Pulmonary Fibrosis - A Medical Dictionary, Bibliography, and Annotated Research Guide to Internet References

PULMONARY FIBROSIS A M EDICAL D ICTIONARY , B IBLIOGRAPHY , AND A NNOTATED R ESEARCH G UIDE TO I NTERNET R E FERENCES

J AMES N. P ARKER , M.D. AND P HILIP M. P ARKER , P H .D., E DITORS

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ICON Health Publications ICON Group International, Inc. 4370 La Jolla Village Drive, 4th Floor San Diego, CA 92122 USA Copyright 2004 by ICON Group International, Inc. Copyright 2004 by ICON Group International, Inc. All rights reserved. This book is protected by copyright. No part of it may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, recording, or otherwise, without written permission from the publisher. Printed in the United States of America. Last digit indicates print number: 10 9 8 7 6 4 5 3 2 1

Publisher, Health Care: Philip Parker, Ph.D. Editor(s): James Parker, M.D., Philip Parker, Ph.D. Publisher's note: The ideas, procedures, and suggestions contained in this book are not intended for the diagnosis or treatment of a health problem. As new medical or scientific information becomes available from academic and clinical research, recommended treatments and drug therapies may undergo changes. The authors, editors, and publisher have attempted to make the information in this book up to date and accurate in accord with accepted standards at the time of publication. The authors, editors, and publisher are not responsible for errors or omissions or for consequences from application of the book, and make no warranty, expressed or implied, in regard to the contents of this book. Any practice described in this book should be applied by the reader in accordance with professional standards of care used in regard to the unique circumstances that may apply in each situation. The reader is advised to always check product information (package inserts) for changes and new information regarding dosage and contraindications before prescribing any drug or pharmacological product. Caution is especially urged when using new or infrequently ordered drugs, herbal remedies, vitamins and supplements, alternative therapies, complementary therapies and medicines, and integrative medical treatments. Cataloging-in-Publication Data Parker, James N., 1961Parker, Philip M., 1960Pulmonary Fibrosis: A Medical Dictionary, Bibliography, and Annotated Research Guide to Internet References / James N. Parker and Philip M. Parker, editors p. cm. Includes bibliographical references, glossary, and index. ISBN: 0-597-84570-0 1. Pulmonary Fibrosis-Popular works. I. Title.

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Disclaimer This publication is not intended to be used for the diagnosis or treatment of a health problem. It is sold with the understanding that the publisher, editors, and authors are not engaging in the rendering of medical, psychological, financial, legal, or other professional services. References to any entity, product, service, or source of information that may be contained in this publication should not be considered an endorsement, either direct or implied, by the publisher, editors, or authors. ICON Group International, Inc., the editors, and the authors are not responsible for the content of any Web pages or publications referenced in this publication.

Copyright Notice If a physician wishes to copy limited passages from this book for patient use, this right is automatically granted without written permission from ICON Group International, Inc. (ICON Group). However, all of ICON Group publications have copyrights. With exception to the above, copying our publications in whole or in part, for whatever reason, is a violation of copyright laws and can lead to penalties and fines. Should you want to copy tables, graphs, or other materials, please contact us to request permission (E-mail: [email protected]). ICON Group often grants permission for very limited reproduction of our publications for internal use, press releases, and academic research. Such reproduction requires confirmed permission from ICON Group International, Inc. The disclaimer above must accompany all reproductions, in whole or in part, of this book.

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Acknowledgements The collective knowledge generated from academic and applied research summarized in various references has been critical in the creation of this book which is best viewed as a comprehensive compilation and collection of information prepared by various official agencies which produce publications on pulmonary fibrosis. Books in this series draw from various agencies and institutions associated with the United States Department of Health and Human Services, and in particular, the Office of the Secretary of Health and Human Services (OS), the Administration for Children and Families (ACF), the Administration on Aging (AOA), the Agency for Healthcare Research and Quality (AHRQ), the Agency for Toxic Substances and Disease Registry (ATSDR), the Centers for Disease Control and Prevention (CDC), the Food and Drug Administration (FDA), the Healthcare Financing Administration (HCFA), the Health Resources and Services Administration (HRSA), the Indian Health Service (IHS), the institutions of the National Institutes of Health (NIH), the Program Support Center (PSC), and the Substance Abuse and Mental Health Services Administration (SAMHSA). In addition to these sources, information gathered from the National Library of Medicine, the United States Patent Office, the European Union, and their related organizations has been invaluable in the creation of this book. Some of the work represented was financially supported by the Research and Development Committee at INSEAD. This support is gratefully acknowledged. Finally, special thanks are owed to Tiffany Freeman for her excellent editorial support.

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About the Editors James N. Parker, M.D. Dr. James N. Parker received his Bachelor of Science degree in Psychobiology from the University of California, Riverside and his M.D. from the University of California, San Diego. In addition to authoring numerous research publications, he has lectured at various academic institutions. Dr. Parker is the medical editor for health books by ICON Health Publications. Philip M. Parker, Ph.D. Philip M. Parker is the Eli Lilly Chair Professor of Innovation, Business and Society at INSEAD (Fontainebleau, France and Singapore). Dr. Parker has also been Professor at the University of California, San Diego and has taught courses at Harvard University, the Hong Kong University of Science and Technology, the Massachusetts Institute of Technology, Stanford University, and UCLA. Dr. Parker is the associate editor for ICON Health Publications.

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About ICON Health Publications To discover more about ICON Health Publications, simply check with your preferred online booksellers, including Barnes&Noble.com and Amazon.com which currently carry all of our titles. Or, feel free to contact us directly for bulk purchases or institutional discounts: ICON Group International, Inc. 4370 La Jolla Village Drive, Fourth Floor San Diego, CA 92122 USA Fax: 858-546-4341 Web site: www.icongrouponline.com/health

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Table of Contents FORWARD .......................................................................................................................................... 1 CHAPTER 1. STUDIES ON PULMONARY FIBROSIS .............................................................................. 3 Overview........................................................................................................................................ 3 The Combined Health Information Database................................................................................. 3 Federally Funded Research on Pulmonary Fibrosis....................................................................... 6 E-Journals: PubMed Central ....................................................................................................... 61 The National Library of Medicine: PubMed ................................................................................ 63 CHAPTER 2. NUTRITION AND PULMONARY FIBROSIS .................................................................. 111 Overview.................................................................................................................................... 111 Finding Nutrition Studies on Pulmonary Fibrosis.................................................................... 111 Federal Resources on Nutrition ................................................................................................. 113 Additional Web Resources ......................................................................................................... 114 CHAPTER 3. DISSERTATIONS ON PULMONARY FIBROSIS .............................................................. 115 Overview.................................................................................................................................... 115 Dissertations on Pulmonary Fibrosis ........................................................................................ 115 Keeping Current ........................................................................................................................ 115 CHAPTER 4. CLINICAL TRIALS AND PULMONARY FIBROSIS ........................................................ 117 Overview.................................................................................................................................... 117 Recent Trials on Pulmonary Fibrosis......................................................................................... 117 Keeping Current on Clinical Trials ........................................................................................... 125 CHAPTER 5. PATENTS ON PULMONARY FIBROSIS ........................................................................ 127 Overview.................................................................................................................................... 127 Patents on Pulmonary Fibrosis.................................................................................................. 127 Patent Applications on Pulmonary Fibrosis .............................................................................. 138 Keeping Current ........................................................................................................................ 153 CHAPTER 6. BOOKS ON PULMONARY FIBROSIS ............................................................................ 155 Overview.................................................................................................................................... 155 Book Summaries: Online Booksellers......................................................................................... 155 Chapters on Pulmonary Fibrosis................................................................................................ 156 CHAPTER 7. PERIODICALS AND NEWS ON PULMONARY FIBROSIS .............................................. 157 Overview.................................................................................................................................... 157 News Services and Press Releases.............................................................................................. 157 Academic Periodicals covering Pulmonary Fibrosis .................................................................. 160 CHAPTER 8. RESEARCHING MEDICATIONS .................................................................................. 161 Overview.................................................................................................................................... 161 U.S. Pharmacopeia..................................................................................................................... 161 Commercial Databases ............................................................................................................... 162 Researching Orphan Drugs ....................................................................................................... 162 APPENDIX A. PHYSICIAN RESOURCES .......................................................................................... 167 Overview.................................................................................................................................... 167 NIH Guidelines.......................................................................................................................... 167 NIH Databases........................................................................................................................... 169 Other Commercial Databases..................................................................................................... 171 The Genome Project and Pulmonary Fibrosis............................................................................ 171 APPENDIX B. PATIENT RESOURCES ............................................................................................... 175 Overview.................................................................................................................................... 175 Patient Guideline Sources.......................................................................................................... 175 Finding Associations.................................................................................................................. 183 APPENDIX C. FINDING MEDICAL LIBRARIES ................................................................................ 185 Overview.................................................................................................................................... 185 Preparation................................................................................................................................. 185

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Finding a Local Medical Library................................................................................................ 185 Medical Libraries in the U.S. and Canada ................................................................................. 185 ONLINE GLOSSARIES................................................................................................................ 191 Online Dictionary Directories ................................................................................................... 191 PULMONARY FIBROSIS DICTIONARY ................................................................................ 193 INDEX .............................................................................................................................................. 271

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FORWARD In March 2001, the National Institutes of Health issued the following warning: "The number of Web sites offering health-related resources grows every day. Many sites provide valuable information, while others may have information that is unreliable or misleading."1 Furthermore, because of the rapid increase in Internet-based information, many hours can be wasted searching, selecting, and printing. Since only the smallest fraction of information dealing with pulmonary fibrosis is indexed in search engines, such as www.google.com or others, a non-systematic approach to Internet research can be not only time consuming, but also incomplete. This book was created for medical professionals, students, and members of the general public who want to know as much as possible about pulmonary fibrosis, using the most advanced research tools available and spending the least amount of time doing so. In addition to offering a structured and comprehensive bibliography, the pages that follow will tell you where and how to find reliable information covering virtually all topics related to pulmonary fibrosis, from the essentials to the most advanced areas of research. Public, academic, government, and peer-reviewed research studies are emphasized. Various abstracts are reproduced to give you some of the latest official information available to date on pulmonary fibrosis. Abundant guidance is given on how to obtain free-of-charge primary research results via the Internet. While this book focuses on the field of medicine, when some sources provide access to non-medical information relating to pulmonary fibrosis, these are noted in the text. E-book and electronic versions of this book are fully interactive with each of the Internet sites mentioned (clicking on a hyperlink automatically opens your browser to the site indicated). If you are using the hard copy version of this book, you can access a cited Web site by typing the provided Web address directly into your Internet browser. You may find it useful to refer to synonyms or related terms when accessing these Internet databases. NOTE: At the time of publication, the Web addresses were functional. However, some links may fail due to URL address changes, which is a common occurrence on the Internet. For readers unfamiliar with the Internet, detailed instructions are offered on how to access electronic resources. For readers unfamiliar with medical terminology, a comprehensive glossary is provided. For readers without access to Internet resources, a directory of medical libraries, that have or can locate references cited here, is given. We hope these resources will prove useful to the widest possible audience seeking information on pulmonary fibrosis. The Editors

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From the NIH, National Cancer Institute (NCI): http://www.cancer.gov/cancerinfo/ten-things-to-know.

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CHAPTER 1. STUDIES ON PULMONARY FIBROSIS Overview In this chapter, we will show you how to locate peer-reviewed references and studies on pulmonary fibrosis.

The Combined Health Information Database The Combined Health Information Database summarizes studies across numerous federal agencies. To limit your investigation to research studies and pulmonary fibrosis, you will need to use the advanced search options. First, go to http://chid.nih.gov/index.html. From there, select the “Detailed Search” option (or go directly to that page with the following hyperlink: http://chid.nih.gov/detail/detail.html). The trick in extracting studies is found in the drop boxes at the bottom of the search page where “You may refine your search by.” Select the dates and language you prefer, and the format option “Journal Article.” At the top of the search form, select the number of records you would like to see (we recommend 100) and check the box to display “whole records.” We recommend that you type “pulmonary fibrosis” (or synonyms) into the “For these words:” box. Consider using the option “anywhere in record” to make your search as broad as possible. If you want to limit the search to only a particular field, such as the title of the journal, then select this option in the “Search in these fields” drop box. The following is what you can expect from this type of search: •

Pulmonary Manifestations of Gastroesophageal Reflux Disease Source: Practical Gastroenterology. 24(4): 27, 30, 34, 42, 46, 49-50. April 2000. Contact: Available from Shugar Publishing. 12 Moniebogue Lane, Westhampton Beach, NY 11978. (516) 288-4404. Fax (516) 288-4435. Summary: Gastroesophageal reflux disease (GERD) is a spectrum of disease best defined as symptoms or signs of esophageal or adjacent organ injury secondary to the reflux of gastric contents (principally acid and pepsin) into the esophagus or beyond (into the oral cavity or lung). This article reviews the pulmonary (lung) manifestations of GERD. The most common presentations are heartburn, a burning sensation behind the breastbone just after meals or on bending over (relieved with antacids or an over the

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counter (OTC) H2 receptor antagonist), or regurgitation (the spontaneous return of gastric contents into the esophagus). Careful study has shown that numerous pulmonary symptoms including asthma, chronic cough, and pulmonary fibrosis amongst others may be due to or exacerbated by GERD. The author focuses on prevalence, clinical presentation, and approach to the diagnosis and therapy of these patients. The patient with pulmonary presentations of GERD often presents a diagnostic challenge for clinicians because of the absence of heartburn or regurgitation. Treatment follows the general principals for management of patients with heartburn and includes use of lifestyle modifications, pharmacological therapy, and antireflux surgery. The author recommends beginning therapy with omeprazole 20 mg twice daily, lansoprazole 30 mg twice daily or rabeprazole 20 mg twice daily before breakfast and dinner for two to three months; this regimen results in 70 percent response rate. Referral for ambulatory pH monitoring to document the effectiveness of therapy and assessment of the presence of continued nocturnal (nighttime) exposure is indicated if patients are refractory to proton pump inhibitors. 1 figure. 1 table. 19 references. •

Extrahepatic Manifestations of Hepatitis C and the Association With Alcoholic Liver Disease Source: Seminars in Liver Disease. 15(1): 101-109. February 1995. Contact: Available from Thieme Medical Publishers, Inc. 381 Park Avenue South, New York, NY 10016. (800) 782-3488. Summary: In this article, the authors consider the extrahepatic manifestations of hepatitis C and the association with alcoholic liver disease. Topics include rheumatoid arthritis; pulmonary fibrosis; lichen planus; salivary gland lesions; porphyria cutanea tarda; Mooren corneal ulcer; aplastic anemia; vasculitis, and essential mixed cryoglobulinemia; glomerulonephritis; generalized vasculitis and miscellaneous vasculitides; and the association between hepatitis C and alcoholic liver disease. The authors conclude that data supporting a link between HCV infection and rheumatoid arthritis and pulmonary fibrosis are not convincing; an association between HCV and lichen planus, Sjogren's syndrome, porphyria cutanea tarda, and Mooren corneal ulceration are more intriguing but far from being established unequivocally. In contrast, data supporting a link between HCV and some vasculitis syndromes, those circumstantial, are convincing. Data also points toward a combination of HCV and alcohol in the pathogenesis of liver injury in patients with alcoholic liver disease. 1 table. 79 references.

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Hepatitis C Virus: What Recent Studies Can Tell Us Source: Postgraduate Medicine. 95(6): 121-122, 125-126, 128-130. May 1, 1994. Summary: In this article, the authors review the hepatitis C virus (HCV), presenting some recent developments in epidemiology and diagnosis, and examining the relationship of HCV infection to other disorders and the effectiveness of current drug therapy. Topics include transmission through blood products and hemodialysis; needlestick in health care workers; the infectivity of bodily secretions; the hepatitis C virus genome; diagnostic considerations; associated medical disorders, including hepatocellular cancer, chronic hepatitis, chronic liver disease, cryoglobulinemia, glomerulonephritis, porphyria cutanea tarda, and idiopathic pulmonary fibrosis; and treatment options, notably interferon alfa-2b and the use of oral ribavirin. 1 figure. 3 tables. 18 references. (AA-M).

Studies

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Respiratory Complications of Gastrointestinal Diseases Source: Gastroenterology Clinics of North America. 27(4): 721-746. December 1998. Contact: Available from W.B. Saunders. 6277 Sea Harbor Drive, Orlando, FL 32887-4800. (800) 654-2452 or (407) 345-4000. Summary: The importance of acid reflux in the pathophysiology of disease of the respiratory tract is becoming increasingly well documented. This article describes the pathogenesis and clinical syndromes affecting the respiratory tract that are commonly encountered in clinical practice. For a substantial percentage of patients with respiratory disorders, gastroesophageal reflux (GER) is the sole cause of these symptoms. Respiratory disorders suspected of being related to GER include asthma, chronic cough, pulmonary fibrosis, pneumonia, and apnea in infants. For each of these disorders, the authors review the pathophysiology, the role of gastrointestinal motility, the interplay of medications, and treatment options. A separate section offers a review of diagnostic techniques in these complications, including radiography, endoscopy, esophageal manometry, and ambulatory pH monitoring. Most important, it has been shown that medical and surgical treatments are available for the effective management of these symptoms and related patient morbidity. 4 figures. 1 table. 115 references.

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New Treatments for Rheumatoid Arthritis Source: Postgraduate Medicine. 106(4): 82-85,88-90,92. October 1, 1999. Summary: This journal article provides health professionals with information on new slow acting antirheumatic drugs that are currently or soon to be available. During the past decade, perspectives on the nature and treatment of rheumatoid arthritis (RA) have changed. RA is now recognized as a serious systemic disorder. Methotrexate, a slow acting agent that inhibits dihydrofolate reductase, has become the gold standard of RA therapy. The result of its pharmacologic effects is impairment in the functioning of inflammatory cells that mediate the rheumatoid process. In addition to its medical effectiveness, methotrexate is well tolerated and relatively inexpensive. However, its potential toxic effects include bone marrow suppression, hepatotoxicity, interstitial pneumonitis, pulmonary fibrosis, increased susceptibility to infection, and pseudo sun sensitivity. Although the immunosuppressive agent cyclosporine has been used to treat RA in Europe for more than a decade, it is rarely used by American and Canadian rheumatologists. It is typically used in combination with methotrexate in patients who do not respond to methotrexate alone. Various cytotoxic drugs have been found to be helpful in the treatment of recalcitrant RA; however, their usefulness is limited by the severity of their toxic side effects. Newly emerging drugs for RA include leflunomide, tumor necrosis factor (TNF)-alpha inhibitors such as infliximab and etanercept, and interleukin-1 (IL-1) inhibitors. Leflunomide, the first antipyrimidine agent to be used to treat RA, inhibits the ability of dihydroorotate dehydrogenase to convert dihydroorotate to orotate. Efficacy studies have demonstrated the effectiveness of the agent. Infliximab contains monoclonal antibodies that bind circulating TNF-alpha, thereby inhibiting the proinflammatory effects of the circulating lymphokine. Although infliximab awaits final approval by the Food and Drug Administration, for use in RA, patients have shown definite clinical improvement after intravenous administration of the agent. However, adverse reactions are common. Etanercept is the first effective biologic antirheumatic therapeutic agent to become available for clinical use. This agent has been effective in relieving RA symptoms and improving function, and its reported adverse effects have been modest. However, its use is limited by the need to administer it parenterally and

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its excessive cost. The most effective IL-1 inhibitor is a human recombinant IL-1 receptor antagonist that is now under investigation. 1 table and 27 references.

Federally Funded Research on Pulmonary Fibrosis The U.S. Government supports a variety of research studies relating to pulmonary fibrosis. These studies are tracked by the Office of Extramural Research at the National Institutes of Health.2 CRISP (Computerized Retrieval of Information on Scientific Projects) is a searchable database of federally funded biomedical research projects conducted at universities, hospitals, and other institutions. Search the CRISP Web site at http://crisp.cit.nih.gov/crisp/crisp_query.generate_screen. You will have the option to perform targeted searches by various criteria, including geography, date, and topics related to pulmonary fibrosis. For most of the studies, the agencies reporting into CRISP provide summaries or abstracts. As opposed to clinical trial research using patients, many federally funded studies use animals or simulated models to explore pulmonary fibrosis. The following is typical of the type of information found when searching the CRISP database for pulmonary fibrosis: •

Project Title: RESPONSE

5-LIPOXYGENASE

PRODUCTS

IN

ASTHMATIC

IMMUNE

Principal Investigator & Institution: Henderson, William R.; Associate Professor; Medicine; University of Washington Grant & Contract Services Seattle, Wa 98105 Timing: Fiscal Year 2002; Project Start 01-APR-1998; Project End 31-MAR-2004 Summary: The 5-lipoxygenase (5-LO) products, the leukotriene (LT)s, are clearly important participants in the pulmonary inflammatory process in patients with asthma. We have developed a protocol for administration of ovalbumin (OVA_ as allergen to induce late-phase allergen-specific pulmonary disease in normal BAL:B/c and C57BL/6 mice. OVA-treated mice display a disease strikingly similar to allergen-induced human asthma. In our mouse model of asthma, we have found that leukotrienes are key mediators of the mucus release and eosinophil infiltration of the airways. With the availability of mutant mice deficient in 5-LO and both isoforms of cyclooxygenase (COX) (together wit specific inhibitors/antagonists of the lipid mediators), we will determine the contribution of the 5-LO pathway to the induction and resolution of allergic airway inflammation and AHR. These studies will be performed in both our standard protocol and a long-term model of allergen-induced lung fibrosis in mice. Our goal will be to define immune mechanisms by which leukotrienes influences the activation and effector functions of T cells and dendritic cells, key cells in the mediation of allergic airway inflammation. Our specifi aims are as follows: Specific Aim 1. To characterize further the role of the 5-LO pathway in allergic pulmonary inflammation and AHR. We will examine these questions: a) Will intrapulmonary 5-LO pathway blockade prevent AHR? b) Will 5-LO pathway blockade resolve ongoing allergic airway inflammation? and c) Wil 5-LO pathway blockade prevent allergen-induced lung fibrosis? Specific Aim 2. To determine the interrelationship of the 5-LO pathway with 2

Healthcare projects are funded by the National Institutes of Health (NIH), Substance Abuse and Mental Health Services (SAMHSA), Health Resources and Services Administration (HRSA), Food and Drug Administration (FDA), Centers for Disease Control and Prevention (CDCP), Agency for Healthcare Research and Quality (AHRQ), and Office of Assistant Secretary of Health (OASH).

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COX-2 pathway and platelet activating factor (PAF) in the mediation of allergic airway inflammation and AHR. The following questions will be studied: a) Is COX-2 pathway activation important in development of allergic inflammation and AHR? and b) Will blockade of secretory phospholipase A2 (sPLA2) and PAF inhibit allergic lung inflammation and AHR? and Specific Aim 3. To determine the mechanisms by which leukotriene inhibition blocks allergic pulmonary inflammation and AHR in the murine model of asthma. We will study these questions: a) Will 5-LO pathway blockade prevent allergen-induced T cell proliferation and/or cytokine generation necessary for airway inflammation and AHR? and b) Is 5-LO pathway activation required for adoptive transfer of AHR b T cells? The more specific our knowledge of the biochemical and immunological changes becomes, the more likely it is that specific interventions producing more benefit than harm in reducing leukotriene-reduced inflammation, will be found. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: A FUNCTIONAL GENOMICS HEART & LUNG DEVELOP. PROGRAM Principal Investigator & Institution: Hoffman, Eric P.; Director, Research Center for Genetic Me; Children's Research Institute Washington, D.C., Dc 20010 Timing: Fiscal Year 2002; Project Start 30-SEP-2000; Project End 31-JUL-2004 Summary: Drs. Eric Hoffman and Dietrich Stephan (whose primary interests are focused on muscle disease and leukemia) are PI and Co-PI on the Program's Expression Array Core and have extensive experience with all aspects of array use and data analysis. They have established collaborations with Dr. Michael Bitmer, Dr. Yidony Chen and the entire NHGRI array community. As a post-doctoral fellow in the laboratory of Dr. Jeffery Trent (member of this Program's External Advisory Committee), and a pioneer in DNA microarray technology, Dr. Stephan developed several alternative approaches to label total RNA for efficient signal detection under varying circumstances which have become the standard protocol used at NHGRI and has become a quite robust and reliable system for detecting signals over several orders of magnitude. In addition to building and using 7K expression arrays, Dr. Stephan was the first at NHGRI to develop genomic DNA microarray systems. Dr. Hoffman's laboratory is particularly interested in systematic assessments of the sensitivity and specificity of the Affymetrix vs. cDNA array approaches, and shows preliminary data pointing out the importance of redundant measurements, and correlative studies. Indeed, the proposed systematic comparison of the Affymetrix and cDNA array experimental platforms should be the first of this type of quality control of expression array data, and should prove highly valuable to both Program investigations and the research community at large. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: A LINKAGE STUDY IN FAMILIAL PULMONARY FIBROSIS Principal Investigator & Institution: Schwartz, David A.; Professor of Medicine and Genetics; Medicine; Duke University Durham, Nc 27706 Timing: Fiscal Year 2002; Project Start 20-AUG-2000; Project End 31-JUL-2005 Summary: (Adapted from Investigator's Abstract) The overall goal of this project is to investigate inherited genetic factors that play a role in the development of pulmonary fibrosis. The overall hypothesis of this investigation is that inherited genetic factors predispose individuals to develop pulmonary fibrosis. The goal of this investigation is to identify a group of genetic loci that play a role in the development of familial

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pulmonary fibrosis. The overall hypothesis is supported by the following observations: familial pulmonary fibrosis is indistinguishable pathologically from idiopathic pulmonary fibrosis and appears to be inherited as an autosomal dominant trait with variable penetrance; pulmonary fibrosis is associated with pleiotropic genetic disorders, such as Hermansky-Pudlak syndrome, neuofibromatosis, tuberous sclerosis, NeimannPick disease, Gaucher's disease, and familial hypocalciuric hypercalcemia; pulmonary fibrosis is frequently observed in autoimmune disease, including rheumatoid arthritis and systemic sclerosis; variable susceptibility is evident among workers who are reported to be exposed occupationally to similar concentrations of fibrogenic dusts; and inbred strains of mice differ in their susceptibility to fibrogenic dust. In conjunction with the exponential growth of human molecular genetics, the investigators state that these clinical observations suggest that a well organized approach to define the genetic determinants of pulmonary fibrosis is scientifically feasible and justified. This project proposes to use standard genetic methodology (linkage analysis) to investigate the distribution of polymorphisms for anonymous genetic markers in families with familial pulmonary fibrosis. The investigators state that their comprehensive genome-wide study, using standard genetic markers, will allow them to identify loci which subsequently may prove to contain novel genes that play a role in the pathogenesis of pulmonary fibrosis. Once genetic loci are defined in familial pulmonary fibrosis, candidate genes can be identified on the basis of both positional and functional criteria. Moreover, they note that this approach will provide basic information on high priority loci that will be applicable to the rapidly evolving dense human transcript map for pulmonary fibrosis in families with two or more cases of pulmonary fibrosis. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: ACID SPHINGOMYELINASE AND NIEMANN-PICK DISEASE Principal Investigator & Institution: Schuchman, Edward H.; Professor &Vice Chairman for Research; Human Genetics; Mount Sinai School of Medicine of Nyu of New York University New York, Ny 10029 Timing: Fiscal Year 2002; Project Start 01-FEB-1992; Project End 31-JAN-2007 Summary: (provided by applicant): Types A & B Niemann-Pick Disease (NPD) are lipid storage disorders resulting from the deficient activity of the lysosomal hydrolase, acid sphingomyelinase (ASM). Type A NPD is a severe neuro-degenerative disorder, which leads to death in early childhood, while patients with Type B NPD have little or no neurological abnormalities and often survive into adulthood. The overall goals of this research are to investigate the underlying causes of the distinct neurological & nonneurological forms of NPD, including the role of ASM in ceramide-mediated cell signaling & disease pathogenesis, and to develop effective treatments for these disorders. Previously, our laboratory: 1) Isolated the full-length cDNAs & genes encoding human & murine ASM, 2) Identified the first ASM mutations causing NPD & developed the first NPD molecular diagnostic program, 3) Constructed a knock-out mouse model for this disorder, 4) Stably over-expressed, purified & characterized recombinant human ASM from CHO cells, 5) Evaluated enzyme replacement, bone marrow transplantation & hematopoietic stem cell gene therapy in the NPD mouse model, and 6) Characterized ceramide-mediated signal transduction in the NPD mouse. In the upcoming funding period we are proposing four specific aims: 1) Investigate the pulmonary disease in NPD mice & develop lung-specific therapies for Type B NPD, 2) Investigate the neurological disease in NPD mice & develop CNS-specific therapies for Type A NPD, 3) Investigate the reproductive biology of NPD mice, including the development of approaches for the selection of normal vs. NPD gametes and

Studies

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preimplantation embryos, and 4) Continue to conduct world-wide ASM mutation analysis & structure/function studies. We believe that this research will provide fundamental insights into the underlying biology of NPD & ASM, and lead to the development of effective treatments for these disorders and/or methods to prevent or minimize NPD births. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: ACTIVATION OF LATENT TGFB BY THE INTEGRIN ALPHA VB6 Principal Investigator & Institution: Munger, John S.; Assistant Professor; Medicine; New York University School of Medicine 550 1St Ave New York, Ny 10016 Timing: Fiscal Year 2002; Project Start 07-FEB-2000; Project End 31-JAN-2004 Summary: Transforming growth factor-beta1 (TGFbeta1) is a widely-expressed cytokine that has major effects on most cell types. TGFbeta1 is anti- inflammatory, pro-fibrotic, and casually linked to fibrotic diseases, e.g. pulmonary fibrosis. TGFbeta1 is secreted in an inactive complex (FTGFbeta1) with its pro-peptide dimer, which is called latencyassociated (LAP). Activation of LTGFbeta1 is a key control point in TGFbeta1 biology, but is poorly understood. Only thrombospondin-1 (TSP1) has been shown previously to active PTGFbeta1 in normal animals. We found that LAP is a ligand for the epitheliumspecific integrin alphavbeta6, and that cells expressing alphavbeta6 bind and activate latent TGFbeta1. This mechanism can explain the heretofore puzzling phenotype of beta6 integrin knock-out mice: inflammation in lung and skin, and protection from bleomycin-induced pulmonary fibrosis. Our results provide the first evidence that dysregulated TGFbeta1 activation causes fibrosis. Our goals are to understand quantitatively the interactions between LTGFbeta and alphavbeta6 that lead to activation, and to develop an animal model an animal model and knowledge to explore fully the biological role of a of avbeta6-mediated LTGFbeta1 activation. In Aim 1 we will analyze the activation mechanism by focusing on alphavbeta6- LTGFbeta1 interactions. We will make TGFbeta1-mull alphavbeta6- expressing cells to which specifically engineered forms of LTGFbeta1 will be added (either by transfection or as recombinant protein). In this system we will then determine the relative activatability of two major forms of LTGFbeta1 (the so-called small and large latent complexes), the relative effects of LTGFbeta1 concentration and alphavbeta6 expression levels of activation, the activatability of soluble and matrix-bound latent TGFbeta1, and the integrin: LTGFbeta1 stoichiometry required for activation. Also, we will assess the influence of integrin/LTGFbeta1 binding affinity on activation. The results will be incorporated into an activation model and related to activation in vivo. In Aim 2, we will create a mouse expressing a mutant form of LTGFbeta1 that cannot be activated by integrins (the RGD integrin binding sites in LAP will be mutated to RGE). The phenotypes of these mice and beta6 integrin null mice will be compared to confirm that the beta6 null phenotype is due specifically to loss of TGFbeta1 activation. To interpret the phenotype will test the ability of other RGD-binding integrins to activate LTGFbeta1. Finally, we will cross RGE-TGFbeta1 mice with TSP1 null mice to assess the summed effects of the two currently known TGFbeta1 activation mechanisms, namely alphavbeta6 and TSP1. The results of these aims will lead to better understanding of alphavbeta6-mediated TGFbeta1 activation in disease. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: ALTERED C/EBPB REGULATION IN IPF LUNG FIBROBLASTS Principal Investigator & Institution: Cruz-Gervis, Roberto A.; Internal Medicine; Meharry Medical College 1005-D B Todd Blvd Nashville, Tn 37208

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Pulmonary Fibrosis

Timing: Fiscal Year 2002; Project Start 01-SEP-2000; Project End 31-JUL-2005 Summary: (Adapted from applicant's abstract) Lung fibroblasts produce large amounts of inflammatory mediators and growth factors that appear to be associated with the development of IPF. The investigators have found that fibroblasts isolated from the lungs of patients with IPF (HF-IPF) produce more IL-6 than do fibroblasts isolated from normal human lungs(HF-NL). Since the IL-6 promoter is under the control of the transcription factors nuclear factor kB(NFkB) and CAAT/enhances binding protein-beta (C/EBPbeta) they measure activation of these proteins. NfkB activation was increased by IL-1beta, but not PGE2, and the response was similar in HF-IPF and HF-NL. In contrast, C/EBPbeta (a.k.a nuclear factor IL-6 [NF-IL6]) activity was increased by both PGE2 and IL-1beta. In addition, there was greater C/EBPbeta activation in HF- IPF compared to HF-NL, which correlated with IL-6 production. C/EBPbeta mRNA contains three inframe translation start sites and, therefore, three different C/EBPbeta isoforms may be produced. The two larger isoforms (p42C/EBPbeta) are transcription activators, while the smallest isoform (p20/EBP beta) lacks a transactivating domain and displays increased DNA affinity, and therefore, is a potent transcription repressor. Recent evidence suggests that C/EBPbeta may have a role in the process of fibrosis. C/EBPbeta has been shown, in liver cells, to stimulate the expression of insulin-like growth factor (IGF)- 1, hepatocyte growth factor (HGF) and collagen-alpha1 genes. Furthermore, transforming growth factor (TGF)-beta induces gene expression of procollagen I via C/EBPbeta. The investigators also have preliminary data showing that, when compared to HF-NL, HF-IPF fail to increase the p20C/EBPbeta:p42C/EBPbeta ratio when stimulated with IL-1beta and PGE2. They hypothesize that phenotypically altered fibroblasts perpetuate the inflammatory and fibrotic response because they have an exaggerated activation of C/EBPbeta, which may be due to a higher p42C/EBPbeta activity, decreased p20C/EBPbeta production, or both. The aim is to establish a relationship between increased C/EPbeta activity, with growth factor (IGF-1, PDGF) production, collagen synthesis and proliferation in primary lung fibroblasts. They will determine whether transfecting primary lung fibroblasts with a plasmid expression vector containing the gene encoding p42C/EBPbeta will result in increased gene expression and protein production of collagen I, and the growth factors TGF- beta, PDGF and IGF-1. In addition, they will determine whether transfecting primary lung fibroblasts with a plasmid expression vector containing the gene encoding p20C/EBPbeta will reduce the stimulated (IL-1beta and TGF-beta) protein production and gene expression of IGF-1, PDGF-1, PDGF and collagen I, as well as fibroblast proliferation. This work could provide a rationale for treatment of IPF with either the p20C/EBPbeta protein or the gene encoding this C/EBPbeta isoform. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: ALVEOLAR EPITHELIUM IN PULMONARY FIBROSIS Principal Investigator & Institution: Mason, Robert J.; Professor; National Jewish Medical & Res Ctr and Research Center Denver, Co 80206 Timing: Fiscal Year 2002; Project Start 01-AUG-2002; Project End 31-JUL-2003 Summary: (Applicant's Abstract) Type II cell hyperplasia is a hallmark of pulmonary fibrosis, but the role of the epithelium on the fibrotic process is unknown. Although there has been numerous studies on the mesenchymal epithelial interactions in the developing lung, there have been relatively few studies in the adult lung. The purpose of this project is to define the epithelial mesenchymal interactions in the adult lung especially as they relate to pulmonary fibrosis. Much of the focus of this proposal will be on the biologic antagonism between KGF and TGF-beta. KGF is an important growth

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factor for type H cells and induces differentiation in vitro, whereas TGF-b inhibits proliferation and antagonizes the effects of KGF on differentiation. Another major question that remains are differences between normal cells and hyperplastic type II cells and if hyperplastic type H cells produced by KGF are different from hyperplastic type H cells seen in fibrotic lung disease. In co-culture normal type II cells inhibit fibroblast growth, but the effect of hyperplastic type II cells from fibrotic lung on fibroblast proliferation is not known. The in vitro studies in this proposal rely on two new culture systems for rat type II cells to maintain differentiated function of type H cells. One has apical access and the other basolateral access. In this proposal we seek to determine (1) if alveolar type H cells can be stimulated to inhibit fibroblast proliferation, (2) the signaling pathways for KGF especially those that lead to differentiation, (3) mechanisms for KGF and TGF-b antagonism, and (4) effects of INFY on production of SP-A and SPD, growth factor receptor expression, and production of chemokines and profibrogenic growth factors. These studies are designed to define the interaction between alveolar epithelial cells and fibroblasts and mechanism of action of KGF. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: ALVEOLAR TYPE II CELL GROWTH IN INJURY Principal Investigator & Institution: Hastings, Randolph H.; Veterans Medical Research Fdn/San Diego Foundation of San Diego San Diego, Ca 92161 Timing: Fiscal Year 2002; Project Start 15-SEP-1999; Project End 31-AUG-2004 Summary: Silica causes lung injury and eventual pulmonary fibrosis. Alveolar type I epithelial cell damage and type II epithelial cell proliferation are prominent features of the injury. Our pilot studies show that type II cells undergo apoptosis afer silica injury also. It is widely believed that type II cell proliferation is an important repair process. However, the functional and structural impact of type II cell growth on recovery from silica induced injury has never been defined and the factors that regulate type II cell growth and death after silica exposure are unknown. Our studies suggest that amino terminal and mid-molecule portions of parathyroid regulate these cell after silicainduced lung injury. PTHrP is a pro- hormone that is processed after translation into different daughter peptides with distinct biologic activities. We have found that lung PTHrP expression falls after silica injury in rats, coincident with the onset of type II cell proliferation and apoptosis. Treating rats with exogenous PTHrP 1- 34, the aminoterminal fragment, reduces pneumocyte division (measured by BrdU uptake), while exogenous PTHrP 67-86, a mid-molecule fragment, decreases type II cell apoptosis (measured by TUNEL staining). We hypothesis that the decrease in amino-terminal and mid-proliferation and apoptosis. Furthermore, we believe that an increase in the type II cell population is beneficial in repairing the epithelium, restoring lung architecture and function, and reducing pulmonary fibrosis. Our specific aims are as follows: 1) We will define the structure-function relationships for the effects of PTHrP peptides on the number of type II cells and fibroblasts in the alveoli after silica injury. 3) We will examine the effect of changes in type II cell number on epithelial repair, progression of pulmonary fibrosis and pulmonary structure after silica-induced lung injury. This project will lead to an understanding of the importance of type II cell growth for epithelial repair after silica-induced lung injury and will assess the role of a novel family of growth factors in regulating pneumocyte growth and death. The project will evaluate the potential use of PTHrP-related therapeutic interventions that might speed or improve recovery following silica-induced lung injury. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Pulmonary Fibrosis

Project Title: ALVEOLIZATION IN FIBRILLIN-1 DEFECTIVE MICE Principal Investigator & Institution: Neptune, Enid R.; Assistant Professor; Medicine; Johns Hopkins University 3400 N Charles St Baltimore, Md 21218 Timing: Fiscal Year 2002; Project Start 13-AUG-2001; Project End 31-JUL-2006 Summary: (provided by applicant): Diseases of impaired gas exchange, such as emphysema and lung fibrosis, are prevalent, clinically burdensome ,and difficult to treat. Because lung transplantation remains the only definitive treatment option for these diseases, much interest has been focused upon understanding the cellular and molecular basis of alveolar formation. As the current knowledge of mammalian lung alveolization is quite limited, this grant is directed towards exploring the mechanism of impaired alveolar septation in mouse models in an effort to understand the requirements of normal septation. The PI has recently observed distal airspace enlargement in two strains of fibrillin-I deficient mice and has found that TGF-beta is a critical mediator of this defect. The first objective is to determine the natural history of impaired septation in three fibrillin-1 defective mouse models. This pursuit should establish whether septation defects may represent important risk factors for the development of emphysema and pulmonary fibrosis. The natural history of the alveolization defects in these models will be correlated with the evolution of aberrant TGF-beta signaling previously observed in two of the models. The second objective is to identify the pulmonary morphologic aberrations which precede the observed septation defects in an effort to reveal critical mediators of septation. Several approaches will be employed to establish whether abnormalities in pulmonary vascular development, matrix composition, or both underlie the disruptions in septation. The final objective is to use chip-based gene expression profiling of murine lung to identify novel and important mediators of septation. The PI of this project is an instructor in the Department of Medicine. She is committed to a career in academic medicine and plans to spend 80 percent of her time in research pursuits. She has had previous training in signal transduction and, more recently, the use of transgenic mouse models to probe human disease. She now wants to expand her research interests to investigating mammalian lung development. To achieve this, she will take sponsored courses on murine development and attend lectures in the developmental genetics department at Johns Hopkins School of Medicine. The environment at Johns Hopkins provides several esteemed scientists who can provide guidance in the use of mouse models to probe lung pathology and developmental aberrations. Their involvement as well as a formal education program in both the Division of Pulmonary Medicine and Institute of Genetic Medicine should facilitate the achievement of her stated research objectives as well as aid in her development into a fully independent investigator. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: ANGIOGENESIS IN HYPEROXIC LUNG FIBROSIS Principal Investigator & Institution: Douglas, Ivor S.; Medicine; University of Colorado Hlth Sciences Ctr P.O. Box 6508, Grants and Contracts Aurora, Co 800450508 Timing: Fiscal Year 2003; Project Start 30-SEP-2003; Project End 31-AUG-2008 Summary: (provided by applicant): This 5-year training program proposes the development plan for a career as an independent biomedical researcher in the area of lung injury and angiogenesis. The principal investigator, has completed Pulmonary and Critical Care Fellowship training through the ABIM Research Pathway. With the sponsors and experienced collaborators he will expand on his scientific skills in preparation for career progression as an independent physician-scientist. The program

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will emphasize skills in molecular biology of angiogenesis and macrophage regulation of fibrosis using a murine hyperoxia model. To advance his knowledge in computational biology he will attend courses in applied statistics. Steven Greenberg M.D. a macrophage biologist, and Paul Rothman M.D., a renowned immunologist in the area of molecular regulation of cytokine signaling, will provide sponsorship. The program will benefit from collaborative expertise of Jan Kitajewski PhD, an expert in angiogenesis and Patty Lee, M.D. who will provide consultative support for the hyperoxia studies. Additionally, George Yancopoulos, a world renowned investigator, will collaborate and serve with the sponsors, Drs. Kitajewski and Lee on an advisory committee every 8 weeks. This committee will review progress and provide close scientific support and career advice. Prolonged hyperoxia results in lung fibrosis in humans and mice. The accompanying vascular remodeling contributes to pulmonary hypertension, right heart failure, and premature death. This program addresses the questions: Do macrophage-derived angiogenic regulatory factors, particularly angiopoietin-2 (Ang-2), contribute to vascular remodeling in response to prolonged sublethal hyperoxia. Do these vascular alterations contribute, independently, to the development of lung fibrosis? And does macrophage-derived Ang-2 directly inhibit endothelial cell survival in response to hyperoxia? The role of macrophage-derived mediators in hyperoxic lung injury remains unclear. We demonstrate in preliminary experiments, increased Ang-2 mRNA and protein in response to hyperoxic exposure in RAW 264.7 and in a mouse model of sublethal hyperoxic lung fibrosis. These mice develop macrophage-predominant cellular infiltration, collagen deposition and pulmonary vascular regression that recapitulates features of subacute lung fibrosis in humans. The following specific aims will be accomplished: 1) The characterization of fibrotic and vascular remodeling responses in hyperoxia-exposed mice by immunohistochemistry, Western and Northern blotting and ELISA to quantify changes in markers of lung fibrosis, angiogenesis and expression of angiogenic regulators and their receptors. The effects on pulmonary vasculature will be evaluated by confocal microscopy. 2) Functional changes in response to altered expression of macrophagederived angiogenic regulators by angiogenesis assays. Macrophage-depleted mice will be used to assess the contribution of lung macrophages. 3) Lung targeted overexpression of Ang-1 or 2 by adenovirus transfection or blocking antibodies prior to hyperoxia to determine the contribution of Ang-2 to pulmonary vascular remodeling and fibrosis during prolonged sublethal hyperoxia. 4) Determine if Ang-2 mediated endothelial survival inhibition is PI3K/Akt dependent. Columbia University has an established record of successful mentorship and training for K08 Career awardees in their preparation for careers as independent. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: APPLIED GENOMICS IN CARDIOPULMONARY DISEASE Principal Investigator & Institution: Haponik, Edward F.; Professor of Internal Medicine; Medicine; Johns Hopkins University 3400 N Charles St Baltimore, Md 21218 Timing: Fiscal Year 2002; Project Start 30-SEP-2000; Project End 31-JUL-2004 Summary: The Clinical Core will serve as the coordinating center for patient recruitment, specimen procurement and data base management. It is the mission of this Core to identify and provide disease-specific candidate genes from patients with such clinical disorders as acute lung injury, COPD, cystic fibrosis, asthma, pulmonary hypertension, pulmonary fibrosis, ischemic heart failure and both lung and cardiac transplant rejection. This core will also be responsible for the establishment of a Cardiopulmonary Tissue Repository that will maintain an archive of cryopreserved

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Pulmonary Fibrosis

tissues and maintain an accurate data base of patient demographic and clinical data for correlation with biological end points produced by cDNA microarray. Following patient consent and registration, tissue samples and peripheral blood are delivered to the Core laboratory. The freezing facility will also serve as a repository for cyropreserved human lung and cardiac specimens. The Core Investigators will meet regularly to review all diagnostic materials on each specimen procured including histochemical stains and biopsies in order to determine a precise diagnosis and relevant demographic and clinical data for entry into the Project's data base which will include critical variables for the analysis of the biological data obtained. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: APPLIED GENOMICS IN CARDIOPULMONARY DISEASE Principal Investigator & Institution: Scott, Alan L.; Physiology; Johns Hopkins University 3400 N Charles St Baltimore, Md 21218 Timing: Fiscal Year 2002; Project Start 30-SEP-2000; Project End 31-JUL-2004 Summary: The overall goal of the Animal/Proteomic Component of the "Applied Genomic Program in Cardiopulmonary Disease" is to define and test the relevance of disease specific gene candidates that predict lung and cardiac remodeling in animal models of cardiopulmonary diseases utilizing gene profiling approaches. Identification of susceptibility genes for human disease is hampered by variability in clinical phenotype, genetic heterogeneity in human populations and the experimental difficulty in addressing the molecular mechanisms underlying complex pathological processes in humans. Thus our strategy is to take advantage of the experimental tractability of murine models of disease to provide high quality of candidate genes underlying remodeling processes in multiple cardiopulmonary disease. To achieve this goal, we have assembled an outstanding group of investigators with broad and overlapping expertise with animal models of cardiopulmonary diseases including asthma, pulmonary fibrosis, cardiac failure, emphysema, hyperoxia-induced lung injury and pulmonary hypertension. Our preliminary data suggest that these models are predictive of human disease and that the gene profiling approach can successfully be used to identify genes important in human disease. The specific aims of this component are l) to define a set of predictor genes for tissue remodeling using Affymetrix 5000 predictor oligonucleotide microarrays (Mu19K) in each of the six animal models of disease; 2) to refine the number of candidate genes and to establish the kinetics of gene expression by constructing custom cDNA arrays for 1000-5000 predictor genes in each model; and 3) to compare and contract gene expression profiles between models and human systems in order to prioritize candidates for further analysis by proteomic and single nucleotide polymorphism (SNPs) approaches; 4) to utilize proteomic approaches to study the consequences of changes in gene expression at the cell and tissue level; and 5) to being to determine the functional relevance of this focused set of genes to remodeling processes by utilizing transgenic and knockout technologies. The combined (mouse and human) approach of this program to the identification of disease specific genes for lung and cardiac remodeling should greatly facilitate future disease discovery. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: BASIS OF VARIABILITY OF LUNG GPCR SIGNALING Principal Investigator & Institution: Liggett, Stephen B.; Professor; None; University of Cincinnati 2624 Clifton Ave Cincinnati, Oh 45221 Timing: Fiscal Year 2003; Project Start 01-SEP-2003; Project End 31-AUG-2008

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Summary: (provided by applicant): Many major signaling events in the lung are carried out by the superfamily of G-protein coupled receptors (GPCRs). These include bronchial smooth muscle relaxation and contraction, mucous secretion, ciliary beat frequency, inflammation, immune cell trafficking, pulmonary vascular tone and permeability, and alveolar fluid and electrolyte transport, as well as many yet to be defined functions. Approximately 75 GPCRs are estimated to be expressed in human lung. Within the next five years, the great majority of all therapeutic agents will target GPCRs. However, the physiologic, pathologic, and pharmacologic behavior of GPCR signaling displays substantial interindividual variability which is thought to be due to common variants (polymorphisms) of the genes encoding these receptors. Such polymorphisms have been estimated to account for as much as 50% of the variability in the response to therapeutic agonists and antagonists targeted to GPCRs. Indeed, with just one GPCR, the (2adrenergic receptor, we have shown that coding and promoter polymorphisms alter receptor expression, function, and regulation in vitro in cells, and in asthmatic patients, they are associated with clinical phenotypes and the response to beta-agonist therapy. The long-term goals of this proposal are to identify polymorphisms of up to 20 pulmonary GPCR genes, and by the use of recombinant expression techniques to delineate their biochemical and pharmacologic impact on cellular signaling relevant to lung homeostasis and pathobiology. In Aim 1, the polymorphisms of these 20 GPCR genes will be delineated in the promoter, 5' untranslated, coding, intron/exon junctions, and 3' untranslated regions from genomic DNA samples from a cohort of 60 ethnically diverse individuals. In Aim 2, the common combinations of polymorphisms (haplotypes) will be delineated in the population. In Aim 3, constructs will be developed and model cell systems utilized for recombinant expression of GPCR haplotypes to determine the biological effects of polymorphisms on receptor expression, signaling, or regulation. These studies will provide the basis for interindividual susceptibility and therapeutic responsiveness, the variability in pathobiology, and the potential for developing new treatment strategies, for a diverse range of lung diseases including asthma, pulmonary hypertension, pneumonia, pulmonary fibrosis, COPD, and pulmonary edema. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: BIODEGRADABLE SEALANT FOR BIOPSY TRACTS IN SOFT TISSUES Principal Investigator & Institution: Hronowski, Lucjan J J.; Biopsy Sciences, Llc 6340 N Pinnacle Ridge Dr Tucson, Az 85718 Timing: Fiscal Year 2003; Project Start 01-MAR-2003; Project End 31-AUG-2003 Summary: (provided by applicant): Percutaneous fine needle biopsy is a common method of determining if a lung nodule is benign or malignant. Using imaging guidance (CT, fluoroscopy, ultrasound or bronchoscopy) a sharp needle is advanced into a suspicious area and cells are aspirated into the needle. These cells are then evaluated under a microscope and are characterized as benign, infectious or malignant. There are over 169,500 cases of lung cancer diagnosed in the U.S. and over 600,000 lung biopsies performed worldwide every year. The most common complication of this procedure is a lung collapse (pneumothorax), which occurs in 20-40% of cases. In greater than 10% of these lung collapses, the patient becomes symptomatic and requires chest tube placement and hospitalization. The development of a novel biopsy needle tract sealant that will reduce or eliminate pneumothorax would be most valuable. The focus of the current investigation is to evaluate the possibility of using an expansile polymer to seal these needle tracts. Further, in the investigation we plan on studying a deployment

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Pulmonary Fibrosis

system that plugs the tract but does not irritate the parietal pleura. Initial Investigation into the tissue compatibility will also be examined. There is presently no commercially available product in the U.S. to prevent or reduce the pneumothorax rate after lung Fine Needle Aspiration Biopsy (FNAB) Procedures. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: PROTEINS

BIOGENESIS

AND

FUNCTION

OF

CONNECTIVE

TISSUE

Principal Investigator & Institution: Bornstein, Paul; Professor; Biochemistry; University of Washington Grant & Contract Services Seattle, Wa 98105 Timing: Fiscal Year 2002; Project Start 01-FEB-1975; Project End 31-JAN-2005 Summary: The transcriptional regulation of the genes encoding the two type I collagen chains, alpha1(I) and alpha2(I), is one of special interest because these genes are expressed at widely different levels that correlate with the tissue specificity of collagen synthesis, and with development and maturation of the organism. Furthermore, the genes for alpha1(I) and alpha2(I) are responsive to cues generated by injury and repair, and by a variety of cytokines, hormones, and pharmacological agents. Finally, the expression of type I collagen genes is disturbed in orders such as pulmonary fibrosis and cirrhosis, and in diseases such as scleroderma. Although post-transcriptional mechanisms undoubtedly play an important role in regulating collagen synthesis, there is good evidence that transcriptional control represents the major means by which this regulation is achieved. A major goal of this grant is to determine how this astonishingly intricate pattern of expression is established and maintained, and how it is altered during development, in response to injury, and in disease. Current studies of gene regulation generally involve the evaluation of mutations in chimeric regulatory/reporter genes in transfection and transgenic experiments. While these approaches represent necessary preliminary steps, it is our contention that definitive results can best be achieved by testing such mutations in the context of the endogenous gene. Gene targeting techniques will therefore be used to create mutations in putative regulatory regions of the Collal gene in mice, and mutant mice will be evaluated for expression of the altered allele and for phenotypic changes. In particular, the proposed experiments will test the hypothesis that modular elements in the Collal gene direct the synthesis of type I collagen selectively to tissues such as skin and bone. It is anticipated that some of the mutations created in mice will generate useful models for human disorders of these tissues, specifically some of the Ehlers-Danlos syndromes and osteoporosis. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: BIOMARKERS IN SPUTUM IN COPD Principal Investigator & Institution: Broide, David H.; Associate Professor of Medicine; Medicine; University of California San Diego 9500 Gilman Dr, Dept. 0934 La Jolla, Ca 920930934 Timing: Fiscal Year 2002; Project Start 23-SEP-2002; Project End 31-JUL-2006 Summary: (provided by applicant): The overall goal of this proposal is to identify biomarkers in the sputum of patients with emphysema that associate with emphysema but not with either healthy controls, smokers without emphysema, or lung diseases associated with airway inflammation and fibrosis such as asthma or idiopathic pulmonary fibrosis. The approach we plan to use to identify sputum biomarkers in emphysema will use both hypothesis driven experiments to explore candidate sputum

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biomarkers of inflammation (LTB4, IL-8, TNF), fibrosis (TGF- beta, PGDF, FGF), metalloproteases (MMP-1, MMP-9, MMP-12, TIMP), elastin degradation (desmosine in urine) as well as an alternative approach using both proteomic analysis of sputum and bronchoalveolar lavage, and genomic studies of airway epithelium and alveolar macrophages to identify potential novel biomarkers of emphysema that correlate with CT scan evidence of emphysema. Levels of biomarkers will be measured over a two year period and be correlated with CT scan extent of emphysema. If such a non-invasive biomarker were identified this could either serve as a biomarker in studying the effects of intervention with anti-inflammatory medications or smoking cessation in subjects with emphysema, or alternatively serve to identify smokers at risk for the development of emphysema. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: FORMATION

CHEMOKINE

RECEPTOR

DYNAMICS

IN

GRANULOMA

Principal Investigator & Institution: Chensue, Stephen W.; Pathology; University of Michigan at Ann Arbor 3003 South State, Room 1040 Ann Arbor, Mi 481091274 Timing: Fiscal Year 2003; Project Start 01-AUG-1998; Project End 31-MAR-2007 Summary: (provided by applicant): Hypersensitivity-type granulomas (GR) are T cellmediated chronic inflammatory lesions observed in a wide variety of infectious and noninfectious diseases often causing serious morbidity and mortality. Understanding the manner in which T cells participate and promote theses lesions will aid the development of interventions. These lesions can be classified as type-1 or type-2 based upon the relative participation of Th1 and Th2 associated cytokines. The current paradigm is that Th1 and Th2 CD4+ helper cells are recruited to inflammatory sites by chemokines (CK) and show selective migration by virtue of differential chemokine receptor expression. Using animal models of synchronized, type-1 and type-2 pulmonary GR formation elicited respectively by protein antigens of Mycobacteria tuberculosis and Schistosoma mansoni, we found indications of both shared and polarized CK receptor expression among type-1 and type-2 CD4+ memory helper cells as well as antigen stimulated regulation. However, the in vivo significance and contribution of these findings to GR formation is unknown. The present proposal will extend this work and test the hypothesis that effector Th1 and Th2 cells are recruited to inflammatory sites by way of innate phase chemokines through polarized postactivation chemokine receptors. The specific aims will employ state-of-the-art laser capture microdissection (LCM) and real time, gene sequence detection technologies to reveal the microenvironmental expression and function of Th cell CK receptors. Aim 1 will use LCM to define the temporal expression and tissue compartmental location of CK transcripts during synchronous Ag-bead and asychronous infectious type-1 and type-2 GR in order to allow correlation to T cell CK receptor expression. Aim 2 will determine the distribution of induced CK receptor transcripts among effector Th1 and Th2 cells generated in vivo. Aim 3 will reveal the potential biologic contribution Aginduced receptors such as CXCR3, CCR4 and CCR8 to selective Th migration using direct chemotaxis of antigen-activated CD4+ T cells. Aim 4 will define the temporal expression of CK receptor transcripts within the tissue microenvironments during synchronous Ag-bead and asychronous infectious type-1 and type-2 GR formation. Finally, Aim 5 will examine the migratory behavior of adoptively transferred CD4+ T cells with targeted CK receptor knockout and concomitant transgenic expression of green fluorescent protein (GFP). These studies will potentially provide novel and

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important information regarding the contribution of CK receptors to CD4+ T trafficking in chronic inflammatory responses. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: CHEMOKINES IN LUNG TRANSPLANTATION Principal Investigator & Institution: Medoff, Benjamin D.; Massachusetts General Hospital 55 Fruit St Boston, Ma 02114 Timing: Fiscal Year 2003; Project Start 04-AUG-2003; Project End 31-JUL-2008 Summary: (provided by applicant): With the proposed Mentored Clinical Scientist Development Award the applicant will continue his investigations into basic mechanisms of lung inflammation. After two productive years in this laboratory the applicant remains firmly committed to a career in academic pulmonary medicine. The proposed research will allow the applicant to master a broad range of laboratory techniques in immunology, cell, and molecular biology. The research experience will be supplemented by a program of study of immunology and medical science. The project focuses on the development of inflammation and fibrosis following lung transplantation and the role of chemokines in these processes. After a lung is transplanted there may be several types of injury to the graft, including ischemia-reperfusion injury, acute rejection, and chronic rejection. These immune mediated injuries contribute to the development of scarring of the airways, so called bronchiolitis obliterans (BO). Over 50% of all lung transplants will develop BO after transplantation, and this remains the major cause of morbidity and mortality after lung transplantation. Neutrophils have been shown to be a prominent component of ischemia-reperfusion injury while T lymphocytes are the primary mediators of both acute and chronic rejection. The proposed project will determine which chemokines are produced after transplantation and their contribution to the development of graft injury and subsequent BO. Further experiments will manipulate chemokine or chemokine receptor expression in animal models of lung transplantation to investigate their role in the development of graft injury and BO. The applicant specifically proposes to: (1) investigate the expression of chemokines and chemokine receptors in the lung following transplantation in patients with and without acute rejection and BO; (2) investigate the role of chemokines in the development of ischemia-reperfusion injury in the airways using the murine tracheal heterotopic model of lung transplantation; (3) investigate the role of chemokines in the development of acute airway rejection and the development of BO in the murine tracheal heterotopic model of lung transplantation; (4) develop a novel murine model of airway rejection and BO. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: CHONDROPROTECTIVE EFFECTS OF SUPEROXIDE DISMUTASE Principal Investigator & Institution: Chu, Constance R.; Orthopaedic Surgery; University of Pittsburgh at Pittsburgh 350 Thackeray Hall Pittsburgh, Pa 15260 Timing: Fiscal Year 2002; Project Start 01-JUN-2000; Project End 31-MAY-2004 Summary: (applicant's abstract): Destruction of articular cartilage through disease processes such as inflammatory arthritis and osteoarthritis causes significant pain and disability. The normal balance between anabolic and catabolic processes is altered so that chondrocyte synthesis of matrix components cannot compensate for the increased rates of degradation found in these conditions. The effects of superoxide dismutase on articular cartilage homeostasis has not been systematically studied. Exploratory studies using extracelluar superoxide dismutase (EC-SOD) knock-out and transgenic mice

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originally developed for evaluating the pathogenesis of pulmonary fibrosis to examine these relationships are proposed. The long term goal is to explore the potential chondroprotective effects of EC-SOD. The working hypothesis is that EC-SOD plays a central role in protecting against cartilage degradation in the arthritic joint. To test the hypothesis that EC-SOD plays a central role in protecting against articular cartilage loss in the arthritic joint by comparing the structural and metabolic responses of articular cartilage following zymosan induced gonarthritis between wild-type, EC-SOD knockout mice which cannot produce EC-SOD, and EC-SOD transgenic mice which produce excess EC-SOD. Studies in experimental models of arthritis have implicated nitric oxide (NO) as an important pathogenic factor. EC-SOD may prevent the reaction of NO with superoxide to form peroxynitrite, a potent oxidant which may account for some of the damaging effects of NO. Interleukin-l (IL-l) inhibition of proteoglycan synthesis has been linked to the presence of NO. To test the hypothesis that prevention of peroxynitrite formation is one mechanism by which EC-SOD protects against IL-1 inhibition of carnage repair, we plan to compare NO production, nitrotyrosine immunoreactivity, matrix metalloproteinases, and proteoglycan synthesis between wildtype, EC-SOD knockout and EC-SOD transgenic mice following intra articular injection of IL-1. IL-l has been associated with development of chondrocyte insensitivity to the anabolic actions of Insulin-like Growth Factor-I (IGF) in both in vitro and in vivo models of arthritis. In contrast to wild-type mice, iNOS knockout mice maintain their anabolic response to IGF-1 following intra articular injection of IL-1. This implicates NO and NO derivatives such as peroxynitrite as critical, factors in the development of chondrocyte insensitivity to IGF-1 in the arthritic joint. To test the hypothesis that peroxynitrite is the reactive species involved in IL-1 induced chondrocyte unresponsiveness to the anabolic effects of IGF-1, we plan to compare NO production, nitrotyrosine immunoreactivity, and proteoglycan synthesis in the presence of IGF-1 between wild-type EC-SOD knockout and EC-SOD transgenic mice following intra articular injection of IL-1. The proposed studies will advance the current knowledge base on the effects of EC-SOD in modulating cartilage degradation in the arthritic joint. These studies will also serve to improve understanding of the relative roles of NO and peroxynitrite on the pathophysiology of arthritis. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: COLLAGEN TRANSCRIPTION AND LUNG FIBROSIS Principal Investigator & Institution: Smith, Barbara D.; Professor; Biochemistry; Boston University Medical Campus 715 Albany St, 560 Boston, Ma 02118 Timing: Fiscal Year 2002; Project Start 01-JUL-2001; Project End 31-MAY-2005 Summary: Certain lung injuries induce large increases in connective tissue content, particularly collagen, resulting in fibrosis. During injury, cells are exposed to molecules such as interferon-gamma (IFN-gamma) and transforming growth factor-beta (TGFbeta), regulating production of matrix components. Clinical trials for interstitial pulmonary fibrosis (IPF) are ongoing. However, very little is understood concerning collagen repression by this mediator. This project focuses on establishing the mechanisms by which collagen transcription is repressed by IFN- gamma. We have recently described a regulatory factor for X-box (RFX) binding site at the collagen transcription start site. RFX1 represses collagen transcription. RFX1 interacts with and activates c-Abl, a non- receptor tyrosine kinase that can phosphorylate itself, RFX1 and the carboxyl domain of RNA polymerase II (CTD). C-Abl interacts with RFX1 at the collagen transcription start site. This proposal determines whether c-Abl participates in signal transduction pathways leading to decreased collagen synthesis. RFX5 forms a

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Pulmonary Fibrosis

complex at the collagen transcription start site when RFX1 is removed. IFN-gamma induces class II transcription activator (CIITA) which interacts with RFX5 forming a complex with two other proteins that activate major histocompatibility class II proteins (MHC-II). Since IFN-gamma represses collagen synthesis, we hypothesize that RFX proteins mediate collagen transcription repression during IFN-gamma treatment. Our overall hypothesis is that IFN-gamma repression occurs on the collagen promoter by cooperative interactions of RFX protein family members at the start site with other proteins binding to the proximal promoter. The specific aims are to; 1) Determine the function and interactions of RFX family before and after IFN-gamma treatment. 2) Examine the functional interactions of c-Abl with RFX proteins at the collagen transcription start site. 3) Examine the localization and kinase activity of c-Abl and RFX in lung fibroblasts under different treatments and in samples of human lung tissue. 4) Use transgenic animals with collagen-promoter-CAT constructs or animals deficient in c-Abl or RFX5 complex to investigate collagen transcriptional regulation during fibrosis and treatment. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: CTGF IN LUNG DEVELOPMENT AND BPD Principal Investigator & Institution: Rosenbloom, Joel; Children's Hospital of Philadelphia 34Th St and Civic Ctr Blvd Philadelphia, Pa 191044399 Timing: Fiscal Year 2002; Project Start 01-AUG-2002; Project End 31-JUL-2003 Summary: (Applicant's Abstract) Although there is increasing evidence that failure of normal lung septation is a component of bronchopulmonary dysplasia (BPD) currently seen in small premature infants, a fibroproliferative response remains responsible for many of the untoward alterations in pulmonary function. While the pathogenic mechanisms underlying this response are complex, it is likely that many of the harmful aspects are mediated by the manifold effects of TGF-B as a final common pathway. Strong evidence suggests that many of the effects of TGF-B on fibroblast proliferation and extracelluar matrix production are mediated by connective tissue growth factor (CTGF). The preliminary data demonstrate that CTGF expression by cultured human fetal lung fibroblasts and airway smooth muscle cells is greatly stimulated by TGF-B1 and that CTGF is expressed in the developing lung. The investigators propose the following hypotheses: (1) A TGF-B superfamily member stimulates the expression of CTGF which acts as a downstream mediator in the regulation of branching and other morphologic events during normal lung development. (2) The signaling pathway by which TGF-B up-regulates CTGF expression involves cellular components in addition to the Smads. (3) CTGF is a major effector molecule in the pathogenesis of pulmonary fibrosis seen in BPD. To test these hypotheses, they will: (1) Determine the temporal and spatial expression of CTGF in the developing mouse and human lung and determine its potential role in lung development by conditional ablation. (2) Define the signaling pathway and mechanisms whereby TGF-B1 up-regulates the expression of CTGF and modulates expression of matrix proteins. (3) Develop strategies for inhibiting the fibrotic response mediated by TGF-B and CTGF. This work will be carried out in close collaboration with Projects 5 & 6, will interact significantly with Projects 1 and 4, will utilize the Tissue Culture Core for the implementation of the mouse lung bud model and will obtain human lung samples from the Clinical Core. Identification of the mechanisms of action of CTGF and the pathways regulating its expression are of considerable importance, since CTGF may play a key role in normal lung development and blocking its abnormal production may ameliorate the fibrotic response seen in BPD. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: DISRUPTION OF PDGF SIGNAL TRANSDUCTION IN LUNG FIBROSIS Principal Investigator & Institution: Lasky, Joseph A.; Associate Professor; Medicine; Tulane University of Louisiana New Orleans, La New Orleans, La 70112 Timing: Fiscal Year 2002; Project Start 30-SEP-2000; Project End 31-AUG-2005 Summary: The formation of scars within the lungs (pulmonary fibrosis) frequently leads to shortness of breath, disability and even death. Occupational exposure to organic or inorganic dusts are well-known to cause pulmonary fibrosis. The etiology of many cases of lung fibrosis remains unknown, or idiopathic, but it is thought that occupational and environmental exposure to particulates may be responsible for upwards of 25% of these idiopathic cases. Unfortunately, current medical treatment is frequently ineffective in halting or reversing lung fibrosis. The overall purpose of this study is to better understand the basic mechanism of lung scarring so that physicians will some day be able to intervene in the pulmonary scarring process and thereby prevent suffering and postpone death. A common structural feature of lung fibrosis is an increase in the number of cells called fibroblasts. A scar is formed when fibroblasts move into a wound and proliferate. Fibroblast proliferation and chemotaxis are mediated by growth factors and cytokines. Platelet-derived growth factor (PDGF) is a potent fibroblast growth factor and chemoattractant, which binds to lung fibroblast surface receptors to generate secondary messengers that stimulate cell growth. Although PDGF expression is upregulated at the site of fibroproliferative lung lesions, it remains to be demonstrated whether or not PDGF is actually directing fibroproliferation during lung fibrogenesis. The hypothesis is that disruption of PDGF receptor signal transduction will significantly inhibit the development of lung fibrogenesis. To test this hypothesis, two strategies directed at disrupting PDGF receptor signal transduction in lung fibroblasts in vitro and in vivo will be employed. The first approach will utilize a newly-developed tyrosine kinase inhibitor that is selective for PDGF receptors, whereas the second approach will use dominant-negative PDGF receptor constructs to inhibit murine lung fibroblast proliferation. It is expected that the information derived from employing these two strategies will be complementary. In concert, these experiments will define for the first time the key role PDGF plays in the pathogenesis of pulmonary fibrosis and help answer whether future research should be directed toward blocking the activity of PDGF to treat lung fibrosis in humans. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: EFFECTS OF IPF AND ITS TREATMENT ON INNATE IMMUNITY Principal Investigator & Institution: Ganz, Tomas; Professor; University of California Los Angeles 10920 Wilshire Blvd., Suite 1200 Los Angeles, Ca 90024 Timing: Fiscal Year 2002; Project Start 01-AUG-2002; Project End 31-JUL-2003 Summary: (Applicant's Abstract) Interferon-gamma (IFN-gamma), a promising agent for the treatment of IPF, is also the most potent known stimulant of host defense against bacterial, fungal and viral infections. We hypothesize that unlike the immunosuppressive drugs prednisone and azathiopnine, IFN-gamma reduces the infectious pulmonary complications of IPF and their adverse effects on the clinical course and respiratory performance. We propose to compare the effects of immunosuppressive regimens vs. IFN-gamma on innate mucosal host defenses and to identify specific mechanisms and molecular effectors that are modulated by the contrasting treatment modalities. We will: 1. Compare the effects of IFN-gamma vs. high dose prednisone on the composition and density of nasal and pharyngeal microbial

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flora. 2. Analyze the effects of IFN-gamma vs. high dose prednisone on known effector proteins of innate host defense in nasal epithelia 3. Analyze the effects of IFN-gamma vs. high dose prednisone on known effector proteins of innate host defense in the lower airways 4. Using proteomics, identify novel proteins (potential effectors and mediators) whose concentrations in nasal or lung fluids are induced or suppressed during treatment with IFN-gamma vs. high dose prednisone As a subsidiary objective, we will sample the azathiopnine-treated patients to test the hypothesis that like high dose prednisone, azathioprine will also have suppressive effects on mucosal host defense compared to IFN-gamma In the aggregate, this study will explore novel concepts of the modulation of mucosal host defenses by immunostimulatory vs. immunosuppressive agents. This is an exciting opportunity to study these mechanisms under well-controlled conditions in humans. The proposal also explores a novel alternative mechanism for the potentially beneficial effect of IFN-gamma in IPF-the avoidance or active prevention of secondary infections that exacerbate the clinical course of conventionally-treated patients with this disease. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: FIBROBLAST ABLATION AS TREATMENT FOR PULMONARY FIBROSIS Principal Investigator & Institution: Henke, Craig A.; Associate Professor of Medicine; Medicine; University of Minnesota Twin Cities 200 Oak Street Se Minneapolis, Mn 554552070 Timing: Fiscal Year 2002; Project Start 01-APR-2002; Project End 31-MAR-2007 Summary: Pulmonary fibrosis is characterized by the progressive accumulation of fibroblasts within the alveolar wall and airspace with subsequent deposition of collagen in the distal airspace. This is manifested clinically by severe shunt physiology and often leads to respiratory failure and death. Unfortunately, therapeutic modalities for fibroproliferative lung diseases, which suppress inflammation, have had limited clinical benefit in terms of halting progressive fibrosis. This suggests therapy targeting fibroblasts is essential. It is well documented that some patients with extensive alveolar fibrosis are capable of full recovery. Experimental evidence indicates that removal of unwanted fibroblasts occurs by apoptosis at a critical timepoint during tissue repair allowing restoration of normal anatomic patterns. Therefore, we hypothesize that selective removal of fibroblasts actively participating in the fibrotic process at a critical time period after lung injury will be beneficial in attenuating pulmonary fibrosis and promoting lung repair. In a proof of principle study we plan to utilize transgenic mice we have developed expressing HSV-TK from the mouse alpha 2 type I [alpha2(I)] collagen promoter to test our hypothesis. To selectively target fibroblasts, an alpha2(I) collagen promoter that has selective expression by fibroblasts will be used. To achieve fibroblast ablation, we will employ the thymidine kinase gene of the herpes simplex virus (HSV-TK). Those fibroblasts actively participating in the fibrotic process (e.g., those actively synthesizing large amounts of type I collagen) will express transgene derived HSV-TK and metabolize the antiviral agent ganciclovir (GCV) to toxic nucleotide analogs which disturb nucleic acid synthesis and induce cell death. To create pulmonary fibrosis, the murine model of bleomycin-induced pulmonary fibrosis will be utilized. In this model of pulmonary fibrosis alpha2(I) collagen gene expression by fibroblasts is markedly increased. Therefore fibroblast ablation will be regulated both regionally by the use of the alpha2(I) collagen promoter to target lung fibroblasts actively synthesizing type I collagen and temporally by when GCV is administered. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: FIBRONECTIN DEPOSITION AND PULMONARY FIBROSIS Principal Investigator & Institution: Hocking, Denise C.; Assistant Professor; Pharmacology-Physiology; University of Rochester Orpa - Rc Box 270140 Rochester, Ny 14627 Timing: Fiscal Year 2002; Project Start 01-APR-1998; Project End 31-MAR-2004 Summary: (Applicant's abstract): Pulmonary fibrosis is a potentially lethal lung disorder characterized by fibroblast proliferation and excessive accumulation of extracellular matrix proteins, including fibronectin and types I and III collagen. As a consequence of matrix deposition, thickening of the alveolar septum and loss of functional alveolar capillaries can lead to impaired pulmonary function. The development of pulmonary fibrosis is a common response to acute injury and inflammation in the lung. Current therapies, however, have little effect on the progression of the disease. Fibronectin expression and deposition are markedly increased in tissues following injury, where it functions as a chemoattractant and adhesive molecule for migrating cells. Excess or inappropriate deposition of fibronectin into the extracellula matrix during the reparative phase of injury has been associated with fibrotic changes. Evidence suggests that polymerized fibronectin may serve as a templat for subsequent collagen deposition. However, the precise relationship between fibronectin polymerization and collagen synthesis and deposition during pulmonary fibrosis is not known. The goal of these studies is to elucidate the role of multimeric fibronectin in regulating cell migration, collagen synthesi and collagen deposition. Recombinant fibronectin fragments, which inhibit fibronectin polymerization in vitro, will be used to determine the relationshi between fibronectin polymerization and collagen deposition, in vitro and in vivo, using an animal model of fibrosis. These studies will provide informatio crucial to understanding the role of extracellular matrix fibronectin in regulating cell activities which contribute to the development of fibrosis. Elucidating the effect of fibronectin matrix assembly on cell behavior is central to designing methods of intervention during abnormal or altered fibronectin deposition, as occurs during fibrosis. As such, the overall aim of this proposal is to identify mechanisms which control and modulate fibronectin deposition in order to define strategies aimed at limiting the excess deposition of extracellular matrix during pulmonary fibrosis. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: FUNCTIONAL ASSESSMENT OF PULMONARY TOXICITY WITH MRM Principal Investigator & Institution: Johnson, G. Allen.; Professor; Radiology; Duke University Durham, Nc 27706 Timing: Fiscal Year 2002; Project Start 30-SEP-1996; Project End 31-AUG-2006 Summary: (Provided by Applicant): We propose continuation of studies of the environmental impact of particulate matter on lung structure and function using in vivo magnetic resonance microscopy. We will expand our previous work with hyperpolarized 3He to support localized three-dimensional structural imaging at 1 x 104 mm3. We will extend our physiologic support system and spiral encoding techniques to quantitative flow measurements and functional imaging at 1 x 10-2 mm3. We will study the impact of particulates in three specific models: 1) elastase-induced injury, a model of emphysema; 2) inflammation and remodeling in the endotoxin sensitive C3H/HeBFeJ mouse, a model of organic dust induced asthma; and 3) inflammation and remodeling in the vanadium pentoxide model of asthma, which is characterized by

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extensive bronchiolar fibrosis. The proposal will refine methods that will be applicable to a much wider range of basic pulmonary studies while obtaining detailed assessment of structural and functional changes in three important models of human disease. By extending the methodologies to the mouse, we will build a critical bridge between man and mouse models of pulmonary disease that will be essential to understanding the pathophysiology of pulmonary disease and the validation of new therapies. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: GENE REGULATION DURING INTERMITTENT HYPOXIA Principal Investigator & Institution: Prabhakar, Nanduri R.; Professor; Physiology and Biophysics; Case Western Reserve University 10900 Euclid Ave Cleveland, Oh 44106 Timing: Fiscal Year 2002; Project Start 30-SEP-2000; Project End 31-AUG-2004 Summary: Episodic or intermittent hypoxia (IH) is associated with many pathophysiological situations, including sleep apneas and lung diseases such as chronic obstructive pulmonary disease (COPD), asthma, or pulmonary fibrosis. Experimental studies on humans, as well as animals, have documented that IH has long term effects on the cardio-respiratory systems leading to pathophysical conditions such as hypertension. Despite its clinical significance little is known about the molecular mechanisms underlying IH. The investigators have devised a technique that exposes cells to alternating cycles of low oxygen simulating IH seen in intact animals. Using this technique, they discovered two features that distinguish IH from SH. First, for a given duration and severity of hypoxia, IH is more potent in activating gene expression. Second, increases in gene expression persisted for hours after terminating IH, a phenomenon resembling long-term facilitation (LTF). The overall goal of the current proposal is to identify the mechanisms underlying activation of gene expression by IH. The application specifically will test the hypothesis that gene activation by IH depends on the duration of oxygenation intervening the hypoxic episodes, and reactive oxygen species (ROS) play a crucial role. The investigators will use an integrated approach to test this hypothesis using both cell culture systems and in vivo experiments on wild type and mutant mice. The current proposal focuses on the regulation of c-fos and tyrosine hydrosylase (TH) genes. The Specific Aims are: 1) To assess the importance of episodic re- oxygenation on gene induction by IH and to determine whether IH-induced elevations in mRNAs are due to increased transcription, stability or both; 2) To determine the role of reactive oxygen species (ROS) and HNE signaling pathways to gene induction by IH; 3) To examine the contribution of Ca2- dependent an HNE signaling pathways to gene expression by IH; 4) To correlate cellular effects of IH on gene expression with chronic systemic responses to IH in intact animals using c-fos knockout mice. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: GENETIC TUBERCULOSIS

CONTROL

OF

PULMONARY

CAVITIES

IN

Principal Investigator & Institution: Hunter, Robert L.; Professor and Chairman; Pathology and Lab Medicine; University of Texas Hlth Sci Ctr Houston Box 20036 Houston, Tx 77225 Timing: Fiscal Year 2002; Project Start 10-SEP-2001; Project End 31-JUL-2006 Summary: (provided by applicant) Since it has no animal or environmental reservoir, M. tuberculosis (MTB) must be aerosolized by diseased individuals in order to propagate. The organism accomplishes this most effectively by forming pulmonary cavities. One of

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us (PMS) has recently observed genetic deficiencies in MTB strains with reduced capacity to cause pulmonary cavities in humans. Another of us (RH) has developed a mouse model that suggests the toxicity of cord factor (trehalose 6,6" dimycolate (TDM)) contributes to the formation of cavities. Together, we now propose to identify and characterize MTB genes that control the formation of pulmonary cavities. We will use: 1) two different DNA microarrays to determine, on a genome wide scale, the presence and expression profile of all MTB genes; 2) a new mouse model that manifests the type of caseating granuloma that gives rise to cavities; and 3) conventional genetic approaches for gene knock-outs and complementation. We will build a matrix database of well characterized clinical isolates that records clinical cavity formation, results of assays of toxic lipids and whole genome DNA microarray identification of gene deletion and expression. Analysis of this database will generate specific hypotheses as to which mycobacterial genes are associated with the production of toxic lipids and/or are responsible for pulmonary cavities. These hypotheses will be tested using conventional genetic approaches and by further application of our mouse model. Specifically, we will use three existing NIH funded collections to identify 30 mycobacterial clones that do and 30 that do not cause cavitation. We will grow these isolates as pellicles and assay the amount, distribution, structure and toxicity of TDM. Next, we will use a Bayesian statistical approach to identify genes whose presence or expression is associated with loss of cavity production in humans and/or with the production of toxic TDM. In parallel, we will validate the relevance of our animal model using strains that do or do not produce cavities in humans. Finally, we will use conventional genetic approaches to knock out or insert genes associated with cavity formation and will test the capacity of these modified strains to produce TDM (or other relevant parameters) and induce cavities in our mouse model. If successful, new insights into the pathogenesis of pulmonary cavities may lead to specific interventions to prevent this highly infectious disease manifestation. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: GENETIC SUSCEPTIBILITY TO CHRONIC BERYLLIUM DISEASE Principal Investigator & Institution: Gordon, Terry; Associate Professor; Physiology and Neuroscience; New York University School of Medicine 550 1St Ave New York, Ny 10016 Timing: Fiscal Year 2003; Project Start 10-APR-2003; Project End 31-JAN-2007 Summary: (provided by applicant): This research proposal will examine the role of host genetic factors in the adverse pulmonary effects of inhaled beryllium. Investigators have clearly demonstrated that only a portion of exposed workers develop chronic beryllium disease (CBD). The reason for the variability in response to inhaled beryllium is not known, but strong evidence suggests that inter-individual differences in the molecular coding for immune system proteins play a major role. Using a gene-by-gene research approach, investigators have shown that one or more nucleotide polymorphisms in HLA are significantly associated with the development of CBD. These molecular epidemiology studies have focused on genetic differences in the major histocompatiblity complex, thus ignoring the potential role of other genes to explain the variability in response to beryllium-induced CBD. Moreover, in addition to modifying genes, environmental factors such as dose and speciation of beryllium are believed to play significant roles in the induction of CBD. Thus, an understanding of the gene environment interaction is critical in clarifying the mechanism(s) of host response to beryllium exposure. A predictive genetic animal model of inter-individual variation for CBD will permit the dissection of the factors of genes and environment in CBD.

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Although genetic studies in mice and guinea pigs have previously suggested inter-strain variability in the response to beryllium, a systematic and complete genome wide search for susceptibility genes for CBD has not been conducted. We will directly address this research gap by examining the response of genetically homogeneous, inbred strains of mice exposed to beryllium using a murine model of beryllium-induced lung granulomas. The primary objectives of this project are: 1) to test the hypothesis that there is a genetic basis for the induction of CBD in response to inhalation of beryllium aerosols; 2) to quantify the contribution of genetic versus environmental factors; and 3) to identify candidate genes that play a critical role in the molecular pathways leading to CBD. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: GENETIC VS ENVIRONMENT IN SCLERODERMA OUTCOMES STUDY Principal Investigator & Institution: Reveille, John D.; Professor; University of Texas Hlth Sci Ctr Houston Box 20036 Houston, Tx 77225 Timing: Fiscal Year 2002; Project Start 01-SEP-2002; Project End 31-AUG-2003 Summary: The hypothesis to be tested in this proposal is that systemic sclerosis (SSC) is a more aggressive disease in non-Caucasians who manifest a higher occurrence of critical organ involvement and a worse prognosis, and that reasons for this may include both genetic factor factors as well as sociodemographic or behavioral determinants. To ascertain this we have established a multi-ethic cohort of 175 patients with SSC of relatively recent onset (