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 qualification programs were a reasonable
mechanism for ensuring product quality. However, many participants voiced
concern over trying to acquire clinical grade material. Many times if a company
finds out that the client is using their product to manufacture a biologic the
company will refuse to supply the product and this impedes the progress of
clinical trials.
The second half of the session discussed the regulatory framework for approving
ancillary products and how such products could be labeled. Mr. Keith Gittermann
of Life Technologies Inc. discussed issues related to media products as
ancillary products.  Under the current regulations, ancillary products would be
regulated as a device but the device regulations for labeling require that if a
manufacturer has knowledge that the product is being used "off label", it is the
manufactures responsibility to provide adequate labeling to include the new use.
This is a liability concern for many manufacturers. Mr. Gittermann presented the
concept paper by the Association of Medical Diagnostics Manufacturers which has
been submitted to the FDA. This involves a very generic labeling claim and
groups related products into product families. A 510(k) would be submitted for
the product family with no clinical data. In addition, the manufacturer would
submit a master file to CBER for each product family. Mr. Mark Gauthier of
Immunex Corporation discussed quality aspects that ancillary products should be
analyzed for in order to maintain the quality of the final therapeutic product.
In addition, he discussed several existing regulatory mechanisms which could
potentially be employed to approve ancillary products. Finally, Mr. Gauthier
presented proposed labeling for ancillary products used in the manufacture of ex
vivo cell products. The last talk was given by Dr. Mark Hukkelhoven of Sandoz
Inc. He discussed the regulatory framework to license cytokines for ex vivo use.
He presented the advantages for sponsors to be able to obtain licensed ancillary
products. Dr. Hukkelhoven suggested that device regulations could be used and
that the labeling would be restricted to the ex vivo use of the product with no
claims on the final cell product.  The discussion focused on what data would be
reasonable for approval of an ancillary product. This intertwined with
discussions on how these products could be labeled to reduce liability for "off
label" use. The final question presented to the group was whether the FDA should
narrow its definition of ancillary products. This question was not discussed but
participants were encouraged to submit responses to the e-mail address for the
conference.
AGENDA
The goals of the Ancillary Product Workshop will be to evaluate the usefulness
of qualification programs for beginning studies and their effects upon the
sponsor. The second half of the workshop examines industry's perspective on the
regulatory framework for ancillary products.
PART I. QUALIFICATION PROGRAMS
    Introduction and Questions
    Joyce L. Frey, FDA/CBER
    Setting up a Qualification Program, Things to Think About!
    Joyce Francis, Ph.D., Genetic Therapy Inc.
    Discussion
PART II. REGULATORY FRAMEWORK AND LABELING
    Overview of Regulations
    Joyce L. Frey, FDA/CBER
    Ancillary Product Manufacturer's Issues and Recommendations,
    Keith D. Gittermann, Life Technologies Inc.
    Regulatory Framework for Ensuring Quality of Products and Potential Labeling
    Mark W. Gauthier, Immunex Corp.
    Sandoz Inc., How to License an Ancillary Product?
    Math Hukkelhoven, Ph.D.
    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
    INTRODUCTION TO THE BREAKOUT SESSION ON ANCILLARY PRODUCTS
    Joyce L. Frey, Ph.D.
    FDA/CBER
INTRODUCTORY QUESTIONS
    Are qualification programs a reasonable mechanism to ensure the quality of
an ancillary products?
    Are there additional tests which need to be performed?
    Are these programs reasonable for single sponsor investigations?
    Are there other alternatives for ensuring the quality of ancillary products
and ultimately of your final therapeutic product?
    REGULATORY FRAMEWORK
    If regulated as a device, 21 CFR 801.4 discusses the labeling for devices
and intended uses.
    If regulated as a device, for most products there is no predicate so a PMA
would have to be submitted.
    If regulated as a biologic, a BLA or PLA and ELA would have to be submitted.
    QUESTIONS FOR THE SESSION
    Are there other regulatory mechanisms for approval of ancillary products?
    Could these products be classified as either a Class 1, 2, or 3 device?
(Labeling regulation)  What would be the major concerns?
    How should an ancillary product be labeled?
    What data would be reasonable for approval of an ancillary product?
    A PLAN FOR CERTIFICATION OF GROWTH FACTORS FOR USE IN EX VIVO GENE THERAPY
    Joyce D. Francis, Ph.D.
    Genetic Therapy, Inc.
    Gaithersburg, MD
USE OF CYTOKINES
    retroviral vector-mediated gene transfer
    expansion of bone marrow cells
AVAILABILITY
    Limited Access
    Length Negotiations
    Potentially Costly
CERTIFICATION PLAN
    Facilitate initiation of clinical trials
    Protect proprietary products
    Less expensive
CERTIFICATION PLAN
    Identification of Vendors
    Handling and Lot Number control
    Lot Release Testing
    Inventory Control
IDENTIFICATION OF VENDORS
    Commercial supplier
    High purity
    Low endotoxin
    Custom production and filling
HANDLING
    Assign Internal Lot Number
    Quarantine
    Lot Release Testing
    QA Audit and Approval
    COA Issued
LOT RELEASE TESTS
    Identity
    Sterility
    Activity
    Purity
    Endotoxin
    Toxicity
    Residuals/Contaminants
INVENTORY CONTROL
    Request for shipment approved by QA
    COA attached
    Complete traceability
SUMMARY
    Plan is acceptable to FDA for Phase I/II Trials\Work on cytokine lots in
progress
    Use in trials scheduled for 1997, if results acceptable
    AN ANCILLARY PRODUCT MANUFACTURER'S ISSUES AND RECOMMENDATIONS
    Keith D. Gittermann
    Director of Regulatory Affairs, Life Technologies, Inc.
ABSTRACT
This talk will focus on the current regulatory issues facing today's
manufacturers of ancillary products for use in various ex vivo clinical
applications. Due diligence considerations restrict the ease of sales of
products for ancillary use in the gene and cell therapy arena. When considered
medical devices, these products cannot be knowingly sold for end uses for which
the manufacturer has not qualified them. Gaining FDA approvals through the PMA
process is not a reasonable option as there are a myriad of potential different
end uses which may need to be qualified. Using cell culture medial as an
example, we will discuss the various regulatory pathways available for a product
to travel down, from research uses to in vivo uses. We will discuss a possible
solution to the issue of off-label use which has been proposed by FDA and by the
Association of Medical Diagnostics Manufacturers.
DEFINITION OF AN ANCILLARY PRODUCT
    "Application of Current Statutory Authorities to human Somatic Cell  Therapy
Products and Gene Therapy Products"
    These products"...are intended to act on cells, rather than to have an
independent effect on the patient. Additionally, the intended action of these
products is not dependent upon incorporation into the somatic cell with
maintenance of the products' structural or functional integrity."
THE ANCILLARY PRODUCT MANUFACTURER'S DILEMMA IS 21 CFR PART 801.4 "MEANING OF
INTENDED USES"
    "...If a manufacturer knows, or has knowledge of facts that would give him
notice that a device introduced into interstate commerce by him is to be used
for conditions, purposes, or uses other than the ones for which he offers it, he
is required to provide adequate labeling for such a device which accords with
such other uses to which the article is to be put."
    In other words, this means a research use product or a simple 510(k) exempt
IVD can suddenly become a medical device requiring a 510(k) or more likely a
PMA.
    This is both impractical and not economically feasible when one considers
the large number of unrelated end uses in which ancillary products are used.
Each use could require a new submission.
    Cell Culture Products are good examples of a product group whose intended
use is largely research, but for which numerous other uses have evolved; some of
which don't meet the requirements of 801.4
    Products include:
    Cell and Tissue Culture Media
    Balanced Salt Solutions
    Human and Animal Sera
    Reagents
        Essential for the growth, survival, and development of cells of human
and animal origin. Some products are used as a wash or holding solution for
cells and tissues, and some products are used as supplements to media. 50-80% of
these product's uses are in research setting for nonmedical purposes which are
not regulated by FDA.
THEY ARE USED IN CLINICAL DIAGNOSTIC PROCEDURES TO SUPPORT CELL CULTURES FOR
VIRAL ISOLATION
    Class 1 medical devices
    Exempt from premarket notification 510(k) procedures
    Device establishment registration and device listing are required
    Must comply with current Good Manufacturing Practices as described in 21 CFR
Part 820
    These are products specifically intended and labeled for sperm
washing/handling procedures and specimen transport procedures.
    These uses are subject to 510(k) clearances.
    Utilized as raw materials for the further manufacture of monoclonal
antibodies, vaccines, and biologic therapeutics.
    The cell culture media is not directly regulated by FDA.
    Drug or device master files are, on occasion, submitted to FDA.
USED BY PHYSICIANS IN BIOMEDICAL APPLICATIONS REQUIRING INVESTIGATIONAL NEW DRUG
(IND) OR INVESTIGATIONAL DEVICE EXEMPTION (IDE) SUBMISSIONS MADE BY THE END
USERS
    These applications include such procedures as ex vivo cell culture,
tissue/organ suspension for transplantation, and hematopoietic stem cell
manipulation.
    For some uses, drug or device master files have been submitted to FDA by the
media manufacturer to support end user's submission.
USED BY PHYSICIANS ON OTHER BIOMEDICAL APPLICATIONS FOR WHICH THE END USER IS
NOT REQUIRED TO SEEK FDA CLEARANCE
    These applications include bone marrow transplants, in vitro fertilization
and related procedures, artificial skin production, and some transplantation
procedures.
    Per 21 CFR Part 801.4, these last two applications could be interpreted to
require the manufacturer of the ancillary product to seek a Premarket Approval
Application (PMA) for each end use oaf which they are aware.
    Over the past few years, the number of new and different types of
"off-label" uses have proliferated while other "off-label" uses have become the
standard of care in some fields.
    Currently each manufacturer must develop its own individual policies for
selling products for these end uses.
    To allow meaningful dialog with end users, to level the playing field
between manufacturers with different policies, and to reduce regulatory and
liability risks to the manufacturer, some sort of reasonable guidelines are
needed.
SOLUTION ZERO
    Consider cell culture media a raw material for use in further manufacture.
It is the end user's responsibility to have appropriate specifications in place
and to qualify the material for his end use. Sale for any application under
current research use or in vitro diagnostic use labeling allowed.
SOLUTION ONE
    Submit master files for product groupings with known off-label uses.
    Notify FDA when sales are made to end users without an IND/IDE
    801.4 still a problem because product will not be labeled or qualified for
the end user's intended application
SOLUTION TWO
    Association of Medical Diagnostics Manufacturers (AMDM) concept paper for
regulation of tissue culture media products
    Tissue Culture Sub-committee comprised of:
        BioWhittaker, Inc.
        Hyclone Laboratories, Inc.
        Irvine Scientific
        JHR Biosciences
        Life Technologies, Inc.
    Provides for a generic label claim which we believe would meet the
requirements of 21 CFR Part 801.4 "For cell and tissue culture processing"
        Related products with known history of "off-label" use grouped into
products families
        Submit master file for each product family
        Notify FDA of sales to known end users with no IND/IDE
        For more specific label claims, submit 510(k) or PMA as appropriate
We are ready to move forward to resolve these issues which have been plaguing
the industry since the mid 1980's. We are prepared to commit resources to work
with FDA in designing a reasonable and equitable solution.
    REGULATORY FRAMEWORK FOR ENSURING QUALITY OF PRODUCTS AND POTENTIAL LABELING
    Mark W. Gauthier
    Immunex Corp.
ABSTRACT
Immunex will present a framework for the regulation of ancillary products, for
ensuring the quality of these products (including the applicability of cGMPs,
safety from infectious agents, specifications, and stability testing), and their
labeling to distinguish them from reagents or drug products that may already be
approved or are under investigational use for in vivo treatment.
REGULATORY FRAMEWORK FOR ANCILLARY PRODUCTS
    Quality Aspects
    Registration
    Conclusions
    Labeling
ANCILLARY PRODUCTS FOR MANUFACTURE OF EX VIVO CELL PRODUCTS
IMMUNEX CORPORATION
    Develops cytokines/growth factors for in vivo therapeutic use
    Provides cytokines/ growth factors for use as ancillary products in the
manufacture of ex vivo cell products
ANCILLARY PRODUCTS FOR USE IN THE MANUFACTURE OF EX VIVO CELL PRODUCTS *
    Not part of the final stem cell product; intended to act solely on the cells
 and not on the patient
    May have an impact on the safety, purity, or potency of the final stem cell
product
    Subject to either drug or device CGMP's
    *From "Draft Document Concerning the Regulation of Peripheral Blood
Hematopoietic Stem Cell Products Intended For Transplantation or Further
Manufacture Into Injectable Products", February 1996
REGISTRATION PROCESS
    Well defined for recombinant DNA products for in vivo therapeutic use
    Not defined for cytokines/ growth factors used as ancillary products
    Issues:
    Quality
    Drugs or device
    Registration process
    Labeling
REGULATORY FRAMEWORK FOR ANCILLARY PRODUCTS
    Quality:
    CGMP's should be applied
        ie: drug vs. device
    Safety from infectious agents should be demonstrated
        ie: viral inactivation/removal
    Specifications should be established
    Product should be stable
    Registration
    Labeling
PRODUCT CONTROL STRATEGY
    Material should be manufactured in compliance with GMPs and includes:
    Process validation
    Raw material controls
    In- process controls
    Bulk drug substance release testing
    Product release testing
    Stability evaluation
SPECIFICATIONS
    Appropriate release specifications should be established to ensure the
safety and quality of the ancillary product. However, these may not need to be
as rigorous as those for a licensed parenteral product
    Cell Culture Reagent --->   Ancillary Product --->Licensed Parenteral
Product
    Bovine Transferrin
    Quality
        Appearance- off white lyophilized powder
        Solubility- soluble at 1% in water
        pH- 4.0 - 5.0
        Residual moisture  5.0%
    Purity
         98%
        Iron content
    Strength
         95%
    Safety
        Endotoxin < 10 EU/mg
        Bioburden < 10 CFU/mg
        Adventitious viral agents - none detected
        Mycoplasma
        Cytotoxicity
    IGF-1
    Quality
        Appearance
    Identity
        MW 9110 daltons (by mass spectrometry)
        NTS > 95% single sequence
    Purity
        >70 %(by HPLC)
    Potency
        ng/ml (by L6 myoblast stimulation)
    Safety
        Endotoxin < 0.1 EU/g
    Sargramostim (Leukine)
    Quality
        Appearance
        pH
        Moisture
        Content uniformity
        Reconstitution Time
    Purity
        Protein content
        SDS-PAGE
        RP-HPLC
        SE-HPLC
     Identity
        Peptide map
        Western blot
    Strength
        BCA
    Potency
        TF-1 Bioassay
    Safety
        Endotoxin
        Sterility
        Particulate Matter
    Extensive testing conducted on the bulk drug substance
PROPOSAL FOR ANCILLARY PRODUCT SPECIFICATIONS
    Identity - Single identity test
    Purity - Some product related impurities may be acceptable
    Strength/potency - in-vitro bioassay required
    Safety - product should not pose safety risk to process or patient
    (ie, sterility, endotoxins, viral contaminants)
    Stability testing: Perform specification tests at 0, 6, 12, 24 months at
intended storage condition.
    Establish minimum testing requirements for release:
    Identity ( One test sufficient)
    Peptide Map, Western Blot, HPLC
    Electrophoresis, N-Terminal Sequence
    Analysis, Mass Spectrometry
    Purity ( One test sufficient)
    Chromatographic methods, SDS - PAGE
    Strength (One test sufficient)
    HPLC, UV Spectroscopy, Amino Acid Analysis
    Colorimetric redox methods
    Potency (one test sufficient
    In vitro Bioassay, Receptor Binding Assay
    Safety
    Bioburden
    Endotoxin
    Host cell DNA*
    Viral agents*
        Demonstrate removal through process validation
SAFETY
    Adventitious viral agents
    Viral contamination could ultimately compromise patient safety
    Viral inactivation/ removal should be demonstrated
    Host cell DNA
    The removal should be demonstrated through the validation of the
manufacturing process
    Endotoxins
    Bioburden
APPROACHES FOR REGULATION
    As in vitro reagents
    As recombinant DNA products for human use
    As "Manufacturing Materials" subject to class I, II, or III device
designation
    As "Bulk Drug Substances" subject to drug regulation (21 CFR 314)
    As "Critical Reagent"
MANUFACTURING MATERIALS - TWO OPTIONS
    CFR 864.2240 cell and tissue culture supplies and equipment
    Devices used to examine, propagate, nourish, or grow cells and tissue
cultures - Class I devices
    CFR 820.60 (d) Manufacturing Material
    Any substance used to facilitate a manufacturing process and which is not
intended to be included in the final device
    Must be removed from the final product or limited to a specified amount,
with written procedures for use and removal
DEVICE REGULATIONS
    Possible mechanisms for registration of ancillary products under the device
regs exist
    GMP's less stringent than for drugs
    Labeling requirements may allow diversion
    May need additional safeguards to assure that patient safety and integrity
of ex vivo cell product will not be compromised
DRUG SUBSTANCES
    Master file required
    Manufacturer subject to FDA inspection
    Not formally subject to CGMP's, but the current trend is toward increasing
applicability
    Change to alternate source of drug substance requires demonstration of
equivalence & prior FDA approval
Master files are not officially "approved" but are reviewed and deficiencies
must be corrected when referenced by an application
CRITICAL REAGENT
    Certificate of analysis needed
    Reference to Master File or IND for method of manufacture
    "Approved" in the context of the product of which the critical reagent is a
component
RESPONSIBILITY OF EX VIVO CELL PRODUCT MANUFACTURER
    Change in ancillary product supplier would require
    In vitro demonstration of equivalence in transduction efficiency
    Prior FDA approval
    Demonstration of removal of ancillary product from cell culture via highly
sensitive assay (ie, ELISA, etc.)
CONCLUSION: PROPOSED REGULATORY FRAMEWORK
FOR ANCILLARY PRODUCTS
    Recommend hybrid system
    Submission of Master File/ IND for method of manufacture
    Subject to drug CGMP's
    Viral inactivation/ removal required
    C of A required
    Other safety tests (sterility, endotoxins)
LABELING
    We recommend use of the following statement for ancillary products:
"CAUTION: Ancillary Product for use in the manufacture of ex vivo cell products.
Not for parenteral use."
ANCILLARY PRODUCTS CURRENTLY MANUFACTURED BY IMMUNEX
    GM-CSF (Sargramostim, LEUKINE) - licensed product
    stimulates macrophages and granulocytes
    PIXY 321 (Milodistim, PIXYKINE) - IND
    stimulates platelets, macrophages and granulocytes
    Flt3 Ligand - IND
    mobilizes peripheral blood progenitor cells.
    LICENSURE OF CYTOKINES FOR EX VIVO USE
    Mark Hukkelhoven, Ph. D.
    Sandoz Research Institute
    East Hanover, NJ
CYTOKINES ARE NECESSARY FOR THE EX VIVO
    Facilitation of retroviral vector-mediated gene transfer
    Expansion of hematopoietic cells
As such they are used in most ex vivo gene therapy protocols and ex vivo
expansion studies
Since they are intended to act on the cells and are not intended to have an
independent effect on the patient, they are regarded as "ancillary products"
HOW TO CURRENTLY OBTAIN AND USE THEM
    Own manufacture (non-GMP) with release specifications
    Buy as reagent, release them internally (develop certification plans)
    If IND exists: obtain product and get cross-reference to IND from sponsor
DISADVANTAGES OF CURRENT PRACTICE
    Companies/investigators need to develop own certification plan
    Access to some growth factors is difficult
    Non-GMP grade material used (adventitious agents)
Potential solution for current practice would be to offer cytokines licensed for
ex vivo use...
ADVANTAGES
    General access to an off-the-shelf product (both in clinical development and
 once a gene or cellular therapy product is licensed for therapeutic use)
    no internal release testing
    consistent GMP grade quality
CONSIDERATIONS
    If available, a licensed cytokine must be the preferred source for ex vivo
protocols, especially in Phase III and certainly once a gene or cellular therapy
product is licensed for therapeutic use
    if not available FDA should allow an internal certification plan
REGULATORY FRAMEWORK
    Medical Device Regulations would be an appropriate, and the preferred,
framework
    If conceptually difficult, licensure as a "well characterized
biotechnological product" could be considered
LABELING
The indication for a licensed cytokine would be restricted to the ex vivo use
of the product without claims on the reinfused cells:
    Cytokine X is indicated for ex vivo use to facilitate:
    retroviral-mediated gene transfer into somatic cells (increase of
transduction efficiency) prior to reinfusion of manipulated cells
    expansion of hematopoietic cells and their precursors prior to reinfusion of
the expanded cell population
    Cytokine X is not approved for direct in vivo use
    The safety and efficacy of the generated gene therapy products or expanded
cell population has not been established
Principle: Allow and promote the availability of licensed cytokines for ex vivo
use without unnecessarily constraining the scientific field
PROPOSED DATA REQUIREMENTS
    CMC
    Full CMC section with some flexibility to be allowed for:
    specs for release of drug substance and drug product (purity)
    stability program
    shorter description of manufacturing and test methods
    In Vitro Pharmacology
    documenting the activity of the cytokine in the ex vivo process
    Pre-clinical safety
    limited amount of pre-clinical tox data, e.g., acute tox study in
appropriate species (also to cover overdoses)
    no clinical data (on cytokine and reinfused cell product
       No ELA
CONCLUSION
Proposal provides easy access to consistently high quality cytokines without
compromising developments in science and medicine in this area
    BREAKOUT SESSION 3:
    FACILITIES AND MANUFACTURING "THE GMP CONTINUUM"
    SUMMARY
    Jay Eltermann, R.Ph., M.S.
The breakout session on facilities and manufacturing was designed to be more
educational rather than address specific problems. In this aspect we were
somewhat different than most of the break-out sessions in this conference. We
welcomed the opportunity to discuss facility and manufacturing issues, and
receive comments to determine where additional guidance is still needed. The FDA
presentations focused on several aspects of the design and operation of gene
therapy facilities. We started with an overview of the regulatory aspects, what
are GMPs and how they could be applied to manufacturing gene therapy products.
We also went over some of the concerns we had for these facilities, with
recommendations for design and operation of transduction and viral vector
facilities. There were presentations regarding the specifics behind the design,
operation and maintenance of a facility designed to manufacture gene therapy
products. We understand that early clinical manufacturing will not be in
compliance with full GMPs,. but attempts should be made to be in the spirit of
GMPs. In keeping with the GMP continuum, we had presentations on academic and
commercial scale facilities. They gave us an excellent overview of their
facilities, important manufacturing issues, and some of the problem areas they
have encountered. It was important to hear the industry's perspectives on these
issues.
In summary, the following points were made during the breakout session.
    We propose and support a sliding scale approach to meeting GMPs for the gene
therapy facilities. We recognize that there are going to be differences in what
a Phase I and commercial facility will be able to achieve with respect to
process control and system validation.
    We extended an invitation to manufacturers to discuss issues early in the
process.
    We request comments regarding the type and scope of guidance documents
needed for gene therapy facilities.
AGENDA
The goal of this breakout session is to provide an overview of the design and
operational features that are recommended for facilities intending to
manufacture gene therapy products. Specific topics will include an overview of
the regulatory framework for the  manufacture of these products, HVAC systems,
environmental monitoring, equipment and validation issues, QC/QA oversight and
appropriate documentation. Speakers from academia and industry will present the
design, operation and manufacturing issues of these facilities, ranging from
small Phase I facilities to commercial scale.
There will be two panel discussions to address questions that arise during the
presentations.
PART I.  FDA PERSPECTIVE
    Introductions
    Jay Eltermann, R.Ph., M.S.
    Regulatory and GMP Considerations for Gene Therapy Facilities.
    Jay Eltermann, R.Ph., M.S., FDA/CBER, Division of Establishment Licensing
    Facility Design Considerations.
    Robert Sausville, FDA/CBER, Division of Establishment Licensing
    Operational Aspects.
    Mary Malarkey, FDA/CBER, Division of Establishment Licensing
    Questions for the Panel.
PART II. ACADEMIC AND INDUSTRY PERSPECTIVES.
    Gene Therapy Initial Manufacturing Facility.
    William R. Tolbert, Ph.D., WR Tolbert & Associates
    University of Michigan Human Application Laboratory, Transition from GLP to
GMP: An Academic Perspective.
    Blake Roessler, M.D.
    Genetic Therapy Inc., Considerations and Strategies in the Planning and
Design of a New Manufacturing Facility.
    Victor Santamarina, Ph.D.
    Questions for the Panel.
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
    REGULATORY AND CGMP CONSIDERATIONS FOR GENE THERAPY FACILITIES
    Jay Eltermann, R.Ph., M.S.
    Division of Establishment Licensing
    CBER/FDA
ABSTRACT
Manufacture of products intended for human use, including products for clinical
trials, should be performed according to current Good Manufacturing Practices
(cGMP). This provides assurance that there is consistency between lots and that
adequate controls are in place to assure the safety, efficacy, and potency of
the product. The principles of cGMPs apply to the manufacturing process as well
as the facilities and equipment in which products are product. This presentation
will be an overview of the applicable regulations and guidance documents, with
emphasis on those relating to equipment and facilities.
REGULATORY AUTHORITY
    Federal Register Notice of 10/14/93, Application of Current Statutory
Authorities to Human and Somatic Cell Therapy and Gene Therapy Products"
    Regulated as biological products (CBER)
    Clinical trials conducted under IND
    Methods of manufacture and facilities conform with CGMP regulations
    Commercial ventures subject to licensure
    RELEVANT GUIDANCE DOCUMENTS
    CFR 211 (facilities, records, personnel)
    CFR 600s (facilities, personnel, live viral vaccines)
    Guidelines (records, validation, testing)
        Aseptic processing
        Preparation of Investigational New Drug Products
    CDC/NIH booklet and RAC guidelines (facility design, control, special
practices)
CURRENT GOOD MANUFACTURING PRACTICES (CGMPS)
    DEFINITION: "A set of current, scientifically sound methods, practices or
principles that are implemented and documented during the development and
production to ensure consistent manufacture of safe, pure and potent products."
    Applies to the process as well as the facilities
GMPS VS GLPS
    Good Laboratory Practices (GLPs) are not Good Manufacturing Practices (GMPs)
    GLPs (21 CFR 58) cover nonclinical facilities - testing, animal facilities
    GMPs cover facilities manufacturing products for human use
    Similar aspects covered by both sets of regulations (records, procedures,
quality control)
    Quality of materials, type of records, manufacturing controls are different
PRINCIPLES ON WHICH CGMPS ARE BASED
    Quality, safety, effectiveness must be designed and built into the product
    Quality can't be inspected or tested into the product
    Each step in the manufacturing process must be controlled to ensure final
product meets specifications
    Documentation provides evidence that compliance with cGMPs has been met
CONTROL OF PRODUCTION STEPS FROM START TO FINISH
    Vector development
     Cell bank characterization (also patient cells)
    Final product characterization
    Production components (ancillary products)
    Testing (analytical, environmental, product)
INFLUENCES ON FACILITY DESIGN AND OPERATION
    Production of vectors (single vs multiple)
    (viral vs non-viral)
    Transduction of patient cells
    Growth/harvest of transduced cells (Cell bank vs Patient cells)
     Multi-use scenarios
    Campaigning vs concurrent production
CONCERNS COMMON TO ALL BIOLOGICALS
    Sterility, endotoxins, identity, purity, potency
    Regulation of both the product and the process
    Reproducibility/consistency between product lots
PRODUCTION CONCERNS
    Characterization of final gene product, whether vector containing
supernatant or transduced cells
    Appropriate test methods & validation
    QC/QA oversight
    Worker/environmental safety
KEY CONCERNS WITH GENE THERAPY FACILITIES
    Cross contamination
    Generation of aerosols
    Spills
    Campaigning within same area
    Simultaneous production
    Potential for mix-ups
    Segregation (clean vs dirty)
    Labeling
        at various production stages
        (n different areas/rooms
    CHANGE OVER PROCEDURES
    Cleaning/inactivation
    Proper area clearance and storage
    Regowning
    Control of the facility/process
    Production area separate
    Equipment dedicated for production
    Restricted access
    Proper maintenance/surveillance
    Documented production/testing
TRANSDUCTION AREAS: OVERVIEW
    Open manipulations performed in a biosafety cabinet (Class 100 conditions)
    HEPA-filtered air/Class 100,000 for manufacturing areas
    No containment features unless steps involve generation of aerosols/viral
vectors
    Hoods cleaned between cell lines
    Personnel appropriately attired
    sterile gloves/mask with open steps
VIRAL VECTOR PREPARATION AREAS: OVERVIEW
    HEPA-filtered air with containment features- modules or facility
    Double-door entrance to facility, usually and air-lock
    One vector per contained area at a time
    Open steps performed in biosafety cabinets
    Clean area between vectors
    Extra gowning in vector prep areas
    Change of gowning between vectors
IN SUMMARY
    CBER has regulatory oversight for gene therapy products
    GMPs are applied, even for clinical trial materials
    Control process from start to finish
    Meet with FDA early to discuss facility, manufacturing, testing or clinical
trials
    FACILITY CONSIDERATIONS
    Robert Sausville
    Division of Establishment Licensing
    CBER/FDA
ABSTRACT
The design of facilities should be based on their intended use. Materials of
construction, Heating Ventilation and Air Conditioning (HVAC) systems; both for
containment and product protection; gowning practices, multi-use issues and
environmental monitoring programs will be discussed, with an emphasis on vector
preparation and ex vivo transduction of patient's cells. Requirements for
maintaining appropriate Biological Safety Level will also be addressed.
GENERAL CONSIDERATIONS
    Designed for aseptic processing
    Smooth surfaces, seamless tile, etc.
    Easily cleanable and impervious finishes
    HEPA filtered air from separate air handling unit
    separation of production from other areas of the facility (often a hospital)
    Class 100,000: general manufacturing areas
    Class 100: all open manipulations
    Separate entrance/gowning area
    Air locks (containment)
    Room(s) for support areas (buffer, media prep.)
STORAGE AREAS (RAW MATERIALS AND CELL BANKS)
    Material and personnel flows designed to maximize efficiency and minimize
mix-ups
    also controlled by procedures
    unidirectional flows where possible
    Concurrent vs. campaigning
    Segregation of:
        HVAC and personnel
    Changeover procedures in place
APPROPRIATE BIOSAFETY LEVEL
    BioSafety Level 2
    Transduction
    BioSafety Level 3
    Vector Preparation
BIOSAFETY LEVEL 2
    access limited
    personnel trained in handling pathogenic agents
    infectious wastes are decontaminated before disposal
    gowning required
    gloves should also be worn if skin contact potential
    Class I or II BioSafety Cabinets to be used:
    for procedures potentially creating aerosols
        aerosol generation should be minimized
    with high concentrations or large volumes of infectious agents
BIOSAFETY LEVEL 3
    Negative pressure or "sink" for containment
    All activities with infectious materials are conducted in biological safety
cabinets
    Class I, II or III may be used
    Passage between two sets of doors is a basic requirement
    An autoclave for decontaminating waste is available
    preferably in the laboratory
    Ducted exhaust provided
    not recirculated
    may be discharged to the outside without being filtered
BIOSAFETY LEVEL 4
    Class III biosafety cabinets
    or personnel in suits with life support systems
    All materials must be autoclaved before leaving BSL4 area
    Exhaust air HEPA filtered
AIR QUALITY
    Recommend that production areas receive single pass air (no recirculation)
    Dedicated air handler where possible
    HEPA-filtered air
    In-line vs. terminal
    Objective: to meet Class 100,000 specifications
    Air pressure differentials between areas
    Positive (aseptic processing)
    Negative (containment) for steps needing greater than BSL-2
    Open steps in biosafety cabinets (Class II)
    Monitored to assure facility is acceptable for production
ENVIRONMENTAL MONITORING
    Demonstrate facility under control
    as part of validation
    Evaluate periodically
    routine monitoring program
    Viable/non-viable particulates and surfaces
    specifications based on desired Class
    Viable particulates
    active air monitoring devices
    settling plates
    Non-viable particulates
    particle counters
        certification of biosafety cabinets and rooms
    Surface "contact" plates or swabs
    monitor cleaning efficacy and personnel asepsis
NASA STANDARDS: VIABLE AIR PARTICLES
    Class 100,000
    2.5 CFU/ ft2
    Class 10,000
    0.5CFU/ ft2
    Class 100
    0.1CFU/ ft2
SETTLING PLATES: EXPOSURE TIMES
    Class 100,000
    1 CFU/ 9cm plate
        0.11 hours
    2 CFU/ 9cm plate
        0.21 hours
    1 CFU/ 14cm plate
        0.04 hours
    2 CFU/14 cm plate
    0.09 hours
    OPERATIONAL ASPECTS
    Mary Malarkey
    Division of Establishment Licensing
    CBER/FDA
ABSTRACT
The "sliding scale" approach to equipment and process validation, with respect
to the GMP continuum, will be discussed. Other operational considerations
including handling and testing of raw materials, personnel training and
responsibilities, functions of the Quality Control/Quality Assurance unit
(QC/QA), and the importance of proper documentation will also be presented.
EQUIPMENT
    Certification
    Calibration
    Validation
EQUIPMENT AND PROCESSES
    Program in place to demonstrate that equipment or process operates as
expected
    Should include periodic monitoring
    Air handlers
    Biological safety cabinets
    pressures
    filter integrity
    airflows; velocities
    leak testing
    Incubators
    uniform temperature
    carbon dioxide
    filters
    Centrifuges
    speed
    vacuum (ultras)
    temperature
    Autoclaves
    temperature
    pressure
    kill cycle
    Lyophilizers
    shelf temperature
    vacuum
    PROCESS
"SLIDING SCALE " APPROACH: AN EXAMPLE
    Autoclave used to prepare sterile materials
    Early (Phase I/II)
        demonstrate proper cycle achieved
        monitor temperature, pressure, and time
        use of biological indicators for verification
        loads not well defined
    Middle (Phase II/III)
        temperature mapping done to determine cold/hot spots
        biological indicators placed to verify cycle at problems points
        loads are somewhat defined
    Late (Phase III+)
        lethality of cycle determined at monitored points
        loads are well defined and standardized
        each load configuration has been mapped or worst-case load has been
validated
PROCESS VALIDATION
    Sterilization
    Decontamination
    Aseptic Processing
    Cleaning
    Inactivation/removal of adventitious agents and other contaminants
RAW MATERIALS
    Critical raw materials - established criteria for acceptance from vendors:
    sterility
    adventitious agents
    activity/purity
    Vendor's Certificate of Analysis
    identity test where possible
    Inventory Control
    proper storage
    FIFO
WATER
    Should meet Water for Injection (WFI) specifications :
    microbials <10 CFU/100ml
    endotoxin <0.25 EU/ml
    chemical tests per USP
    WFI for all product contact surfaces and formulations
    May be purchased
PERSONNEL
    Designated person in charge of facility
    Responsible for:
    limiting access
    training
    maintenance/safe operations
    writing and enforcing procedures
    Production personnel are trained (periodic retraining)
    Appropriately gowned for production step
WRITTEN PROCEDURES
    Develop and implement standard operating procedures (SOPs)
    use of equipment
    decontamination and cleaning
    proper handling
    Documentation in the form of
    batch records
    log books/recording charts
    quality control test records
QUALITY ASSURANCE AND QUALITY CONTROL
    QC "unit" to pass or reject:
    raw materials
    in-process materials
    final "product"
    QA "unit" to review
    batch records
    test results
    environmental results
    evaluate effectiveness of current process/SOPs
FINAL THOUGHTS
    Design facility for worst-case = maximum flexibility
    Consider filing a Master File for facilities handling several IND products
Meet with FDA to discuss your Phase I/II (or III!) facility.
    GENE THERAPY INITIAL MANUFACTURING FACILITY
    William R. Tolbert, Ph.D.
    WR Tolbert & Associates
ABSTRACT
New gene therapy advances most often take place in academic institutions and
early stage biotech companies. Transformation of these research break-throughs
into approved new therapies for the treatment of human disease is a long an
arduous task. One of the first road blocks is the bringing of research processes
and facilities into compliance with Current Good Manufacturing Practices
(CGMPs). Change, creativity and continual improvement are hallmarks of basic
research, while safety, consistency, emphasis on process control, documentation
and resistance to change are essential to ensure comparability of products from
the clinic to commercial production. There is a real difference in philosophy
that makes conversion of research process to biopharmaceutical manufacturing
particularly difficult for research based institutions and scientists.
One of the first steps in the course of CGMP conversion is design of an initial
manufacturing facility for preparation of clinical material to be used in human
trials. This presentation will describe come of the basic facility requirements
for production of viral vectors for either in vivo or ex vivo applications.
Support facilities will also be addressed.
GENE THERAPY FACILITY
    Research vs. CGMP Requirements
    Required Functions
    Facility Planning
    Aseptic and Contained Processing
    Facility Design
    Flow Patterns
RESEARCH VS. CGMPS
    Philosophical Approach
    Research - Change, Improved Methods, New Knowledge
    CGMPs - Purity, Safety, Efficacy, Consistency
    Practical Considerations
    Research - Decision at Level of Researcher
    CGMPs - Regulation Compliance, Documentation, No Major Operator Decisions
REQUIRED FUNCTIONS AND AREAS
    Receiving / Quarantine Storage
    Released Materials Storage
    Quality Control Laboratory
    Cell Bank
    Aseptic / Contained Processing Laboratories
    Mechanical Space
    Quality Assurance / Documentation Area
FACILITY PLANNING
    Process Definition / Scale of Operations
    Vector Production - In vivo / Ex-vivo
    Patient Cell Transduction
    Number of Products / Patients
    Vector Type / Level of Containment
    Clinical Phase I, II, III / Initial Commercial
    Internal Institutional Applications or External Sale of Products
    Limitations of Space, Personnel, Financial Resources - Maximize Flexibility
    Renovation of Existing Laboratories Areas or New Construction
    Autoclave and WFI or Pre-sterilized Single Use Disposables and Prepared
Medium
    Stick-Built or Modular Construction
    Schedules for Construction/Validation/Use
ASCEPTIC/CONTAINED PROCESSING AREAS
    Class 100,000 HEPA Filtered, Single Pass Air - Recommend Low Filtered Air
Returns
    Defined Air Flow and Pressure Differentials
    Aseptic Areas - Positive Air Pressure
    Contained Areas - Negative Air Pressure
    Class II, Type B Biological Safety Hoods
    Seamless Welded Flooring with Covering
    Solid Ceiling with Sealed Lighting
    Epoxy Resin Wall and Ceiling Coating
    All Penetrations Sealed (Silicone Caulk)
    No Personnel Access Between Vector Production and Transduction Areas
    Controlled Access - Recommend Card Sys
    Emergency Power for Critical Systems
    Recommend Interlocking Gowning Room Doors and Interlocking Pass Throughs
    Recommend Stainless Steel Work Surfaces and No Cabinets, Drawers, Sinks or
Drains
    Recommend Use of Dedicated Rolling Carts for Material Transfer
    Recommend Full Coverage Single Use Disposable Gowning
FACILITY DESIGN EXAMPLE
    Two Vector Production Laboratories
    Two Transduction Laboratories
    No Autoclave - Pre-Sterilized Disposables
    Access from One Side - General Use Hall
    Mechanical Space Available in Penthouse
    Manufacturing Space - 1800 Sq. Ft.
    Support Space - 700 Sq. Ft.
SUMMARY
    Conversion of Research Advances to CGMP Production is Challenging
    A Well Designed Facility Provides a Controlled Environment for Manufacturing
    Facility Design Should Enhance Process Consistency by Defining Flow Patterns
    Facility - Procedures - Personnel Must All Be Combined for Successful
Manufacturing
WR Tolbert & Associates Consultants to the Biopharmaceutical Industry
Cypress Woods Drive
San Diego, CA 92131-3535
    TRANSITION FROM GLP TO GMP: AN ACADEMIC PERSPECTIVE
    Blake Roessler, M.D.
    University of Michigan Human Applications Laboratory
ABSTRACT
The purpose of The Human Applications Laboratory (H.A.L.) At the University of
Michigan Hospital is to provide a physical plant for medium scale production of
biological reagents and culture of human cells for autologous transplantation as
required for Phase I/II investigator-initiated studies.
The University of Michigan H.A.L. is currently nearing completion of a major
renovation project that will transform the H.A.L. from a single room facility
into a multi-room, multi-use biological production facility. An overview of the
transition process from a smaller to a medium sized facility will be discussed.
Discussion will also include an overview of problems encountered in the design,
construction, start up, and daily operation of a medium sized facility in an
academic environment.
LIMITATIONS:
    Suboptimal GMP
    One room, one time, one project
    Materiel storage
    Product storage
LABORATORY GOALS:
    Performance of multiple projects
    Vector production
    Autologous cell culture
    'Not too big, not too small'
LABORATORY REDESIGN:
    Modular floorplan
    Efficient utilization of available space
    Worker safety and convenience
    Ease of cleaning
    Quality air
CONSTRAINTS:
    Existing building
    Fixed floorspace
    Shared HVAC
    Total cost
SPECIFICATIONS:
    750 square feet
    3 laminar flow hoods
    3 pair stacking incubators
    Class 10,000 air
    One technician
PRODUCTION STATISTICS:
    Retroviral supernatants
    Batch size 1-5 liters
    5 batches per annum
    Contract validation
SPECIFICATIONS:
    1500 square feet
    4 production rooms
    Class 100,000 air
    Class 100 hoods
    Two technicians
    CONSIDERATIONS AND STRATEGIES IN THE PLANNING AND DESIGN OF A NEW FACILITY
    Victor Santamarina, Ph.D.
    Genetic Therapy, Inc.
THE MOST CRUCIAL BUILDING COMPONENTS ARE:
    Physical isolation of steps
    Cleaning and decontamination
    HVAC
TIME TO MARKET MUST BE SHORTENED, SEVERAL
 CONSTRUCTION-RELATED OPTIONS EXIST:
    Initiate construction of a purpose built facility during phase II/III
Clinical Trials
    Enter into contract manufacturing agreement.
    Construct an interim limited facility
CONSIDERATIONS AND STRATEGIES IN THE PLANNING & DESIGN OF A NEW MANUFACTURING
FACILITY
    Issues
    Timing - Schedule
    Scope - Design Requirements
    Scope
    Process Definition
    Defines process requirements
    Process flow diagrams
    Defined by Phase III
    Marketing Requirements
    Forecast determines production levels
    Plant capacity and construction schedule
    Plant Definition
    Process
    Capacity
    Floor plan/Building components
    Plant Definition
        Process Flows
        Material Flows
        Equipment Flows
        Personnel Flows
PLANT DEFINITION
    Market Assessment
    Initial
    Market Increase
    Pre-launch Inventory
    Initial Market Needs
    Stability
        Post Launch Production Requirements
PLANT DEFINITION
Process Areas   Physical Isolation
Consistent with process     Process Areas
steps
Unidirectional flows    Process - Non process areas
Support Activities  Utilities
QC  Environment control - HVAC
Materials Handling  Sanitary & non sanitary systems
Final product storage &
shipping
    HVAC
    Determined by floor plan and HVAC design
    Room Classification
        Processing Areas
        Airlocks
        Corridors
    Pressure differences & flows
    Multiple vs dedicated air handlers
    Recycle vs one-pass-thru
    Isolation (Barrier) Technology
SCOPE-DESIGN REQUIREMENTS
    Room Classification
    Class 100,000
        Corridors processing areas airlocks
    Class 10,000
        All processing areas & airlocks between processing areas
    Class 1,000
        Filling room if required by scope
    Class 100
        All open operations
    Air Flows
        Clean room vs containment issues compromise
        Clean room positive toward access/exit airlocks
        Airlocks slightly negative toward the outside or
        access corridors to provide containment
    Air Handlers
        Preferably dedicated in critical areas
        Required dedicated air handler for filling room
    HVAC Design Options
        One-pass-thru recommended as permitted by economy of design
ENGINEERING
    Schematic design should start and be updated as process develops into
clinical trials. Complete when Phase III starts
    Development design should parallel Phase III followed by detailed design
    Validation
    The validation issues must be addressed in the schematic design and should
parallel the design development
CHARACTERISTICS OF HVAC OPTIONS
Single Pass Recirculated
Less Mechanical Equipment   More Mechanical Equip (Blowers, Ductwork)
Expensive to Operate    Cost Effective Operation
Low risk of cross contamination     Potential for cross contamination
of system   (Exhaust air recycled)
Burnett, M.B., Santamarina, V.G., Omstead, D.R. (1991).Design of a Multiple
Purpose Biotech Pilot Plant and Production Facility. Recombinant DNA Technology
I. Annals of the New York Academy of the Sciences, Vol. 646, pp. 357-366. New
York Academy of Sciences, New York.
SUMMARY
    Timing
    Conceptual design should start with clinical trials
    Full process design and conceptual plant design should be in place by phase
III
    Scope
    Decision for plant size should be in place with market data by phase III.
    Design should incorporate as much as possible the following:
        Unidirectional flows
        One-pass-thru and dedicated air handlers
        Combination of positive and negative airflow to airlocks to accommodate
clean room requirements and containment to the outside
    Support activities such as QC labs, materials handling of raw materials and
finished products and shipping should be included in the envelope of a
manufacturing plant design
    BREAKOUT SESSION 4:
    GETTING STARTED IN GENE THERAPY VECTOR DEVELOPMENT
    SUMMARY
    Angus J. Grant, Ph.D.  FDA/CBER
During this session, we heard a series of presentations that were intended to
help investigators bring gene therapy ideas from the research bench to phase I
clinical trials. This endeavor has challenges unique to gene therapy. To make
matters more difficult, as in any emerging technology, the challenges keep
shifting. For example, from a regulatory perspective, review by the NIH-ORDA-RAC
has been changing and may soon be a thing of the past. Special considerations
must still be taken into account for the societal and human subject issues
involving the  utilization of this technology.  The IRBs, the proposed
NIH-ORDA-GTPC specific subject conferences, and the FDA  will  oversee the
development and use of gene therapy technologies. As an investigator confronts
the challenges in bringing gene therapy protocols to the clinic, a great number
of skills and resources will be tested. The presentations within this session
were designed to provide insight and information to make these challenges more
attainable.
The first set of presentations pointed out the importance of evaluating the
strengths and weaknesses of ones particular gene therapy program and how to use
this information in the early planning process. It was pointed out how good
planning can avoid costly and time consuming mistakes. While experience in the
strategic planning of gene therapy vector development can be of great utility,
so can the experience of a variety of contract services. For investigators
unfamiliar with the IND process, there are resources that can be turned to, some
for a fee, that can reduce or eliminate fatal flaws in your program development.
 There are a variety of consulting and contracting  companies that can cover the
spectrum of tasks involved in gene therapy program and product development,
including support for overseeing the legal ramifications in other corporate
interactions. In addition, there are companies that in addition to providing
specific services not immediately available in every institution, can provide
valuable advice on a variety of related issues for the overall gene therapy
program due to their experience with other programs. Outsourcing support for
this fast moving technology can provide established experience and cutting edge
technologies, that are in place, that may be especially important for the
smaller institutions.
The second set of presentations centered more on the production of the gene
therapy vectors. The GLP and GMP continuum was discussed for investigators
planning on producing the product themselves or for investigators that need to
understand what is required or necessary when they discuss outsourcing the
production. Finally, we heard from a company that, for a fee, can produce the
entire gene therapy vector, ready for use in the clinical trial.
In all of the presentations, it was made clear that bringing a gene therapy idea
from the research stage to the clinic is an arduous undertaking and requires a
tremendous number of skills and resources. One recurrent theme was the need for
good communication. It usually requires a large number of people and
organizations to successfully bring a product to the clinic, and good
communication is essential. Also, while there may be a substantial cost to
outsourcing for strategic planning, for regulatory and legal support, for
outsourcing quality control, or for outsourcing the entire manufacturing
process, the savings in time and mistakes may easily make up for the cost. Good
communication is also important for interactions with the FDA. By frequent and
open communication with the FDA, an investigator can avoid costly and time
consuming delays and can get a lot of free advice. It is clearly important for
each investigator to evaluate their own resources, expertise, facilities and
goals so that they can determine the best course of action to bring their ideas
to the clinic.
During this conference and during this session we have heard a sample of the
support that can be utilized in the field of gene therapy, but it was not an
exhaustive collection. We hope that through these presentations we have helped
provide investigators with useful information that will bring their ideas from
the bench to the bedside more effectively and that these therapies will
eventually lead to improvements in the treatment of human diseases.
AGENDA
The goals of the session center on enabling investigators to bring their ideas
from the bench to the bedside. It is intended that this session will build on
the lessons learned from he first day of the conference, where the FDA IND
considerations and some NIH resources were presented. However, there are a
considerable number of additional challenges that must be dealt with when
planning to bring new therapies into human subjects. It is not unusual for an
institution, organization, or corporation to lack the capabilities and expertise
necessary to accomplish all the tasks involved in bringing a gene therapy
product to the clinic. In such cases, the investigator leading the gene therapy
project must turn to outside resources to provide the necessary expertise,
support or services.
During this breakout session, there will be a series of presentations that  will
cover strategic planning of product and protocol development, important
regulatory and legal considerations when preparing to submit an IND, resources
to support IND submissions, and resources to support production of the gene
therapy vectors and the quality control characterization of the gene therapy
product.  In addition to the presentations, there will be an opportunity for
questions and discussion.
    Introduction.
    Angus J. Grant, Ph.D., FDA/CBER
    Starting a Clinical Gene Therapy Program: Planning & Implementation.
    Bruce Merchant, M.D., Ph.D., Merchant-Taylor International, Inc.
    Key Issues and Factors in Developing the Gene Therapy IND.
    Gary E. Gamerman, M.S., J.D., Fenwick & West, LLP
    Contract Services Supporting Gene Therapy Development and Clinical Trials.
    Jeffrey M. Ostrove, Ph.D.. Microbiological Associates, Inc.
    Implementation of a Phase I Gene Therapy Program in a GLP-GMP Continuum.
    James A. Taylor, Ph.D., Merchant-Taylor International, Inc. -
    Monoclonal Antibody and Recombinant Protein Production Facility (soon to add
viral vectors), National Cancer Institute, Frederick Cancer Research &
Development Center.
    Steven Creekmore, Ph.D., M.D., National Cancer Institute -
    Considerations for Process Development and Manufacturing.
    Shawn L. Gallagher, MAGENTA Inc.
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
    STARTING A CLINICAL GENE THERAPY PROGRAM: PLANNING AND IMPLEMENTATION
    Bruce Merchant, M.D., Ph.D.
    MTI Inc., La Jolla, CA   (tel. 619-546-4334)
ABSTRACT
Developing a new Clinical Gene Therapy program is a daunting project. In all
likelihood, it will involve seeking objectives, marshaling resources, and
confronting tasks that you have not confronted before. It will challenge your
coalition-forming, team-building and communication skills in ways that they have
not been previously challenged. It will set you face-to-face with new concepts,
new frustrations, new opportunities, and new rewards.
Gene Therapy offers the potential to intervene in chronic diseases at a primary
"first cause" level. Optimally, it will make available therapies for many
diseases which currently lack effective treatments. By spearheading the
development of a Clinical Gene Therapy program in your organization, you can
take leadership in bringing "closure" between cutting-edge basic Molecular
Biology and meaningful applications of this core science in the clinic. The
whole program can be "tracked" by a special road map called a Master Project
Schedule which can be used as an extremely effective tool both in planning and
in implementation.
Making your Clinical Gene Therapy program a successful reality, will depend upon
your being well informed, highly motivated, and adequately resourced. Included
among essential resources are crucial funding, skilled and talented clinical and
scientific colleagues, and a well motivated, team-oriented staff. You will need
to develop a mind set that is open to new ideas, that values both afferent and
efferent communication, and that espouses the principals of good project
management. There will be much to learn as you proceed, including new scientific
concepts, special clinical considerations, basic regulatory principles, good
manufacturing practices, and patient-centered ethical sensitivities.
STARTING A CLINICAL GENE THERAPY PROGRAM: PLANNING AND IMPLEMENTATION
    The Incentives are to:
    Intervene in chronic diseases at a primary "first cause" level.
    Provide important new "cutting edge" therapies to patients.
    Make available therapies for many diseases lacking currently available
treatment.
    Effect "closure" between basic science and meaningful clinical applications.
PLANNING: THE INITIAL STEPS+COMMIT TO WRITING
    Specific Objectives of the Program.
    Early - Intermediate - Long Term
    What constituencies will be served?
    What resources are available?
    Financial+Institutional/Funding Sources
    Space/Facilities
    Human+Scientific/Clinical
    Human+Managerial/Manufacturing/Quality Control
PLANNING: THE INFORMATION GATHERING STEPS
    What internal resources are available and committed?
    What solid external resources can be counted on?
    What "mix" of these resources is right for your organization?
    How is that "mix" likely to evolve over the next several years?
    What contingencies are available in case the program evolves differently
than expected?
PLANNING: THE DECISION MAKING STEPS
    Who the Clinician/Scientists will be.
    What Departments/Divisions will be involved.
    What the Budget will be (& Funding Sources).
    Who the Project Leader will be (Accountability).
    What Space/Facilities will be available (When).
    What human talent will be provided.
    Internal
    External
    What contingencies will be provided for.
IMPLEMENTATION
    Determine what the first clinical study will be.
    Determine if you will develop a Vector Production Facility.
    Develop a Master Project Schedule (Importance of cGMPs).
    Recruit Key Personnel.
    Production Facility Manager
    QA/QC Manager
    Design a Production Facility (Spirit of GMP).
IMPLEMENTATION+FACILITY & DESIGN
    Decide What Equipment will be required.
    Get good advice on the basic Facility Design.
    From colleagues "who've been there"
    From qualified outside experts
    From Workshops/Forums
    From the FDA. (Pre-construction Package and Meeting)
    Engage an Architectural Firm.
    Settle on the final Plan.
    Conduct the bidding process.
    Obtain Permits.
    Do the Construction.
    Perform Validation on the Facility, the Equipment and on Key Production
Processes.
IMPLEMENTATION+PROCESS DEVELOPMENT
    Determine what vector(s) will be used.
    Determine which labs will be dedicated areas.
    Determine which labs will be "campaign" supporting.
    Develop Probable Production Scenarios and Flow Patterns.
    Determine a list of 25-50 Key Production Activities and develop SOPs for
each.
    Understand that SOPs will evolve (both in content and in numbers).
IMPLEMENTATION+CLINICAL/REGULATORY MANAGEMENT
    Move Clinical Plans from Outlines to Finished Protocols.
    Augment Dialogue with CBER (e.g. Pre-IND Meeting).
    Gain IRB and IBC approvals.
    Prepare and Submit an IND (Possibly also a separate DMF).
    Register Study with NIH ORDA.
    Initiate Gene Therapy Clinical Trials.
PITFALLS TO BE AVOIDED
    Trying to do everything yourself. Teamwork is Essential.
    Assuming that Scientific Excellence assures Manufacturing Reliability (Two
District Disciplines).
    Assuming that there will be no major glitches (Develop contingency Plans).
    Assuming that all your colleagues are marching to the same drummer.
    Failing to communicate thoroughly and frequently.
    Failing to recognize team accomplishments.
    Failing to develop follow and modify (as required) a Master Project
Schedule.
REWARDS
    Bringing cutting edge gene therapies to bear on problematic genetic &
chronic diseases.
    Learning practical "nuts and bolts" realities about quality product
Manufacture.
    Working as a team with skilled, and dedicated Scientists, Clinicians,
Regulators and Outside Experts
Effecting "closure" between basic Molecular Biology and those Diseases which be
favorably treated by gene
    KEY ISSUES AND FACTORS IN DEVELOPING THE GENE THERAPY IND
    Gary E. Gamerman, MS, JD
    Fenwick & West LLP, 1920 N St., NW Suite 650
    Washington, DC 20036, 202-463-6300
    (fax) 202-463-6250
ABSTRACT
Developing a gene therapy IND requires consideration of a number of preclinical
and clinical issues, some of which are common to all biotherapeutics, others are
unique to this technology. This presentation will identify the most important
issues and strategies for addressing them in the IND and product development
plan. The discussion will be highlighted with examples of the types of problems
that can sidetrack a gene therapy IND.
DEVELOPMENT PROCESS MANAGEMENT AN INCREMENTAL DYNAMIC APPROACH
    Many Points for Binary/Multivariable Decisions During Development Cycle
    Go/No Go
    Process Selection
    Early Choices Channel Later Options and Selection
    Development Candidate
    Manufacturing Technology
    Clinical Development Path
    Cell Substrate
    Formulation
QUESTIONS TO GUIDE IND DEVELOPMENT & PLANNING
    Regulatory Science v. Bench Science
    Questions to Address Before Any Testing
    What is the Product
    How to use It
    How to Test It
    What will it Do
    How to Make It
    How to Sell It
WHAT IS THE PRODUCT?
    Product Characteristics
    Identity
    Composition
    Inherent Impurities
    Activities
        Direct
        Collateral
    Process Characteristics
    Product Variability & Heterogeneity
    Process Limitations On Purity & Composition
WHAT WILL IT DO?
    Intended Biological Effects
    Expected Collateral Effects
    Possible Risk Events
    Indicators of Activity
    Early Low Dose Treatment
    Higher Doses
    Indicators of Toxicity
    Traceability of Event Product
HOW TO USE IT?
    Indication
    Population
    Dose
    Amount, Schedule, Route, Duration
    Formulation
    Setting
HOW TO MAKE IT?
    Safe
    Consistent
    Validatable
    Scaleability
    Robust/Controllable
    Efficient/Cost-Effective
    Upstream/Downstream Considerations
HOW TO TEST IT?
    Good Science Not Matrix Repetition
    Follow the Product's Nature
        Preclinical: Laboratory Testing
    Product Characterization
    Process Characterization
        Preclinical: Animal Testing
    Model Human Biology
    Model Human Protocols
        Human Testing
    Relevant, Goal-Oriented Design
    Consider Practical Issues
CONTRACTS IN GENE THERAPY DEVELOPMENT
    Types of Contracts
    Contract Manufacturing Organizations (CMO's)
    Contract Research Organizations (CRO's)
    Research Agreements:
        Investigators, Institutions, Others, Patient Consent Forms
    Material Transfer Agreements
    Consultants/Employees
SELECTING CONTRACTORS
    CRO's, Contract Manufacturers, Testing Labs:
    Do They Have Relevant Expertise
    Will They Service YOU
    How Will Problems Be Resolved
    Do They "Unsell" or Let You Fly Blind
    What/How Have They Done Lately
    To Them It's a Contract, To You It's the Company:
    You Cannot Surrender Control or Responsibility
    You Need to Understand What Must Be & Will Be Done
    Trust, But Audit!
    Contracts:
    Everything is Negotiable
    Regulatory Control is Essential
MAJOR CONTRACTING ISSUES
    Respective Obligations
    Intellectual Property
    Regulatory Compliance
    Dispute Resolution & Enforcement
    Warranties, Indemnification's & Remedies
RESPECTIVE OBLIGATIONS
    Need to Detail Mutual Responsibilities:
    Who, What, Where, When & How
    What Standards Will be Used for Performance
    Task & Payment Schedule
    Procedures for Charges & Deviations
    Resource Allocation
    Procedures for Remedies If Faulty Performance:
    Discussion, Arbitration, Litigation
    Refund/Rework
    Limitation of Liability
REGULATORY & PERFORMANCE COMPLIANCE
    Qualify Contractors
    Performance Guaranty & Certification
    Contractor's QA Efforts
    Your Monitoring & Auditing
    Disclosure of FDA Inquiries & Inspections
    Non-FDA Obligations
    EPA, OSHA, Employment, State, Local & Foreign Law
INTELLECTUAL PROPERTY
    Primary Rights
    Patents
    Trade Secrets
    Copyrights
    Derivative Rights
    Product Related
    Study Related
    Other
    Use & Disclosure
    Technology & Data
    Publicity/Status
DISPUTE RESOLUTION & ENFORCEMENT
    High Level Discussion
    Most dispute can be resolved if addressed early
    Evaluate the reason and significance for discrepancies
    Economic Leverage
    Payment Schedules & Termination
    Arbitration
    Litigation
    A very last resort
    Keep good documentation
    Foreign Contractors?
WARRANTIES, INDEMNIFICATIONS & REMEDIES
    Warranty = Promise on Current or Future Status
    Patent
    Regulatory Compliance
    Performance
    Capabilities
    Indemnifications = Coverage Against Third Party Actions
    Standards, Causes
    Respective Duties
    Remedies = Rights in Events of Breach
    Injunctions
    Money
    Performance reprocess, rework)
    Other (facility take over)
    Limitations of Rights
    types & amounts
CONTRACT MANUFACTURING ORGANIZATIONS
    Duties & Timing
    GMP & Process Definition Compliance
    Subcontracting
    Auditing
    Document
    Intellectual Property
    Warranties/Indemnification/Remedies
CONTRACT RESEARCH ORGANIZATIONS
    Detailed Description of Tasks
    What is Included
        Monitoring & Audits
        Data Handling & Timing
        Report Generation
        GDEA Certification
    Who will be assigned
    Modifications
    Extension & Alteration of Protocol
    Ongoing, Interim & Final Reports
    Inspections & Documentation
    Payment Schedules
CLINICAL RESEARCH AGREEMENTS
    Investigators
    Compliance: Reference Investigators Brochure, Audits/Inspections
    IP & Publications
    Conflicts & Time
    Payment Basis & Schedule
    Patients & Study coordinators
    Institutions
    IRB Compliance
    Costs
    Indemnifications
    Accreditations
    Employees
CONSULTANTS & EMPLOYEES
    Intellectual Property
    Regulatory Compliance
    Future Actions
    Not debarred/disqualified
    Confidentiality
    Conflicts/Exclusivity
    Non-Compete Issues
    General Employment Concerns
MATERIAL TRANSFER AGREEMENT
    Determine How Material Can Be Used
    Will there be grant back of data rights?
    Will use be consistent with regulatory objectives?
    State Who Owns Derivative IP
    Limit Who the Material Be Transferred To
    Include Warranty Disclaimers
    Ensure Proper FDA Labeling and/or End-User Certifications
PATIENT CONSENT FORMS
    Only Protects Against Study Risks
    Does Not Protect Against
    Unmentioned risks
        associated with investigational product
        associated with procedures
    Negligence or wrongful act by company or investigator
    Breach of regulatory of other obligations
    Must be Drafted and Executed In Compliance with FDA & Other Regulations
    Need to Update As New Risks are Discovered
    Do Not Minimize or Mischaracterize Risks or Benefits
KEY CONTRACTING PRINCIPLES
    Avoid Mumbo-Jumbo
    Clear, Concise, Complete, Correct
    Check the Boilerplate
    Sign in Haste, Suffer in Leisure
    Ask Questions Before, Not After
    Verify Obligations and Remedies
    Communicate Regularly
    Know What Your Contractor is Doing
    Let Them Know What You Expect
    Check Project Status
    CONTRACT SERVICES SUPPORTING GENE THERAPY DEVELOPMENT AND CLINICAL TRIALS
    Jeffrey M. Ostrove, Ph.D.
    Microbiological Associates, Inc., 9900 Blackwall Road
    Rockville, MD. 20850, Ph.: 301-738-1000, FAX: 738-1036,
    E-mail: jostrove@microbio.com
ABSTRACT
This presentation will focus on what is required to move a gene therapy project
from the research laboratory into the clinic. The first step in this process is
to ensure that the cell substrates that are being used to produce viral-based
vectors for gene therapy are free of common, adventitious contaminants such as
mycoplasma and replication-competent viruses. The next step is the establishment
of a Master Cell Bank (MCB) and its characterization according to the FDA
"Points to Consider" documents. The safety tests that are required to be
performed on each lot of clinical supplies manufactured will also be described.
Data on the incidence of common microbial and viral contaminants that have been
detected in mammalian cell substrates used to produce gene therapy vectors and
the incidence of replication-competent retroviruses (RCR) and adenoviruses (RAC)
will be presented. Perhaps most important, the above process will be described
with the context of the timing required for each step in the process. Planning
and execution to a timetable are critical factors in reaching the clinic in a
timely fashion.
Other services available for scientists developing gene therapies such as the
biodistribution of gene therapy vectors administered to animal models and
associated toxicology studies will be presented. Microbiological Associates has
also developed assays for the monitoring of patients undergoing
retroviral-mediated gene transfer. Western Blots have been qualified that can
detect the presence of antibodies against amphotropic retrovirus-based vectors
as well as PCR-based assays to detect the presence of retroviral sequences in
clinical samples.
Microbiological Associates (MA) is a contract research and development
organization which provides a full range of laboratory-based services to clients
developing gene therapies. Our facilities and services are maintained according
to the principles of Good Laboratory Practices (GLP) and current Good
Manufacturing practices (cGMP). MA services span the spectrum of pre-clinical
development of viral-based vectors through the manufacturing of clinical
supplies and patient monitoring. MA laboratories are continually developing new
assays to support the development of gene therapies.
CONTRACT SERVICES SUPPORTING GENE THERAPY DEVELOPMENT AND CLINICAL TRIALS
PROVIDED BY MICROBIOLOGICAL ASSOCIATES, INC.
    Provides GLP/GMP - compliant biological safety testing services to:
    Pharmaceutical, biotechnology, chemical, cosmetics, medical device, and
personal care products industries worldwide
CONTRACT SERVICES
    PRECLINICAL DEVELOPMENT
    REGULATORY CONSULTANTS
    FACILITIES CONSULTANTS
    VALIDATION SERVICES
    PRECLINICAL TOXICOLOGY
CONTRACT SERVICES
    SAFETY TESTING OF CELL SUBSTRATE
    CONTRACT MANUFACTURING
    LOT RELEASE TESTING
    CLINICAL TRIALS MANAGEMENT
    PATIENT MONITORING
REPLICATION COMPETENT RETROVIRUS TESTING BY MICROBIOLOGICAL ASSOCIATES (5/96)
    Amplification in Mus    %
    dunni   Positive
        mL (n=129)  0
            > 100 mL (n=101)    11
                Cocultivation (n=173)   4
PATIENT MONITORING
    PCR FOR RCR
    Rapid DNA extract is prepared from PBMC
    PCR is performed using primers for amphotropic envelope
    Products are viewed on an agarose gel
    Positives confirmed by RE digest or hybridization
    Negatives confirmed by spiking with known positive DNA
PCR FOR RCR RESULTS
    Dilutions of Positive control (plasmid) DNA show a 3-10 copy sensitivity in
.5 g cell DNA.
    Negative control DNA does not give a signal.
    DNA from amphotropic virus infected cells are positive.
    DNA from numerous cell lines are negative.
    The assay has been performed by 3 operators.
PATIENT MONITORING: WESTERN BLOT ASSAY
    Murine amphotropic virus, 4070A provides antigen.
    Strips are incubated with positive control, negative control, or test serum.
    Secondary anti-Ig conjugated to peroxidase are incubated with the strips.
    Colored precipitate (band) of appropriate MW indicates positive.
PATIENT MONITORING: WESTERN BLOT RESULTS
    Positive and Negative controls work
    Test serum spiked with positive control serum scores positive
    Nearly 100 patient samples tested
CONTRACT SERVICES: IN VIVO STUDIES
    Preclinical Efficacy
    Toxicology
    Biodistribution
CONTRACT SERVICES: ADVANTAGES
    Reduction in Time for Available Services.
    Utilization of Scientific Expertise Resident in Contract Testing Company.
    GLP, GMP and Regulatory Considerations.
    Better Utilization of Internal Resources.
CONTRACT SERVICES: DISADVANTAGES
    Perceived Loss of Control
    Economic Implications
HISTORICAL PERSPECTIVE ON QC TESTING ASSAY
    Mycoplasma
    Research
        Cells  (421/4412)
        Products (6/416)
    Regulatory
        Cells (53/1483)
        Products (42/5921)
CGMPS - QUALITY
    Quality, Safety, and Effectiveness Must be Designed and Built into Products.
    Quality Cannot be Tested into the Finished Product.
    Each Step of the Manufacturing Process needs to be Controlled.
HUMAN CELL BANK CHARACTERIZATION
    Sterility
    Mycoplasma
    TEM
    In Vivo & In Vitro Adventitious viruses
    Isoenzyme & Cytogenetic Analysis
    Tumorigenicity (in vivo or in vitro)
    Hep B, EBV, HIV, HTLV, AAV
PRODUCT FLOW : PRELIMINARY TESTING
    Research Lab Producer Cell Line
    Preliminary Testing
    Mycoplasma
    Sterility
    Replication Competent
    Retrovirus
MYCOPLASMA
    Continuous Cell Cultures - Frequently Contaminated
    (>3,000) Contaminated of >20,000 Cell Cultures Tested
    ) of 2.800 Isolated were Bovine Species
    Mycoplasma Arginini - 26%
    Acholeplasma Laidlawii - 8.5%
    Pirum - 7%
    ) of 2.800 Isolated Were Human Species
    Mycoplasma orale - 29.5%
    of 2.800 Isolated Were of Porcine Species
    Mycoplasma Hyorhinis
PRODUCT FLOW - MCB
    Research Lab Master Cell Bank (MCB)
    (Produced Under GMPs)
    Preliminary Testing
    Master Cell Bank Cell Line Characterization (CLC)
    Mycoplasma
    Sterility
    In Vitro Adventitious Virus
        Co-Cultivation with Mus dunni
        Extended XC Plaque Assay
    Isoenzyme Analysis
    In Vivo Adventitious Virus
        Mouse Antibody Production (MAP)
PRODUCTION RUN
    Archive Samples
    Supernatant
    End of Production Cells (1% or 10e8, whichever is less)
LOT RELEASE TESTING (VECTOR-CONTAINING SUPERNATANTS)
    Potency (titer)
    Identity
    Sterility
    General Safety
    Endotoxin
    Tests for Virus/Adventitious Agents
    Includes Replication-Competent Viruses
FOOD AND DRUG ADMINISTRATION REQUIREMENTS: REPLICATION-COMPETENT RETROVIRUS
TESTING
    Current FDA Recommendations - 10/93
    Master Cell Bank (MCB)
         Cells - Co-Cult with Mus dunni/PG4 Endpoint
         Volume tested - 1% or 108 cells - whichever is less
         Supernatant - 5% Amplification on Mus dunni/PG4
        endpoint
    Manufacturer's Working Cell Bank (MWCB)
        Either supernatant or cells (as above)
    Qualification Run Including Transduction
EX VIVO TREATMENT
    Transduced Cells (1%)
    Co-Cult Mus dunni/PG4 Endpoint
    General Consideration:
    Tests should be complete prior to patient administration, if possible. If
not, start testing
    IMPLEMENTATION OF A PHASE I GENE THERAPY PROGRAM IN A GLP/GMP CONTINUUM
    James A. Taylor, Ph.D.
    Taylor Associates, San Diego, CA  (tel. 619-675-0808)
ABSTRACT
Gene therapy product development, perhaps more than any area of therapeutic
research, has arisen from academic institutions and newly-formed biotechnology
companies. Implementation of early gene therapy programs, therefore, generally
requires a large leap through the regulatory continuum from research to an
environment of FDA-mandated Current Good Manufacturing Practice (CGMP)
requirements established to produce investigational clinical supplies.
General concepts that impact the regulatory continuum from research to "Good
Academic Research Practice" to "Good Laboratory Practice" to the "spirit" and
"letter" of CGMPs will be discussed. Specific operational considerations that
apply to gene therapy facilities will be addressed beginning with a facility
design which can accommodate transduction and vector production laboratories.
Emphasis will be placed on development of a Master Project Schedule and key
Standard Operating Procedures as well as the importance of documentation and
validation.
THE SCIENTIFIC-REGULATORY CONTINUUM
ACADEMIC RESEARCH
       Scientific rigor
       Peer review
       Publication format
GOOD ACADEMIC RESEARCH PRACTICES
       Reliance on protocols
       Enhanced documentation
       Facility impact
       Training
       Certification/calibration
GOOD LABORATORY PRACTICES
       Standard operating procedures (SOPs)
       Quality assurance
       Qualification
       Release Criteria
       Product characterization
GOOD MANUFACTURING PRACTICES
       Batch records
       Quality control
       Validation
       Specifications
THE GLP-GMP CONTINUUM
    Spirit vs letter of GMP compliance
    Concepts differ by degree
    Degree of SOP preparation
    Degree of validation
    Degree of product characterization
MASTER PROJECT SCHEDULE KEY FACILITY PROCEDURES
    Facility
    Restricted access
    Facility maintenance
    Systems
    Environmental monitoring
    Controller operations
    Systems maintenance
    Equipment
    Equipment calibration
    Preventive maintenance
    Use of dedicated equipment
    Use of disposable labware
    Use of preformulated media
    Manufacturing
    Gowning
    Product/waste transfer
    Product changeover
    Computerized records
    Personnel
    GMP training
    Employee training
    Health & safety program
    Quality Control/Quality Assurance
    Quarantine & release
    Product specifications
    Assay development
    Document preparation
    Audit function
    Validation
    Cleaning validation
    Systems validation
    Assay validation
    Equipment validation
    Computer validation
    Process validation
    MONOCLONAL ANTIBODY AND RECOMBINANT PROTEIN PRODUCTION FACILITY NATIONAL
    CANCER INSTITUTE-FREDERICK CANCER RESEARCH & DEVELOPMENT CENTER
    Stephen Creekmore, M.D., Ph.D.
    Biologic Resources Branch, NCI-FCRC, Frederick, MD
ABSTRACT
The Monoclonal Antibody and Recombinant Protein Production Facility (MARP) is
an NCI-sponsored facility established at the NCI Frederick Cancer Research and
Development Center to perform rapid and cost-effective production of
clinical-grade biologicals for NIH-supported Phase I and Phase II clinical
trials. Operating in temporary quarters for the past 18 months, MARP staff have
manufactured several clinical-grade products: more than 500 gm of monoclonal
antibodies, 5 recombinant vaccines, and 1 recombinant immunotoxin. The MARP has
also vialed and released over 20 peptide vaccines. Recently, the MARP was
relocated to a specially-designed and constructed building at NCI-FCRDC. When
fully operational, the new facilities will triple the capacity to produce,
purify, and vial clinical-grade biologics using mammalian, bacterial, or insect
cell systems. Current projects include work on monoclonal antibodies,
recombinant proteins, immunotoxins, peptides, and DNA and viral vectors. Working
in close collaboration with the requesting scientific laboratories, MARP staff
develop and validate production and purification procedures appropriate for
scale-up from the laboratory models. MARP staff discuss the proposed approaches
in pre-IND meetings with the FDA and the requesting scientific laboratory.
Production, purification, formulation, and vialing are performed in dedicated
suites according to current Good Manufacturing Practices (cGMPs). The MARP also
provides cGMP documentation (Chemistry, Manufacturing, and Control) for
incorporation in the IND. Cost accounting of MARP projects allows fair recovery
of costs from users, and aids cost control measures. MARP projects have been
supported by funds from several sources: NCI projects are authorized by the
Decision Network Committee of the Division of Cancer Treatment, Diagnosis, and
Centers (DCTDC). Projects from other governmental agencies or departments (e.g.,
NIAID) are funded through interagency agreements. Some projects are funded
through budgeted costs in extramural grants. Other outside sources of funding
may also be considered. Projects are prioritized by the Biological Response
Modifiers Operating Committee (BOC) whose membership is drawn from NCI, NIAID,
FDA, and other agencies.
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.
PRODUCTION OF CLINICAL AND LABORATORY-GRADE REAGENTS: SUMMARY OF CAPABILITIES OF
BIOLOGICAL RESOURCES BRANCH, NCI
NCI MONOCLONAL ANTIBODY/RECOMBINANT PROTEIN PRODUCTION FACILITY (MARP)
    A contractor (SAIC)-operated and dedicated facility at NCI-FCRDC
    Purpose: Small-to-Medium scale sterile/cGMP production facility
    Monoclonal antibodies
    Recombinant proteins
    Peptides
    10-50 grams for NIH-sponsored Phase I and Phase II clinical trials and
preclinical studies
    A method resource in support of NIH-sponsored clinical research
APPROACH TO CLINICAL DEVELOPMENT
    Provide a range of services in support of investigator-initiated
translational research
    Keep costs down
    Assist as many scientifically meritorious, approved projects as possible
    identified by peer review
    Decision Network Committee prioritizes projects requiring significant NCI
resources
    Identify promising candidates which may then be carried into full clinical
development by CTEP and/or industry
    pilot clinical data help decide which projects to pursue
ASSUMPTIONS
    Success rate of agents entering clinical trials 5% or less
    Lowering barriers to earlier clinical testing may:
    increase total number of successes
    stimulate interest in further development by industry and other
investigators by "proof of concept"
    permit earlier refocusing of efforts by rapid "disproof of concept"
    Trials by "inventor-clinicians" particularly need support, and represent an
appropriate target of BRB efforts
    Increasing demand for support investigator-initiated, peer-reviewed clinical
research projects
    Increased proportion from standard R01 pool
    Continued but limited use of set-asides and other "targeted" approaches
Availability of low cost, cGMP production, IND filing, toxicology can determine
whether a project is feasible (and fundable)
PROJECT FLOW FROM THE INVESTIGATOR TO ITS INCORPORATION INTO THE MARP SCHEDULE
       Originator contacts the NCI-DCTDC-BRB
       The BRB chief submits proposal to MARP head
       Cost estimate of M&M, personnel, and opportunity costs is made
       Cost estimates are confirmed with originator+this takes about one week
       Project is reviewed by the Biological Operating Committee (BOC), and if
necessary, the Decision Network (DN), prior to placement on the MARP calendar
       Once project is initiated, a particular technical services scientist is
assigned and detailed discussions with originating laboratory begin
WHO USES THE STERILE/CGMP BIOLOGICS MADE AT THE MARP/FCRDC?
    Grantees
    Intramural investigators
    Contract clinical studies
Production priorities are set through the BOC
EXAMPLES OF SERVICES AVAILABLE
    Mammalian cell products
    Suspension culture
    Monoclonal antibodies
    Packaging cell lines
    DNA vaccines
    Viral production
    Large Scale Production of monoclonal antibodies
    8 hollow fiber pathways
    hybridoma, CHO cells
    monoclonal antibodies 10-200 gm
    Fermentation products
    Recombinant proteins
    immunotoxins
    recombinant vaccines
    cytokines
    Protein purification/conjugation/vialing/release
    conjugation of antibodies with chelator
    peptide vaccines
    Large-scale fermentation capacity
    300/3000 gallon fermentors (bulk pharmaceutical grade)
    natural products
    Quality assurance and regulatory compliance
    pre-IND meetings with FDA
    process validation
    documentation
    preparation of CMC section of IND
    formulation and vialing
    testing and release
    toxicology
BRB PRODUCTION PROJECTS
    Finishing and vialing peptide vaccines (DCB, DCS)
    Anti-lymphoma antibodies for immunotoxin production (extramural/DCS)
    Anti-Hodgkin's (CD30) antibody (DCS)
    k-ras proteins for vaccines (DCS)
    Anti-ID vaccines for lymphoma and myeloma (DCS)
    Anti-EGFR antibodies (extramural)
    Chimeric anti-myeloma antibody (extramural)
    Anti-colon antibody (DCB)
    Anti-HIV antibody (NIAID)
    Anti-Tac immunotoxin (DCB)
    Anti-bombesin antibody (DCS)
    IL-7 cytokines (DCS/extramural)
    Retesting clinical grade cytokines: IL-2, IL-1, TNF (DCS, extramural)
    Chelation of monoclonal antibodies
    Preclinical repository for cytokine standards, selected reagents (all)
For Further Information, Call:
Stephen Creekmore
Chief, Biological Resources Branch
DTP. DTCDC, NCI
NCI-FCRDC
Frederick, MD 21702-1201
(301) 846-1098
Creekmor@FCRFV1.NCIFCRF.gov
or
George Robertson
Head, Monoclonal Antibody/Recombinant Protein Facility
SAIC, Building 459
NCI-FCRDC
Frederick, MD 21701
(301 846-5474
Robertso@FCRFV1.NCIFCRF.gov
or Any staff member of the Biological Resources Branch or Monoclonal
Antibody/Recombinant Protein Facility staff
    GETTING STARTED IN GENE THERAPY VECTOR DEVELOPMENT - CONSIDERATIONS FOR
    PROCESS DEVELOPMENT AND MANUFACTURING
    Shawn L. Gallagher, MAGENTA Corp., Rockville, MD
    Ph.: 301-738-3936, FAX: 738-1605, E-mail: sgallagher@microbio.com
ABSTRACT
Once a transgene of interest is identified and the appropriate vector system for
delivery is chosen, the process development phase of a gene therapy project must
begin in earnest. During this exciting time period many decisions must be made
without the benefit of complete information and missteps are inevitable.
However, if consideration is given to clinical manufacturing issues early
enough, major project delays can be avoided. Among the critical issues which
must be addressed are the following:
    Clarification of regulatory environment
    Choice of host cell line (packaging, complementing, etc.)
    Selection of optimum virus strain
    Identification of appropriate facilities
    Optimization of production process
    Choice of in-house production or contract manufacturing option
    Development of appropriate formulation
    Selection of container/closure system
    Initiation of stability studies
CONSIDERATIONS FOR PROCESS DEVELOPMENT AND MANUFACTURING
OUTLINE
        Process Considerations
        Production Options
        Key Factors for Evaluation of Alternatives
    Cost Analysis
        Some Thoughts Regarding Partnering for Production
        Recommendations
KEY STEPS FOR PROCESS DEVELOPMENT EFFORTS
    Cell Line Development
    Virus Development
    Production System
    Purification Strategy
    Formulation
    Container/Closure
OPTIONS FOR ACQUIRING CLINICAL MATERIAL
    In-House Manufacturing
    National Gene Vector Laboratories
    Corporate Partnering
    Academic Partnering
    Contract Manufacturing
GLOBAL VARIABLES FOR A COMPARATIVE ANALYSIS
    Technology
    Time
    Regulatory Environment
    Product Development Strategy
    Money
FIXED COSTS
    Depreciation
    Facility Operation
    Personnel
FACILITY SCHEMATIC ADVANTAGES OF PARTNERING FOR MANUFACTURING
    Captive Technical and Regulatory "Consultants"
    Reduced Timeline to Product Availability
    Available Capacity / Optimized Technology
    Experienced Manufacturing Team
    No Need to Purchase Capital Equipment
    Full Quality Assurance Systems
    Validated Facilities
BENEFITS OF IN-HOUSE PRODUCTION
    Flexibility to Select Technology
    Direct Control of Quality
    Reduced Cost
    Ability to Shift Resources to Accommodate Unforeseen Requirements
    Need for Control
RECOMMENDATIONS FOR USE OF EXTERNAL RESOURCES
    Pilot Studies
    Special Projects
    Formulation and Filling
    Transient Needs
    Second Source
IMPORTANT CHARACTERISTICS OF A MANUFACTURING PARTNER
    cGMP Experience
    Biologics Master File in Place
    Adequate Capacity
    Corporate Commitment
    Willingness to Transfer Technology
THE COMMUNICATION BASICS
    Initial Information Exchange
    Establishing Confidentiality
    Project Definition
    Technology Transfer
    Use of Project Management Tools
    Production
RELATED DOCUMENTS
    Points to Consider in:
    Production and Testing of New Drugs and Biologics Produced by Recombinant
DNA Technology, 1985 (Plus 1992 Supplement)
    Human Somatic Cell Therapy and Gene Therapy, 1991 (Plus 1/96 Draft Addendum)
    Characterization of Cell Lines Used to Produce Biologicals, 1993
RELATED DOCUMENTS
    Cooperative Manufacturing Relationships for Biologicals (FDA Policy
Statement - Federal Register 11/92)
    FDA Guidance Document Concerning Use of Pilot Manufacturing Facilities for
the Development and Manufacture of Biological Products; Availability (Federal
Register 6/95)
SUMMARY RECOMMENDATIONS
    Spend the Time to Develop Robust Processes
    Define Release / Acceptance Specifications Early
    Develop a Change Control System
    Establish Project Review Checkpoints (Audits)
    Use Project Managers
These topics will be discussed with an emphasis placed on the resources
available to assist in the successful completion of this challenging process.
    BREAKOUT SESSION 5:
    DEVELOPMENT OF NEW VECTOR SYSTEMS
    SUMMARY
    Neal DeLuca, University of Pittsburgh; Robert Anderson, FDA/CBER; Parris
Burd,
    FDA/CBER
Part I. Herpes virus-based Systems
Four different HSV vector systems being developed for use in clinical trials
were described. Neal DeLuca described his efforts towards constructing mutant
Herpes viruses which do not express critical regulatory proteins (ICP4) and
other IE proteins (ICP27 and ICP0) and described the biologic effects of the
deletions. Elizabeth Rollinson described the construction, manufacture, and
biologic properties of a 'non-infectious', disabled Herpes virus which does not
express gH (DISC). This virus, when produced in a GH-complementing cell line, is
able to introduce itself into a cell, but cannot complete a replication cycle.
Alfred Geller described his studies of a virus encapsidated HSV-1 plasmid vector
developed for the treatment of Parkinson's Disease. Robert Martuza described the
development of replication-competent attenuated Herpes viruses for the purpose
of tumor therapy. These viruses are able to grow in transformed cells, but do
not replicate efficiently in normal cells.
Part II. AAV-based Vectors
The potential use of AAV vectors for gene transfer and stable expression in
hematopoietic progenitor cells, neuronal and muscle cells was discussed.  Jude
Samulski gave a brief overview of AAV biology, described his efforts at
producing AAV in the absence of Adenovirus and presented results of ongoing
studies of long-term viral expression in brain and muscle. Saswati Chatterjee
described efforts aimed at stable gene transfer into primitive hematopoietic
progenitor/stem cells. Samuel Wadsworth described AAV development undertaken by
Genzyme Corp and Gary Kurtzman described development of a helper-free AAV
production system.  Tom Reynolds discussed quantitation of infectious vector for
titering and issues related to patient monitoring.
AGENDA
PART I. HERPES SIMPLEX VECTORS
This session will discuss:
1.) the potential use of AAV vectors for gene transfer and stable expression  in
different tissues,
2.) production of AAV vectors in the absence of helper adenovirus, and
3.) quantitation of infectious vector for titering and patient monitoring.
    Herpes Simplex Virus Mutants as Gene Therapy Vehicles
    Neal DeLuca
    DISC-HSV a Novel Vector System
    Elizabeth Rollinson
    Modifying Neuronal Physiology with a Helper Virus-free HSV-1
    Alfred Geller
    Replication Competent Herpes Virus for Tumor Therapy
    Robert Martuza
    Discussion
PART II. ADENO-ASSOCIATED VIRAL VECTORS
This session will discuss and compare three different HSV vector systems:
1.) vectors with modified regulatory and immediate early genes,
2.) HSV-1 plasmid vectors (amplicons), and
3.) vectors with late gene deletions.
    AAV Vectors In Vivo
    Jude Samulski
    Adeno-Associated Virus Vectors for Transduction of Hematopoietic Progenitor
Cells
    Saswati Chatterjee
    Development of AAV Vectors for Gene Therapy
    Samuel Wadsworth
    AAV Vectors Can Be Efficiently Produced Without Helper Virus
    Gary Kurtzman
    Development of Assays for the Detection and Quantitation of Recombinant
AAV-CFTR Vector
    Tom Reynolds
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
    HERPES SIMPLEX VIRUS MUTANTS AS GENE THERAPY VEHICLES
    Neal DeLuca
    University of Pittsburgh School of Medicine, Pittsburgh, PA
Herpes simplex virus (HSV) is an extremely lytic virus that encodes
approximately 75 gene products. However, given its capacity to establish latency
in neurons, its broad host range, and the ability to obtain high infectious
titers, HSV may be useful as a gene transfer vector. The cascade of viral gene
expression and the infectious cycle of the virus can be blocked by deleting an
essential regulatory protein of the virus, ICP4. Mutants in ICP4 only express
five other viral genes (IE genes). The activities of the IE proteins all affect
aspects of host cell metabolism, resulting in rapid cell death. The theme of our
research in this area is to delete the genes that are expressed in the absence
of ICP4 in order to help determine how the products of these genes function, and
to eliminate the observed cytotoxicity, providing a more efficacious vector.
Three of the five IE genes are essential for virus replication (ICP4, ICP27, and
ICP0). Therefore, it was necessary to construct cell lines that would
simultaneously complement all three. Using such cell lines it was possible to
construct viruses deficient in different subsets of IE genes, efficiently
propagate them, and determine their effects on host cell survival and gene
transfer. Details on how these viruses were constructed, their phenotypes, and
their usefulness as vectors will be discussed.
    DISC-HSV A NOVEL VECTOR SYSTEM
    Dr. Elizabeth A. Rollinson et al
    Cantab Pharmaceuticals Research Ltd., Cambridge, England, CB4GN.
DISC-HSV is a Disabled Infectious Single Cycle Herpes Simplex virus from which
the gene encoding gH, which is essential for virus entry, has been removed. A
complementing producer cell line has been generated which allows for the
production of phenotypically complete, infectious virions, but because there are
no overlapping sequences between the cell line and virus, no replication
competent virus is generated during manufacture. The virus is, therefore,
capable of delivering its genome and heterologous genes into host cells but
cannot spread or cause disease. The excellent safety profile of DISC-HSV has
been demonstrated by extensive preclinical testing, including demonstration of
non-neurovirulence/neuroinvasiveness, safety in immunocompromised animals,
negligible probability of recombination, inability to reactivate from the latent
state, antiviral drug sensitivity, absence of injection site reactivity and
general safety in primates and laboratory animals. The DISC-HSV vector will
enter Phase I/II trials shortly for genital herpes.
As well as this excellent safety profile, DISC-HSV offers additional advantages
as a gene delivery vehicle, including: capacity to accommodate large amounts of
foreign DNA (30 kb compared with 4-7 kb for AAV/AV/RV), broad host cell range
(dividing and non-dividing, lymphocytic epithelial, neuronal, tumour cells),
high transduction efficiency (for example, 60-100% transduction in primary
leukaemia cells, compared with around 4% for an equivalent retroviral or
adenoviral vector), and the added safety feature that incorporation of the
heterologous gene into the disabling locus within the viral genome minimises the
risk of gene transfer to wildtype HSV. In a murine tumor model, administration
of a DISC-HSV expressing mGM-CSF resulted in inhibition of tumour growth.
In addition, Cantab is exploring means of expanding the range of possibilities
for gene delivery with DISC-HSV, namely using HSV amplicons, which would allow
delivery of DNA fragments up to 150 kb in the absence of HSV-encoded gene
products, thereby minimising the potential problems associated with
immunogenicity of the vector, and allowing long-term gene expression without the
cytolytic effect of HSV replication. The DICS-HSV provides an excellent helper
virus system for this purpose. The amplicon gene delivery technique is
potentially highly versatile; because the DISC-HSV vector and producer cell line
have already been extensively characterized, a new gene therapy application
would require only a new plasmid construct. The establishment of a manufacturing
process and a regulatory framework for the clinical use of one amplicon-vectored
gene would provide a very powerful basis for applications involving other genes.
    MODIFYING NEURONAL PHYSIOLOGY WITH A HELPER VIRUS-FREE HSV-1 PLASMID VECTOR
    SYSTEM
    Alfred I. Geller
    Division of Endocrinology, Children's Hospital, Boston, MA 02115
We have developed a helper virus-free packaging system for HSV-1 plasmid
vectors. After cotransfection into cultured fibroblast cells of a HSV-1 vector
and a HSV-1 cosmid set lacking the DNA cleavage/packaging signals, the modified
cosmid set can provide all the trans-acting functions required for the
replication and packaging of the HSV-1 plasmid vector. The resulting vector
stocks are free of detectable helper virus, and consequently, gene transfer into
the rat brain is more efficient and causes reduced cytotoxic effects and immune
responses compared to vector stocks prepared using a helper virus.
We are using this vector system to explore the potential for gene therapy of
Parkinson's Disease (PD). Injection of a HSV-1 vector which expresses tyrosine
hydroxylase into the partially denervated striatum in the 6-hydroxydopamine rat
model of PD resulted in long-term (1 year) biochemical and behavioral recovery.
Gene expression was maintained for this period and persistence of vector DNA was
demonstrated.
HSV-1 vectors may also have potential for studying the molecular and cellular
basis of specific behaviors. We have found that a HSV-1 vector that contains the
tyrosine hydroxylase promoter can be used to target expression to substantia
nigra pars compacta (SNc) neurons. A constitutively active protein kinase C
(PKC) was delivered into Snc neurons, and resulting changes in dopaminergic
neurotransmission caused changes in rat motor behavior. Thus, the PKC pathway in
Snc neurons regulates motor behavior.
    REPLICATION COMPETENT ATTENUATED HERPES VIRUS FOR TUMOR THERAPY
    Martuza RL, Mineta T, Hunter W, Todo T, Todo M, Rabkin S.
    From The Georgetown Brain Tumor Center, Molecular Neurosurgery Laboratory,
and
    the Department of Neurosurgery, Georgetown University, 3800 Reservoir Rd.
NW,
    Washington, DC 20007 USA (Fax: 202-687-3744)
In order to overcome some of the limitations of replication-incompetent
retrovirus and adenovirus systems for tumor therapy, we have explored the
progressive development, construction, and testing or replication-competent
herpes virus for the purpose of tumor therapy. Our goals have been to develop a
virus that is replication-competent in tumor cells and therefore can spread
within the tumor, yet is replication-incompetent in the normal brain so that
pathological effects will be minimized. We have progressively explored various
single mutations in the herpes simplex virus including mutations or deletions of
the thymidine kinase, DNA polymerase, ribonucleoside reductase, and gamma 34.5
genes. Using both in vitro and in vivo  data from these studies, we have now
constructed a viral vector with multiple mutations. This virus has been called
G207 and contains deletions at both gamma 34.5 loci and a lac Z gene disruption
of the ribonucleoside reductase locus. These deletions confer upon G207 a series
of properties that are potentially useful for testing against human glioma and
other tumors: 1, G207 can replicate in and kill human glioma (and other tumor)
cells; 2, G207 is not able to replicate in rodent neurons or astrocytes in
culture; 3, G207 is not toxic to mouse or primate brain upon intracerebral
inoculation; 4, G207 is hypersensitive to ganciclovir; 5, G207 is temperature
sensitive. Toxicity studies have been done or are in progress in mice by the
intracerebral, intraventricular, and intravenous routes. Sub-human primate
toxicity has been done or is in progress utilizing high titer inoculation 107
and 109 pfu) intracerebrally in aoutus, a species known to be exquisitely
sensitive to herpes virus.
     AAV VECTORS IN VIVO
    R. Jude Samulski, Ph.D.
    Gene Therapy Center, University of North Carolina at Chapel Hill, CB#7352,
    Chapel Hill, NC 27599-7352 USA
We have utilized the parvovirus AAV as a vector in initial tests aimed at
targeting neuronal and muscle cells for long term gene transduction and
expression. Efforts aimed at characterizing gene transduction in non-dividing
tissue has been achieved in both the rat and mouse model. Successful
transductions with reporter gene and various therapeutic gene cassettes have
been tested. Analysis from these studies suggest that AAV transduction in the
brain demonstrated differential and persistent expression patterns for over a
year. In no case were obvious signs of toxicity noted. Further studies suggest
long term gene expression in muscle of immuno-competent animals. Molecular
analysis of the AAV vector suggest integration as a mechanism for persistence.
Continuing efforts in improving vector production has allowed rAAV production in
the complete absence of Adenovirus infection. These studies and the important
implications they have on AAV vectors in gene therapy will be discussed.
    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
    Divisions of Pediatrics, Hematology/BMT, and Surgery, City of Hope National
    Medical Center, 1500 East Duarte Road, Duarte, CA 91010
We are exploring the potentials of the use of adeno-associated virus (AAV)
vectors for stable gene transfer into primitive hematopoietic progenitor/stem
cells. AAV vectors encoding both RNA and protein based transgenes under the
control of heterologous viral or cellular housekeeping gene promoters were
constructed and encapsidated into transducing vectors. Transduction with AAV
vectors was resistant to heat, DNase, and chloroform treatment but was abrogated
by pretreatment with anti-AAV antibodies. Analysis of AAV transduction of human
CD34+ cells revealed equivalent levels of transgene expression in both freshly
isolated and cytokine pre-stimulated cells. In short term transduction assays,
transgene expression was observed in 40-95% of cells from different donors,
suggesting efficient entry, nuclear localization, and second strand synthesis of
AAV vector sequences. Transduction of human bone marrow and cord blood-derived
primitive myeloid progenitor cells tested in long term cultures (LTBMC) and long
term culture-initiating cells (LTCIC) assays resulted in gene transfer at a
frequency of 0.2-1 copy/cell at 6-8 weeks post-transduction in the absence of
selective pressure. Fluorescent in situ hybridization (FISH) and Southern
analyses indicated the presence of integrated AAV vector sequences in CD34
chromosomal DNA in transduced cells within one week after transduction and were
detectable at comparable levels six weeks after transduction. These results
indicated that AAV vectors are capable of stable integration into primitive
hematopoietic progenitors. Transcription of the vector-encoded transgene was
detectable in LTBMC at 6-8 weeks post-transduction, indicating the presence of
transcriptionally active intracellular vector sequences. Cells from different
donors transduced in parallel with the same batches of vector showed variability
in transduction frequencies, possibly indicating a genetic basis for AAV
attachment/entry. Hematopoietic stem cell transduction was evaluated in a murine
model of serial marrow transplantation. Lethally irradiated primary recipients
of AAV vector-transduced marrow cells showed evidence of long term,
multi-lineage reconstitution. Transgene-expressing cells were detected in the
periphery at 6 months post transplantation. Serial transplants performed with
marrow cells from primary recipients resulted in the reconstitution of secondary
hosts with transduced cells. Transgene expression was detected in
lympho-hematopoietic tissues at 6 months after secondary transplantation and
12.5 months after initial transduction. Interestingly, in a few mice, cells of
both myeloid and lymphoid lineages revealed identical restriction fragment
length polymorphisms, suggesting vector integration in a common progenitor cell,
most likely a pluripotent hematopoietic stem cell. The ability to reconstitute
animals long term with AAV-vector transduced cells and to detect vector-specific
transcripts and integrated vector sequences in multiple lineages with similar
RFLP patterns suggest the utility of this vector for stem cell gene therapy.
    DEVELOPMENT OF AAV VECTORS FOR GENE THERAPY
    Samuel C. Wadsworth
    Genzyme Corporation
Gene therapy vectors based on adenoassociated virus (AAV) offer potential
advantages over other viral and non-viral vectors. AAV vectors have a relatively
simple protein coat and in general are deleted for all viral genes suggesting
that they may be less immunogenic than other viral vectors. The parent virus can
integrate into the human genome, apparently mediated by viral REP activity
suggesting a mechanism for long-term vector gene expression.  In addition, the
parent virus is not associated with any known disease and there are reports that
AAV vectors are less inflammatory. However, clear assessment of potential AAV
advantages is clouded by the difficulty in producing sufficient quantities of
vector for extensive preclinical studies at increasing dosage levels. Moreover,
AAV vector preparations are frequently contaminated with adenovirus and
wild-type AAV further complicating interpretation of gene transfer experiments.
To address these limitations we have made modifications to the AAV helper
plasmid used in vector production and in the production process itself. These
changes allow increased vector production without contamination with wild-type
AAV. More recently we have established a method for producing AAV vector in
which Ad helper virus is replaced with isolated Ad early genes, eliminating
adenovirus contamination. Progress in AAV vector production and characterization
will be reviewed as well as progress in improving AAV vector gene transfer.
    AAV VECTORS CAN BE EFFICIENTLY PRODUCED WITHOUT HELPER VIRUS
    P. Colosi, S. Elliger, C. Elliger and G. Kurtzman
    Avigen, Inc., Alameda, CA.
Vectors derived from adeno-associated virus (AAV) are being actively
investigated for several gene therapy applications. Current methods for
producing AAV vectors require that producer cells be infected with a helper
virus, most commonly adenovirus. Adenovirus can be separated from AAV vectors by
density gradient centrifugation and any residual infectious adenovirus
inactivated with heat. However, the requirement for a helper virus in the AAV
vector production process poses limitations for the production of vectors for
clinical use. By identifying the adenoviral genes that are necessary for AAV
vector replication, we developed an efficient method for producing AAV vectors
that does not rely on helper virus infection. Human 293 cells, which stably
express the adenovirus E1a and E1b genes, were systematically co-transfected
with a plasmid containing the AAV vector and helper sequences and combinations
of plasmids containing the adenoviral early genes. This analysis revealed that
the adenovirus E2a, E4orf6 and VA RNA genes were necessary and sufficient to
achieve levels of AAV vector production equivalent to those obtained with
adenovirus infection. No infectious adenovirus was generated. The AAV vectors
produced by transfecting 293 cells with these adenoviral early genes ("Ad-Free"
method) were as efficient at transducing cells in vitro as those produced with
adenovirus infection. We are currently evaluating AAV vectors produced by the
Ad-Free method in animals. Balb/c mice treated with a single intramuscular
administration of an AAV vector containing the gene for human erythropoietin
(hEpo) controlled by the CMV immediate early promoter show persistent elevation
of serum levels of hEpo and concomitant increases in red blood cell production.
The in vivo response to the administration of vector produced by either Ad-Free
method or with adenovirus was identical. The Ad-Free method greatly simplifies
AAV vector production and purification and may ultimately contribute to their
acceptability for clinical use in gene therapy.
    DEVELOPMENT OF ASSAYS FOR THE DETECTION AND QUANTITATION OF RECOMBINANT
    AAV-CFTR VECTOR
    Daryn Debeiak, Molly Fee-Maki, Teresa Johnston, Tom Reynolds
    Targeted Genetics Corporation, Seattle
AAV is a non-pathogenic human parvovirus that is being tested as a gene therapy
vector in the setting of cystic fibrosis. AAV vectors are incapable of
replication in the absence of either virally encoded rep genes or helper
functions usually provided by adenovirus. We have developed an assay to
quantitate infectious vector using a cell line inducible for AAV rep and cap
functions. This assay has been further modified to rapidly detect the presence
of infectious vector that could be shed by patients following vector
administration.
    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
The goals of the session were to discuss the recommendations for detection of
replication-competent retrovirus during retroviral vector production and patient
follow-up, in order to provide a basis for revision of the current FDA
recommendations for RCR resting.
Regarding recommendations on RCR detection during retroviral vector production,
our goal is to move toward the development and definition of RCR detection
systems which are scientifically based, can be standardized, and will allow a
balance between sensitivity of RCR detection and risk of patient exposure. The
FDA is working towards development of a standardized stock of RCR which would be
titrated and distributed to all investigators performing RCR detection assays.
This would allow investigators to "calibrate" their assay sensitivity. In
particular, we had a discussion of our current recommendation of testing of 5%
of the supernatant for RCR. Statistical predictions would suggest that RCR
detection assays should be based on the sensitivity of the assay, and how many
replicate samples need to be tested to achieve a limit concentration of RCR. The
question of what the limit concentration or tolerated dose should be (i.e., 1
RCR/ml; 0.1 RCR/mL, etc.) was raised but not answered. It was suggested that the
risk of RCR administration be compared to known risk factors for certain patient
populations (i.e., cancer patients, relative risks from chemotherapy and
radiation therapy). In a review of over three years of RCR testing by one
contract laboratory, 3/384 samples were positive for RCR, and that each of these
were due to contamination with the positive control. An additional issue which
was raised was the current recommendation to perform both co-cultivation and
supernatant testing at several stages during production+whether this dual
testing adds value. Everyone agreed that this was an important question to
address, but no one had data to support one test over the other. The presence of
an endogenous virus in the Mus dunni cell line used for amplification of RCR was
discussed. This virus is activated under circumstances where hydroxycortisone is
present (1 mM). No instances of spontaneous activation have been reported,
suggesting that this cell line, although not appropriate in indications where
hydroxycortisone is present, is still a useful reagent in RCR detection assays.
Regarding data accumulated to date with patient monitoring for RCR, the
available evidence suggests that no patients have received detectable levels of
RCR. We had presentations which summarized the data from three different
clinical trials, representing over 400 patients (with multiple time points
tested for many patients). All patient samples examined by polymerase chain
reaction for RCR sequences in their peripheral blood mononuclear cells were
negative. Some of the patients who received either direct injection of vector or
vector-producing cells had evidence of murine retrovirus-specific antibodies by
Western blot or ELISA. Patients had received greater than one injection of
material before developing antibody reactivity. The consensus expressed was that
the antibody reactivity represented reactivity to vector, rather than authentic
RCR, and that in cases where direct injection of vector and/or vector-producing
cells was used, that serological assays could be misleading. Further, there was
consensus that serological assays represented a convenient and simple
alternative assay to PCR for sponsors administering retroviral vectors ex vivo.
Data from an in vivo study of RCR infection of monkeys demonstrated that
healthy, immuno-competent monkeys can be infected with RCR. The dynamics of
infection were examined in the four monkeys on the study. In general infectious
virus was isolated from the PBMCs only at early time points post-inoculation
(2-6 weeks). Assays for RCR-specific antibodies and PCR assays detecting
envelope sequences in the PBMCs were positive early (by 4 weeks) and remained
positive in most animals for at least one year.  Several proposals for altering
the frequency of patient monitoring were made, but no consensus was reached.
Proposals ranged from having quarterly testing for the first year to having a
one-time test 12 months after administration, and one proposal suggested linking
patient monitoring to the type of vector production system used (i.e., rate of
recombination and RCR generation from different packaging cells). Alternative
proposals to our current recommendation of life-long patient monitoring were
also made: patient follow-up for two years or as clinically indicated, and/or
development of a patient tracking system which would report adverse events to
the FDA and is linked to a patient identification card to allow for notification
of the patient's participation in a gene therapy clinical trial.
The results presented in the workshop will provide the basis for a continuing
dialog both within the FDA and with the gene therapy community to develop new
recommendations for RCR testing. We encourage and welcome any additional data,
comments, or ideas which might be pertinent to such discussions. Please submit
all comments to the e-mail account GTINFO@A1.CBER.FDA.GOV by close of business
September 30, 1996
AGENDA
The goals of the session are to discuss the recommendations for RCR detection
during retroviral vector production and patient follow-up in order to provide a
basis for discussion of the current FDA recommendations for RCR testing.
    Specific questions for discussion include the following:
    A.  Regarding recommendations on RCR detection during retroviral vector
production, our goal is to move toward the development and definition of RCR
detection systems which are scientifically based, can be standardized, and will
allow a balance between sensitivity of RCR detection and risk of patient
exposure. In particular,
1) What limit concentration of RCR should be achieved in RCR detection assays?
2) Should volume tested be linked to lot size, and, if so, in what way?
3) What virus(es) should be used as standards for determination of relative
assay sensitivity?
B. Regarding our current recommendations for patient monitoring,
1) What assays are most appropriate/sensitive? Should patient population and/or
mode of vector administration be considered in deciding which assays should be
applied?
2) How often should patients be examined for RCR? Is our current recommendation
for life-long monitoring reasonable and rational? RCR testing during retroviral
vector production and in human clinical trials
    FDA/CBER Overview and goals of the session
    Andra Miller, Ph.D.
    Statistical models for testing for replication-competent retrovirus
    Tie Hua Ng, Ph.D., FDA/CBER
    Experimental issues in testing for replication-competent retrovirus
    Carolyn Wilson, Ph.D., FDA/CBER
    Viral safety of biotechnology and gene therapy products-proposed conventions
for sampling and testing for RCR
    John Spaltro, Ph.D., Microbiological Associates, Inc.
    Sensitive assays for detection of replication competent retrovirus
    Joe Hughes, Ph.D., Quality Biotech, Inc.
    Somatix Therapy Corporation, Replication competent retrovirus testing for
manufacturing and gene monitoring of patients in clinical trials
    Lawrence Cohen, Ph.D. and Joe Rokovich, Ph.D.
    Targeted Genetics, A novel murine retrovirus identified during testing for
helper virus in human gene transfer trials
    Tom Reynolds, M.D., Ph.D.
    Infection of normal rhesus monkeys with a murine replication-competent
retrovirus (RCR)
    Arifa Khan, Ph.D., CBER
    RCR testing in human clinical trials of a directly injected retroviral
vector in patients with HIV: methods, results , and impact on frequency of
patient sampling
    Dale Ando, M.D., Chiron Corporation
    Monitoring of cancer patients receiving retroviral vector gene therapy
    Gerard J. McGarrity, Ph.D., Genetic Therapy, Inc.
    Summary and Discussion
    Estuardo Aguilar-Cordova, Ph.D., Baylor College of Medicine
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
    TESTING FOR REPLICATION-COMPETENT RETROVIRUS
    Tie-Hua Ng1, Carolyn Wilson2, Andra Miller3 and Peter Lachenbruch1
    Divisions of 1Biostatistics and Epidemiology, 2 Cellular and Gene Therapies,
    and 3Application Review and Policy, CBER, FDA
Retroviral vectors are replication-defective viruses and are used in gene
therapy protocols. In the process of producing these vectors,
replication-competent retroviruses (RCR) sometimes arise, causing a public
health concern. Therefore as part of the production process, the FDA requests
all sponsors to perform a series of cell culture experiments to ensure that
their material is RCR-free prior to administration in the clinic.
How much material needs to be tested in order to provide a reasonable confidence
that the material administered will be RCR-free? Several years ago, this
question was discussed with an advisory committee. It was decided that 5% of the
cell supernatant and 1% (or 108) of the producer cells would be tested. We have
chosen to examine our current recommendations in response to concerns from
sponsors especially as they are producing larger lots of material. For example,
for a 200 liter lot 5% of the supernatant is 10 liters of material for testing.
To determine the testing requirements, we begin with the following three
characteristics.
(1) The concentration of RCR in the production lot; c (RCR/ml).
(2) The volume of the sample to be tested: vt (ml).
(3) The sensitivity of the assay: s (ffu/ml) (ffu: focus forming unit; represent
detectable RCR).
The basic assumption regarding the sensitivity of the assay is as follows. The
test on a sample volume vt will be positive if the number of RCR in the sample
is greater than or equal to s vt. For example, if s = 3 ffu/ml and vt = 2 ml,
then a single test of 2 ml will be positive if and only if the sample contains
at least 6 RCR. The detection of RCR in the infectivity assay is therefore
concentration dependent.
With this assumption, it is recommended that vt = 1/s be tested. The probability
of detection (POD) with the recommended volume may be computed based on the
Poisson distribution with parameter l = c vt = c/s, and is given by 1- exp
(-c/s). This probability is very low when c<<s. To increase the POD to a desired
level, say (1-b) for some small b (e.g., 0.05), it is necessary to perform k
tests, where k is the smallest integer greater than or equal to (-s/c)ln(b)
(i.e., rounding up) and ln denotes the natural log.
What value of c can we tolerate? For s = 1 and b = 0.05, if c = 0.1, then k =
30, which is manageable. However, if we cannot tolerate concentration higher
than 0.001 (c = 0.001), then k = 2996, which becomes impractical. If we limit k
to a reasonable number, say 50, and b = 0.05, then s/c  -50 ln(b) = 16.69.
Therefore, the sensitivity has to be no larger than 16.69 times the
concentration or the concentration has to be no smaller 1/16.69 of the
sensitivity.
The last question may be answered by specifying the patient risk of exposure to
the RCR. For a patient dose of vd, if we specify that the risk of exposure to
RCR is a, for some small a (e.g., 0.1), then c = -ln(1 - a)/vd. For example, if
a =0.1 and vd = 100 ml, then c = 0.0010536. With the limitation of the assay
sensitivity, there should be a balance between the desired limit concentration
and the number of replicates which are reasonable to perform.
    EXPERIMENTAL ISSUES IN TESTING FOR REPLICATION-COMPETENT RETROVIRUS
    Carolyn A. Wilson*, Katie Whartenby, and Andra Miller
    Divisions of Cellular and Gene Therapies* and Application Review and
Policy,
    CBER, FDA
Retroviral vectors can occasionally be contaminated with replication-competent
retrovirus (RCR) due to rare recombinational events which occur in packaging
cell lines used during production. Presence of RCR represents a safety concern
for patients because of their ability to integrate into the host genome and
potentially have a role in tumorigenesis, as well as the documented ability of
murine retroviruses to pseudotype human retroviruses, such as human
immunodeficiency virus. For these reasons, the FDA has made recommendations for
RCR testing at several stages during retroviral vector production and in ex vivo
transduced cells, where appropriate. We ask that 5% of the supernatant and 1% or
108 cells be examined in RCR detection assays.
We are currently evaluating our RCR testing recommendations. Data will be
presented from experiments which are being done to test assumptions underlying
our current recommendations, as well as to test predictions from a statistical
model which has been developed (see abstract by Tie-Hua Ng). Using the PG-4 S+L-
assay, we are examining several parameters affecting assay sensitivity, such as
number or replicate samples, high titer retroviral vector-containing
supernatant, and amplification on Mus dunni cells prior to detection on PG-4
cells.
    VIRAL SAFETY OF BIOTECHNOLOGY AND GENE THERAPY PRODUCTS-PROPOSED CONVENTIONS
    FOR SAMPLING AND TESTING FOR RCR
    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 retrovirus (RCR) in
retroviral gene therapy products of the possible introduction of viral
contaminants from virus/cell banks, components of animal origin, or adventitious
introduction during processing require that viral safety assurance testing be
performed prior to product evaluation in human clinical trials.
Testing programs to detect the presence of RCR should require the use of
sensitive and well-characterized methods for RCR detection, coupled with
strategic sampling of production volumes in order to support viral safety
assurance. Current requirements for RCR testing of gene therapy products
requires the sampling of five percent of the total production volume. With
increasing production volumes, test volume requirements are increasing and
testing methodologies have been developed to allow for economical testing of
larger test volumes. However, alternate methods have different sensitivities for
the detection of RCR, and the extent of viral safety assurance for the product
may be uncertain.
Results characterizing the sensitivities of a variety of amplification and
end-point test procedures developed for RCR determination at Microbiological
Associates will be presented. Test configurations including roller bottle, well,
and dish techniques will be presented and statistical methodology used in the
determination of the sensitivity or Probability of Detection (P.O.D.) for the
tests will be discussed.
Using the assay P.O.D., a new strategy for the viral safety assessment of large
volume production batches will be presented. Sampling issues will be addressed,
as well as the impact of new methodologies with small test volume requirements
and high throughput capability. Several examples of the proposed strategy
implementation will be presented and discussed.
    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
    Quality Biotech, Inc., Camden NJ 08104
Amplification of Mus dunni cells followed by a sensitive infectivity assay such
as the PG4 S+L- focus forming assay has become one of the standard procedures
for the detection of replication competent retroviruses (RCRs). This combined
system is routinely used in many laboratories for testing gene therapy vectors
and other biopharmaceuticals (i.e., murine hybridomas). In our laboratory this
system has proven to be one of the most sensitive assays for the detection of
infectious or recombinant murine leukemia viruses (MuLV). The 14 day test is
used either as a co-cultivation for cells or an amplification for supernatants
from cell and production lots; these assays have two subpassages supplying fresh
cells at each passage. Additional passages (up to 5) have been performed on some
Master and Working Cell Banks; validation studies have demonstrated that only a
low number of passages is needed to achieve maximum sensitivity for many
retroviruses.
We have demonstrated in validation experiments that the Mus dunni co-cultivation
assay will enhance the detection of known amount of RCR by a least one log 10
unit. This enhanced detection of RCR has been demonstrated for small volumes as
well as large volumes of sample. For these tests up to 20 ml or 100 to 125 ml of
test material is plated on each T150 flask or roller bottle of Mus dunni cells,
respectively. The PG4 S+L- assay has also been validated for a number of tests.
The data from these studies show very good linearity for titrated virus (r2 of
0.95 to 0.99), reproducibility within or between tests of 0.35 and 0.45 log10
units respectively, and very good sensitivity of 2 to 3 focus forming units
(FFU)/ml.  Large volume sampling versus multiple small volume testing for the
PG4 S+L- assay has also been examined. Introduction of a method to fix and stain
the PG4 S+L- has significantly improved the recording of infectious retrovirus
foci.
We have examined a variety of gene therapy products from a number of sources for
the detection of RCRs. The tests have either included the amplification on Mus
dunni cells followed by the PG4 S+L- assay or just the PG4 S+L- assay (on
materials already amplified by the client). Current regulatory guidelines
suggest that 1% of the producer cells (108 minimum) and 5% of a harvested
supernatant are required for RCR testing. In our experience, the typical number
for co-cultivations has ranged from 107 to 108 and vector lots have ranged from
a few milliliters to 3 liters. Cell banks, production lots, and all supernatants
have been routinely negative for the presence of RCR. The results from several
hundred PG4 S+L- assays of client-amplified material demonstrated only three
positive tests for RCR. These samples were found to be RCR positive due to
cross-contamination with a retrovirus used by the sponsor company for
co-cultivation controls.
In addition to evaluation of gene therapy vectors for RCR, we have examined a
number of other products for evidence of infectious MuLV. Evaluations of other
biopharmaceutical products, such as murine hybridomas, often demonstrate a small
number of infectious units associated with high levels of retroviral particles.
Initial sampling from the unamplified products may show undetectable to 10
FFU/ml. Upon amplification of these same lots, high levels of PG4 S+L- -positive
retroviruses (typically 104 to 105 FFU/ml) are expressed. Thus the Mus dunni/PG4
S+L- system still remains a very sensitive assay for detection of infectious
MuLV retroviruses in a variety of biopharmaceuticals. Highly sensitive PCR-based
assays (QC-PCR, PERT) can also be used for the detection of specific RCR in
recombinant viral preparations. The strategy for these tests would involve
customizing the PCR protocol for the most likely RCR resulting from rare
recombination events with the packaging cell line DNA. The development of
PCR-based assays is ongoing for a variety of gene therapy vectors. Such
PCR-based tests should provide a rapid and highly sensitive RCR detection assay,
which can be used alone or in combination with scaled-down co-cultivation
studies.
    REPLICATION COMPETENT RETROVIRUS TESTING FOR MANUFACTURING AND MONITORING OF
    PATIENTS IN CLINICAL TRIALS
    L. Cohen and J. Rokovich
    Somatix Therapy Corporation, Alameda, CA
Somatix Therapy Corporation has developed the MFG-S retroviral vector with the
Y-CRIP packaging cell line as a delivery system for the huGM-CSF transgene. The
MFG-S-huGM-CSF transducing particles have been employed to create genetically
modified tumor cell vaccines for use in clinical protocols since late 1993. For
each patient autologous tumor tissue is cultured ex vivo, transduced with
MFG-S-GM-CSF, irradiated, and administered to the patient.
Replication competent retrovirus (RCR) testing is employed at two distinct
stages of manufacture+at the end of transducing particle manufacture and at the
end of the tumor tissue processing. The development of the RCR cocultivation and
supernatant testing methods will be described, the controls identified, and the
sensitivity of the assays discussed. Recommendations will be made as to the
appropriate RCR testing samples and methods (cocultivation vs. supernatant) at
various stages of manufacture, and the potential standardization of this
testing.
On September 20,1993 the FDA requested that all IND sponsors using a retroviral
vector monitor clinical trials subjects for evidence of replication competent
retroviruses by three different methods. Somatix will relate its RCR monitoring
experience with over 20 patient-years of experience. Additionally,
recommendations for future monitoring will be presented including the type of
testing and sampling frequency.
    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
    Fred Hutchinson Cancer Research Center1, Department of Pathology2, and
Medical
    Scientist Training Program4 and Targeted Genetics Corporation3, Seattle,
    Washington
Testing of retroviral vector supernatants and transduced cells used in human
clinical trials for the presence of replication-competent retrovirus (RCR) is
required. We have detected a novel inducible retrovirus produced under certain
conditions during sensitive marker rescue assays using a Mus dunni  cell line.
The implications of this result and impact on RCR testing of retrovirus vectors
and transduced cells will be discussed.
    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
    1Division of Viral Products, CBER, FDA, Bethesda, MD; 2Microbiological
    Associates, Inc., Rockville, MD; 3Laboratory of Immunopathology, NIAID, NIH,
    Bethesda, MD.
Replication-competent retroviruses (RCR) can arise in come cases unexpectedly
and spontaneously from retrovirus-based packaging cell systems used in human
gene therapy products. RCR containing an amphotrophic murine leukemia virus
(MuLV) envelope possess a broad host range and can infect a wide variety of
primate cells including human. To evaluate the potential of RCR infection in
humans, four normal juvenile rhesus monkeys were inoculated with a vector virus
preparation (S3A) which contained RCR (provided by Genetic Therapy, INC.,
Gaithersburg, MD). The monkeys were monitored regularly for virus infections by
1.) isolation of infectious MuLV from the PBMC; 2.) Western blot analysis for
antibody response; and 3.) DNA PCR for persistence of viral sequences in the
host. Clinical changes were monitored by serum chemistry and hematology. The
data indicate that all four animals were infected with RCR; although virus
replications was early and transient, persistence of viral sequences was seen.
Each animal responded uniquely with respect to the kinetics of infections and
antibody response. The results of a three year study will be presented.
    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
PCR assays capable of detecting a single copy of target sequence in the
background of >11 x 105 cell equivalents have been developed for monitoring
HIV-positive clinical trial subjects for the presence of RCR in PBMC and for the
presence of vector in semen samples. These tests have been used to monitor the
297 patients enrolled in the Phase I and Phase II clinical trials for HIV-IT(V),
an immunotherapeutic recombinant retroviral vector expressing the HIV-1 env/rep
genes. Patients have been treated with cumulative doses of up to 180 million
cfu/pt (Ph II). In Phase I studies, 79/81 patients have been tested for RCR at a
minimum of three time points. In Phase II studies, all patients (216) have been
tested for RCR. Tests were uniformly negative. Semen samples from 119 patients
in the Phase II trial were analyzed for the presence of HIV-IT(V) vector
sequences using PCR.
We have recently developed an ELISA that can detect antibodies to amphotrophic
MLV. We have used this assay to screen serum samples from 46 Phase I clinical
trial patients that had previously tested negative for RCR by PCR. Only one
subject showed a positive change in anti-MLV titer after HIV-IT(V) with negative
RCR by PCR. The ELISA test may be more appropriate for monitoring patients
enrolled in ex vivo product trials, since ex vivo products should not express
amphotrophic MLV proteins.
There has been no evidence of RCR infection in the HIV-infected Phase I and II
clinical trial patients treated with HIV-IT(V). This is consistent with results
from the Quality Control testing of the product; cocultivation of 108
post-production cells and direct testing of bulk HIV-IT(V) product with an MLV
replication-permissive cell line, were negative. Semen analysis did not reveal
any evidence of germline transduction. These data show that RCR infection and
germline transmission have not been detected in patients treated with the
HIV-IT(V). As such, a reduced testing frequency for RCR and germline
transduction in patients seems reasonable.
    MONITORING OF CANCER PATIENTS RECEIVING RETROVIRAL VECTOR GENE THERAPY
    Gerard J. McGarrity, Ph.D.
    Genetic Therapy, Inc., Gaitherburg, Maryland 20878
In 1993 CBER presented recommendations for monitoring of patients that have
received retroviral vectors for gene therapy. These recommendations have
included the type and frequency of assay for this patient population. Here we
present the results of monitoring of 57 patients enrolled in brain cancer
protocols involving the use of murine vector producer cells that deliver a
retroviral vector encoding for Herpes simplex thymidine kinase. These 57
patients were enrolled in six different protocols that involved the delivery of
approximately 1 x 109 vector producer cells to patients that had recurrent
glioblastoma multiforme and other neoplasms in the CNS. Patient assays have
included the use of methods to detect antibodies to the mouse producer cells and
to vector components as well as polymerase chain reaction (PCR) assays on
peripheral blood lymphocytes to detect the presence of vector and replication
competent retrovirus (RCR). RCR has not been detected in any of 263 specimens
from 57 different patients by PCR. In addition, six different patient specimens
were also negative in biological assays for PCR. Presence of vector was detected
in certain patients. The majority of these, however, received two or more
inoculations of producer cells. The methods and implications of these results
will be discussed.
    BREAKOUT SESSION 7:
    PHARMACOLOGY & TOXICOLOGY
    SUMMARY
    Anne M. Pilaro and Joy Cavagnaro
    FDA/CBER
 Speaker Presentations
Five speakers provided presentations, demonstrating how preclinical studies in
animals have facilitated clinical development of adenoviral, retroviral, or
adeno-associated viral vectors as therapeutic approaches in the treatment of
cancer, cystic fibrosis, AIDS, or inborn errors of metabolism. Brendan Lee
(Baylor College of Medicine) discussed how changes in the route of
administration of adenovirus can affect the toxicity profile of the vector; it
was concluded that the limiting toxicity of adenoviruses by any route was
induction of a dose-related, inflammatory response. Additionally, a sharp
threshold in the dose-related toxicity of adenovirus was noted after intravenous
administration and was accompanied by a clinical picture resembling disseminated
intravascular coagulation, which may limit the use of these vector by this route
at high doses in the clinic. Using a murine model of oyster glycogen-induced
peritonitis, Jeff Holt (Vanderbilt University) demonstrated that the presence of
underlying disease could enhance the toxicity and dissemination of a retroviral
vector used for treatment of peritoneal cancers. A similar, reversible toxicity
(elevation of hepatic transaminases and infiltration of inflammatory cells) was
noted in 1/6 patients treated with this vector, suggesting that the mouse model
provided a relevant means to assess the potential clinical toxicities associated
with this regimen. By contrast, Ron Crystal (Cornell Medical Center and GENVEC)
discussed how the clinical toxicity noted in one cystic fibrosis (CF) patient
treated with an adenoviral vector encoding the corrected CFTR gene was not
predicted by studies conducted in Rhesus monkeys before the onset of the phase 1
trial. The toxicity observed in the patient resulted in a change in the strategy
of dosing for future trials, such that the volume and the method of
administration were vastly altered. These changes have resulted in successful
administration of the vector to 12 CF patients without the clinical sequelae
noted in the first trial, and effective re-treatment has been accomplished
without toxicity. Studies in the Rhesus monkey with an adeno-associated viral
vector (AAV) were presented by Tom Reynolds (Targeted Genetics), and focused on
the mechanism of vector dissemination after intra-bronchial administration and
the potential clinical and pathologic consequences of infection with AAV-based
vectors.  Data demonstrated that the low-level spread of the vector was most
likely due to inadvertent inoculation of hematologic cells during the treatment
procedure, was detected only transiently, and was without pathologic
consequence. Assays to attempt to "rescue" and generate replication-competent
AAV vector displayed only limited viral recovery in monkeys carrying latent
infection with wild-type AAV and subsequent infection with very high levels of a
host-range mutant, wild-type adenovirus. These parameters were identified from
the preclinical studies as considerations to be followed during the clinical
trial. Finally, Mike Blaese (NCHGR, NIH) presented clinical data from patients
treated with retrovirally-modified, gene-transduced cells for several different
indications, including adenosine deaminase deficiency and AIDS. Long-term
follow-up (out to 6 years in one case) has demonstrated in some cases the
persistence of the gene-modified cells, improvement in functions associated with
the transduced genes, and no evidence of significant clinical toxicities or
generation of replication-competent retrovirus recombinants, helper viruses, or
pseudotypes. There was a wide degree of variability in response between
patients, suggesting that as the animal models were not predictive of some
clinical responses, so humans may not always predict other humans' responses
either.  In the absence of suitable animal models to study these issues,
long-term safety of gene-modified cells will only be determined by clinical
evidence, and will require careful clinical monitoring.
Discussion Session
Five questions were put to the audience for open discussion, and are presented
individually below. A general conclusion was that for each of the issues
discussed, the route, the dose, and the indication for which the vector was
intended should be carefully considered and balanced with the risks to determine
what would be acceptable.
1.  How much RCA is "safe" to administer to patients? What preclinical data are
useful to address the safety of RCA present in adenoviral vector preparations,
especially after alternate routes of administration?
Conclusion: Regarding the administration of doses of adenoviral vector
containing higher amounts of replication-competent virus (RCA) than the current
standard allows, it was concluded that the risks may be more acceptable by some
routes (e.g. subcutaneous) or in some indications (e.g. cancer) than in others
(e.g. intra-bronchial in cystic fibrosis). However, increased amounts of RCA may
be permitted in these situations if data are provided by the sponsor from
experiments designed specifically to address the safety of RCA administration in
the presence of the vector, and can provide an adequate, quantitative estimate
of the risks. Preclinical data were cited by sponsors demonstrating that
administration of up to 106 RCA in the lung in an excess of
replication-defective particles was acceptable, while in the clinic, doses of
vector which included RCA levels of 10 or 100 were administered without adverse
effects. The group also concurred that RCA may no longer be as great a concern
in the future, as further improvements in the way these vectors are produced and
in the gene constructs themselves decrease the likelihood of recombination
events.
2.  What preclinical studies should we consider to demonstrate the
delayed/long-term consequences of gene therapy? What animal models will be most
useful for each vector class?
Conclusion: Discussion concurred that more preclinical data may be needed,
depending on the duration and expected consequences of the proposed therapy.
Animal models may be useful to study long-term effects, especially if predictive
results of general organ toxicity are obtained in early studies. Current
clinical monitoring practices in humans will also contribute to long-term safety
assessment.
3.  What safety studies should be required to demonstrate a lack of
toxicity/developmental effects to germ line cells when positive gene transfer is
seen.
after inadvertent distribution of vector to the gonads?
after intended targeting to germ cells?
Conclusion: Intended targeting of vectors to germ cells was not discussed at the
present time. If a positive result of gene transfer to the gonads is obtained in
an animal study, it was agreed that further preclinical studies need to be
carefully defined to better assess the risks to the patient population. Study
designs should employ the best available techniques, and explain or justify the
limitations of detection for the method employed. These studies should initially
focus on identification of the cell type in the gonadal tissue in which the
signal is positive, with follow-up studies conducted if "positive" gene transfer
is obtained, i.e. is there integration into the germ cell.  Current practice is
to include monitoring for potential targeting to the germ line (i.e. analysis of
semen samples from male patients receiving gene vectors for the inserted
construct) as part of the clinical protocol.
4.  At what phase in development should full-scale reproductive toxicity studies
of gene therapy constructs be considered?
Conclusion: Studies to define the effects of gene therapy on reproductive
capacity and/or outcome will be based on the gene product, as well as the
patient population intended for treatment. It was concluded that traditional,
reproductive toxicity study designs and timelines during drug development may
not be appropriate for gene therapy vectors. An example was cited from the area
of DNA vaccines, in which a problem-specific approach to the design of a
reproductive study to support use of the vaccine in pregnant women was employed.
By conducting a specific, targeted preclinical study designed to answer the
safety of this product in pregnant animals, data should be generated that will
facilitate its use in the intended population.
5.  Is there a mechanism for "data-sharing" or conducting "investigator-neutral"
studies to support various, recurrent questions about the safety of gene
therapy, and if so, which areas would be most useful to focus on?
Conclusion: The group concluded that there was an overall willingness to share
and/or publish information from preclinical studies, but acknowledged that there
were differences between corporate and academic institutions' ability to make
data publicly available. As products move into phase 2 and ultimately, phase 3,
there was some concern that data would become less available to the field. The
general consensus was that focus should be on the total "product", and not on
individual vectors. Annual scientific conferences, such as are sponsored by the
Cystic Fibrosis Foundation yearly in Williamsburg may be useful as a means to
open communication in the field; alternatively, forums or conferences sponsored
through ORDA/ODAC may achieve this end as well.
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
AGENDA
I. INTRODUCTION
    Anne Pilaro
II. SPEAKER PRESENTATIONS - 20 MIN EACH
    How changes in route of administration of adenoviral vectors affect the
toxicity in vivo
    Brendan Lee, Baylor College of Medicine
    How a disease state may modify the distribution and safety of retroviral
vectors in vivo, using a newly developed murine model system
    Jeff Holt, Vanderbilt University
    Distribution studies in support of adeno-associated viral vector safety
after administration to the lung
    Tom Reynolds, Targeted Genetics
    Using in vitro data to support the safety or retrovirally-transduced cells
in the absence of a suitable animal model for in vivo safety testing
    Eric Poeschla, UCSD
    Preclinical studies with adenoviral vectors to define safety parameters for
clinical monitoring after targeted administration
    Ron Crystal, GenVec/Cornell Medical Center
    Long-term follow-up of patients receiving retrovirally-transduced cells:
What have we learned?
    Mike Blaese, NCHGR
III. PANEL DISCUSSION, JOY CAVAGNARO, CHAIR
    Topics for panel discussion are as follows (approximately 5-10 min each):
    1. How much RCA is "safe" to administer to patients? What preclinical models
are useful to address the safety of RCA present in adenoviral vector
preparations, especially after alternate routes of administration?
    2. What preclinical studies should we consider to demonstrate the
delayed/long-term consequences of gene therapy? What animal models will be most
useful for each vector class?
    3. What safety studies should be required to demonstrate a lack of
toxicity/developmental effects to germ line cells when positive gene transfer is
seen+after inadvertent distribution of vector to gonads?--after intended
targeting to germ cells?
    4. At what phase in development should full-scale reproductive toxicity
studies of gene therapy constructs be considered (if ever)?
    5. Is there a mechanism for "data sharing" or conducting "investigator
neutral" studies to support various, recurrent questions about the safety of
gene therapy, and if so, which areas would be most useful to focus on?
IV. SUMMARY AND CLOSING REMARKS+ANNE PILARO AND JOY CAVAGNARO
    The goal of this session is to understand how preclinical studies have
helped to identify common areas of concern regarding the safety of gene therapy
for each of the different vector types. Specific questions include:
    How changes in the route of administration of a vector may modify its
toxicity.
    How the presence of a disease state may modify the safety of a given gene
therapy and whether normal animals serve as good predictors of the clinical
experience.
    When can pre-clinical studies be minimized?
    How preclinical data have identified parameters to follow in the clinic.
    What pharm/tox studies should be considered in support of future phase 2 and
 phase 3 gene therapy trials?
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
    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
    Department of Cell Biology, Vanderbilt University, Nashville, TN, and the
    University of Washington.
Interperitoneal infusion of retroviral vectors is a model system we have
employed for initial preclinical and early clinical studies. Mouse studies
demonstrate that spread of retroviral vector from the peritoneal cavity to blood
and tissues (identified by PCR) is dependent on vector dose and enhanced by
oyster glycogen-induced peritonitis. Although XM6:antifos and LXSN-BRCA1
retroviral vector infusion show similar spread to tissues, LXSN-BRCA1 produces a
transient peritonitis and focal hepatocellular necrosis which is dose-dependent
and enhanced by oyster-glycogen induced peritonitis. Clinical studies are just
beginning, but 1 of 6 patients treated with intraperitoneal LXSN-BRCA1 for
ovarian cancer developed a transient temperature (102.4) and focal peritoneal
signs eight hours after infusion. Fluid cytology showed increased neutrophils
consistent with a limited peritonitis. PCR/Southern studies show that vector can
be detected in peritoneal fluid for 24 hours after infusion, and plasma studies
show no development of antibody to retroviral envelope proteins in two patients
(we have seen antibodies develop in one patient treated with intrapleural
infusion of XM6: antifos vector). Dose escalation studies in patients are
continuing along with concurrent animal studies aimed at further understanding
of the mechanism of LXSN-BRCA1 peritonitis.
    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
    Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, TX
    77030.
As part of preclinical studies for adenoviral mediated gene therapy,
toxicological studies of E1 and E1/E2 deleted adenoviruses were carried out with
different routes of delivery in several animal models. Up to 1.5 X 109 p.f.u. E1
deleted adenovirus expressing the Herpes virus thymidine kinase gene (Ad.RSV-tk)
was delivered via intracerebral injection in nonhuman primates and cotton rats.
Cotton rats were studied with and without preimmunization to wild-type
adenovirus. Histologic inspection showed inflammation of the ependyma and
choroid plexus and at the injection site. No differences were noted between the
doses analyzed, gancyclovir (GVC) treatment, or status of preimmunization.
Similarly, injection of higher doses up to 1.5 X 109 p.f.u. of Ad. RSV-tk with
and without GVC treatment was carried out in baboons. Adverse clinical symptoms
were only seen in the group injected with the highest dose and who were treated
with GVC. However, MRI and necropsy revealed cystic changes at the higher dose
and necrotic foci in all groups. These experiments showed toxicity associated
with direct intracerebral injection of Ad.RSV-tk and GVC treatment.
Similar studies were carried out via injection into dorso-lateral prostates of
C57BL/6 mice. No differences in local inflammation between the doses analyzed
were noted in the groups without GVC. However, a greater degree of leukocytic
infiltration was noted at higher doses treated with GVC. Vector sequences were
localized primarily to the dorsolateral and ventral prostate and detected
outside of local inflammatory response.
The effect of systemic delivery of up to 109 infectious units (i.u.) of
Ad.RSV-tk via intracardiac injection with and without GVC treatment was studied
in cotton rats. Primary distribution was in the heart, lung and thymus.
Microscopic lesions were noted in the heart and spleen of animals given the two
higher doses with greater splenic hemosiderosis noted in the GVC treated group.
Alternative systemic delivery via intravenous injection was studied in baboons
and C3H/HeOuJ mice. Injection of 1.1 X 1013 particles/kg of an E1 deleted
adenovirus expressing b-galactosidase (Ad.PGK-b-Gal) resulted in lethargy and
petechial lesions which progressed to dyspnea by 48 hours. Elevated liver
functions and abnormal hematological values were noted. Pathology analysis
showed severe and diffuse vasculopathy.
Intravenous administration of E1 and E1/E2 deleted adenoviruses expressing human
a-1-antitrypsin at up to 9 X 1013 and 1 X 1013 particles/kg, respectively, was
analyzed in C3H/HeOuJ mice. Dose-related toxicity was noted primarily in the
liver and spleen. A dose-independent transient thrombocytopenia was also
detected. Potential safety advantage of E2 deleted adenovirus was evaluated by
measurement of serum liver function markers and by histopathology.
An overview of these experiments will be presented and their cumulative
significance to the safety of adenoviral vectors in gene therapy will be
discussed.
    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
    Eudowood Division of Pediatric Respiratory Sciences1 and the Departments of
    Physiology2, Genetics3, and Comparative Medicine4, Johns Hopkins University
    School of Medicine, Baltimore, MD, and Targeted Genetics Corporation5,
    Seattle, WA.
AAV is a nonpathogenic human parvovirus that has potential application as a gene
therapy vector. Preclinical studies were conducted to support human trials of an
AAV-CFTR vector in the setting of cystic fibrosis. In the first study, vector
was administered to the right lower lobe of rhesus macaques at several doses. No
toxicity was observed at any dose level using a variety of clinical and
laboratory measures. Vector DNA and RNA were detected in the lungs of treated
animals for up to 6 months and no RNA expression was observed. Additional
studies showed that low levels of vector DNA could be detected transiently in
the peripheral blood lymphocytes of animals following vector administration.
Vector shedding was not observed following bronchoscopic administration to the
lung. The low level vector rescue was only observed following high dose anatomic
co-administration of vector and wild-type AAV in the presence of adenovirus.
These results indicate that in the rhesus model, AAV vectors are capable of
mediating efficient and persistent gene transfer in the absence of toxicity, and
that the risks of vector shedding and rescue appear to be low.
    IN VIVO ADENOVIRUS VECTOR PREDICTABILITY OF SAFETY STUDIES IN EXPERIMENTAL
    ANIMALS
    Ronald G. Crystal, MD
    Division of Pulmonary and Critical Care Medicine, The New York
    Hospital-Cornell Medical Center, New York, New York
The demonstration of the ability of replication-deficient adenovirus vectors to
transfer genes in vivo led to the first human trials of adenovirus-mediated gene
transfer in 1993. Since then, adenovirus vectors have been used to transfer
genes to more than 120 humans, in trials directed toward the therapy of cystic
fibrosis, colon cancer metastatic to liver, malignant mesothelioma and
bronchogenic carcinoma. Safety studies have been carried out in experimental
animals in order to evaluate the type and extent of possible adverse events that
might be expected in humans. Now that there has been more than three years
experience with adenovirus vectors in humans, it is useful to look back at the
animal safety studies that have been carried out, and to ask the question: are
in vivo adenovirus vector safety studies in experimental animals predictable of
adenovirus vector administration to humans?
The bulk of the experience with adenovirus vectors in humans is focused on
cystic fibrosis (CF), a common recessive disorder caused by mutations of the
cystic fibrosis transmembrane conductance regulator (CFTR) gene, a 250 kb, 27
exon gene on chromosome 7. The major manifestations are on the airway epithelial
surface, with purulent mucus, opportunistic infections, chronic inflammation and
progressive loss of lung function. There is compelling logic underlying the
concept that the respiratory manifestations of CF could be prevented by gene
therapy. First, there is overwhelming evidence that the respiratory
manifestations of CF result from insufficient CFTR function in the airway
epithelium. Second, the deficiency in CFTR function in the airway epithelium can
be corrected in vitro by transfer of the normal CFTR cDNA into the airway cells.
Finally, a variety of strategies have been used to successfully transfer the
normal human CFTR cDNA to the airway epithelium of experimental animals in vivo.
Collectively, the initial gene therapy studies using adenovirus vectors have had
three goals: (1) to demonstrate the feasibility of gene transfer and expression
in the respiratory epithelium; (2) to evaluate the ability of the transferred
gene to function; and (3) to define the safety of the gene transfer process. Two
respiratory epithelial targets have been evaluated: the airway epithelium (the
site of the clinical abnormalities) and the nasal epithelium (a surrogate site
used to evaluate function). Several important lessons have been learned. First,
it is feasible to transfer the CFTR cDNA to the human respiratory epithelium in
vivo, with expression of the CFTR cDNA at the mRNA and protein levels. Second,
although not observed in all studies, trends toward functional correction of the
electrophysiologic biologic abnormality have been observed following CFTR cDNA
transfer to the nasal epithelium. Third, it is now possible to quantify
vector-derived CFTR gene expression in bronchial epithelial samples at the mRNA
level, and to determine the extent of vector-derived CFTR mRNA expression
compared to endogenous CFTR mRNA expression, thus providing a quantitative goal
to measure success of gene transfer to the airways. Finally, the "safety window"
of doses and volumes has been approximated for administration of adenovirus
vectors to the airways. Adverse events have been minimal, but they have been
observed in both the nasal epithelium and the lung. In the nasal epithelium,
mild, reversible inflammation has been observed at high doses. In the lung,
parenchymal inflammation has been observed at high doses when the vector
inadvertently reached the alveoli, rather than remaining in the bronchi (the
target of the therapy).
Interestingly, although dose-dependent inflammation is observed in the lung
parenchyma in animals receiving adenovirus vectors, studies with high doses in
several species (mouse, cotton rat, and non-human primates) have failed to
predict the adverse events observed in humans. From this experience, the
following conclusions can be drawn. First, if toxicity is observed in
experimental animals, human studies must be approached with caution. However, if
no toxicity is observed in animals, the same caution must be applied to human
studies, i.e., the animal studies may not predict the human responses. Second,
no experimental animals are identical to humans in regard to adenovirus vectors,
and there are no experimental animal models that exactly mimic human disorders,
such as CF. Third, non-human primates do not seem to offer any advantages over
rodents in regard to safety predictability for these vectors. Finally, despite
the need to carry out studies in experimental animals before initiating human
studies, it is only by studying humans that true safety profiles of biologic
reagents, such as adenovirus gene transfer vectors, can be determined.
    APPENDIX
    Overview of Regulatory Documents Available by CBER FAX BACK
 FDA GUIDANCE DOCUMENTS
The Center for Biologics Evaluation and Review (CBER) of the Food and Drug
Administration (FDA) provides specific guidance to sponsors for the purposes of
aiding the development and evaluation of products. Specific guidance is based
upon scientific principles and scientific experimentation derived from in-house
as well as academic and industry sources. FDA does not intend that guidance
documents be all-inclusive. Alternative approaches could be warranted in
specific situations, and certain aspects might not be applicable in all
situations. If an applicant believed the application procedure described in a
guidance document was inapplicable to a specific situation for a particular
product, the applicant could provide, for CBER's consideration, information
supporting an alternative process. If an applicant chooses to use alternative
processes, the applicant may wish to discuss the matter further with the agency
to prevent expenditure of money and resources on activities that later might be
determined to be inappropriate to FDA. Guidance documents do not create or
confer any rights for or on any person, do not operate to bind FDA or the
public, and represent the agency's current thinking.
    Points to Consider (PTC): Recommendations to follow or consider for areas
where there is limited product experience or rapidly changing technology.
    Letter to Sponsor :A guidance document sent to sponsors, sometimes
proprietary, describing the current regulatory approach to a specific issue.
     Other Guidance Documents Include guidelines and memoranda to
manufacturers and sponsors.
INTERNATIONAL CONFERENCE ON HARMONIZATION (ICH)
    Guidelines developed by joint agreement of the US, Europe, Japan, and their
industry representatives.  Upon final acceptance, ICH documents supersede FDA
guidelines and PTC where applicable.
LAW/UNITED STATES CODE
    Code of Federal Regulations (CFR) (CFR Title 21- Food and Drugs) A
compilation of all effective government regulations and interpretations of the
law. A regulation is defined in 21 CFR 10.3 (a) as "an agency rule of general or
particular applicability and future effect issued under a law administered by
the Commissioner or relating to administrative practices and procedures".
Regulations are usually subject to public comment before finalization. The CFR
is binding upon both FDA and sponsors.
    Federal Register Notice A procedural announcement in the Federal Register of
contemplated actions or policy decisions and which provides the opportunity for
public comment.
    Food, Drug and Cosmetic Act and the Public Health Service Act Two of the
major laws that mandate the responsibilities of the Food and Drug
Administration. Specific interpretations of these laws are contained in the CFR.
FDA and sponsors are bound by these laws.
    DOCUMENTS AVAILABLE FROM THE CBER
NEW TELEPHONE NUMBER!
1-888-CBER-FAX
FAX INFORMATION SYSTEM - 1-301-827-3844
July 19, 1996
Document    Document Name
Number
1000    Guideline for Quality Assurance in Blood Establishments,
        7/11/95, 36 pages
1001    OELPS, Advertising and Promotional Labeling Staff Procedural
        Guidance Document (Draft), 8/94, 6 pages
1002    Points to Consider - Characterization of Cell Lines Used to
        Produce Biologicals, 7/12/93, 42 pages
1003    Points to Consider in Human Somatic Cell Therapy and Gene
        Therapy, 8/27/91, 21 pages
1004    Draft Guideline for the Validation of Blood Establishment
        Computer Systems, 9/28/93, 32 pages
1005    Points to Consider in the Production and Testing of New Drugs
        and Biologics Produced by Recombinant DNA Technology, 4/10/85,
        14 pages
1006    Supplement to the Points to Consider in the Production and
        Testing of New Drugs and Biologicals Produced by Recombinant
        DNA Technology: Nucleic Acid Characterization and Genetic
        Stability, 4/6/92, 9 pages
1007    Draft Points to Consider in the Manufacture and Testing of
        Monoclonal Antibody Products for Human Use (8/94), 43 pages
1008    Points to Consider in the Production and Testing of Interferon
        Intended for Investigational Use in Humans, 7/28/83, 21 pages
1009    Guideline for Adverse Experience Reporting for Licensed
        Biological Products, 10/15/93, 56 pages
1010    ICH Guideline Quality of Biotechnological Products: Analysis
        of the Expression Construct in Cells Used for Production of
        r-DNA Derived Protein Products, 2/23/96, 3 pages
1011    Draft ICH Document-Quality of Biotechnological Products:
        Stability Testing of Biotechnological/Biological Products,
        8/21/95, 19 pages
1012    ICH Guideline for Industry, Clinical Safety Data Management:
        Definitions and Standards for Expedited Reporting, 3/1/95, 16
        pages
1013    Points to Consider in the Manufacture and Testing of
        Therapeutic Products for Human Use Derived from Transgenic
        Animals, 8/22/95, 20 pages
1014    Draft Points to Consider for the Evaluation of Combination
        Vaccines: Production, Testing, and Clinical Study (1995),
        10/3/95, 28 pages
1015    Guidance for Industry; Content and Format of Investigational
        New Drug Applications (INDS) for Phase 1 Studies of Drugs,
        including Well-Characterized, Therapeutic,
        Biotechnology-derived Products, 11/95, 17 pages
1016    Draft Guidance, Changes to an Approved Application, 1/96, 19
        pages
1017    Draft Guidance, Changes to an Approved Application for a
        Well-Characterized Therapeutic Recombinant DNA-Derived and
        Monoclonal Antibody Biotechnology Product, 1/96, 14 pages
1018    Draft Addendum to The Points to Consider in Human Somatic Cell
        and Gene Therapy, 1/96, 18 pages
1019    ICH Final Guidance on the Need for Long-Term Rodent
        Carcinogenicity Studies of Pharmaceuticals, 3/1/96, 4 pages
1020    ICH Guideline on Detection of Toxicity to Reproduction for
        Medicinal Products: Addendum on Toxicity to Male Fertility,
        4/5/96, 3 pages
1021    FDA Guidance Concerning Demonstration of Comparability of
        Human Biological Products, Including Therapeutic
        Biotechnology-derived Products, 4/26/96, 10 pages
1022    Draft ICH Guideline on Viral Safety Evaluation of
        Biotechnology Products Derived From Cell Lines of Human or
        Animal Origin, 5/10/96, 11 pages
1023    Guidance for Industry: The Content and Format for Pediatric
        Use Supplements, May 23 1996, 7 pages
1024    Guidance on Applications for Products Comprised of Living
        Autologous Cells Manipulated Ex Vivo and Intended for
        Structural Repair or Reconstruction, May 24, 1996, 10 pages
2000    Betaseron Approval Information, 7 pages
2001    1993 Biologic & Drug Approvals, 9 pages
2002    Advisory Committee Information Line Instructions, 1 page
2003     Baxter IGIV Talk Paper, 2/25/94, 2 pages
2004    How to Obtain a Certificate to Foreign Governments, 1 page
2005    How to Make a Freedom of Information Act Request, 1 page
2006    How to Use a Fax Modem with the CBER Fax Information System, 1
        page
2007    How to Obtain CBER Documents by Internet Mail, 1 page
2008    How to Obtain CBER Documents by File Transfer Protocol (FTP),
        1 page
2010    1994 CBER Approval Actions, 1/10/95, and FDA Talk Paper - 1994
        Medication Approvals, 1/17/95, 7 pages
2011    Press Release and Approval Letter, Varicella Vaccine, 3/17/95,
        5 pages
2012    Information about Export Applications and Requirements, 6/95,
        21 pages
2013    1995 CBER Approval Actions and FDA Talk Paper - FDA 1995
        Approvals, 1/16/96, 6 pages
*2014   CBER User Fee Performance Goals, Actions as of 6/30/96, 15
        pages
2015    Press Release and Approval Letter, Respiratory Syncytial Virus
        Immune Globulin Intravenous (Human), 1/19/96, 5 pages
2016    Press Release and Backgrounder, HIV Home Test Kit, 5/14/96, 6
        pages
2017    Talk Paper, FDA Approves Second Interferon for Multiple
        Sclerosis, 5/17/96, 2 pages
3000    FEDERAL REGISTER notice - Alternatives to Lot Release,
        7/20/93, 3 pages
3001    FEDERAL REGISTER notice - Application of Current Statutory
        Authorities to Human Somatic Cell Therapy, 10/14/93, 3 pages
3002    FEDERAL REGISTER notice - Interim Rule - Human Tissue Intended
        for Transplantation, placed on public display 12/10/93, 9
        pages
3003    FEDERAL REGISTER notice - Final Rule - Adverse Experience
        Reporting Requirements for Licensed Biological Products,
        10/27/94, 12 pages
3004    FEDERAL REGISTER notice - Proposed Rule - Adverse Experience
        Reporting Requirements for Human Drug and Licensed Biological
        Products, 10/27/94, 20 pages
3005    FEDERAL REGISTER notice - Biological Products; Allergenic
        Extracts Classified in Category IIIB; Final Order; Revocation
        of Licenses, 11/16/94, 10 pages
3006    FEDERAL REGISTER notice - FDA's Policy Statement Concerning
        Cooperative Manufacturing Arrangements for Licensed Biologics,
        11/25/92, 3 pages
3007    FEDERAL REGISTER notice - Home Specimen Collection Kit Systems
        Intended for Human Immunodeficiency Virus (HIV-1 and/or HIV-2)
        Antibody Testing; Revisions to Previous Guidance, 2/23/95, 2
        pages
3008    FEDERAL REGISTER notice - Changes to be Reported for Product
        and Establishment License Applications; Guidance, 4/6/95, 4
        pages
3009    FEDERAL REGISTER notice - FDA Guidance Document Concerning Use
        of Pilot Manufacturing Facilities for the Development and
        Manufacture of Biological Products; Availability, July 11,
        1995, 4 pages
3010    FEDERAL REGISTER notice - July 27, 1995 - Subcommittee Meeting
        of the National Task Force on AIDS Drug Development on Drug
        Development Issues, September 13 & 14, 1995, 2 pages
3011    FEDERAL REGISTER notice - August 21, 1995 - International
        Conference on Harmonisation, Draft Guidelines, 11 pages
        1) Analysis of the Expression Construct in Cells Used for the
        Production of r-DNA Derived Protein Products
        2) Conditions Which Require Carcinogenicity Studies for
        Pharmaceuticals
        3) Detection of Toxicity to Reproduction: Addendum on Toxicity
        to Male Fertility
        4) Stability Testing of Biotechnological/Biological Products
3012    FEDERAL REGISTER notice - September 27, 1995 - Prominence of
        Name of Distributor of Biological Products, 6 pages
3013    FEDERAL REGISTER notice - October 25, 1995 - Characterization
        of Biological/Biotechnology Pharmaceutical Products; Notice of
        Public Workshop, 2 pages
3014    FEDERAL REGISTER notice - December 8, 1995 - Interim
        Definition and Elimination of Lot-by-Lot Release for
        Well-Characterized Therapeutic Recombinant DNA-Derived and
        Monoclonal Antibody Biotechnology Products, 6 pages
3015    FEDERAL REGISTER notice - December 8, 1995 - Advertising and
        Promotion; Draft Guidances, 2 pages.
3016    FEDERAL REGISTER notice - January 3, 1996 - Statement of
        Organization, Functions, and Delegations of Authority, 2 pages
3017    FEDERAL REGISTER notices - January 29, 1996 - pages 2733-2750,
        1) Well-Characterized Biotechnology Products; Elimination of
        Establishment License Application; 2) Changes to an Approved
        Application; 3) Draft Guidance; Changes to an Approved
        Application for Well-Characterized Therapeutic Recombinant
        DNA-Derived and Monoclonal Antibody Biotechnology Products;
        Availability; 4) Changes to an Approved Application; Draft
        Guidance; Availability, 18 pages
3018    FEDERAL REGISTER notice - February 26,1996 - Draft Document
        Concerning the Regulation of Placental/Umbilical Cord Blood
        Stem Cell Products Intended for Transplantation or Further
        Manufacture into Injectable Products; Availability of draft
        document, 2 pages
3019    FEDERAL REGISTER notice - March 7, 1996 - Notice of Public
        Meeting March 15, 1996; Commissioner's Roundtable-Regulatory
        Approach to Products Comprised of Living Autologous Cells
        Manipulated Ex Vivo and Intended for Structural Repair or
        Reconstruction, 2 pages
3020    FEDERAL REGISTER notice - May 3, 1996 - Current Good
        Manufacturing Practice; Proposed Amendment of Certain
        Requirements for Finished Pharmaceuticals, 13 pages
3021    FEDERAL REGISTER notice - May 14, 1996 - Final Rule,
        Elimination of Establishment License Application for Specified
        Biotechnology and Specified Synthetic Biological Products, 7
        pages
3022    FEDERAL REGISTER notice - May 28, 1996 - Guidance on
        Application for Products Comprised of Living Autologous Cells
        Manipulated Ex Vivo and Intended for Structural Repair or
        Reconstruction; Availability, 2 pages
*3023   FEDERAL REGISTER notice - July 10, 1996 - International
        Conference on Harmonisation; Final Guidelines on Stability
        Testing of Biotechnical/Biological Products; Availability;
        Notice, 5 pages
*3024   FEDERAL REGISTER notice - July 17, 1996 - International
        Conference on Harmonisation; Guideline on Structure and
        Content of Clinical Study Reports; Availability; Notice, 25
        pages
4000    Memorandum to Registered Blood Establishments, Deferral of
        Blood and Plasma Donors based on Medications, 7/28/93, 4 pages
4001    Memorandum to Registered Blood Establishments, Recommendations
        Regarding License Amendments and Procedures for Gamma
        Irradiation of Blood Products, 7/22/93, 20 pages
4002    Memoranda to Registered Blood Establishments, Revised
        Recommendations for Testing Whole Blood, Blood Components,
        Source Plasma and Source Leukocytes for Antibody to Hepatitis
        C Virus Encoded Antigen (Anti-HCV), 8/5/93 and 8/19/93, 9
        pages
4003    Memorandum to Registered Blood Establishments, Guidance
        Regarding Post Donation Information Reports, 12/10/93, 3 pages
4004    Memorandum to Registered Blood Establishments, Donor
        Suitability Related to Laboratory Testing for Viral Hepatitis
        and a History of Viral Hepatitis, 12/22/93, 4 pages
4005    Memorandum to Registered Blood Establishments, Recommendations
        for the Invalidation of Test Results When Using Licensed Viral
        Marker Assays to Screen Donors, 1/3/94, 5 pages
4006    Memorandum to Registered Blood Establishments, Recommendations
        for Deferral of Donors for Malaria Risk, 7/26/94, 2 pages
4007    Memorandum to Registered Blood Establishments, Use of an FDA
        Cleared or Approved Sterile Connecting Device (STCD) in Blood
        Bank Practices, 8/5/94 with Transmittal Memo, 8/12/94 (This
        corrects Memorandum dated 7/29/94), 9 pages
4008    Draft Recommendations for the Quarantine and Disposition of
        Potentially HBV, HCV or HTLV-I Contaminated Units from Prior
        Collections from Repeat Donors with Repeatedly Reactive
        Screening Tests. (Intended for discussion at the meeting of
        the BPAC on 12/15/94.), 11/22/94, 5 pages
4009    Memorandum to Registered Blood Establishments, Recommendations
        to Users of Medical Devices That Test for Infectious Disease
        Markers by Enzyme Immunoassay (EIA) Test Systems, 12/20/94, 17
        pages
4010    Memorandum to Registered Blood Establishments, Timeframe for
        Licensing Irradiated Blood Products, 2/3/95, 1 page
4011    Draft Discussion Points for Screening and Testing Donors of
        Human Tissue Intended for Transplantation and Human
        Reproductive Tissue, and for Establishment Registration,
        intended for discussion at the FDA workshop to be held 1:00 to
        5:30PM on 3/24/95, 3/25/95, 8 pages
4012    Draft Background Information for Discussion at the Workshop on
        Leukoreduced Products on March 27, 1995, 3/9/95, 20 pages
4013    Memorandum to Registered Blood and Source Plasma
        Establishments, Revision of FDA Memorandum of August 27, 1982:
        Requirements for Infrequent Plasmapheresis Donors, 3/10/95, 3
        pages
4014    Memorandum to Licensed Establishments Performing Red Blood
        Cell Immunizations, Revised Recommendations for Red Blood Cell
        Immunization Programs for Source Plasma Donors, 3/14/95, 6
        pages
4015    Recommendations for the Deferral of Current and Recent Inmates
        of Correctional Institutions as Donors of Whole Blood, blood
        components, Source Leukocytes, and Source Plasma, 6/8/95, 5
        pages
4016    Draft Document Concerning the Screening and Testing of Donors
        of Human Tissue Intended for Transplantation, 6/12/95, 9 pages
4017    Recommendations for Donor Screening with a Licensed Test for
        HIV-1 Antigen, 8/8/95, 14 pages
4018    Precautionary Measures to Further Reduce the Possible Risk of
        Transmission of Creutzfeldt-Jackob Disease by Blood and Blood
        Products, 8/8/95, 6 pages
4019    Disposition of Products Derived from Donors Diagnosed with, or
        at Known High Risk for, Creutzfeldt-Jakob Disease, 8/8/95, 3
        pages
4020    Recommendations for Labeling and Use of Units of Whole Blood,
        Blood Components, Source Plasma, Recovered Plasma or Source
        Leukocytes Obtained from Donors with Elevated Levels of
        Alanine Aminotransferase (ALT), 8/8/95, 2 pages
4021    Guidance for Blood Establishments Concerning Conversions to
        FDA-Reviewed Software Products, November 13, 1995, 6 pages
4022    Memorandum to Registered Blood and Source Plasma
        Establishments, Donor Deferral Due to Red Blood Cell Loss
        During Collection of Source Plasma by Automated
        Plasmapheresis, 12/4/95, 2 pages
4023    Draft Document Concerning the Regulation of
        Placental/Umbilical Cord Blood Stem Cell Products Intended for
        Transplantation or Further Manufacturing Into Injectable
        Products, December, 1995, 18 pages
4024    Draft Document Concerning the Regulation of Peripheral Blood
        Hematopoietic Stem Cell Products Intended for Transplantation
        or Further Manufacture Into Injectable Products, February
        1996, 15 pages
4025    Memorandum to Registered Blood Establishments, Additional
        Recommendations for Donor Screening With a Licensed Test for
        HIV-1 Antigen, 3/14/96, 6 pages
4026    Memorandum to Registered Blood and Plasma Establishments,
        Additional Recommendations for Testing Whole Blood, Blood
        Components, Source Plasma and Source Leucocytes for Antibody
        to Hepatitis C Virus Encoded Antigen (Anti-HCV), 5/16/96, 3
        pages
4027    Memorandum to Registered Blood Establishments, Recommendations
        and Licensure Requirements for Leukocyte-Reduced Blood
        Products, 5/29/96, 12 pages
*4028   Memorandum to Registered Blood and Plasma Establishments,
        Recommendations for the Quarantine and Disposition of Units
        from Prior Collections from Donors with Repeatedly Reactive
        Screening Tests for Hepatitis B Virus (HBV), Hepatitis C Virus
        (HCV) and Human T-Lymphotropic Virus Type I (HTLV-I)
5000    CBER Organizational Listing, January 1996, 13 pages
5001    CBER Organization Charts, January 1996, 28 pages
5002    CBER Mission and Vision Statements, 8/14/95, 2 pages
5003    CBER Telephone and Mail Routing Directory, May 21, 1996, 22
        pages
5004    CBER Strategic Plan for 2004, 8/14/95, 23 pages
5005    CBER Annual Report FY94, 33 pages
5006    CBER Annual Report FY95, 52 pages
6000    FDA December 17, 1993, Letter to Manufacturers: Bovine Derived
        Materials (BSE), 3 pages
6001    CBER December 28, 1993, letter describing preliminary position
        regarding Interim Rule - Human Tissue Intended for
        Transplantation, 3 pages
6002    Dr. Zoon's Slides - 1/27/95, The State of CBER 1994: The Year
        of Reinvention, 17 pages
6003    CBER Refusal to File (RTF) Guidance for Product and
        Establishment License
6004    CBER March 31, 1994, letter to Blood Establishment Computer
        Software
6005    Letter to Sponsors of INDs using Retroviral Vectors - 9/20/93,
        2 pages
6006    Letter to Sponsors of INDs for Human Immunoglobulin Products -
        5/23/94, 4 pages
6007    Review Performance Statistics, September, 1995, 27 pages
6008    Letter to Manufacturers of Licensed Anti-HIV Test Kits -
        5/26/94, 4 pages
6009    Letter to IVIG Manufacturers, October 3, 1994, 3 pages
6010    Dear Colleague Letter, Update on the status of efforts to
        improve the efficiency and effectiveness of the biologic
        product review and approval program, 11/15/94, 17 pages
6011    Memorandum, OTRR, Alternatives to Lot Release Policy, October
        27, 1994, 6 pages
6012    CBER Trends and the Managed Review Process, Dr. Zoon's Slides,
        9/19/94, 16 pages
6013    CBER's International Activities, 9/23/94, 6 pages
6014    CBER Update, Dr. Zoon's Slides, 11/2/94, 3 pages
6015    The Role of Science in the Regulation of Biological Products,
        Dr. Zoon's Remarks at the Dedication of Building 29B,
        11/22/94, 7 pages
6016    Statement on FDA Approval of AIDS Virus Test System Based on
        Oral Fluid Samples, 12/23/94, 2 pages
6017    Letters to Manufacturers of Immune Globulin (Human) - Testing
        for Hepatitis C Virus RNA, 12/27/94, 2 pages
6018    Letter to Blood Establishment Computer Software Manufacturers,
        2/10/95, 5 pages
6019    Letter to Manufacturers of Immune Globulin (Human), 3/3/95, 2
        pages
6020    Letter to Manufacturers of Immune Globulin (Human), 3/13/95, 3
        pages
6021    Letter to Health Professionals - Immune Globulin (Human),
        3/14/95, 7 pages
6022    Background Information - Poison Ivy/Poison Oak Extracts,
        3/16/95, 14 pages
6023    Letter to Health Professionals - Market Withdrawal - Plasma
        Products Produced from Blood Derived from a Donor with
        Probable CJD, 3/29/95, 5 pages
6024    Addendum to March 29, 1995 Letter - Plasma Product Withdrawal
        Associated with Probable CJD Donor, 3/31/95, 4 pages
6025    Memorandum to Licensed Manufacturers - Extended Expiration
        Dating of U.S. Standard Pertussis Vaccine, Lot No. 11, 4/6/94,
        1 page
6026    Error and Accident Reports: Summary for First Quarter FY 95
        3/22/95, 5 pages, Summary for FY 94, 10/26/94, 22 pages (Total
        : 27 pages)
6027    Dr. Zoon/Mr. Beatrice Slides, FDA Workshops on Regulatory
        Policy Issues in the Development & Manufacture of
        Biopharmaceuticals & Other Biotechnology-Derived Products,
        5/95, 6 pages
6028    Dr. Zoon Slides, Drug Information Association, 6/26/95, 6
        pages
6029    Error and Accident Reports: Summary for Second Quarter FY-95,
        9/22/95, 17 pages
6030    Error and Accident Reports: Summary for Third Quarter FY-95,
        9/22/95, 17 pages
6031    Letter to Specific Sponsors, Changes in Lot Release
        Requirements for Well-Characterized Therapeutic Recombinant
        DNA-derived and Monoclonal Antibody Products, 11/9/95, 2 pages
6032    Reinventing the Regulation of Drugs Made from Biotechnology;
        President Bill Clinton, Vice President Al Gore, 11/95, 12
        pages
6033    Dr. Zoon Slides, Well-Characterized Biotechnology Products:
        Evolving to Meet the Needs of the 21st Century, 12/11/95, 6
        pages
6034    Dear Colleague Letter, Regarding Reverse Transcriptase
        Activity in Viral Vaccines Produced in Chicken Cells, 1/4/96,
        2 pages
6035    Dr. Zoon Slides, BioEast, January 16, 1996, 6 pages
6036    Dr. Zoon Slides, Northwest Biotech, January 18, 1996, 12 pages
6037    Error and Accident Reports: Summary for Fourth Quarter FY-95,
        1/17/96, 18 pages
6038    Meeting Notice; Gene Therapy Conference: Development and
        Evaluation of Phase I Products; July 11, 1996 and Workshop on
        Vector Development; July 12, 1996, 4 pages
6039    Reinventing the Regulation of Cancer Drugs - Accelerating
        Approval and Expanding Access; President Bill Clinton and Vice
        President Al Gore, March 1996, 9 pages
6040    Error and Accident Reports - Annual Summary for FY95, April
        17, 1996, 42 pages
6041    Letter to Manufacturers of FDA-Regulated
        Drug/Biological/Device Products, Bovine Spongiform
        Encephalopathy (BSE), May 9, 1996, 2 pages
7000    Individual Sponsor/Investigator IND Packet, 38 pages
7001    Commercial IND Packet, 73 pages
7002    Application for Establishment License for Manufacture of
        Biological Products, Form FDA 3210 (5/94), 18 pages
7003    Review Checklists from FDA/CBER Workshop for Licensing Blood
        Establishments, January 30 & 31, 1995, 27 pages
7004    Establishment & Product License Applications for Manufacturers
        of Blood and Blood Components [Forms 2599, 2599(a), 2600,
        3098, 3098(a), 3098(b), 3098, 3098(d), 3098(e), and General
        Instructions], 28 pages
7005    FDA/CBER Workshop for Licensing Blood Establishments, January
        30 & 31, 1995, Slides describing request to use computer
        crossmatch, 9 pages
7006    Reviewer Guidance, Computer Software, 4/26/95, 4 pages
7007    Draft Reviewer Guidance, Changes in Personnel, 10/95, 1 page
7008    Draft Reviewer Guidance, Informed Consent for
        Plasmapheresis/Immunization, 10/95, 4 pages
7009    Draft Reviewer Guidance, Disease Associated Antibody
        Collection Program, 10/95,  2 pages
7010    Blood Registration Information and Form FD 2830 (08/95), 4
        pages
7011    Application to Market a New Drug, Biologic or an Antibiotic
        Drug for Human Use, FDA3439, 5/14/96, 2 pages
7012    Draft Reviewer Guidance for a Premarket Notification
        Submission for Blood Establishment Computer Software, 4/12/96,
        7 pages
8000    Guide to Inspections of Blood Banks, Division of Field
        Investigations, Office of Regional Operations, Office of
        Regulatory Affairs, September, 1994, 18 pages
8001    Methods of the Allergenic Products Testing Laboratory,
        October, 1993, 120 pages
8002    Guide to Inspections of Source Plasma Establishments, Division
        of Field Investigations, Office of Regional Operations, Office
        of Regulatory Affairs, December, 1994, 17 pages
8003    Computer Assisted Product License Application (CAPLA) Guidance
        Manual, March 1996, 88 pages
*8004   Guide to Inspections of Infectious Disease Marker Testing
        Facilities, 6/1/96, 11 pages
9001    Guidelines, Points-to-Consider, and other guidance documents
        for Human Biological Products available from the Division of
        Congressional and Public Affairs, 6/96, 16 pages
9002    Memorandum and Related Documents Pertaining to Human Blood &
        Blood Products, 7/5/96, 7 pages
9003    List of Guidelines available from the Center for Drug
        Evaluation and Research, 20 pages
9999    Documents Available from the CBER FAX Information System, 10
        pages
*Items added since 7/10/96
	Who Do You Call to...
REQUEST AN IND PACKET?
	Office of Communication, Training, and Manufacturers Assistance
301-827-1800
SET UP A PRE-IND MEETING?
	Joyce Frey Vasconcells,  acting Chief, Cytokine and Gene Therapy Branch,
CBER 301-594-0830
DISCUSS PRODUCT-SPECIFIC QUESTIONS?
	Cell and Gene Therapy Branch, CBER 301-594-0830
DISCUSS PRE-CLINICAL PHARMACOLOGY/TOXICOLOGY ISSUES?
	Pharm/Tox Branch Division of Clinical Trial Design and Analysis, CBER
301-594-5599 or 301-594-5600 fax. 301-594-0513
DISCUSS CLINICAL TRIAL DESIGN ISSUES?
	Division of Clinical Trial Design and Analysis 301-594-5600
INQUIRE ABOUT CENTER JURISDICTION ISSUES?
	Joy Cavagnaro, acting Director, Quality Assurance, CBER 301-827-0379
(tel. and fax.) e-mail: CAVAGNARO@A1.cber.fda.gov
	Eugene Berk, CDRH 301-594-1190 301-594-0379 (fax.)
	James C. Morrison, CDER 301-594-5443 301-594-5298 (fax.)
DISCUSS PROPOSED CHANGES TO AN EXISTING IND?
	Specific IND reviewers, contact CSO assigned to the IND (see IND
acknowledgment letter)
ASK REALLY WEIRD QUESTIONS?
	John Bishop, Assistant to the Director, Division of Cellular and Gene
Therapies, 301-827-0687