inflammation associated pro-cancer signaling in human lung cancer, especially in K-ras mutant human lung tumors with high NE content. Our preliminary data show that NE-mediated degradation of IRS-1 enhances growth of lung tumors. Interruption of NE function may inhibit K-ras mutant lung cancers that are resistant to other targeted therapies. The long-term goal is to inhibit NE as a novel therapy for treating lung cancer. To achieve this goal we propose three Specific Aims: 1. To establish the clinical relevance of the K-ras/NE/IRS-1 signaling pathway in non-small cell lung cancer (NSCLC) by interrogating tumor tissue from well-annotated cases from our lung cancer SPORE tissue bank. We hypothesize that neutrophil and NE content are inversely correlated with patient outcome, while IRS-1 is directly correlated with patient outcome. Moreover, we hypothesize that K-ras mutant lung tumors have the greatest neutrophil/NE and the least IRS-1 content. Emphysema or presence of airflow obstruction in patients may also associate with elevated neutrophil and NE tumor content and reduced IRS-1 content. 2. We will establish the efficacy of alpha-1-antitrypsin (α1-AT, a natural NE inhibitor) as well as synthetic low MW NE inhibitors in relevant preclinical models of lung adenocarcinoma. To test efficacy, we will use the Kras transgenic model of adenocarcinoma and a model employing NNK with and without cigarette smoke to induce lung tumors that are a mixture of K-ras mutant and K-ras wild type tumors. Proportion of induced tumors that are K-ras mutant and K-ras wild-type genotype in vehicle-treated and NE inhibitor-treated animals will be compared. Biomarkers relevant to the NE/IRS-1 pathway as well as inflammatory biomarkers and presence of emphysema will also be evaluated in preclinical models. 3. By the fourth year of this project, we will initiate clinical trials using an NE inhibitor for adjuvant treatment of surgically-resected NSCLC. We hypothesize that NE inhibition will improve disease-free survival followed adjuvant cisplatin-based chemotherapy. A phase II trial design is proposed that could be used with α1-AT or a small molecular weight NE inhibitor. We hypothesize that NSCLC with K-ras mutations will benefit from treatment with an NE inhibitor. Depending on the results of NE inhibitors on K-ras wild type tumors in pre-clinical models, this class of drugs may also be predicted to have value in patients with K-ras wild-type genotype, possibly those with emphysema. Biomarkers found to be modulated by NE inhibitor treatment in preclinical models will also be examined in patients undergoing NE inhibitor treatment.
Core A: Administrative Core
Core Director:
Jill M. Siegfried, Ph.D.
Core Co-Directors:
Marc Socinksk, M.D.
Joel L. Weissfeld, M.D., Ph.D.
The Administrative Core will oversee the scientific, fiscal, and organizational activities of the UPCI Lung Cancer SPORE, including oversight of scientific and translational progress, oversight of expenditures, scheduling regular meetings of the SPORE investigators, organizing our participation in annual SPORE meetings and interactions with the National Cancer Institute and other translational investigators, facilitating collaborations with other SPOREs and translational groups, and developing and preparing annual reports. The Administrative Core will also coordinate meetings with the Internal/External Advisory Board members, interact with our patient advocates, facilitate team science between investigators of different disciplines, undertake periodic evaluations of progress, and change direction of the SPORE Projects as necessary. The Administrative Core will use the SPORE flexibility provision to discontinue projects that fail to meet translational goals if needed. The Administrative Core will coordinate travel of SPORE investigators to the annual NCI-sponsored Translational Workshop, and other translational meetings, and will assist SPORE investigators in the preparation of manuscripts. The Core will oversee the Developmental Research and Career Development Programs to ensure smooth functioning of these components. The Administrative Core will coordinate SPORE Research Core activities with the UPCI to avoid redundancy and ensure that joint activities between the UPCI Lung and Thoracic Malignancies Program and the Lung Cancer SPORE are carried out efficiently, and that the two Programs act to complement and synergize with each other. The Administrative Core will also ensure that the SPORE Research Cores provide outstanding service to SPORE investigators. The Administrative Core will also communicate with the NCI Program Office to ensure that SPORE guidelines are followed and that the SPORE mandate is carried out. The Administrative Core will work with lung cancer Patient Advocates and lung cancer advocacy groups to raise lung cancer awareness and will work with the Development Office of UPC to secure philanthropy to supplement the SPORE budget.
Core B: Clinical Core
Core Co-Directors:
Joel L. Weissfeld, M.D., M.P.H.
Rodney Landreneau, M.D.
The Clinical Core for the University of Pittsburgh Cancer Institute (UPCI) Lung Cancer SPORE supports the infrastructure needed to satisfy the clinical research needs of the individual SPORE translational research projects. Fulfillment of SPORE translational research aims requires access to human subjects and associated clinical outcome information and biological materials (blood and tissue). In addition, successful completion of SPORE clinical trials and other interventions requires human research conducted according to exacting standards designed to protect research subjects from research risks. The Clinical Core provides the expertise and resources needed 1) to design and complete clinical trials, 2) to recruit, characterize, and follow human subjects for translational studies of laboratory-based cancer biomarkers, and 3) to obtain blood-based and tissue-based materials in support of clinical trials and biomarker research projects. To meet these objectives, the Clinical Core articulates the following four specific aims: Specific Aim 1: Design and implement clinical trials and clinical studies, Specific Aim 2: Identify, solicit, and enroll subjects into SPORE clinical trials, patient registries, and high risk cohorts, Specific Aim 3: Collect, manage, and store high quality risk factor and clinical outcome information, and Specific Aim 4: Deliver protocol-directed interventions and collect blood and tissue samples. The Clinical Core designs and implements clinical trials through the active participation of experienced clinical investigators. To enroll subjects into SPORE clinical studies, the Clinical Core integrates with local lung cancer treatment programs and with the Pittsburgh Lung Screening Study (PLuSS), an established high risk cohort containing over 3600 current and ex-cigarette smokers. In the renewal period, the Clinical Core proposes continued serial blood and sputum collections and additional computed tomography (CT) lung cancer screenings, restricted to the high risk subset in the PLuSS Extension Study. To collect and manage research data, Clinical Core investigators receive database management support through the Biostatistical/Bioinformatics Core and organize the activities of approved research protocols pertaining to subject recruitment, data and biological sample collection, and follow-up.
Core C: Tissue and Blood Bank Core
Core Co-Directors:
Rajiv Dhir, M.D.
Sanja Dacic, M.D.
The Tissue and Blood Bank Core for the Lung Cancer SPORE will serve as a shared resource for the main Research Projects and for the Career Development and Developmental Research Programs. The Core will collect, process, store and distribute tissue and body fluid specimens from patients diagnosed with lung cancer or with suspected lung cancer, and from subjects who are members of the PLuSS cohort and the PLuSS High-Risk Sub-Cohort. Triage and distribution of all specimens will be prioritized according to a plan established with all SPORE investigators, and approved by the Tissue and Blood Bank Core Pathologists. The Core will procure and triage fresh human lung tissue, including tumor, adjacent uninvolved and normal tissues distal from the tumor, and biopsies of the airway, from lung cancer patients undergoing resections or bronchoscopies, as well as individuals undergoing these procedures for reasons other than lung cancer. After triage under sterile conditions, tissues designated by the Core Pathologist as normal or abnormal will be either immediately distributed to investigators as needed for experimentation or stored for future use. Lymphocytes, serum, and plasma will be separated from other blood components and used immediately or stored for future analysis. Sputum will also be collected and processed. Tissues will also be formalin fixed for paraffin embedding. Tumor blocks from patients enrolled in clinical trials will be obtained and stored by the Core. Paraffin embedded specimens will be either sectioned for use in staining protocols or used for the preparation of tissue microarrays (TMAs). The Core will also carry out Kras and EGFR mutation analyses on microdissected tumor specimens and will utilize tumor tissue sections for EGFR amplification analyses by fluorescence in situ hybridization (FISH). The Core will also carry out routine and special pathology such as immunohistochemistry, morphometry, digital imaging and photography. Modifications will be made as necessary to meet any changes in SPORE research goals. All tissues will be collected through IRS- approved protocols on which Tissue and Blood Bank Core pathologists will be co-investigators. The Tissue Core will take advantage of infrastructure at UPCI for procurement of tissue, but will not duplicate it.
CORE D: BIOINFORMATICS AND BIOSTATISTICS CORE
Core Co-Directors:
Vanthi Gopalakrishnan, Ph.D.
Daniel P. Normolle, Ph.D.
This Core brings together biostatisticians and bioinformaticians who are experienced in the scientific areas covered by the SPORE and who are already close collaborators with SPORE lead investigators. The Specific Aims of the Biostatistics and Bioinformatics Core are 1. To contribute statistical expertise to the design and perform the analysis of data from in vitro and in vivo experiments and clinical trials proposed in the SPORE, in developmental research projects, and by career trainees funded by this SPORE; 2. To work with the Project and Core investigators, including the Clinical Core and Tissue and Blood Bank Core, to ensure that data collection and database development are appropriate for the requisite statistical analyses while ensuring patient confidentiality and the integrity of all SPORE tissue banks; and to provide PLuSS and other SPORE data for approved research by local researchers and collaborators in other institutions; 3. To collaborate with investigators in writing and preparing progress reports, abstracts, manuscripts, and presentations, and 4. To conduct methodological research motivated by the challenges of SPORE research.
To achieve these aims, Core members are fully integrated into the research teams to reach study. They maintain frequent communication both within the Core and with other SPORE researchers. The projects of the University of Pittsburgh Lung SPORE are designed to share information and resources. For example, several projects will make use of the same sets of subjects (retrospective UPCI Lung cancer patients and PLuSS participants). In addition, the Pittsburgh SPORE has proposed to share data and patient samples with the UCLA Jonnson Comprehensive Cancer Center and the University of Texas - Southwestern. The Core will ensure that resources are used efficiently, and each project builds on the information gained from other Pittsburgh SPORE projects and other institutions. Core D will facilitate the smooth transfer of techniques, data, and patient tissues among projects and institutions, while ensuring patient confidentiality.