FORUM 1996: GENE THERAPY DEVELOPMENT AND EVALUATION OF PHASE I PRODUCTS VECTOR DEVELOPMENT SPONSORED BY CENTER FOR BIOLOGICS EVALUATION AND RESEARCH FOOD AND DRUG ADMINISTRATION and NATIONAL CANCER INSTITUTE NATIONAL INSTITUTES OF HEALTH Editors: Parris R. Burd, Ph.D., Philip D. Noguchi, M.D. and Angus J. Grant, Ph.D. JULY 11-12, 1996 NATCHER CONFERENCE CENTER NIH CAMPUS BETHESDA, MD GENE THERAPY CONFERENCE SCIENTIFIC STEERING COMMITTEE Angus J. Grant , co-chair Parris R. Burd, co-chair Robert Anderson Joy Cavagnaro Stephen Creekmore Jay Elterman Joyce Frey-Vasconcells Andra Miller Philip Noguchi Anne Pilaro Craig Reynolds Sharon Risso George Robertson Carolyn Wilson Published by: The Division of Cellular and Gene Therapies HFM-518 Center for Biologics Evaluation and Research Food and Drug Administration 1401 Rockville Pike Rockville, MD 20852 U.S.A. tel. 301-827-0681 fax. 301-827-0449 e-mail: GTINFO@A1.cber.fda.gov Note from the Organizers: This conference originated from Pre-IND meetings which suggested that some sponsors had misunderstandings of how the FDA went about the business of evaluating gene therapy products. We first started formulating ideas of a training session describing the FDA review process in September of 1995. Coincident with this were discussions with NCI concerning its new biotherapuetic production facility and their interest in producing gene therapy vectors. Shortly thereafter, the National Gene Vector Laboratories (NGVL) program was announced, and suddenly a two day conference emerged out of the mist. We never anticipated that this conference would turn out to be so large, but are delighted nonetheless. Lessons learned from this year's conference will be used to make next year's conference even more useful and productive. OVERVIEW OF THE CONFERENCE AND GOALS Entry of gene therapy products into the clinic requires overcoming many barriers. Foremost among them are funding and regulatory concerns. One of the most useful aspects of gene therapy product development to date has been the free exchange of information common to specific products and product areas. This open environment has been critical for keeping gene therapy regulation in pace with developments in the field. Moreover this information exchange has been instrumental in facilitating public acceptance and support of this rapidly developing area. FDA and NIH share a common goal of promoting the development of useful, safe, and effective therapies for human disease. This conference was designed with two primary goals in mind. The first was to bring together scientists developing gene therapy products with the intention of explaining to them what kinds of information are appropriate and necessary for entry into Phase I clinical trials and what sources of funding the NIH has made available to facilitate early product development and evaluation. The second goal of the conference was to provide a forum for the open exchange of scientific, manufacturing, and clinical experiences as they relate to FDA regulatory requirements. ANNOUNCEMENT OF SECOND ANNUAL GENE THERAPY CONFERENCE This conference is the first of what is hoped to be an annual event. The Second Annual FDA/NIH International Gene Therapy Conference is scheduled for July 15-18, 1997 at the Natcher Conference Center on the NIH campus in Bethesda, MD. Watch the FDA-CBER website for updates: http://www.FDA.GOV/CBER/CBERftp.html Topics for discussion at the conference are solicited. Please send suggestions to our gene therapy conference e-mail account: GTINFO@A1.cber.fda.gov or call the Division of Cellular and Gene Therapies/FDA at 301-827-0681. Watch the CBER web site for updates. TABLE OF CONTENTS OVERVIEW OF THE CONFERENCE AND GOALS July 11, 1996 Development and Evaluation of Phase I Products GENE THERAPY CONFERENCE - INTRODUCTION Kathryn C. Zoon, Ph.D. RAC UPDATE Lana Skirboll, Ph.D. INTRODUCTION TO THE FDA AND NIH ROLES IN GENE THERAPY CLINICAL TRIALS Philip D. Noguchi, M.D. OVERVIEW OF THE FDA IND REVIEW PROCESS Andra Miller, Ph.D. REGULATORY PRINCIPLES FOR SOMATIC CELL AND GENE THERAPY U.S. FDA PERSPECTIVE Dr. Robert W. Anderson ANCILLARY PRODUCTS Joyce L. Frey, Ph.D. PRECLINICAL SAFETY AND ACTIVITY TESTING OF CELLULAR AND GENE THERAPIES Anne M. Pilaro, Ph.D. Joy A. Cavagnaro, Ph.D. CONSIDERATIONS IN CLINICAL TRIAL DESIGN FOR GENE THERAPY PHASE I TRIALS David Wilde, M.D. INTERNATIONAL REGULATORY CONSIDERATIONS, EXPORT AND IMPORT ISSUES Elaine C. Esber, M.D. OVERVIEW OF FUNDING MECHANISMS AVAILABLE FROM THE NIH FOR THE SUPPORT GENE THERAPY PROJECTS Craig Reynolds, Ph. D. TECHNOLOGY TRANSFER (OTD/OTT)FUNDING MECHANISMS FOR NCI SPONSORED GENE THERAPY RESEARCH Diane Bronzert NIAID SUPPORT FOR HUMAN GENE THERAPY Nava Sarver, Ph.D. NATIONAL HEART, LUNG, AND BLOOD INSTITUTE: SUPPORT FOR GENE THERAPY RESEARCH Sonia I. Skarlatos, Ph.D. NIDDK SUPPORT FOR GENE THERAPY RESEARCH Catherine McKeon, Ph.D. THE NATIONAL GENE VECTOR LABORATORIES Richard A. Knazek, M.D., Ph.D. NCI SPONSORED SMALL SCALE MANUFACTURE OF GENE THERAPY PRODUCTS George A. Robertson, Ph.D. CURRENT AND SPECIAL NIH RESOURCES FOR THE DEVELOPMENT OF GENE THERAPY PRODUCTS Elizabeth C. Lovoy, J. D., M. P. H. ELECTRONIC ACTIVITIES AND REGISTRIES UNDER THE FDA SMART PROGRAM Mary A. Buesing M.D. JULY 12 VECTOR DEVELOPMENT PLENARY TALK Dr. Alan E. Smith BREAKOUT SESSION 1: ADENOVIRAL VECTOR DEVELOPMENT SUMMARY Kathleen Hehir, Genzyme Corporation; Parris Burd, FDA/CBER; Robert Anderson/FDA/CBER CONSIDERATIONS FOR THE DETERMINATION OF ADENOVIRUS VECTOR PARTICLE CONCENTRATION AND INFECTIVITY Bruce C. Trapnell, M. D. RAPID IDENTIFICATION OF ENCAPSIDATED VIRAL DNA AND ASSESSMENT OF VIRAL INFECTIVITY Paul Shabram NEW. MORE SENSITIVE METHOD TO ASSESS ADENOVIRAL VECTOR INFECTIVITY Beth Hutchins, Ph.D. VIRAL SAFETY OF BIOTECHNOLOGY AND GENE THERAPY PRODUCTS+STATISTICAL REVIEW OF RCA TEST METHODS John Spaltro, Ph.D. MANUFACTURING AND REGULATORY ISSUES FOR DEVELOPING ADENOVIRUS VECTORS AS A PRODUCT Dominic Vacante, Ph.D. SMALL SCALE PRODUCTION OF ADENOVIRUS VECTORS INTENDED FOR PHASE I CLINICAL STUDIES Cassandra Nyberg and Estuardo Aguilar-Cordova DEVELOPMENT OF ADENOVIRAL VECTORS FOR CLINICAL TRIALS AT THE UNIVERSITY OF PENNSYLVANIA James M. Wilson, M.D., Ph.D. SINGLE SPONSOR DEVELOPMENT OF ADENOVIRUS VECTORS FOR PRODUCT LICENSURE Kathy Hehir BREAKOUT SESSION 2: ANCILLARY PRODUCTS SUMMARY Joyce L. Frey-Vasconcells, FDA/CBER INTRODUCTION TO THE BREAKOUT SESSION ON ANCILLARY PRODUCTS Joyce L. Frey, Ph.D. A PLAN FOR CERTIFICATION OF GROWTH FACTORS FOR USE IN EX VIVO GENE THERAPY Joyce D. Francis, Ph.D. AN ANCILLARY PRODUCT MANUFACTURER'S ISSUES AND RECOMMENDATIONS Keith D. Gittermann REGULATORY FRAMEWORK FOR ENSURING QUALITY OF PRODUCTS AND POTENTIAL LABELING Mark W. Gauthier LICENSURE OF CYTOKINES FOR EX VIVO USE Mark Hukkelhoven, Ph. D. BREAKOUT SESSION 3: FACILITIES AND MANUFACTURING "THE GMP CONTINUUM" SUMMARY Jay Eltermann, R.Ph., M.S. REGULATORY AND CGMP CONSIDERATIONS FOR GENE THERAPY FACILITIES Jay Eltermann, R.Ph., M.S. FACILITY CONSIDERATIONS Robert Sausville OPERATIONAL ASPECTS Mary Malarkey GENE THERAPY INITIAL MANUFACTURING FACILITY William R. Tolbert, Ph.D. TRANSITION FROM GLP TO GMP: AN ACADEMIC PERSPECTIVE Blake Roessler, M.D. CONSIDERATIONS AND STRATEGIES IN THE PLANNING AND DESIGN OF A NEW FACILITY Victor Santamarina, Ph.D. BREAKOUT SESSION 4: GETTING STARTED IN GENE THERAPY VECTOR DEVELOPMENT SUMMARY Angus J. Grant, Ph.D. FDA/CBER STARTING A CLINICAL GENE THERAPY PROGRAM: PLANNING AND IMPLEMENTATION Bruce Merchant, M.D., Ph.D. KEY ISSUES AND FACTORS IN DEVELOPING THE GENE THERAPY IND Gary E. Gamerman, MS, JD CONTRACT SERVICES SUPPORTING GENE THERAPY DEVELOPMENT AND CLINICAL TRIALS Jeffrey M. Ostrove, Ph.D. IMPLEMENTATION OF A PHASE I GENE THERAPY PROGRAM IN A GLP/GMP CONTINUUM James A. Taylor, Ph.D. MONOCLONAL ANTIBODY AND RECOMBINANT PROTEIN PRODUCTION FACILITY NATIONAL CANCER INSTITUTE-FREDERICK CANCER RESEARCH & DEVELOPMENT CENTER Stephen Creekmore, M.D., Ph.D. GETTING STARTED IN GENE THERAPY VECTOR DEVELOPMENT - CONSIDERATIONS FOR PROCESS DEVELOPMENT AND MANUFACTURING Shawn L. Gallagher, MAGENTA Corp., Rockville, MD BREAKOUT SESSION 5: DEVELOPMENT OF NEW VECTOR SYSTEMS SUMMARY Neal DeLuca, University of Pittsburgh; Robert Anderson, FDA/CBER; Parris Burd, FDA/CBER HERPES SIMPLEX VIRUS MUTANTS AS GENE THERAPY VEHICLES Neal DeLuca DISC-HSV A NOVEL VECTOR SYSTEM Dr. Elizabeth A. Rollinson et al MODIFYING NEURONAL PHYSIOLOGY WITH A HELPER VIRUS-FREE HSV-1 PLASMID VECTOR SYSTEM Alfred I. Geller REPLICATION COMPETENT ATTENUATED HERPES VIRUS FOR TUMOR THERAPY Martuza RL, Mineta T, Hunter W, Todo T, Todo M, Rabkin S. AAV VECTORS IN VIVO R. Jude Samulski, Ph.D. ADENO-ASSOCIATED VIRUS VECTORS FOR TRANSDUCTION OF HEMATOPOIETIC PROGENITOR CELLS Saswati Chatterjee, Elizabeth Shaughnessy, Grace Fisher-Adams, Di Lu, Greg Podsakoff, and K. K. Wong DEVELOPMENT OF AAV VECTORS FOR GENE THERAPY Samuel C. Wadsworth AAV VECTORS CAN BE EFFICIENTLY PRODUCED WITHOUT HELPER VIRUS P. Colosi, S. Elliger, C. Elliger and G. Kurtzman DEVELOPMENT OF ASSAYS FOR THE DETECTION AND QUANTITATION OF RECOMBINANT AAV-CFTR VECTOR Daryn Debeiak, Molly Fee-Maki, Teresa Johnston, Tom Reynolds BREAKOUT SESSION 6: RETROVIRAL VECTORS SUMMARY Estuardo Aguilar-Cordova, Ph.D., Baylor College of Medicine; Andra Miller, Ph.D., FDA/CBER; Carolyn A. Wilson, Ph.D., FDA/CBER TESTING FOR REPLICATION-COMPETENT RETROVIRUS Tie-Hua Ng1, Carolyn Wilson2, Andra Miller3 and Peter Lachenbruch1 EXPERIMENTAL ISSUES IN TESTING FOR REPLICATION-COMPETENT RETROVIRUS Carolyn A. Wilson*, Katie Whartenby¡, and Andra Miller¡ VIRAL SAFETY OF BIOTECHNOLOGY AND GENE THERAPY PRODUCTS-PROPOSED CONVENTIONS FOR SAMPLING AND TESTING FOR RCR John Spaltro, Ph.D., SENSITIVE ASSAYS FOR DETECTION OF REPLICATION COMPETENT RETROVIRUSES Joseph Hughes, Lisa M. Rudderow, Teresa M. Byrne, Kim Bowers, Dilip Patel, Michael Burnham, Ingo Georgoff, Richard Metz, Fredrika McDevitt, and Michael D. Lubeck REPLICATION COMPETENT RETROVIRUS TESTING FOR MANUFACTURING AND MONITORING OF PATIENTS IN CLINICAL TRIALS L. Cohen and J. Rokovich A NOVEL MURINE RETROVIRUS IDENTIFIED DURING TESTING FOR HELPER VIRUS IN HUMAN GENE TRANSFER TRIALS A. Dusty Miller 1,2, Lynn Bonham1, John Alfano1, Hans-Peter Kiem1, Tom Reynolds3, and Greg Wolgamot1,2,4 INFECTION OF NORMAL RHESUS MONKEYS WITH A MURINE REPLICATION-COMPETENT RETROVIRUS . Arifa S. Khan1, Muhammad Shahabuddin1, Teresa A. Galvin1, Jeffrey Ostrove2, Exeen M. Morgan2, and Janet Hartley3 MONITORING HUMAN CLINICAL TRIAL SUBJECTS RECEIVING DIRECT ADMINISTRATION OF A RECOMBINANT RETROVIRAL VECTOR FOR THE PRESENCE OF REPLICATION COMPETENT RETROVIRUS (RCR) AND FOR THE INADVERTENT TRANSDUCTION OF GERMLINE CELLS: METHODS, RESULTS, AND IMPACT ON TESTING FREQUENCY Dale G. Ando and Chiron HIV Immunotherapy Team MONITORING OF CANCER PATIENTS RECEIVING RETROVIRAL VECTOR GENE THERAPY Gerard J. McGarrity, Ph.D. BREAKOUT SESSION 7: PHARMACOLOGY & TOXICOLOGY SUMMARY Anne M. Pilaro and Joy Cavagnaro PHARMACOLOGY/TOXICOLOGY STUDIES FOLLOWING INTRAPERITONEAL INFUSION OF RETROVIRAL VECTORS. J.T. Holt, D. Tait, P. Obermiller, S. Redlin-Frasier, R.A. Jensen, C.L. Arteaga, and M.C. King TOXICOLOGICAL STUDIES OF ADENOVIRUS MEDIATED GENE TRANSFER VIA A) INTRACEREBRAL AND INTRAPROSTATIC INJECTION IN NON HUMAN PRIMATES AND COTTON RATS, AND B) SYSTEMATIC DELIVERY INTO NONHUMAN PRIMATES, COTTON RATS, AND MICE B. Lee, T.L. Timme, H.D. Shine, W. O'Neal, N. Carey, G.B. Hubbard, D.A. Carrier, C. Nyberg, R. Barrios, S. Rajagopaian, P.R. Wyde, A. Rojas-Martinez, C.A. Montgomery, S.L.C. Woo, A. Beaudet, T.C. Thompson, and E. Aguilar-Cordova DISTRIBUTION AND RESCUE STUDIES OF AN AAV-CFTR VECTOR IN RHESUS MACAQUES S.A. Afionel, C.K. Conrad1, W.G. Kearns3, S. Chunduru1, R. Adams4, T.C. Reynolds5, W.B. Guggino2, G.R. Cutting3, B.J. Carter5, and T.R. Flotte1 IN VIVO ADENOVIRUS VECTOR PREDICTABILITY OF SAFETY STUDIES IN EXPERIMENTAL ANIMALS Ronald G. Crystal, MD Overview of Regulatory Documents Available by CBER FAX BACK DOCUMENTS AVAILABLE FROM THE CBER Who Do You Call to... July 11, 1996 Development and Evaluation of Phase I Products Goals: To acquaint investigators with the necessary information required by CBER for evaluation of products intended for phase I clinical trials and the resources currently available to investigators through the NIH. 8:00 Welcome - Kathryn C. Zoon, Ph.D. - Director, CBER 8:05 Opening Remarks - Jay P. Siegel, M.D. - Director, Office of Therapeutics, CBER/FDA 8:10 RAC Update - Lana R. Skirboll, Ph.D. - Associate Director for Science Policy, NIH CHAIR FOR THE MORNING SESSION: PHILIP NOGUCHI, M.D., FDA 8:20 Introduction - Philip Noguchi, M.D. - Director, Division of Cellular and Gene Therapies, CBER/FDA 8:30 Overview of the FDA IND review process. Andra E. Miller, Ph.D., CBER 9:00 Recommendations for products intended for Phase I clinical trials. Robert Anderson, Ph.D., CBER 9:30 Specific considerations for the manufacture of gene therapy products. Ancillary products, non-licensed products used in the manufacture of a therapeutic. Joyce L. Frey-Vasconcells, Ph.D., CBER 10:00 Break 10:30 The role of pre-clinical studies in support of phase I clinical trials: Proof of concept and safety studies. Anne M. Pilaro, Ph.D., CBER 11:00 Considerations in clinical trial design for gene therapy phase I trials. David. Wilde, M.D., CBER 11:30 International regulatory considerations, export and import issues. Elaine C. Esber, M.D., CBER CHAIR FOR THE AFTERNOON SESSION, CRAIG REYNOLDS, Ph.D., NCI 1:30 Overview of NIH resources available for gene therapy product development. Craig Reynolds, Ph.D., NCI 1:50 National Cancer Institute, Diane Bronzert 2:00 National Institute of Allergy and Infectious Diseases, Nava Sarver, M.D. 2:10 National Heart, Lung, Blood Institute, Sonia Skarlatos, Ph.D. 2:20 National Institute of Diabetes and Digestive and Kidney Diseases, Catherine McKeon, Ph.D. 2:30 Discussion, Break 3:00 National Center for Research Resources - National Gene Vector Laboratories, Richard Knazek, M.D. 3:30 NCI-sponsored small scale manufacture of gene therapy products, George A. Robertson, Ph.D. 4:15 NCI-Office of Technology Development, Elizabeth C. Lovoy, J.D., M.P.H. 4:40 Electronic Activities & Registries under the FDA SMART Program, Mary A. Buesing, M.D., CBER 5:00 Closing Remarks GENE THERAPY CONFERENCE - INTRODUCTION Kathryn C. Zoon, Ph.D. Director Center for Biologics Evaluation and Research Food & Drug Administration There has been tremendous growth in investigational new drug (IND) applications for gene therapy. One IND was submitted in 1989 whereas in 1996, 41 were submitted. The current number of gene therapy INDs through June 30, 1996 is 142. THREE PRINCIPLES UNDERLIE THIS EMERGING FIELD: Sound science is the foundation for decision making. The development and regulation of gene therapy products are collaborative and iterative processes among government, industry, public, and academia. Development of gene therapy policy and regulation needs to be a public process which allows ethical, social, scientific, and other issues to be debated and discussed in an open and public forum. THREE CURRENT AREAS OF CONCERN IN GENE THERAPY Replication competent retrovirus (RCR) testing Original policy was implemented because of Nienhuis (NIH) experiment in monkeys Now that sponsors are increasing the size of production lots, this testing policy is becoming logistically burdensome. Solution: Data should be (and has been) publicly discussed; Statistical models developed with preliminary testing validation; Standards should be made available and Further testing and results should be shared. Source and supply of "clinical" grade vector With the growth in gene therapy products, the supply of vector suitable for clinical use is becoming a limiting factor. Solutions: The NIH National Gene Vector Laboratories (NGVL), with FDA as a liaison, are providing financial support for the production and distribution of vectors; FDA meets with corporate and academic sponsors to help plan small scale production facilities; NIH/NCI opens a GMP production facility that includes vectors (with the assistance of FDA scientists). Education of new investigators: As the field expands, issues of how to effectively help new investigators through the IND process arise. Solutions: FDA/NIH sponsored workshops on the nuts and bolts of gene therapy regulation; The RAC process allows publication of clinical protocols; Increased emphasis by FDA on guidance documents & their availability (FDA-CBER & NIH Guidance documents are available on the World Wide Web); FDA will be implementing electronic IND submissions. FUTURE ISSUES AND NEW CHALLENGES IN THE FIELD Development of new vector systems These will require novel safety testing & pharmacology/toxicology studies. There is a desire/need to coordinate studies to reduce unnecessary duplication of testing, and to be able to collect and correlate data from multiple sponsors for a given product area. Some vectors present new challenges such as herpes and lentivirus vectors. A major challenge will be how to deal collectively with the issues prospectively, rather than on a case by case basis. NEW APPROACHES AND NEW INDICATIONS FOR GENE THERAPY In utero gene therapy. Germ line gene therapy. Enhancement gene therapy. Ethical and societal issues need to be discussed openly and decisions made. Establish a forum for public discussion to help guide FDA decisions. License approval of gene therapy products Develop a scientifically sound approach to regulate these novel products. Gene therapies are complex products and often use multiple facilities to support their production. RAC UPDATE Lana Skirboll, Ph.D. Associate Director for Science Policy, NIH ENHANCED NIH OVERSIGHT OF GENE THERAPY ACTIVITIES Notice of Intent To Propose Amendments to the NIH Guidelines Regarding Enhanced Mechanisms for NIH Oversight of Recombinant DNA Activities Federal Register, July 8, 1996 (61 FR 35774) STRUCTURAL COMPONENTS OF NIH PROPOSAL ORDA ADVISORY COMMITTEE (OAC) Public forum Chartered advisory committee Notice of scheduled meetings will published in the Federal Register and convened 4 times/year Standing membership of 6 to 10 individuals representing the scientific, legal, ethical, and public advocacy communities. Maintain public accountability for gene therapy research In-depth gene therapy data analysis Through OAC, the NIH Director will continue to promulgate necessary amendments to the NIH Guidelines Facilitate timely public reporting of significant clinical events Identify and prioritize proposed GTPC topics GENE THERAPY POLICY CONFERENCES (GTPC) Public forum Numerous participants who possess significant scientific, ethical, and legal expertise and/or interest that is directly applicable to a specific recombinant DNA research issue. Notice of scheduled meetings will be published in the Federal Register and convened 4 times/year Discussion topics may include: (1) basic research on novel gene delivery vehicles, (2) novel applications of gene transfer, and (3) relevant ethical and societal implications of a particular application of gene transfer technology Discussion topics do not preclude the use of novel protocols as a focus for conference discussion Topics will be identified through OAC recommendations, interagency communications, ORDA, the scientific community, patient advocates, ethicists, public health professionals, and the public Where appropriate, findings and recommendations will be submitted to the NIH Director for dissemination to DHHS components and the public OFFICE OF RECOMBINANT DNA ACTIVITIES (ORDA) Continue to provide administrative support relevant to recombinant DNA activities Enhance public awareness of human gene therapy research through data management activities Identify, coordinate, and disseminate relevant information as novel technologies emerge Provide administrative support to both OAC and GTPC Optimize interagency communication and collaborative efforts to streamline procedures and facilitate the flow of relevant information FUNCTIONAL ANALYSIS OF NIH PROPOSAL RAC OAC GTPC Public Forum Yes Yes Yes Standing Members Yes Yes No Chartered Yes Yes No Broad Membership Yes Yes Yes - But ad hoc Representing Scientific, expertise as Ethical, Legal, and Public required by Health Concerns discussion topic Terms 4 3-4 years No terms years Number of Meetings per Year 3-4 Policy Recommendations Yes Yes Yes Protocol Review and Yes *No *No Approval/Disapproval Minutes Yes Yes Yes Data Management and Adverse Yes Yes Yes Event Reporting Review Individual Protocol Yes No - But may No - But can provide Informed Consent Documents propose Informed forum for public consent issues discussion of for GTPC informed consent issues *Does not preclude a specific protocol used as topic for OAC or GTPC discussion. RECOMMENDATIONS OF THE RAC AD HOC REVIEW COMMITTEE (VERMA COMMITTEE) The RAC should no longer carry out case-by-case review of every clinical gene transfer protocol. Review of protocols by the RAC in an open public forum should continue in several areas of concern in which a particular protocol or new technology represents a significant degree of departure from familiar practices. The RAC should define the criteria and work out procedures for identifying specific protocols requiring public review. The RAC should continue to provide advice on policy matters revolving around gene therapy and other recombinant DNA issues to the NIH Director, individual members of the research community, institutional review boards, and the public. This critical function should be extended, enabling RAC explicitly to provide advice and recommendations on policy matters to the Food and Drug Administration (FDA). Hence, the committee recommends that the NIH Director urge the Commissioner of the FDA to exempt the broad area of gene therapy from many of the proprietary restraints reserved for ordinary therapeutic drug products and biologics that come under FDA review. NIH IMPLEMENTATION RAC is eliminated and Consolidated Review will not be maintained GTPC preserves such a forum and provides for more in-depth discussion of both the science and ethical issues related to a specific gene therapy issue Not applicable to the proposed new structure, since this proposal cedes review and approval to the FDA Fully met by GTPC. Each GTPC will focus on a single issue; therefore, policy advice will be substantially augmented under this new mechanism Fully responds to the recommendation regarding the continued need for data monitoring and adverse event reporting INTRODUCTION TO THE FDA AND NIH ROLES IN GENE THERAPY CLINICAL TRIALS Philip D. Noguchi, M.D. Director, Division of Cellular and Gene Therapies OTRR, FDA This morning's session will be devoted to those who would like to know how to conduct a gene therapy trial. Human gene therapy represents the publicly visible portion of a rapidly evolving medical revolution. Gene therapies could not be contemplated without the ready availability of purified cytokines to allow expansion and differentiation of cells. Likewise, monoclonal antibody technology has allowed the purification of a variety of cells from a number of tissues that can then be transduced with vectors. The investment by NIH in basic biomedical research studies has created the ability to isolate medically relevant genes and to create the vectors used in gene therapies. The Food and Drug Administration (FDA) derives its authority from the Food, Drug and Cosmetic Act (revised) (FD&C Act) and Section 351 of the Public Health Service Act (PHS Act). Both Acts allow regulations to be promulgated that require federal oversight over clinical trials using experimental products. The authority used for the regulation of gene therapy had its origin in 1902 as the Biologics Control Act, which mandated that things biologic, such as viruses and serums, could not be shipped, sold or bartered unless it was prepared in a licensed and inspected establishment. In 1986, FDA announced in a policy statement that gene therapies would be considered biological products subject to licensure by FDA. The Center for Biologics Evaluation and Research (CBER) of the FDA is currently responsible for the regulation of the manufacturing and labeling of biologic products. Investigational clinical trials in humans with gene therapy products are subject to the general requirements for drugs and biologics in Title 21 of the Code of Federal Regulations (CFR). This includes 21 CFR Part 312, the Investigational New Drug Application (IND) which has a number of requirements, including extensive documentation of methods of production and preclinical testing. In general, as products move from phase 1 through phase II and phase III, other portions of the 21 series pertain, including the 200 series on Current Good Manufacturing Practices (GMPS), the 300 series on new drug applications, the 600 series on biologics establishments and licensing. This level of regulation is the same for all biological products that are considered to be drugs, and focus on safety, purity, potency and efficacy considerations. In 1975, the Recombinant DNA Advisory Committee (RAC) was formed to create guidelines that would allow recombinant DNA experiments to be conducted in laboratories under safe conditions. Over the past two decades, the RAC has evolved its guidelines and oversight to where only clinical protocols involving gene transfer are reviewed in public. FDA and NIH have interacted on an increasingly frequent basis during the past four years. The RAC meetings allow for public discussion of both accomplishments and adverse findings associated with gene therapy. Because these meetings are public, the discussions of safety concerns can be immediately communicated to the entire industry and a consensus to resolve the concerns can be reached with industry that would otherwise be restricted by the trade-secret regulations. An instructive example occurred in June, 1993. The RAC and FDA received notice of an adverse reaction in the third patient to participate in a gene therapy protocol for cystic fibrosis using an adenoviral vector. Because of the public nature of gene therapy protocols, the RAC and FDA were able to communicate the details of this adverse event to other investigators. This allowed for appropriate modification of the protocol that allowed not only the original trial to continue with close FDA oversight, but also allowed four other protocols to be approved with modified dosing. OVERVIEW OF THE FDA IND REVIEW PROCESS Andra Miller, Ph.D. Division of Application Review and Policy CBER, FDA OVERVIEW INVESTIGATIONAL NEW DRUG (IND) APPLICATION Regulatory Approach Pre-IND stage When to file Content of the IND submission Review and notification processes Types of submissions to the IND SOMATIC CELL THERAPY DEFINITION The prevention, treatment, cure, diagnosis or mitigation of disease or injuries in humans by the administration of autologous, allogeneic or xenogeneic cells that have been manipulated or altered ex vivo. Includes cells that have been: Propagated Expanded Selected Pharmacologically treated Altered in biological characteristics GENE THERAPY DEFINITION A medical intervention based on modification of the genetic material of living cells. Cells may be modified ex vivo for subsequent administration or altered in vivo by gene therapy products given directly to the subject. Ex vivo manipulation of cells is also a type of somatic cell therapy. REGULATORY AUTHORITY Federal Register Notice of Oct. 14, 1993 (Application of Current Statutory Authorities to Human Somatic Cell Therapy Products and Gene Therapy Products) Compliance with part 312 of the Code of Federal Regulations at the investigational stage. Clinical Trials conducted under IND Subject to establishment and product licensure PHS Act Points to Consider documents REGULATORY STRATEGIES TRADITIONAL APPROACH Evaluate and identify safety concerns Quality Control Sound Scientific Principles Case by Case approach NEW APPROACH: PUBLIC PROCESS PHASES OF INVESTIGATION (21 CFR 312.21) Phase I Investigational Studies Designed to evaluate safety and side effects Phase II Investigational Studies Designed to evaluate efficacy and dose ranging Phase III Investigational Studies Expanded study, additional information on efficacy and safety FDA'S OBJECTIVES To assure safety and rights of subjects in all phases of investigation To assure in phase II and III, that scientific quality of the investigations will support licensure Safeguard the public health while promoting development of new products BEFORE FILING AN IND Sponsor may request a pre-IND meeting or teleconference in writing Request should include: Description of the product Proposed development plan Outline of specific issues to be addressed Agenda/attendees list Written requests submitted to Chief of Cytokine and Gene Therapy Branch, DARP: Dr. Joyce Frey Acting Chief Cytokine and Gene Therapy Branch Division of Application Review and Policy OTRR/CBER HFM-591 1401 Rockville Pike Rockville, MD 20852 301-594-0830 PRE-IND MEETING REQUEST IS EVALUATED AND MEETING DATE SCHEDULED Meeting package due 2 weeks before meeting, including: Preclinical data Manufacturing summary Clinical protocol Issues to be discussed PRE-IND MEETING The sponsor should present: Product manufacturing scheme Data regarding the product characterization Proposed specifications for lot release data or actual lot release data Pre-clinical in vitro and in vivo data identifying the product's activity, efficacy and toxicity Data supporting the product's clinical use Proposed phase 1 clinical protocol IND SUBMISSION Sponsor must generate enough preclinical data to: Ensure the safety of clinical trials Determine the parameters for human studies Establish the scientific rationale Request CBER IND Packet: Office of Communication, Training and Manufacturers Assistance (OCTMA) HFM-40 301-827-1800 Submit INDs to: Director, DARP HFM-99 1401 Rockville Pike Rockville, MD 20852-1448 FDA/CBER/OTRR CONTENT AND FORMAT OF THE IND (21 CFR 312.23) Cover Sheet - Form FDA-1571 Provides basic information to FDA Identifies sponsor and investigational drug Identifies phase of clinical investigation Identifies parties responsible for monitoring conduct of the trial Table of contents Introductory Statement and General Investigational Plan Description of the drug Summary of previous human experience Overall plan for investigating the drug Investigator's Brochure Required if product is supplied to clinical investigators other than the sponsor Description of the drug Summary of pharm/tox effects Pharmacokinetic and biological disposition Summary of safety and effectiveness in humans from prior studies Description of possible risks and precautions Protocols Protocol for each planned study including: Statement of objectives and purpose of study Inclusion & exclusion criteria Doses to be administered Description of clinical measurements to be used to monitor effects Investigator data (Form FDA 1572) Chemistry, Manufacturing and Control Information Description of composition, manufacture and control of the investigational product Physical, chemical or biological characteristics Method of manufacture Analytical methods used Stability data/ Initial specifications Description of placebo Labeling Pharmacology and Toxicology Information Adequate information to support the safe conduct of the proposed clinical study IRB approved Consent Form Previous Human Experience with the Investigational Drug Additional Information MASTER FILE SUBMISSION (VS IND) CFR 314.420) Alternative mechanism for submission of product and manufacturing information Does not include clinical protocol Permits holder to incorporate the information by reference when submitting an IND or To authorize other persons to rely on the information, without disclosure Five types I-V Type II most useful for biological product development MASTER FILE SUBMISSION Often contains: Product manufacturing and purification schemes Lot release protocols Tissue culture media Other proprietary information needed to support IND application Example: retroviral vector supernatants FDA staff access to MF via cross reference letter Filed to MF and the new IND CBER staff review and comment on contents of MF Neither approved or disapproved IND REVIEW PROCESS Upon receipt of IND or MF, FDA issues: Acknowledgment letter IND or MF number IND application reviewed within 30 days Review Team: Product reviewer Pharmacology/Toxicology reviewer Clinical reviewer Consult reviewer, as needed Emphasis of review is on data to support: Manufacturing and quality control issues Product characterization Rationale Sound Scientific Principles Pre-clinical studies Product development Clinical protocol Communication of Review Decision Within 30 days (proceed or hold) Clinical Hold Unreasonable risk to the patient Insufficient information provided to assess risk Inadequate Investigator's Brochure Clinical investigator not qualified Letter issued detailing hold issues, comments and requests In order to proceed with clinical study: Correct deficiencies Submit additional information as amendment to IND Receive notification to proceed by FDA SUBMISSIONS TO THE IND Amendments (21 CFR 312.30) Protocol New protocol Change in protocol New investigator Information New toxicology , product or other technical information Discontinuance of a clinical study IND Safety Reports Written reports of ADRs within 10 days Telephone reports of fatalities within 3 days Annual Reports Within 60 days of IND anniversary Status of studies in progress Summary information General investigational plan for next year REGULATORY PRINCIPLES FOR SOMATIC CELL AND GENE THERAPY U.S. FDA PERSPECTIVE Dr. Robert W. Anderson Cytokine and Gene Therapy Branch Division of Application Review and Policy Center for Biologics Evaluation and Research Food and Drug Administration REGULATORY CONCERNS COMMON TO ALL BIOLOGICALS Safety, identity, purity, potency Regulation of both the final product and the manufacturing, process Reproducibility/consistency of product lots CONCERNS RELEVANT FOR SOMATIC CELL AND GENE THERAPY PRODUCTS Product issues Characterization of cells/cell lines Adventitious agent testing Replication competent virus Pre-clinical/clinical issues Toxicity secondary to exogenous gene expression Insertional mutagenesis Effects on germ cells In vivo recombination and pseudotyping CONTROL OF PRODUCTION PROCESS Vector development Cell and virus bank establishment and characterization Final product characterization and lot release Products used in manufacture (Ancillary Products) VECTOR DEVELOPMENT AND CHARACTERIZATION Derivation of vector Source, modifications and function of component parts Description of regulatory elements Molecular characterization of the final vector All components of the vector required for its biological function should be verified by molecular methods of analysis Sequence analysis may be limited to the insert, flanking regions and any regions of the vector which are modified is acceptable for early phases of clinical development MASTER VIRUS BANK CHARACTERIZATION History-generated from molecularly cloned and characterized constructs Identity Genetic stability and integrity Potency Expression of gene product Biological activity Safety-freedom from adventitious agents Bacteria Fungi Mycoplasma Virus, including RCV (Replication Competent Virus) VECTOR MODIFICATIONS Previously Change in vector = New Product = New IND Presently Related vectors + New Product + New IND Minor Modifications IND amendment Abbreviated testing-focusing on specific safety concerns Case by case MASTER CELL BANK CHARACTERIZATION History Culture and storage conditions Identity testing Genetic stability and integrity Protein product Cellular Morphology/Phenotype Cellular Isoenzymes Safety testing-freedom from adventitious agents Bacteria Fungi Mycoplasma Virus, including RCV WORKING CELL BANK CHARACTERIZATION Freedom from adventitious agents Limited testing for RCV Limited routine identity testing Validation that aliquots can consistently be used for final product production VECTOR-CONTAINING SUPERNATANTS Purity Residual cellular DNA, RNA, protein Non-infectious virus (particle/PFU [IU]) Production materials Endotoxin Potency Biological activity Rate and regulation of gene expression Identity Genetic stability and integrity Safety Sterility (bacterial and fungal) Mycoplasma General Safety Tests for Adventitious Virus (vector system dependent) EX VIVO TRANSDUCED CELLS Description of cell source, isolation and culture conditions Purity Viability Endotoxin Potency Biological activity Rate and regulation of gene expression Identity Patient's identity label Phenotypic and genetic markers Safety Sterility (bacterial and fungal) Mycoplasma General Safety Tests for Production Organisms PRODUCT-SPECIFIC SAFETY CONSIDERATIONS Retroviral vectors Primarily ex vivo gene transfer Adenoviral vectors Primarily in vivo gene transfer Plasmid vectors Both ex vivo and in vivo Often in the presence of liposomes RETROVIRAL VECTOR SAFETY CONSIDERATIONS Replication competent retrovirus (RCR) Can arise from recombination events in retroviral vector packaging cell lines used during production Presence of RCR is a safety concern: Retroviruses integrate into the genome Murine retroviruses can pseudotype HIV, resulting in expanded host range Immunosuppressed monkeys exposed to RCR develop lymphomas within 200 days RCR TESTING Testing performed during production and for each lot: Analysis of cells and supernatant Master Cell Bank Cells (1% or 108) and supernatant (5%) Production Lot Ex vivo transduced cells (preferable to have results before patient administration) Analysis of cells or supernatant Working Cell Bank RCR ASSAY DESIGN STEP 1: Amplification Culture cells or supernatant with permissive cell line, e.g., Mus dunni cells for 18-20 days STEP 2: Viral detection PG4 S+L- focus forming assay Validated alternative assays are acceptable (marker rescue) PATIENT MONITORING Perform periodic monitoring for evidence of RCR infection Assays Serological assays for evidence of antibody to retroviral envelope protein Direct assays for viral nucleic acid in peripheral blood leukocytes using PCR Assays for reverse transcriptase Frequency Monthly during treatment Monthly for the first three months following completion of treatment Every three months for the remainder of the year following treatment Yearly thereafter Identification of RCR via direct culture of patient PBL should be attempted Submit written IND safety report Submit results of monitoring in annual progress report ADENOVIRAL VECTOR SAFETY CONSIDERATIONS Administration of high doses of adenoviral vector to humans can result in both an acute viral toxicity and a vigorous immune response resulting in inflammation. It has been suggested that viral toxicity may be due to induction of a cytokine cascade. Animal testing of adenoviral vectors has shown that the immune response is CD8+ T cell mediated (CTL response). For some applications, target cells may have a rapid turnover requiring repeat administrations, resulting in potentially greater immune response each time. ADENOVIRAL VECTOR SAFETY CONSIDERATIONS Patient dose based on particle to infectious unit ratio Specification < 100:1 particle: PFU (IU) Detection of replication competent adenovirus Specification < 1 RCA/ patient dose Demonstration of prior immunity to adenoviruses Consent form should reflect risk of adenovirus infection Shedding of RCA as well as vector should be determined If RCA present, culture and characterize Risks of adenoviral vector administration may differ for different populations (e.g. cystic fibrosis vs cancer patients) PLASMID VECTOR SAFETY CONSIDERATIONS Removal of potential contaminants: Bacterial RNA, Protein and DNA Avoid use of CsCl and EtBr Liposome preparations Residual solvents used in production Potential toxicity of liposome preparation PRODUCT MANUFACTURE AND CHARACTERIZATION AN EVOLVING PROCESS The method of product manufacture should be appropriate for the investigational phase of the study cGMP continuum Consult DEL (301-827-3031) concerning facilities issues The degree of product characterization should be appropriate for the investigational phase of the study Safety, identity, purity and potency Phase I Safety (Product Specific) Consult Product Reviewer COMMENTS AND QUESTIONS Robert W. Anderson, Ph.D. Cytokine and Gene Therapy Branch, Division of Application Review and Policy Center for Biologics Evaluation and Research (HFM-591) Food and Drug Administration 1401 Rockville Pike Rockville, MD 20852-1448 594-0525 (FAX) ANCILLARY PRODUCTS Joyce L. Frey, Ph.D. Acting Chief, Cytokine and Gene Therapy Branch Division of Application Review and Policy FDA/CBER/OTRR ANCILLARY PRODUCTS Definition Regulatory Framework FDA's Concerns and Recommendations Qualification Program DEFINITION Components used during manufacturing that should not be present in the final product Examples Growth Factors Cytokines Monoclonal Antibodies Cell Separation Devices Media and Media Components REGULATORY FRAMEWORK As critical components with defined set of specifications As a drug or biologic As a device REGULATORY FRAMEWORK: FDA'S CONCERN Ancillary products can affect Safety Potency Purity of the final therapeutic product FDA RECOMMENDATIONS Clinical Grade Material Licensed Product Establish qualification programs and set Specifications Adequate characterization Adherence to GMPs QUALIFICATION PROGRAM What is an adequate qualification program? Questions to consider What's the ancillary product? animal derived ? purified protein ? cell supernatant ? What's the process used to produce the ancillary product? GENERIC QUALIFICATION PROGRAM Safety testing adventitious agents sterility endotoxin mycoplasma Activity (functional) Purity (consistency) Verification of residual levels in the final therapeutic product ANCILLARY PRODUCTS: PROBLEM Ancillary Product: Growth Factor called "ZIP" Problem Clinical grade form of ZIP is no longer available FDA recommendation ? CERTIFICATION OF ANCILLARY PRODUCT - ZIP sterility adventitious agents activity (functional) purity (consistency) endotoxin MECHANISM FOR CERTIFICATION Obtain a Certificate of Analysis from the manufacturer Sponsor can initiate testing Manufacturer can supply information in a Masterfile which would be cross referenced Combination of the above POTENTIAL SOLUTION TO THE PROBLEM Step 1: Set minimum specifications Step 2: Identify suppliers Step 3: Purchase samples Step 4: Verify specifications and initiate additional tests Step 5: Purchase quantities for the trial Step 6: If revial ancillary product, the sponsor should certify the sterility, activity, and purity SOURCES OF INFORMATION Federal Register, Application of Current Statutory Authorities to Human Somatic Cell Therapy Products and Gene Therapy Products, October 14, 1993, Vol 58, No. 197, page 53248-53251 Joyce L. Frey, Cytokine and Gene Therapy Branch, Division of Application Review and Policy, (301)594-0830 PRECLINICAL SAFETY AND ACTIVITY TESTING OF CELLULAR AND GENE THERAPIES Anne M. Pilaro, Ph.D. Joy A. Cavagnaro, Ph.D. FDA/CBER DEFINITION OF GENE THERAPY Introduction into the human body of genes or cells containing genes foreign to the body for the purposes of prevention, treatment, diagnosis or curing disease CANDIDATE DISEASES FOR GENE REPLACEMENT THERAPY Disease Target Cells Gene Defect thallassemia erythrocytes B-hemoglobin SCID lymphocytes ADA deficiency CGD cytochrome b monos/neutrophil s LAD CD18 B-subunit monos/neutrophil s Gaucher's macrophages glucocerebrocida se Hunter's macrophages iduronate desulfase cystic respiratory epi CFTR fibrosis GENE THERAPY PROTOCOLS REVIEWED BY FDA IN 1995 Indication or Disease Submissions State cancer 27 cystic fibrosis 3 bone marrow marking 5 AIDS 3 inborn errors of 5 metabolism infectious diseases 2 TYPES OF VECTORS FOR GENE TRANSDUCTION plasmid DNA retroviruses adenoviruses adeno-associated viruses others METHODS OF GENE INTRODUCTION direct administration into host (e.g. tumor, s/c depot) transduced somatic cells transduced hematopoietic cells CONCERNS RELEVANT TO PRECLINICAL SAFETY TESTING OF VECTOR-BASED THERAPIES aberrant localization or trafficking level and persistence of viral gene expression inappropriate expression of gene product replication of viral vector germ-line transfer of vector gene insertional mutagenesis POTENTIAL SAFETY CONCERNS FOR GENE-MODIFIED SOMATIC CELL THERAPIES phenotype/activation state of effector cell population aberrant localization or trafficking level and persistence of viral gene expression inappropriate expression of gene product inappropriate immune activation germ-line transfer of vector gene insertional mutagenesis GOALS OF PRECLINICAL SAFETY EVALUATION For All Therapeutic Products: Recommend initial safe starting dose and safe dose-escalation scheme in humans Identify potential target organ(s) of toxicity Identify appropriate parameters for clinical monitoring Identify "at risk" patient populations (inclusion/exclusion) APPROACHES TO PRECLINICAL SAFETY/ACTIVITY STUDIES creative, problem-solving data-driven no one "right" way to conduct studies should be based on best available science, technology to date careful design and judicious use of animals should allow for early initiation of clinical studies should allow for uninterrupted clinical development CRITICAL PARAMETERS IN PRECLINICAL STUDIES OF GENE THERAPY SPECIES SELECTION vector dissemination vector/transgene toxicity INDICATION host immune response ROUTE intrinsic pathology vector/RCA replication DOSE SELECTION WHEN ARE SPECIFIC IN VIVO SAFETY STUDIES NEEDED? novel therapies, not amenable to in vitro models novel route, schedule of administration potential for aberrant host response, not evaluable in efficacy model CASE STUDY- PLASMID VECTOR ENCODING HLA/B7 AND B2MG How can route affect activity/toxicity of gene therapy products? introduction of foreign gene expression activates host immune response intra-tumor expression leads to expression in tumor, other organs but no toxicity Nabel et al., Human Gene Ther., 3:649-656, 1992 Stewart et al., Human Gene Ther., 3:267-275, 1992 "worst-case" scenario - iv injection - uptake/expression in all highly perfused tissues in the absence of organ pathology Stewart et al., Human Gene Ther., 3:267-275, 1992 human data show tumor uptake, expression only, no ADE's reported Nabel et al., Proc. Natl. Acad. Sci. USA, 90:11307-11311, 1993 SELECTION OF SPECIES FOR IN VIVO TESTING relevant to biology/mechanism of action relevant to route and method of delivery animal models of disease may provide useful safety information non-human primates are not required for all studies alternative models (transgenics, homologous tumors/species, SCID) CASE STUDY - REV M10-TRANSDUCED CD4 T CELLS IN HIV INFECTION Using alternative animal models to answer specific questions In vitro data demonstrate inhibition of HIV replication, strong "proof of concept" Malim et al., J. Exp. Med., 176:1197-1201, 1992. question of whether transduction alters tumorigenicity, safety, of CD4 cells in vivo? SCID/hu xenograft with RevM10 cells, no organ pathology, tumors no changes serum chemistries, clinical picture CASE STUDY - CUSTOM-DESIGNED RAS OR P53 PEPTIDES Using a murine homologue to determine the safety of a therapy no suitable animal model for human mutated ras or p53 no safety data on repeated administration of vaccine, vector distribution, or potential for autoimmune response recommended use of mouse homologue prototypes to test safety of: multi-dose administrations trafficking of peptide-pulsed MNC, induction of inappropriate immune activation potential for autoimmune response to irradiated autologous cells histopathology of target organs EXAMPLES OF ANIMAL MODELS OF DISEASE USED IN PRECLINICAL EFFICACY/SAFETY EVALUATION Animal Model Disease MRL^^^Document Error^^^/lpr mice Lupus cotton rats , +/- RSV; cystic fibrosis infection W/Wv mice Fanconi's anemia Wobbler mice ALS db/db mice Diabetes Streptozocin [mice/rats] Diabetes SIV [primates] HIV Canine E. coli fibrin clot Sepsis Galactosamine [dogs] Hepatic failure Woodchuck , chimp HBV infection hepatitis MPTP [primate] , 6-OHDA Parkinson's disease [rat] Aged primates Alzheimer's disease EAE [primates, rodents] Multiple Sclerosis WHEN CAN PRECLINICAL SAFETY STUDIES BE MINIMIZED? strong efficacy model rationally designed to answer specific questions optimizing schedule and conditions previous human experience similar product, dose, regimen CASE STUDY - PBL TRANSDUCTION WITH HAIRPIN RIBOZYME GENE How can in vitro data satisfy safety and efficacy requirements? In vitro "proof of concept", safety studies in vitro evidence of positive gene transduction, resistance to HIV-1 infection in HeLa, Jurkat, Molt-4 cells Ojwang et al., Proc. Natl. Acad. Sci. USA, 89:10802-10806, 1992 Yamada et al., Gene Ther., 1:38-45, 1994 in vitro evidence of transduction, resistance in primary PBL no inhibition of cell proliferation, altered phenotype no detection of viral "escape" or resistant clone development in long-term culture Leavitt et al., Human Gene Ther., 5:1115-1120, 1994 CASE STUDY - RETROVIRAL TRANSDUCTION OF HEMATOPOIETIC STEM CELLS WITH MDR-1 GENE FOR CANCER CHEMOTHERAPY How can "efficacy data" satisfy safety requirements? In vitro "proof of concept" studies in vitro evidence of positive gene transduction, resistance to taxol in both human, murine stem cells Hegewisch-Beckeret al., Brit. J. Haematol., 90:1876-883, 1995 Hanania et al., Gene Ther., 2:285-294, 1995 no inhibition of cell proliferation, altered phenotype by FACS to 11% increase in Rh123 efflux in transduced human cells by FACS (index of MDR-1 function) CASE STUDY - RETROVIRAL TRANSDUCTION OF HEMATOPOIETIC STEM CELLS WITH MDR-1 GENE FOR CANCER CHEMOTHERAPY How can "efficacy data" satisfy safety requirements? In vivo "efficacy" model murine transplant system ; serial transplants + taxol, 10 mg/kg no formal toxicology testing, HOWEVER, monitored mice: no adverse effects on engraftment, reconstitution beneficial effect on survival conferred resistance to escalating doses of taxol support from literature that overexpression of MDR-1 had no adverse effect on engraftment, differentiation Mickisch et al., Cancer Res., 51:5417-5424, 1991 Mickisch et al., Proc. Natl. Acad. Sci. USA, 88:547-551, 1991 CASE STUDY - IDS-TRANSDUCED PBL FOR HUNTER'S SYNDROME Consideration of available information (e.g. SCID-ADA gene therapy) no relevant animal model strong in vitro "proof of concept" data Braun et al., Proc. Natl. Acad. Sci. USA., 90:11830-11834, 1993 methodology, vector, dosing regimen well characterized little associated toxicities, ADE in humans treated to date Blaese et al., Human Gene Ther., 4:512-527, 1993 THE NEXT GENERATION-WHAT DO WE AS REGULATORS NEED TO SEE? first generation vectors/new molecular entities, change in route of administration pharmacologic profiles, dose/response relationship, proof of concept full toxicology profiles, vector distribution focus is on SAFETY; rationale is secondary, but still important vectors - the next generation(s), minor changes in non-control regions abbreviated toxicology; bridging studies abbreviated pharmacology - does it work better? the same? CASE STUDY - ADENOVIRAL VECTOR FOR CYSTIC FIBROSIS Comparison of first and second generation vectors in a "bridging" study question of biology, immune/inflammatory response to different constructs baboons; first/second generation vectors instilled into contralateral lung lobes Goldman et al., Human Gene Ther., 6:1839-851, 1995 methodology, vector, dosing regimen well characterized gene stability prolonged, inflammation decreased with second generation vector marked decrease in hexon protein levels in tissues receiving second generation vector THE NEXT GENERATION-WHAT DO WE AS REGULATORS NEED TO SEE? key points to define NOAEL, toxic doses; target organs, key pathology shape/steepness of the dose/response curve (both pharm & tox) ED, EC50 - may be in vivo or in vitro points to monitor in the clinic THE CHALLENGE OF ANIMAL STUDIES How have data from preclinical studies facilitated clinical development of gene therapy? Example: Using preclinical data to recommend dose escalations for adenoviral vectors for CF dose-escalation studies of CFTR vectors in cotton rats show a 2 to 10-fold difference between NOAEL and threshold doses for toxicity consistent between 2 different vectors, 2 serotype backbones similar studies in Rhesus monkeys, baboons show 10-fold difference in dose between NOAEL and threshold toxic doses clinical data demonstrated first ADR after 100-fold escalation; hypoxia, fever, pneumonia - some evidence of these effects in primate models at high doses Based upon these data, recommendation to sponsors was to consider dose-escalations in 1/2 log (3-fold) increments between cohorts, and to tighten up stopping rules, patient monitoring to include these events. QUESTIONS TO BE ANSWERED BY PRECLINICAL STUDIES What is the relationship of the dose to the biologic activity? What is the relationship of the dose to the toxicity? Does the route and/or schedule affect activity/toxicity? What risks can be identified for the clinical trial? THE DILEMMA OF PRECLINICAL STUDIES FOR GENE THERAPIES Reversibility of the toxicity may be missed in in vitro models Adverse events in animals: may not be relevant to clinical trials may be acceptable in more seriously ill populations Long-term toxicities or events: may not be determined in preclinical animal studies may be missed in phase 1 trials; may require long-term follow-up SUMMARY Preclinical studies for gene therapy should be rational, scientifically-designed, based on best available technology, methods to date Efficacy studies for gene therapy can also provide rationale, safety data for phase 1 trial entry Animal studies are dependent on body of information available, question being asked Maximal information may be obtained in alternative animal models; primates are not a priori necessary Sponsors are encouraged to conduct, publish experiments for further advancement of the field FURTHER QUESTIONS? WHEN IN DOUBT, TALK TO US! Anne Pilaro Lauren Black Mercedes Serabian Dave Green (301) 594-5599 or 594-5600 (301) 594-0513 (FAX) Acknowledgments and thanks to Dr. Joy Cavagnaro, Office of the Director, Center for Biologics Evaluation & Research (301) 827-0372(301) 827-0440 (FAX) CONSIDERATIONS IN CLINICAL TRIAL DESIGN FOR GENE THERAPY PHASE I TRIALS David Wilde, M.D. Division of Clinical Trial Design & Analysis OTRR/CBER/FDA CLINICAL TRIAL DESIGN Scientific rationale Objectives (endpoints)) Experimental design Selection of subjects Informed consent Compliance with therapy Assessment of trial results Tolerability of therapy Data collection Statistical analysis Monitoring of study SPECIFIC ISSUES OF GENE THERAPY TRIALS Size of trial (phase 1 OR 1/2) Novel product Unknown adverse reaction profile Unpredicted host response Functional lifespan of product Informed consent Access to therapy SIZE OF GENE THERAPY TRIAL Safety concerns are paramount preclinical data using animal models or in vitro test data to support safety or efficacy expose fewest number of people to risk Exploratory concepts limits the scope of hypothesis testing limits allocation of resources Need for rapid feedback of information technical performance of gene product in vivo if in vivo results do not match expectations, need flexibility to alter vector production or experimental design USE OF A NOVEL PRODUCT Vectors or gene products may change Commonly used vectors contain a variety of different gene products Vectors "evolve" due to ongoing technical progress Natural gene product may be modified as a fusion protein or may contain point mutations Gene products using consensus sequences reduce genetic polymorphism, but do not represent naturally occurring products UNKNOWN ADVERSE REACTION PROFILE Special concerns about adverse reactions Lack of long-term safety data from historical controls Potential for atypical presentation of adverse reaction prior to its recognition Unknown frequency of adverse reaction secondary to genetic manipulation (RCR, insertional mutagenesis, gene migration, etc.) Do some vectors have a higher incidence of adverse reactions? Unknown reversibility of adverse reaction (unlike drug toxicity) Interaction of patient medications with vector (e.g., retroviral vectors) Duration of safety monitoring required, due to chronic nature of genetic defect Cost of monitoring for adverse reactions UNPREDICTABLE HOST RESPONSE How effective is the route of vector delivery (direct versus indirect inoculation)? After delivery, does the vector localize and remain in the desired site? Does gene transcription occur (use of vector-only control population)? Does the gene product have the expected functional activity? What is the host immune response to gene therapy? autologous cells manipulated ex vivo expression of viral products on cells reactivity to antigen depots (plasmid DNA) FUNCTIONAL LIFESPAN OF PRODUCT Sparse data on kinetics of clearance or marked cells Immune clearance ADCC antigen-specific cytolysis Sequestration (deficient homing) Cell types (e.g., lymphocyte, fibroblast, stem cell) Host resistance Ablation (suicide gene) INFORMED CONSENT Intent of trial Risks (known or potential) Proposed benefit Duration and methodology of therapy Duration and methodology of follow-up Provision for autopsy "Recycling" of study subjects ACCESS TO THERAPY Need well-defined patient population to offset small sample size Gender balance Patient drop-out and cross-over Commitment to long-term monitoring INTERNATIONAL REGULATORY CONSIDERATIONS, EXPORT AND IMPORT ISSUES Elaine C. Esber, M.D. REGULATORY CONSIDERATIONS Export of Gene Therapy Products Import of Gene Therapy Products Communication with Foreign Government Officials Memoranda of Understanding (MOU)/Memoranda of Agreement (MRA) MOUs enlist aid of counterpart governments in assuring imports meet our requirements, e.g., GMPs, GLPs MRAs generally mean either reliance upon one another's conformity assessment system or, where such reliance is not practicable, exchange of the results of conformity assessments EXPORT OF GENE THERAPY PRODUCTS Products approved (licensed) in the US fully meeting FDA requirements Must meet requirements of importing country No restrictions or controls to exportation Products that are unapproved in the US Can be shipped under IND regulations (21 CFR 312), or Subject to the Drug Export Act Amendments of 1986 and 1996 DRUG EXPORT ACT AMENDMENTS OF 1986 AND 1996 Drug Export Act Amendments of 1986 Pub L. 99-660; November 14, 1986 Created New Section 802; three tracks FDA Export Reform and Enhancement Act of 1996 Pub L. 104-134; signed by President Clinton April 26, 1996 Revises Sec. 801 and 802 of FD&C Act and Subsection (h) of Sec. 351 of PHS Act FDA EXPORT REFORM AND ENHANCEMENT ACT OF 1996 Exports of products for investigational use in importing country Exempted from regulation by FDA if shipped to one of 25 countries listed [€ 802 ©] Australia, Canada, Israel, Japan, New Zealand, Switzerland, and South Africa, or any country "in the European Union or a country in the European Economic Area" FDA approval of export required for shipment of investigational product to unlisted countries (21 CFR 312.110) Review includes scientific review of protocol, safety data and a letter from importing country approving import Export of products for commercial use in importing country Authorizes shipment to "listed country" Authorizes exportation to "unlisted country" if approved by "listed country" Eliminates need for FDA approval for export; replaces with FDA notification Eliminates need for investigations in US Others FDA may restrict exports, if: Not manufactured in GMP facility Adulterated Not meeting basic export requirements Imminent hazard Inadequate labeling Imports for Export [€ 801 (d)(3)] Statutory requirement is recordkeeping Stages in Manufacture Final Drug Product Bulk - Drug Substance Mixture] Partially Processed Raw Material Partially processed biologics [€351(h)] may be exported if: not in a form applicable to prevent or treat disease not intended for sale in the US is intended for further manufacture into final dosage form outside the US must be manufactured using GMPs and meet basic export requirements Still undergoing interpretation Regulations are being considered for clarification Agency encourages questions if unclear IMPORT OF GENE THERAPY PRODUCTS INTO US FOR HUMAN USE Investigational New Drug Regulations apply [21 CFR 312.1] if a biologic/drug Investigational Device Exemption Regulations apply to device components [21 CFR 812] Marketing applications will depend on product - BLA, NDA, or PMA COMMUNICATIONS WITH FOREIGN GOVERNMENT OFFICIALS Formal relationships with governments: ICH with EU and Japan Bilateral relations, e.g., Tripartite, Trilateral WHO Formal communication with government officials, e.g. Information exchange of public information Harmonization JOINT REVIEWS Regulations codified in 1993 and 1996 to enable information exchange while maintaining confidentiality [21 CFR 20.89] Applicable to exchange of any document , e.g., draft proposed rules and predecisional documents; excludes trade secret information concerning manufacturing methods and controls Foreign government agency signs statement that: affirms its ability to maintain confidentiality commits not to disclose FDA determines either: sponsor of product has provided written authorization, or disclosure would be in the interest of public health, or disclosure is to a foreign visiting scientist who must sign confidentiality commitment OVERVIEW OF FUNDING MECHANISMS AVAILABLE FROM THE NIH FOR THE SUPPORT GENE THERAPY PROJECTS Craig Reynolds, Ph. D. NCI/FCRDC MECHANISMS AVAILABLE FROM THE NIH FOR THE SUPPORT OF GENE THERAPY PROJECTS Preclinical Basic Research Clinical Trials Production Other PRECLINICAL BASIC RESEARCH SUPPORT R01 - research grants R03 - small research grants (RFA only) R41/R42 - Small business technology transfer (STTR) grants R43/R44 - small business innovation research (SBIR) grants P01 - program project grant P30 - core center grant (NIDDK) P50 - specialized centers of research U19 - research program cooperative agreement (RFA) only U43/U44 - SBIR cooperative agreement CLINICAL TRIALS SUPPORT R01 - research grant R03 - small research grant (RFA only) R21 - exploratory/developmental grant P01 - program project grant U01-U10 - cooperative agreement clinical research PRODUCTION SUPPORT NGVL - National Gene Vector Laboratories (NCRR) MARP - MoAb/Rec. Protein Production Facility (NCI) OTHER SUPPORT Toxicology (nonprimate/primate) Pharmacology and formulation Preclinical animal models TECHNOLOGY TRANSFER (OTD/OTT)FUNDING MECHANISMS FOR NCI SPONSORED GENE THERAPY RESEARCH Diane Bronzert Cancer Therapy Evaluation Program, Division of Cancer Treatment Diagnosis and Centers, National Cancer Institute, NIH, Bethesda, MD (Tel: 301-496-8866) The National Cancer Institute (NCI) has a diverse program in gene therapy with approximately $28 million in extramural research project grants (RPG) awards for fiscal year 1995. The principal mechanisms used for funding gene therapy throughout NIH are investigator initiated research grants (RO1, R29), Small Business Innovation and Technology Transfer grants (SBIR, STTR), and program project grants (P01). Program project grants have proven to be a very effective mechanism to fund the multi-disciplinary efforts that are necessary to translate basic research discoveries to the clinic. Areas of special interest include the development of new delivery systems, animal models, and new therapeutic approaches. In addition , NCI has published Program Announcements using the small grant (R03) and exploratory grant (R21) mechanisms for the support of therapeutic clinical trials. The primary categories for oncology gene therapy studies are the following: (1) new vaccines or studies to increase/create immunity to cancer cells (e.g., production of cytokines); (2) studies to specifically kill or prevent proliferation of cancer (e.g., suicide genes, prodrugs, antisense); or (3) studies to increase the dose of chemotherapy agents administered to patients by protecting the hematopoietic system. NIAID SUPPORT FOR HUMAN GENE THERAPY Nava Sarver, Ph.D. Division of AIDS, NIAID For HIV induced disease and immune dysregulation, gene-based therapies can potentially be used with other current therapies to further enhance the therapeutic benefit to patients. Gene therapy approaches for treating HIV are designed to enhance immune function or to block specific functions critical to viral replication, thereby limiting or curtailing virus dissemination. HIV gene therapy approaches involve the introduction of therapeutic or protective genes into lymphocytes (CD4; CD8 CTL) or their precursor stem cells (CD34; others?), including those mobilized to the peripheral blood and those derived from cord blood. Antiviral genes include those encoding for (1) proteins (transdominant mutants, single-chain antibodies, toxin genes); (2) RNAs (ribozymes; RNA decoys; antisense); and (3) cytokines or regulatory factors (IFN; eIF-5A). For a therapeutic outcome, these strategies require efficient delivery of genes to target cells, homing to target cells, long term expression, localization to the proper cellular compartment, and fulfillment of other parameters common to most other human gene therapies. The NIAID supports studies in gene-based therapies to address identified problem areas and to improve efficiency and predictable use in clinical setting. NIAID's Division of AIDS (DAIDS) evaluates gene therapy as a molecular-based approach to add to the existing arsenal of therapies to treat HIV disease antiviral drugs, adoptive immune therapies, cytokine therapy). Among these support programs are: (I) investigator initiated R01-type awards (R01, R29, R37, others); (2) the National Cooperative Drug Discovery groups for the Treatment of HIV Infection (NCDDG-HIV) which focuses on preclinical studies); (3) the Strategic Program for Innovative research on AIDS (SPIRAT) which focuses on translational research from advanced preclinical to pilot clinical studies; (4) Small Business Innovative Research grants (SBIR) and Small Technology Transfer Research grants (STTR). Research directions encouraged under these programs, eligibility, and scope will be discussed. Nava Sarver, Ph.D. Chief, Targeted Interventions Branch Division of AIDS, NIAID TEL: 301-496-8197 FAX: 301-402-3211 email: ns18p@nih.gov NATIONAL HEART, LUNG, AND BLOOD INSTITUTE: SUPPORT FOR GENE THERAPY RESEARCH Sonia I. Skarlatos, Ph.D. NIH, NHLBI In 1992, the National Heart, Lung, and Blood Institute (NHLBI) convened a "Working Group on Gene Therapy Approaches and Resources for Heart, Lung and Blood Research" to provide a framework for the scientific evolution of human gene therapy in a suitable and systematic manner. The Working Group articulated scientific needs and identified broad avenues for the NHLBI to foster gene therapy. Strategies to facilitate the development of human gene therapy included: (1) encouragement of research efforts for the basic tasks involved in gene therapy; (2) development of animal models; (3) stimulation of collaboration among scientists from different disciplines; (4) establishment of shared support facilities; and (5) recruitment of established investigators and training of new investigators. Using the Working Group report as a guide, the NHLBI presently supports and will continue to support a diverse array of research activities to advance gene therapy of heart, lung and blood diseases. The NHLBI currently supports six clinical studies. Three are for cystic fibrosis, which will test the clinical efficacy and safety of adenoviral vectors, adeno-associated vectors and liposomes. The other three are for the treatment of ADA deficiency with ADA-transformed peripheral blood stem cells, for gene therapy of Gaucher's disease with hematopoietic stem cells transformed to express glucocerebrosidase, and for gene therapy of Fanconi's anemia type C. The NHLBI will be releasing, shortly, a request for applications (RFA) on "Fundamental Biological Principles for Gene Transfer for Cardiovascular, Pulmonary and Hematologic Diseases". This RFA invites Program Project grant applications for support of research efforts to advance gene transfer technology and its potential application to cardiovascular, pulmonary and hematologic diseases. In addition, the NHLBI, through a program announcement on "Gene Therapy Vector Design and Development", will be encouraging collaboration between academia and industry to design and develop vectors and delivery systems for cardiovascular, pulmonary and hematologic diseases. Support for this program will be provided by the Small Business Technology Transfer Program. For further information on any of these programs, please contact: Sonia I. Skarlatos, Ph.D. NIH, NHLBI Division of Heart and Vascular Diseases Two Rockledge Center, Suite 10186 6701 Rockledge Drive, MSC 7956 Bethesda, MD 20892-7956 Phone: 301-435-0550 FAX: 301-480-2858 E-Mail: skarlats@gwgate.nhlbi.nih.gov NIDDK SUPPORT FOR GENE THERAPY RESEARCH Catherine McKeon, Ph.D. Metabolic Diseases and Gene Therapy Research Program, National Institutes of Diabetes and Digestive and Kidney Diseases, NIH, Bethesda, MD 20892 (Tel: 301-594-8810) The NIDDK supports research in diabetes, endocrinology, metabolic diseases, nutrition, kidney, digestive, urologic and hematologic diseases. NIDDK support for gene therapy research has been predominantly to develop treatments for genetic metabolic and hematologic diseases, however, other applications of interest to NIDDK can be envisioned as the technology develops. Most of our support for gene therapy research is through Investigator Initiated research grants (RO1). In addition, the NIDDK supports program projects (P01), Core Centers for Gene Therapy Research on Cystic Fibrosis and Other genetic Disease (P30), Centers of Excellence in Molecular Hematology (P50), and co-sponsors the National Gene Vector Laboratories. Approximately $40 million was spent on basic and applied gene therapy research in 1995. Other mechanisms for funding of gene therapy research are the Small Business Innovation Research Grant (SBIR) and the Small Business Technology Transfer Grant (STTR). The NIH is Congressionally mandated to expend 2.5% of it's budget on the STTR program. These programs are designed to promote collaborations between academic scientists and small business concerns to develop and market new technologies. Because of the unique collaboration between academic researchers and industry in the field of gene therapy, this mechanism should be considered. One example of a commercial product developed with NIDDK funds provided through the SBIR program is the liposome component, Lipofectamine. Special emphasis topics published by NIDDK include improving vector production methods; development of new and improved vectors; development of packaging lines with new or altered viral envelope proteins; and engineering tissue specific promoters which provide stable expression. THE NATIONAL GENE VECTOR LABORATORIES Richard A. Knazek, M.D., Ph.D. Clinical Research, NCRR, NIH ABSTRACT The National Gene Vector Laboratories (NGVL) provide shared resources to facilitate the production of clinical grade adenoviral, DNA plasmid and retroviral vectors for use by eligible U.S. investigators in Phase I or II experimental human gene therapy protocols. This is an NIH resource, with laboratories located at Indiana University, University of Michigan and University of Pennsylvania, that produces vectors under GMP conditions and provides them, at no cost, to the requesting investigator. The NGVL is supported by cooperative agreement awards from the National Center for Research Resources, co-sponsored by the National Cancer Institute, National Institutes of Diabetes and Digestive and Kidney Diseases, and National Heart, Lung and Blood Institute, and with co-funding from the Office of AIDS Research. Applications received by the coordinating NGVL must contain detailed descriptions of both the requested vector and the clinical protocol in which it is to be used. Such vectors must have completed pre-clinical testing. Proposals will be evaluated by an external Scientific Review Board and the Steering Committee of the NGVL with the ultimate selection being based upon scientific merit, feasibility and availability of NGVL resources. Priority for vector production will be given to protocols that have received peer-reviewed grant support. Preference will be given to NIH-sponsored research. Application packets can be obtained from: Kenneth Cornetta, M.D. Richard A. Knazek, M.D. NGVL Coordinating Center Clinical Research Indiana University National Center for Research Resources/NIH IB442, 975 West Walnut Street 6705 Rockledge Drive, Room 6030 Indianapolis, Indiana Bethesda, Maryland 20892-7965 46202-5121 Tel: 317-274-0843 Tel: 301-435-0790 Fax: 317-274-0396 Fax: 301-480-3661 e-mail: e-mail: Richardk@ep.ncrr.nih.gov Ken_cornetta@iucc.iupui.edu The next deadline for submission of applications is September 3, 1996. NATIONAL GENE VECTOR LABORATORIES An interactive group of laboratories whose purpose is to provide clinical grade adenoviral, DNA plasmid, and retroviral vectors to eligible clinical investigators for gene therapy protocols. DISEASES TARGETED BY GENE THERAPY PROTOCOLS APPROVED BY THE RAC Rheumatoid arthritis HIV Storage Diseases Cystic Fibrosis Monogenic Diseases Arterial Diseases Cancer FIRST ONE-FOURTH OF PROTOCOLS REVIEWED BY THE RAC Adenovirus Non-Virus Retrovirus LAST ONE-FOURTH OF PROTOCOLS REVIEWED BY RAC Adenovirus AAV Non-Virus Retrovirus VECTOR MILESTONES Basic Research Initial Vector Generation Preclinical Studies Safety Studies Large-Scale Vector Production Pharmacology/Toxicology Testing Patient Studies NATIONAL GENE VECTOR LABORATORIES Indiana University-Coordinating Center Retroviral Vectors University of Pennsylvania Adenoviral Vectors University of Michigan Non-viral Vectors ELIGIBILITY REQUIREMENTS Qualified Investigators Qualified Domestic Institutions or Government Agencies Capability for Both Clinical Research and Scientific Support Submit Request to coordinating NGVL Follow Guideline of Oversight Committees IRB approval and initial discussions with FDA Agree to Post-distribution monitoring STEERING COMMITTEE Directors and Associate NGVL Directors Indiana University University of Pennsylvania University of Michigan Outside Experts University of Chicago Salk Institute St. Jude Children's Research Hospital University of Pittsburgh NIH Representatives NCRR NHLBI NIDDK NCI SCIENTIFIC REVIEW BOARD Scientists with expertise in gene therapy and relevant fields COMPETITIVE REVIEW Pre-clinical laboratory and animal data Current status of the proposed vector Suitability and technical feasibility of the proposed gene therapy Clinical Protocol Design Qualifications of the Investigator Qualifications of the Laboratory Qualifications of the Clinical Facility Ethical Implications Resources Required Likelihood of Success relative to other clinical trials and other gene therapy approaches APPLICATION Letter of Agreement Abstract Biographical Sketches of Investigators Resources Summary of Proposal Clinical Protocol with Consent Form IRB and IBC Approval Letters Sequence Data (if available) Toxicology Data (if available) Relevant Publications Letters of Collaboration FDA Contact Information Vector Distribution Agreement "REQUEST FOR VECTOR PROCEDURE" Submit application to coordinating NGVL Initial Review by 2 outside reviewers and 1 SC Member Reviewers request additional information through the coordinating NGVL Reviews returned to coordinating NGVL for distribution to the Steering Committee Members Availability of NGVL resources determined Notification of Applicant UPON APPROVAL OF 'REQUEST FOR VECTOR' Biological Materials are submitted to NGVL Batch of vector is generated at NGVL Samples of batch are returned to investigator Safety Testing Toxicology Studies upon final approvals from regulatory groups clinical vector is shipped to investigator DEADLINES FOR SUBMISSION OF 'REQUESTS FOR VECTOR' September 3, 1996 April 7, 1997 September 8, 1997 Kenneth Cornetta, M.D. NGVL Coordinating Center Indiana University School of Medicine IB442, 975 West Walnut Street Indianapolis, Indiana 46202 tel. (317) 274-0448 NCI SPONSORED SMALL SCALE MANUFACTURE OF GENE THERAPY PRODUCTS George A. Robertson, Ph.D. The Monoclonal Antibody/Recombinant Protein Production Facility (MARP) provides biopharmaceutical manufacturing support for NIH-supported investigators. Biologics are manufactured under FDA Good Manufacturing Practices (GMP) for Phase I/II human clinical trial, or advanced preclinical animal testing. The MARP is operated under contract by Scientific Applications International Corporation (SAIC), which provides operations and technical support to the Frederick Cancer Research and Development Center for the NCI. The NCI Project Officer of the MARP is the Chief of the Biological Resources Branch of the National Cancer Institute. A newly renovated production facility, located in building 459, has just been occupied by the MARP. This facility contains laboratories capable of producing cell banks, monoclonal antibodies using hollow-fiber bioreactors, complete biochemistry laboratories for product purification and characterization, aseptic areas for purification and vialing and a BL3-designed production suite equipped to support animal cell and microbial propagation. Projects for gene therapy and/or DNA based therapeutics can be products in one of several areas, depending on the level of biocontainment required. Infectious agents and those requiring BL3 containment can be produced in the bldg. 459 containment laboratory, on a campaign basis. Packaging cell banks can be prepared in the animal cell culture laboratories and characterized in house or by utilizing contract testing laboratories. DNA can be fermented in one of several laboratories, depending on quantity and containment requirements. Aseptic purification and vialing can be conducted in the aseptic production suite in bldg. 459. Critical spaces include three class 100 rooms. The MARP staff, in conjunction with the Quality Assurance group, is able to produce the Chemistry, Manufacturing and Control (CMC) section of INDs as a regular part of every project destined for the clinic. For further information, contact Dr. Stephen Creekmore, Biological Resources Branch, DTP, DCTDC, NCI-FCRDC, Frederick, MD 21701-1201, (tel) 301-846-1098, (fax) 301-846-5429, (e-mail) Creekmor@ncifcrf.gov. CURRENT AND SPECIAL NIH RESOURCES FOR THE DEVELOPMENT OF GENE THERAPY PRODUCTS Elizabeth C. Lovoy, J. D., M. P. H. Office of Technology Development, NCI (301) 496-0477 ABSTRACT Among the traditional sources of funding for NIH scientists are: funds appropriated by Congress; royalties received from the licensing of NIH patents (the use of which was previously restricted to "technology transfer-related" activities, and more recently made available for "mission-related" activities); gift funds received from the private sector; and funds received from biotechnology and pharmaceutical industry collaborators under Cooperative Research and Development Agreements (CRADAs). The NCI Office of Technology Development (OTD) provides an array of services to NCI scientists, helping them to combine and leverage the resources of both the NCI and the private sector to speed the development and commercialization of technology arising in NCI's laboratories for the overall benefit of the public health. One of the major services offered by the OTD is the negotiation of transactional agreements between the NCI and outside parties (including universities, research institutes, and pharmaceutical and biotechnology companies) which allow resources to flow both to and from the NCI. These transactional agreements enable the exchange of research materials under Material Transfer Agreements (MTAs); the provision of experimental drugs and biologics to the NCI under Clinical Trail Agreements (CTAs); and the contribution of staff, facilities, equipment, supplies and intellectual property (as well as funds to the NCI, but not to the collaborator ) under CRADAs. In addition, there are an array of "hybrid" agreements (such as MTA-CRADAs) which can extend NCI scientists' access to various companies' propriety materials and other technology for use in gene therapy, as well as other research. The objective of these agreements and the manner in which they are negotiated by the NCI OTD are to foster partnerships between NCI scientists and outside institutions, and to do so as expeditiously and as easily as possible. Each of these agreements offers unique opportunities to NCI scientists to establish such partnerships and to broaden their access to necessary resources, including in some cases, funding. Any NCI scientist who wishes to explore these avenues should contact the NCI Office of Technology Development an (301) 496-0477. In addition, information on these opportunities may be obtained from our Website at http://www.nci.nih.gov/hpage/ttrans.htm TRADITIONAL SOURCES OF FUNDING FOR NIH SCIENTISTS Funds appropriated by Congress Royalties from the licensing of NIH patents Gift funds from the private sector Funds received from biotechnology and pharmaceutical industry collaborators under a Cooperative Research and Development Agreement (CRADA) FORMAL COLLABORATION Material Transfer Agreements (MTAs) Clinical Trial Agreements (CTAs) Cooperative Research and Development Agreements (CRADAs) Confidentiality Agreements (CDAs) MATERIAL TRANSFER AGREEMENT (MTA) Provides for the transfer of research material Research purposes only+not for commercial purposes such as screening, production or sale Material may not be used in human subjects No option or grant of future intellectual property rights. No funds may be received by PHS UBMTA (Uniform Material Transfer Agreement) has been developed to expedite transfer of research materials among non-commercial entities CLINICAL TRIAL AGREEMENT (CTA) Similar to an MTA but provides for the transfer of material for use in human subjects under an approved clinical protocol No option or grant of future intellectual property rights No funds may be received by PHS Companies may provide a "Guest Researcher" to NCI. Guest Research is paid directly by the company. COOPERATIVE RESEARCH AND DEVELOPMENT AGREEMENT (CRADA) Agreement between one or more federal laboratories and one or more non-federal parties Federal laboratories may provide personnel, services, facilities, equipment (but not funds) Collaborator may provide funds, personnel, services, facilities, equipment Areas of research may include basic research, pre-clinical studies, clinical research, and/or any miscellaneous areas of interest to the NCI and the Collaborator CONFIDENTIALITY AGREEMENT Used to transfer information (data) between parties Restrict the disclosure of information to third parties Prohibit commercial use of any information shared Define the boundaries as to what is confidential information require that confidential information be clearly indicated "CONFIDENTIAL" Limited duration of confidentiality CRADA DEVELOPMENT CRADAs can develop from A collaborative research project or Soliciting CRADA proposals in the Federal Register and followed up by marketing through trade journals or direct company contact CRADA For the NIH the CRADA: Makes private sector resources (reagents or drugs, facilities, expertise, personnel and funds) available for research, development, or clinical testing Acts as a Strategic Business Plan for product development or commercialization which adds value to NIH science, technology, or inventions For the private sector the CRADA: Makes government resources (facilities, intellectual property and expertise) available for research, development or clinical testing Addresses present and future intellectual property rights Allows an option to negotiate and exercise non-exclusive or exclusive licensing rights CRADA Both parties must make an intellectual contribution to the collaboration Confidentiality obligations are provided Exclusive access to NCI-generated data An option to exclusively license NCI or joint inventions arising from the collaboration NCI may receive funding from the collaborator to support CRADA research Extensive review process CRADA LETTER OF INTENT (LOI) Permits collaborative research to begin prior to the final approval and execution of the CRADA When the CRADA is signed, the effective date of the CRADA is retroactive to the date of the LOI for purposes of confidentiality of information exchanged between the parties and for purposes of intellectual property developed between the date of execution of the LOI and the date of execution of the formal CRADA Effective for one year+may be extended for an additional year if the collaboration is still active and if the CRADA is still under negotiation PHS CRADA POLICY CRADAs are authorized when the collaborator will make significant intellectual contributions to the research project, or will contribute essential research materials or technical resources, not otherwise reasonably available Outside organizations must have fair access to the collaborative opportunities. Special consideration is given to small business and preference to those that are located in the U.S. and agree to manufacture in the U.S. products developed under the CRADA CRADAs are not intended to be a general funding mechanism to support research in a PHS laboratory CRADA FUNDS CAN BE USED FOR: Supplies Equipment Travel Expenses Salary for Non-FTE personnel Contractor Services CRADA DEVELOPMENT AND APPROVAL PROCESS A written CRADA should be developed as soon as the federal scientist and their counterpart(s) negotiate the research plan (written description of the research and development project, including each party's contribution to the planned research and development The CRADA process consists of two stages. The first step is a "Drafting and Negotiation" stage where a "Draft Agreement" is created The second stage of the CRADA process the "Approval" stage HYBRID AGREEMENTS AND OTHERS MTA-CRADAs CTA-CRADAs Letter of Intent (CRADAs) MATERIAL TRANSFER CRADA (MTA-CRADA) Form of CRADA in which PHS may obtain a unique research resource for use in the conduct of specified research which is consistent with the missions of the laboratory Thirteen month term+includes a 30 day period for PHS to disapprove of the CRADA in which case it is returned for review by the NIH CRADA Subcommittee No funds may be received by PHS under the MTA-CRADA Streamlined approval process CLINICAL TRIAL CRADA CRADA with additional terms appropriate for collaboration involving human clinical studies As with any CRADA, the parties have certain rights to CRADA subject inventions. The federal laboratory may accept funds Exclusive access by the collaborator to the clinical data and to the Investigational New Drug Application may be granted LETTER OF INTENT (LOI) FOR CRADA Can be used to permit collaborative research to begin pending final approval and execution of the CRADA Patentable inventions may be made by NCI employees and employees of the collaborator in the course of this joint research and confidential information exchanged NCI agrees that should a CRADA be approved and executed, it will be retroactive to the date of the Letter of Intent The Letter of Intent is not an irrevocable commitment on the part of NCI to enter into a CRADA with the collaborator SUMMARY The NCI has a variety of tools available to foster collaboration and facilitate protection and transfer of information and technology ELECTRONIC ACTIVITIES AND REGISTRIES UNDER THE FDA SMART PROGRAM Mary A. Buesing M.D. CBER SMART Project Officer AGENDA Two main focuses in 1996: Workflow and document management RMS IND Internal Regulatory Correspondence Executive Document Exchange Regulatory Management System: the targeted integrated information system (electronic database) Ongoing efforts in: CHAPEL Lot release Gene Therapy Information network IND INTERNAL REGULATORY CORRESPONDENCE (ICOM) Scope Generate, store and retrieve internally generated documents such as meeting minutes, telecons, review comments, letters to sponsors Link internally generated documents to specific submissions Phase 1: Provide ad hoc routing for some key documents ICOM - PURPOSE Enable CBER staff to perform better reviews Manage documents critical to IND review process Generate and retrieve internally generated documents Link related documents Query data pertinent to the review Enable CBER to perform faster reviews Route documents quickly Track review status User's In-Box Letter Generation ICOM - IMPLEMENTATION Conduct user acceptance testing with representative users Deliver to 75 users this year RMS: EXECUTIVE DOCUMENT EXCHANGE Scope Route correspondence: response to congressional inguiries, points to consider, federal register notices, all guidelines, ICH/GRP docs Between the Center Director, Office Directors, and Division Directors Create, import, route, comment upon, and consolidate comments Champion : Mark Elengold, Dir. Office of Communication, Training and Manufacture's Assistance EDE - PURPOSE Enable CBER management to review and respond quickly to high priority requests Replace "Pony Express" system Route and comment upon documents Improve current search capabilities Utilize advanced tools to facilitate rapid response Conduct acceptance testing with 3 representative users Deliver to 35 users this year COMMON CAPABILITIES FOR ICOM & EDE Document Management Routing Organization and Navigation Viewing and Modifications STATUS TRACKING RMS - ANNOTATIONS WITH RE:MARK LOT RELEASE IMAGING SYSTEM (LRIS) LRIS PURPOSE Allow parallel reviews of Lot protocols Provide Guidance to Industry on submissions of Lot Release Data electronically INTEGRATE COMMERCIALLY AVAILABLE SOFTWARE ( ACROBAT) TO SUPPORT SUBMISSIONS AND REVIEW OF IMAGES LRIS - VALUE Reduces review time; improves quality of review process Enables all manufacturers to submit electronic protocols using existing tools Helped pioneer use of Adobe Acrobat technology within CBER Imaging technology can be used in many CBER initiatives (e.g., CAPLA), and is being used for RMS Helped define resolution standards capturing images and graphics Facilitates archiving - photos deteriorate quickly CAPLA - PURPOSE Manage incoming CAPLAs Provide guidance to industry Develop procedures for handling CAPLAs Build system to track CAPLAs CAPLA - Guidance for Industry Specifies preferred COTS software Provides guidance on who to contact and when Describes CBER technical environment Supports move from sponsor-submitted hardware/software to CBER-owned equipment Supports move to network CAPLAs CAPLA - Internal Procedures Documents procedures for CBER to follow in advance of and upon receipt of CAPLA submission Defines roles and responsibilities Establishes time-frames for key activities CAPLA Guidance Manual Availability of the CAPLA Guidance Manual -FR 3/21/96 Internet email requests for the manual: FTP : "FTP.FDA.GOV", or "CDVS2.CDER.FDA.GOV" Browsers: "HTTP:^^^Document Error^^^/www.fda.gov/cber/cberftp.html" CAPLA Guidance Manual For questions about CBER FTP contact Mark Elengold at: or elengold@A1.cber.fda.gov GENE THERAPY - BACKGROUND NIH Recombinant DNA Advisory Committee (RAC) Public forum for discussion of gene therapies Collected and published clinical and product data Congress Mandated FDA build registry for longitudinal follow-up CBER Currently 130+ INDs, over 600=0 patients PLAs in the near future GTIN - Objectives Subject registry to meet Congressional mandate Permits longitudinal analysis to addresses concerns on long-term effects of gene therapy Provides prototype of functional system to drive FDA policy on data collection requirements Prototype populated with Cystic Fibrosis Foundation (CFF) data GTIN - Accomplishments Delivered Registry (Alpha prototype) to OTRR/DCGT in Spring 1996 Laptop PC Delivered capabilities that provide information on gene therapy INDs Delivered capability for NIH to generate reports to support RAC GTIN- Value Satisfied Congressional mandate Serves as a catalyst to establish policy necessary to implement Novel Therapy registries Confidentiality Who collects the data Who enters the data Length of follow-up Future Steps: Address policy issues of data gathering from "womb to tomb"(e.g. CRF modification), linking to other registries such as the cystic fibrosis registry Await new CBER database completion Budgetary constraints re:longitudinal follow-up & data integrity (?PDUFA renewal) Implementation of a xenotransplantation registry OTHER AUTOMATION EFFORTS CBER Electronic IND Pilot CBER working group forming chair: Fred Miller M.D. every office represented Pilot to begin within next few months to review Gene Therapy electronic INDs INTERCENTER EFFORTS CDER/CBER Electronic Submissions Committee formed May 1996 charter: electronic format for CFRs and line listings (draft by fall 1996, industry workshop fall 1996, FR notice DEC 1996) chair: Kaye Fendt (CDER) CARS (computer review of safety) charter : data model for review of safety chair: Kaye Fendt (CDER) JULY 12 VECTOR DEVELOPMENT Goals: To present and discuss issues concerning the development, production, and use of viral vectors for gene therapy. 8:00 Overview and Goals, Philip Noguchi, M.D., Director Division of Cellular and Gene Therapies, CBER 8:15 Plenary Talk, Alan E. Smith, Ph.D., Senior Vice President, Genzyme Corporation 9:00 Morning Breakout Sessions 1. Adenoviral Vectors 2. Ancillary Products 3. Facilities & Manufacturing 4. Getting Started in Gene Therapy Vector Development 1:30 Afternoon Breakout Sessions 5. Development of New Vector Systems 6. Retroviral Vectors 7. Pharmacology & Toxicology 5:00 Conclusions and Summaries of the Breakout Sessions PLENARY TALK Dr. Alan E. Smith Senior Vice President Genzyme Corporation GENE THERAPY-WHAT IS IT? Direct use of nucleic acid vectors for any therapeutic purpose HOW IS IT DONE? Many applications -each is different ADA, cancer, CF, HIV, etc. In Vivo or Ex Vivo Various vectors retrovirus, adenovirus, AAV, cationic lipids Many genes wild type genes, cytotoxic genes, tumor suppresser genes, molecular decoys, ribozymes, etc. WHY ARE WE EXCITED BY IT? Simple, rational theory Broad application Potentially very powerful Initial, preliminary successes HOW IS IT GOING IN THE CLINIC? Adenosine Deaminase/SCID Retrovirus mediated transfer to T-cells - Sept. 1990 Long term improvement in patient immune function Selective advantage of transduced cells? Patients still on protein replacement therapy Very fortunate first choice Cancer Many successful experiments in animal models Animal data often hard to reproduce in humans Various studies in progress Mobilization of immune system Cytotoxic drugs with bystander effect Direct injection Primarily safety studies to date Cystic Fibrosis Adenovirus and lipid trials reported Aerosolization and repeat dosing in progress Safety data generally satisfactory but doses low Efficacy data modest Although the basic principles of gene transfer and gene function are established in several model systems, there is no unequivocal case of efficacy yet established in humans WHAT ARE THE SCIENTIFIC ISSUES? Identity of and access to target tissue and cells Efficacy of gene transfer and expression Inflammation associated with vector administration Duration of expression Efficacy of repeat dosing Clear cut clinical end points IDENTITY OF AND ACCESS TO TARGET TISSUE AND CELLS Identity and isolation hematopoietic stem cells - CD34+? Accessibility of airway cells/submucosal glands, mucocillary clearance, mucus layer Systemic targeting of tumor cells, organs Diseased tissue Knowledge of and access to target cells essential but often problematic EFFICACY OF GENE TRANSFER AND EXPRESSION Retrovirus vector transduction of human hemopoietic stem cells Adenovirus vector transduction of differentiated human airway epithelia Cationic lipid mediated gene transfer AAV transduction of most human cell types The potency of vectors, in terms of i.u./cell or molecules/cell required, needs to be improved INFLAMMATION Associated with liver, muscle and lung administration adenovirus Mediated by inactivated vector Associated with lung administration of cationic lipids Involves innate immune system (macrophage, neutrophils, NK cells, cytokines) Toxicity associated with repeated administration of both viral and non viral vectors may well become dose limiting DURATION OF EXPRESSION Retroviruses transcriptional shut off Adenovirus influenced by vector construct CTL responses )beware of transgene) Cationic lipids mechanism unknown Muscle, perhaps brain, appear exceptions Transient gene expression is caused by a variety of different molecular mechanisms REPEAT DOSING Efficacy of adenovirus re-administration limited Probably mediated by neutralizing Ab Likely barrier for all proteinaceous vectors Is possible using cationic lipid vectors Cationic Lipids and integrating vectors less problematic CLINICAL END POINTS CF - No obvious, readily measured clinical end point Cancer - Studies often involve terminal patients ADA - Additional ongoing therapies complicate interpretation Urgent need for better clinical measures of success and meaningful surrogate markers WHAT CAN BE DONE? Establish whether these are barriers in humans Fine tune amount and frequency of dosing Improve vector Co-administer other agents WHAT NEXT? Basic Science Enhance our knowledge of virology, immunology, gene expression, cell biology, etc. Novel vectors, cell culture and animal models Clinical Studies Animal data cannot predict human Human clinical studies essential Will drive formulation of future hypotheses WHAT ARE THE REGULATORY ISSUES? RAC review of individual applications discontinued FDA in pivotal role Safety paramount Facilitate collection of scientifically sound, interpretable human data Crucially important role in ensuring safety and timely accumulation of human data to test and to formulate hypotheses leading to eventual clinical success HOW IS IT GOING AT FDA? Regulations are rigorous, but not onerous Many issues negotiable-driven by data Review is timely Approach agency in spirit of collaboration not confrontation, staff as colleagues not bureaucrats CF ADENOVIRUS GENE THERAPY: GENZYME CORPORATION'S EXPERIENCE Vector/Trial Type Submitted Time for "Approval" Ad2/CFTR-1 (Nasal Trial) IND 4/2/93 3.5 months Ad2/CFTR-2 (Repeat Dose IND 12/10/93 2.5 months Nasal Trial) Ad2/CFTR (Lobar/Aerosol Major 11/10/94 <30 days Trial) Amendment Ad2/CFTR-8 (Vector Switch) Major 7/26/95 2 weeks Amendment CF CATIONIC LIPID GENE THERAPY Vector/Trial Type Submitted Time for "Approval" #67:pCF1 IND 12/5/95 <30 days 30 days included: Christmas New Year Governmental shut down Washington snow emergency GENE THERAPY - JULY 1996 Unusual case of drug development involving especially close collaboration between public and private sectors and between investigators and FDA Many constituencies: patients, scientists, clinicians, regulatory bodies, private sector, investors, media, advocacy groups, politicians Barriers to gene transfer identified in animals, more basic science and clinical studies essential Despite hurdles, great promise with undoubted contributions to medicine in next century Time frame of ultimate and widespread success hard to predict, important to manage expectation BREAKOUT SESSION 1: ADENOVIRAL VECTOR DEVELOPMENT SUMMARY Kathleen Hehir, Genzyme Corporation; Parris Burd, FDA/CBER; Robert Anderson/FDA/CBER Part I. Determination of Adenovirus Particle Number and Infectivity Bruce Trapnell discussed considerations for the determination of adenovirus vector particle concentration and infectivity. Enumeration of total virion concentration by optical absorbance was found to be very precise, but accuracy was dependent upon on physical disruption of the virion to eliminate artifacts of light scattering and was affected by the value used for the extinction coefficient. Biologic assays for enumerating infectious or functional virions, however, were highly dependent upon the conditions under which the assay was performed. Paul Shabram discussed an alternate method for particle determination which does not require a highly purified test article. He also presented data from infectivity modeling experiments which suggest that the true rate of Adenovirus infectivity may approach unity. This notion was reinforced further by data presented by Beth Hutchins who described a FACS-based infectious titer assay which showed that infectivity rates are consistently higher than conventional plaque assays would suggest. John Spalatro brought us back to earth and reminded us that no matter what methods are used, one must be mindful of the statistical considerations and limitations of the assay employed. Part II. Production Issues for Phase I and Beyond Our ultimate goal is to bring adenoviral-based products to licensure. A great many issues remain to be solved for this to occur. This session presented three approaches towards bringing a product to market and attempted to provide concrete examples of what we mean by the GMP continuum. Dominic Vacante presented an overview of the regulatory and manufacturing issues associated with developing Adenoviruses as products. These included selection and characterization of the cell substrate and seed virus, qualification of raw materials, methods of cell expansion, infection, harvest and purification as well as considerations for product formulation, filing, stability and final product characterization and lot release specifications. How far one must develop these concepts was then put into the context of different schemes of product development. Estuardo Aguilar described the approach of the Baylor College of Medicine Gene Vector Lab has taken to develop products intended only for proof of concept/safety studies in Phase I clinical trials. Jim Wilson of the University of Pennsylvania then described his approach for developing products through Phase II and early Phase III studies. A key component of this program is the emphasis upon the toxicology of vector development. Kathy Hehir described Genzyme Corporation's single sponsor approach for bringing Adenovirus-based products to licensure. The take home question is: WHEN THE TIME COMES WILL YOU BE READY FOR SUCCESS? AGENDA The goals of the session are to explore the different approaches for quantitating virus particles and their infectivity and to explore sponsors's approaches to the production and scale-up issues associated with moving products towards phases ii and iii and licensure. Questions to be addressed include: Is viral plaque formation the only way to evaluate virus infectivity? How does production scale-up affect particle infectivity? What are the hurdles to large scale Adenovirus production? PART I. DETERMINATION OF ADENOVIRUS PARTICLE NUMBER AND INFECTIVITY Considerations for the Determination of Adenovirus Particles and Infectivity Bruce Trapnell Rapid Identification of Encapsidated Viral DNA and Assessment of Viral Infectivity Paul Shabram New, More Sensitive Method to Assess Adenoviral Vector Infectivity Beth Hutchins Statistical Considerations for the Quantification of Adenovirus Infectivity John Spaltro Discussion and Summary PART II. PRODUCTION ISSUES FOR PHASE I AND BEYOND Manufacturing and Regulatory Issues for Developing Adenovirus Vectors as a Product Dominic Vacante Small Scale Production of Adenovirus Vectors Intended for Phase I Studies Estuardo Aguilar-Cordova Development of Adenovirus Vectors for PhaseII/III Studies in a University Setting James Wilson Single-Sponsor Development of Adenovirus Vectors Intended for Product Licensure Kathy Hehir Discussion Questions, comments, and requests for further information about this session may be submitted after the conference by e-mail to: GTINFO@A1.CBER.FDA.GOV CONSIDERATIONS FOR THE DETERMINATION OF ADENOVIRUS VECTOR PARTICLE CONCENTRATION AND INFECTIVITY Bruce C. Trapnell, M. D. Genetic Therapy, Inc., Gaithersburg, Md. Development of adenoviral vectors as therapeutic agents for a number of applications of in vivo human gene therapy has resulted in numerous preclinical and clinical studies. However, lack of standardization of the methods for quantifying the physical concentration and functionally active fraction of virions used in the various studies has often made comparison of data between studies difficult or impossible. In this context, several of the common procedures for quantifying adenoviral vectors have been extensively evaluated to define the variables which affect quantification of adenoviral vector concentration and bioactivity. The methods of evaluation of total virion (particle) concentration included electron microscopy and optical absorbance. The methods for evaluation of the concentration of functional (infectious) virions included detection of adenovirus-mediated gene transfer (transgene transfer and expression) and plaque assay on 293 cell monolayers. Enumeration of total virion concentration by optical absorbance was found to be very precise, but the accuracy was dependent on physical disruption of the virion to eliminate artifacts from light scattering as well as on the value used for the extinction coefficient. Both biological assays for enumerating functional or infectious virions were highly dependent on the conditions under which the assay was performed. Virion adsorption time and volume were particularly important; target cell density and amount of serum present during adsorption did not affect results significantly and multiplicity of infection was not important independent of its effect on vector concentration. Under optimal conditions, the bioactivity of the vector, defined as the fraction of total virions which leads to detected target cell infection, was determined to be 0.10 using the plaque assay and 0.29 using the gene transfer assay. This difference is most likely due to the fact that detection by gene transfer requires only measurement of levels of transgene expression in the infected cell while plaque formation is dependent on a series of biological events of much greater complexity. These results show that the exact conditions for determination of infectious virion concentration and bioactivity of recombinant adenoviral vectors are critical and must be standardized for comparability of data and for accurate assessment of bioactivity. These observations may be very useful in comparison of data from different preclinical and clinical studies, and may have important implications for how adenoviral vectors can optimally be used in human gene therapy. RAPID IDENTIFICATION OF ENCAPSIDATED VIRAL DNA AND ASSESSMENT OF VIRAL INFECTIVITY Paul Shabram Canji, Inc. Analytical Anion Exchange High Performance Liquid Chromotography of Recombinant Ad-5 Particles The expanding use of adenoviral vectors for gene therapy has brought about the need for new analytical tools. We have developed an Anion Exchange High Performance Liquid Chromatography (AEHPLC) method to analyze recombinant adenovirus serotype 5 samples. Before this assay available analytical methods consisted of either long term biological assays or required highly purified test articles. These methods were inadequate for optimizing adenovirus production and purification. The AEHPLC assay can be used to enumerate virus particles in either crude lysates or highly pure samples because it selects for virus by virtue of its surface characteristics. It can be used to assess particles in both dilute and concentrated samples over a wide dynamic range. Moreover, the population of virus particles eluted in the peak contains most of the infectious virions. The AEHPLC assay is a sensitive technique that overcomes the limitations of previous methods and provides an essential tool to accomplish process optimization. Since this assay can quantitate particles in a complex medium it can be used for studies in cell culture. In the infectivity study below AEHPLC was used to control for particle concentration. Are Recombinant Ad-5 Particles Mostly Infective? It is commonly held that the majority of adenovirus particles are non-infective. For gene therapy this belief has led to concerns that a dose of adenovirus would consist of mostly inactive and potentially toxic material. The industry standard is currently set to at least 1 active particle in 100 particles. Serial infection experiments, performed by others and ourselves, indicated that infectious titer assays used to calculate the "particle to infectious particle ratio" were misleading. Many events must occur before an infection can be detected. The first event, the collision of a particle with a cell, can be predicted on the basis of thermodynamic properties. Using Fick's Laws of diffusion a collision rate can be predicted; but the assay conditions that are typically used (plaque assay or TCID50 assay) do not allow for simple data analysis. Flow cytometry, however, is perfectly suited to allow experiments to be set up in such a manner that "semi-infinite solution" conditions apply. Under these conditions an equation can be derived that will predict the number of cells that have experienced a collision with an Ad-5 particle. The results we obtained using this scheme was surprising. Our data suggests that material possessing a particle to infectious unit ratio (TCID50) of @ 50:1 is no more than 2:1 after considering diffusion limitations. From this data we conclude that most particles are infective. NEW. MORE SENSITIVE METHOD TO ASSESS ADENOVIRAL VECTOR INFECTIVITY Beth Hutchins, Ph.D. Canji, Inc. A challenge in the delivery of a gene by an adenoviral vector is the preparation and accurate characterization of clinical dosage forms. Quality of dosage forms is evaluated using both biological and physical measurements. One critical biological property of a replication incompetent adenovirus is the infectious titer. This measure is used in combination with the particle number to assess the specific activity of an adenoviral preparation (particle number to infectious titer ratio). In addition, dosing has frequently been conducted using the infectious titer measurement. Two types of infectivity assays are traditionally used to determine the infectious titer of adenovirus preparations. The plaque-forming unit assay (pfu) is a quantitative procedure that scores viral plaques (complete cytopathic effect) as a function of dilution. The tissue-culture infective dose procedure TCID50) is a quantal assay that assesses infectivity as a function of intracellular staining for a viral antigen by direct immunofluorescence. Both methods suffer from limitations including a high degree of inter-assay variability and are affected by factors such as virus replication status, vector characteristics, and virus-293 cell interactions. We have recently developed a new, flow cytometry-based methods (FACS) to assess adenovirus infectious titer. This assay appears to be more sensitive and reproducible than the more traditional methods. The FACS method is a quantitative procedure that assesses infectivity as a function of the number of cells staining positively for a viral antigen. The FACS method is more sensitive to the events used to monitor evidence of infection than is either the pfu or TCID50 method. This results in infectious titers that are consistently higher than the titers determined using the pfu or TCID50 assays when assessing the same material. Our experiences also indicate that none of the assays determines the total amount of infectious material present. We performed serial infection experiments during which sample was transferred to fresh 293 cells after 15 minute incubations. Each set of exposed cells was assessed for evidence of adenovirus infection 48 hours post infection, with infectious titers calculated for each well. When these individual well titers were summed, the total infectious titer was greater than would be predicted using any of the infective titer assays discussed previously. The subsequently calculated particle number to infectious titer ratio was lower than the previously calculated ratio using the TCID50 or pfu assay titers. All of these results indicate that more particles are infectious than the traditional assays would suggest and that infectious titer should only be used as a secondary test for quality. Physical based methods, such as OD 260 nm in SDS, total DNA , or the Resource Q HPLC method, are powerful and reliable measures of virus concentration. Particle number measurements have low inter-assay variability and have been shown to be accurate through comparison with direct particle counts using electron microscopy. As a result, particle numbers to infectious titer ratio specifications are based on two methods with very different precision. The ability to set a specification for this ratio must be based on the reliability of the less precise method. Because of the limitations of infectious titer assays, the measure of adenovirus infectivity is best used, not as an absolute indicator of active virus, but as a relative assessment of virus batch quality. Given that clinical dosage forms are characterized by a variety of means, including specific activity based on transgene function and/or expression, the use of infectivity as a relative assessment of batch quality makes sense. By applying this strategy and understanding the limitations of infectious titer assays, a consistent yet reliable quality dosage form can be produced using set specification ranges by which the product can be evaluated for release. VIRAL SAFETY OF BIOTECHNOLOGY AND GENE THERAPY PRODUCTS+STATISTICAL REVIEW OF RCA TEST METHODS John Spaltro, Ph.D. Microbiological Associates, Inc. Viral safety and the risk of viral contamination are major issues in the manufacturing and control of biotechnology and gene therapy products. The potential for the occurrence of replication competent adenovirus (RCA) in replication defective gene therapy products requires that RCA testing be uniform. Statistical analysis of adenovirus titration and RCA test method data will be presented. Procedures required to determine the probability of detection (P. O. D.) for the RCA method, as well as, current limits of detection for the RCA methods will be described. Findings will be discussed in terms of a new perspective, using the assay P. O. D., to identify and compare allowable limits to currently defined 1 RCA per dose specification. MANUFACTURING AND REGULATORY ISSUES FOR DEVELOPING ADENOVIRUS VECTORS AS A PRODUCT Dominic Vacante, Ph.D. MAGENTA Corporation Ph: (301) 738-3938 Fax: (301) 738-1605 E-Mail: Dvacante@microbio.com Gene therapy clinical trials using adenoviral vectors were initiated in early 1993. Production involved the relatively straight-forward transfer of research processes to a GMP laboratory for small-scale manufacture of the vector. As adenovirus vectors move from phase I clinical trials to phase III and licensure, the challenges for manufacturing and regulatory groups will increase dramatically. An overview of current methodologies and regulatory guidelines will be presented with a view to the future. Specific areas of discussion will include: Selection and characterization of the cell substrate and seed virus Raw materials Cell Expansion Infection/harvest Purification Formulation, filing, and stability Product characterization and release specifications SMALL SCALE PRODUCTION OF ADENOVIRUS VECTORS INTENDED FOR PHASE I CLINICAL STUDIES Cassandra Nyberg and Estuardo Aguilar-Cordova Gene Vector laboratory, Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, TX 77030 In biologics, lot-to-lot identity is essential since the final product is complex and may not be tested in its entirety. Therefore, when considering the production of viral vectors for clinical studies two predominate variables must be taken into account: where and how the vector will be produced. The first refers to the physical characteristics of the production facility. The facility must be of sufficient quality to minimize potential irregularities from equipment or environmental sources. The "how" addresses the technical and regulatory procedures of the process. The technical aspects include the biological and physical variables of production which might affect the uniformity of the final manufactured product. In the case of adenoviral vectors these variables include the amplification process (cells and seed vector utilized, multiplicity of infection, volume of transaction, time of harvest, etc.), the purification process (isolation from cellular components), the storage conditions (buffer and temperature), and the vector characterization (quantity and potency). Understanding the effects of these variables of production can lead to increased yields and more predictable end-product characteristics. The regulatory aspects of the "how" include the quality assurance and quality control procedures established to ensure process and product purity, potency, and identity. We will discuss the approach the Baylor College of Medicine Gene Vector Laboratory has taken to address the issues mentioned above towards production of adenoviral vectors utilized in Phase I clinical studies. DEVELOPMENT OF ADENOVIRAL VECTORS FOR CLINICAL TRIALS AT THE UNIVERSITY OF PENNSYLVANIA James M. Wilson, M.D., Ph.D. University of Pennsylvania, Philadelphia, PA A fully integrated program from basic discovery to clinical trials has been established within the Institute for Human Gene Therapy at the University of Pennsylvania. The scientific foundation for this Institute lies in its faculty who participate in basic research relevant to the ultimate development of effective gene therapies. Over 150 faculty participate in research programs focused on cystic fibrosis, genetic diseases, cancer, cardiovascular diseases, and infectious diseases. An independent program in translational and clinical research has been established to facilitate the translation of basic discovery to human pilot experiments. Within the translational research program is our vector manufacturing laboratory, called the Human Applications Laboratory, as well as a program in toxicology. Critical to the implementation of appropriate preclinical studies in animals was the establishment of an animal service unit, which includes primate and rodent facilities, managed by the Institute staff. A clinical research program supports the investigator in the design and conduct of a clinical trial and interfaces with those support units necessary for the study to develop. Quality assurance and quality control for all translational and clinical trials is provided through the Regulatory Affairs, which is run by the Deputy Director of the Institute for Human Gene Therapy, Dr. Nelson Wives. We have learned that the production of clinical grade vector is only one part of a successful program in the development of human gene therapy. The characterization of the product and development of the protocol is highly dependent on the performance of a diverse array of preclinical studies to assess safety. During the last two years we have received IND approval for five clinical trials. Our experience has reinforced the importance of toxicology, not only in the initial application process, but during the conduct of the trial, when adverse events emerge or modifications in the vector are made. Several programs are rapidly proceeding through Phase I clinical studies with plans for Phase II studies underway. In anticipating the eventual success of one or more of these programs, we have considered issues relevant to the eventual licensure of the resulting products, if improvements clearly need to be made in the manufacturing and purification process of the vector. Our current experience suggests an anticipated cost of approximately $1,000,000 for vector production and safety testing through the preclinical phase to the completion of a phase I trial. Over 50% of this cost is in the manufacturing of the vector which clearly can be reduced with improved technologies. We are also addressing the role of contaminating chromosomal DNA in the final prep and the impact this will have on the eventual safety of the product. Finally, we believe it is essential that issues of long-term safety and the reproductive toxicology be addressed at an early phase before we are confronted with the licensure of a product. SINGLE SPONSOR DEVELOPMENT OF ADENOVIRUS VECTORS FOR PRODUCT LICENSURE Kathy Hehir Genzyme Corporation What are the key issues for the production of Adenovirus vectors for Phase III clinical trials and toward product licensure? New production scale-up and purification methods must be coupled with assays that are able to assess product activity, function, purity and equivalence to CsCl centrifugation methods. Batch sizes may be limited by testing requirements or assay sensitivity. Facility design that accommodates single products must also allow flexibility during product development. Formulation must address shelf-life stability as well as modes of drug administration. Product release specifications that pose challenges include RCA and host cell protein and DNA contamination. RCA assays must be able to detect low levels of RCA in a background of high levels of vector and the assays must be able to be validated. RCA is a patient safety concern but product specific limitation or allowances can be addressed if appropriate pre-clinical and clinical data is available. Current production and assay methods as well as planned development will be discussed. BREAKOUT SESSION 2: ANCILLARY PRODUCTS SUMMARY Joyce L. Frey-Vasconcells, FDA/CBER The first half of the breakout session discussed the feasibility and the type of testing that constituted a good qualification program for ancillary products. Dr. Joyce Francis presented a qualification program had been established at Genetic Therapy, Inc. and the rational behind the program. The general consensus after much discussion was that qual