 Thoracic Surgery Research LaboratoryThe Thoracic Surgery Research Laboratory consists of approximately 1,400 square feet of research space in Medical Science Research Building II. The major areas of research focus include thoracic oncogenesis and tumor biology, lung transplantation, lung reduction surgery for emphysema, and development of an implantable artificial lung. The Thoracic Surgery Tumor Biology Program is defining unique genetic markers in thoracic malignancies which may ultimately have both prognostic and therapeutic implications. This collaborative effort involves DNA, RNA and protein analysis of freshly harvested lung and esophageal cancer specimens and concurrent correlation between clinical tumor stage, the presence of particular genetic markers, and prognosis. Current research has focused on the analysis of intestinal-type Barrett's metaplasia and the potential genetic and biochemical properties of this epithelium which render it at risk for the development of adenocarcinoma. Molecular studies of both esophageal and lung tumors using the technique of two dimensional genome scanning is identifying regions of DNA which are altered by either amplification or loss. The subsequent identification of the genes associated with these alterations detected in these tumors represents a major research effort. An additional line of investigation involves the characterization of potential chemopreventatives which may be useful in reducing cancer risk in patients with premalignant Barrett's metaplasia. Many of the above basic research projects are being performed in collaboration with other departments including Pathology, Internal Medicine, Radiation Oncology, Pediatrics, Human Genetics, and Pharmacology. Techniques
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Location/ContactB560 MSRB II/0686 Active ProjectsMolecular Studies of Esophageal AdenocarcinomasInvestigators: David G. Beer, Ph.D., Surgery; Mark Orringer, M.D., Surgery; Mark Iannettoni, M.D., Surgery; Sam Hanash, M.D., Ph.D., Pediatrics and Thomas Glover, Ph.D., Pediatrics and Human Genetics Most of the important cancer-related genes associated with the development and progression of esophageal adenocarcinomas are unknown. Utilizing the technique of Landmark Genome Scanning, which involves the two-dimensional separation of radiolabeled restriction fragments of genomic DNA and computer image analysis, we have identified the presence of amplified DNA in these tumors. Amplification is an important mechanism underlying the overexpression of cancer genes in tumors. The amplified DNA fragments are directly isolated from the gels, cloned and sequenced. The chromosomal location of the amplified DNA is determined using Fluorescence In situ Hybridization. The identification of the amplified and overexpressed gene(s) associated with these fragments is continuing to demonstrate the genes involved in the development and progression of this deadly disease. Recent studies have utilized oligonucleotide arrays to examine gene expression changes during progression of Barrett's mucosa to adenocarinoma. Toward Molecular Classification of Tumors: Lung Cancer ProjectInvestigators: David G. Beer, Ph.D., Surgery; Mark Orringer, M.D., Surgery; Mark Iannettoni, M.D., Surgery; Sam Hanash, M.D., Ph.D., Pediatrics Lung cancer is the major cause of cancer death in the United States. Although progress has been made in our understanding of the cause of lung cancer as well as in the treatment of some types of this disease, the prognosis for most patients with lung cancer has remained poor. There is a significant need to understand why some patients who present with early stage I lung cancers do very poorly while others with apparently the same type and tumor stage do well. Advances in current technologies which allow molecular assessment of a large number of genes from individual tumors, may provide answers to this important question as well as potentially identify new approaches for therapeutic intervention or early diagnosis. We have examined both the mRNA expression profile using cDNA arrays and the protein expression profiles using quantitative two-dimensional gel electrophoresis (2D gels) in non-small cell lung cancers. Specifically, stage I lung adenocarcinomas from patients demonstrating either a good or a poor clinical outcome have recently been uncovered (Nature Med., 2002). Biomarker Development LaboratoryInvestigators: David G. Beer, Ph.D., Surgery; Mark Orringer, M.D., Surgery; Sam Hanash, M.D., Ph.D., Pediatrics The major goal of this project is the identification of novel cancer biomarkers using serum from lung cancer patients. A major focus is to utilize the serum as a probe of 2D-protein Western blots to identify autoreactive proteins present in lung cancer patients' serum. The protein spots which are revealed by the Western blots are then identified using mass spectroscopy. The Role of Tumor Necrosis Factor Alpha in Rat Lung Allograft RejectionInvestigators: Mark D. Iannettoni, M.D., Surgery; Douglas Arenberg, M.D., Internal Medicine and Steven L. Kunkel, M.D., Pathology Tumor necrosis factor alpha (TNF - a) is a cytokine of pleiotropic effects on an array of cells. Although TNF -a was first recognized as a product of mononuclear phagocytes, it is now known to be synthesized by activated T lymphocytes. T lymphocytes are the cells which are involved in the immunological response, and therefore we feel that there should be a marked association of TNF - a with rejection. Recently, elevated circulating levels of TNF - a have been associated with heart, liver, and kidney allograft rejection and therefore we are studying the role of TNF - a for lung transplant rejection. Our goal is to demonstrate with a model of rat lung transplantation the time dependent expression of TNF - a messenger RNA from a transplanted lung undergoing rejection. We will use inbred strains of RT1-incompatible, specific pathogen free rats, which are known to reject lung allografts at 5 to 6 days post transplant. Single left lung transplantation will be performed through a left thoracotomy, with the rat intubated and under halothane general anesthesia. A pneumonectomy will be performed on the recipient and vascular as well as bronchial continuity will be established in the donor lung using end to end microvascular anastomosis of the pulmonary artery vein and bronchus. Following simultaneous reperfusion and ventilation the chest will be closed and the rats will be intubated. In a time dependent fashion at postoperative days one, two, four, five and six, the rats will be sacrificed, the lungs perfused free of blood and bronchial alveolar lavage in both the native and transplanted lungs will be performed prior to the removal of the lungs. The native and transplanted lungs will then be immediately frozen in liquid nitrogen and homogenized with isolation of total cellular RNA. Northern blot analysis of TNF - a messenger RNA will be performed and associated with the time dependents. A second group of transplants will be performed and the animals will be sacrificed in likewise fashion but they will undergo histiologic and EM studies to document rejection. We will then correlate the levels of TNF - a messenger RNA in association with the pathologic findings of rejection. If there is a strong association of levels of TNF - a messenger RNA with rejection, we will then take an additional group of rats and give them passive immunization with anti-TNF antibodies and repeat the experiments looking for histologic rejection as well as TNF - a messenger RNA levels. If there is an association between TNF - a messenger RNA levels with rejection and the antibody immunization will prolong graft survival and decrease the levels of TNF - a messenger RNA, then we can make an association of the cytokine with transplantation rejection. Functional Improvement in Patients Following Lung TransplantationInvestigators: Mark D. Iannettoni, M.D., Fernando J. Martinez, M.D., Internal Medicine and Joseph P. Lynch III, M.D., Internal Medicine The functional improvement in patients following lung transplantation is not well characterized. Pulmonary function testing, exercise testing and questionnaire techniques are being used to evaluate the functional improvement in patients following lung transplantation. This also is being used to improve selection criteria for the procedure. Lung Reduction Surgery for EmphysemaInvestigators: Mark D. Iannettoni, M.D., Surgery and Fernando J. Martinez, M.D., Internal Medicine The efficacy and biomechanics of lung reduction surgery (reduction pneumoplasty or pneumectomy) is being studied using pulmonary function testing, diaphragm fluoroscopy, and exercise function testing in patients with severe emphysema. The Role of C-X-C Chemokines in Angiogenesis in Lung CancerInvestigators: Mark D. Iannettoni, M.D., Surgery and Douglas Arenberg, M.D., Internal Medicine CXC chemokines are known angiogenic factors. The role of these molecules in the progression of lung cancer and their relationship to prognosis are being studied. Patients with adverse prognostic characteristics may be offered adjuvant therapy for their tumors. The Development of a Totally Implantable Artificial Lung and its effects of Physiology and Biochemical Activity in a Sheep ModelInvestigators: Mark D. Iannettoni, M.D., Robert Bartlett, M.D., Ronald Hirschl, M.D., Patrick Montoya, M.D., Scott Merz, Ph.D., Jonathan Haft, M.D. This project involves the development of a totally implantable artificial lung to be used for mechanical ventilatory support as well as in pulmonary transplantation as for a bridge to transplantation. The project involves the use of an extracorporal device placed in parallel with the native circulation in acute and chronic animal models. The effects on pulmonary physiology and biochemistry of this device are currently under investigation. Cancer Center Core GrantInvestigators: Mark B. Orringer, M.D., Surgery and Max Wicha, M.D. (P.I.), Director, Cancer Center This is a Core Grant for the University of Michigan Cancer Center. It consists of nine interdisciplinary clinical research programs and six basic science research programs. Dr. Orringer is the Director of the Thoracic Oncology program which is involved with a number of clinical and basic science multidisciplinary projects, e.g., evaluating the efficacy of preoperative chemotherapy and radiation therapy in the treatment of esophageal carcinoma and selected lung cancers, identifying prognostic oncogenes in thoracis malignancies, etc. |