THE NIH RECOMBINANT DNA GUIDELINES Bob Hashimoto The University of Louisville April 2001

INTRODUCTION The National Institutes of Health (NIH) Guidelines for Research Involving Recombinant DNA Molecules is the reference document for research compliance with recombinant DNA molecules.

SCOPE AND APPLICABILITY SECTION I OF THE RECOMBINANT DNA GUIDELINES

REGULATORY OVERVIEW If your institution receives Federal funding, then it must comply with the NIH Guidelines for recombinant DNA research. Even if a project is privately sponsored, that research experiment must still be conducted in accordance with the NIH Guidelines.

DEFINITIONS Recombinant DNA (rDNA): Defined as either: 1. Molecules constructed outside living cells by joining natural or synthetic DNA segments to DNA molecules that can replicate in a living cell; or 2. DNA molecules that result from the replication of those described above.

DEFINITIONS, CONT. Risk Levels: I. Low Risk: risk level of agents and/or operations having minimal effect on personnel, other animal or plants under ordinary use. This classification is restricted to all etiologic agents designated as Biosafety Level 1 by the CDC.

DEFINITIONS, CONT. Risk Levels: II. Moderate Risk: risk level of agents and/or operations requiring special conditions for control or containment because of (a) known pathogenicity to personnel, other animals or plants; (b) concentration; or c) genetic alteration (synergistic effect) with other materials.

DEFINITIONS, CONT. Risk Levels: III. High Risk: risk level of agents and/or operations requiring additional control measures beyond those for moderate risk. This classification includes all etiologic agents designated Class 4 or 5 by the CDC and oncogenic viruses classified as high risk by the NCI.

SOMETHING TO REMEMBER Non-compliance to the NIH Guidelines may result in: forfeiture of funding suspensions or other limitations a requirement for NIH review and approval for all recombinant DNA research

SAFETY CONSIDERATIONS SECTION II OF THE RECOMBINANT DNA GUIDELINES

RISK GROUPS CLASSIFICATION OF ORGANISMS Risk Group 1: not known to cause disease 2: rarely serious disease, with therapeutic intervention 3: serious, lethal disease with therapeutic intervention 4: serious, lethal disease with no therapeutic intervention

RISK ASSESSMENT Review classification of organism Review research procedures to be performed Assess available facility/physical barriers (biosafety levels) Potential for inadvertent release Other factors, such as volume, concentration, replication competency

CONTAINMENT Standard practices Special procedures, equipment Available facility/facility design Biological barriers

CLASSIFICATION OF EXPERIMENTS PART III OF THE NIH RECOMBINANT DNA GUIDELINES

SECTION III-A Major Actions Requires: IBC, RAC, NIH Director Review and Approval prior to the initiation of work Drug resistance to organisms Prevent compromise to agriculture/medicine

SECTION III-B Requires: NIH/OBA, IBC Review and Approval Before Initiation of Work Containment determined by NIH/OBA Example: Deliberate Cloning of Toxin Molecules Lethal to Vertebrates at an LD50 of Less Than 100 Nanograms/Kg of Body Weight (e.g., Botulinum Toxin)

SECTION III-C Deliberate Transfer of Recombinant DNA into Research Participants Requires: RAC Review, IBC, and IRB Review and Approval Before Participant Enrollment Requires RAC Review prior to local institutional review. Must be reviewed by NIH prior to research participant enrollment.

SECTION III-D Requires IBC Review Prior to the Initiation of Work Requires: IBC Review and Approval prior to the initiation of work Involves RG 2-4 agents, host/vector system Cloned DNA from RG 2-4 agents into non- pathogenic prokaryotes RG 2-4 agents into whole animals, usually transgenic Recombinant plants

SECTION III-E IBC notification at the initiation of work Requires: IBC notification at the initiation of work (BL-1 containment) DNA Contains Less than 2/3 viral genome Transgenic Rodents with ABSL1 containment Whole Plants-minimal containment required

SECTION III-F Exempt Exempt from the NIH Guidelines and Does Not Require IBC Review and Approval Recombinant DNA that is: Not in Organisms Not a risk to the environment

ROLES AND RESPONSIBILITIES PART IV OF THE NIH RECOMBINANT DNA GUIDELINES

RESPONSIBILITIES UNIVERSITY OF LOUISVILLE (SECTION IV-B) Ensure compliance with NIH Guidelines Establish IBC Appoint a Biosafety Officer Ensure IBC has expertise in the research that is reviewed Establish a medical surveillance program as needed Report all accidents to the NIH

IBC MEMBER THE BIOSAFETY OFFICER When the institution conducts recombinant DNA research at BL3, BL4, or Large Scale (greater than 10 Liters), a Biological Safety Officer is mandatory and shall be a member of the Institutional Biosafety Committee (see section lV-B-3, Biological Safety Officer).

RESPONSIBILITIES IBC(SECTION IV-B-2) The Institutional Biosafety Committee must be composed of no fewer than five members so selected that they collectively have experience and expertise in recombinant DNA technology and the capability to assess the safety of recombinant DNA research and to identify any potential risk to public health or the environment. ( IV-B-2-a-(1)

IBC MEMBERSHIP COMMUNITY MEMBERS At least two members shall not be affiliated with the institution (apart from their membership on the IBC) and who represent the interest of the surrounding community with respect to health and protection of the environment (e.g. officials of state or local public health or environmental protection agencies, members of other governmental bodies, or persons active in medical, occupational health, or environmental concerns in the community). lV-B-2-a-(1)

IBC: CONFLICT OF INTEREST No member of an Institutional Biosafety Committee may be involved (except to provide information requested by the IBC) in the review or approval of a project in which he/she has been or expects to be engaged or has a direct financial interest. lV-B-2-a-(4)

RESPONSIBILITIES IBC(SECTION IV-B-2-b-1) PURPOSE OF THE IBC Reviewing recombinant DNA research conducted at or sponsored by the institution for compliance with NIH Guidelines as specified in Section III, Experiments Covered by the NIH Guidelines

RESPONSIBILITIES IBC This review shall include: (i) independent assessment of the containment levels required by the NIH Guidelines (ii) assessment of the facilities, procedures, practices, and training and expertise of personnel involved in recombinant DNA research. ( IV-B-2-b-(1))

IBC COORDINATION WITH OTHER COMMITTEES NIH RECOMBINANT DNA GUIDELINES

RESPONSIBILITIES- IBC, IRB sponsored Phase 1 clinical trials If the University of Louisvile is part of a non- NIH funded Phase 1 Clinical Trial protocol and receives NIH funding for other research, it is obligated to review the non-NIH sponsored research and follow the NIH Guidelines or possibly lose NIH funding for all research.

RESPONSIBILITIES IBC-IRB INTERACTION The University of Louisville Institutional Review Board (IRB) must become familiar with the provisions of Section III-C-1, the introduction of recombinant DNA molecules to research participants. The IRB and IBC must jointly review gene therapy applications.

IBC GENE THERAPY EVALUATION It is the responsibility of the IBC: to evaluate the potential of risk of the vector/ biohazard agent to the patient, family members or the environment and determine controls as appropriate. to evaluate the efficacy and the possibility of potential benefits of the therapy versus the concomitant biohazard risk of the vector with regard to the available therapy to the patient.

IBC GENE THERAPY EVALUATION The IBC shall evaluate: adverse events in previous clinical trials and animal studies to predict the potential of similar events in future trials the appropriate level of monitoring for potential microbial shedding

IRB GENE THERAPY EVALUATION It is the responsibility of the IRB to determine: the risks to subjects (research participants) the anticipated benefits to subjects the importance of the knowledge that may reasonably be expected to result the informed consent process to be employed

IBC-IACUC INTERACTIONS Certain experiments with animals require IBC review and approval: Section III-D-4, whole animals Section III-E-3, transgenic rodents Appendix Q: Containment requirements for large mammals Any introduction of biohazardous agents(shedding)

IBC-IACUC INTERACTIONS The University of Louisville Institutional Animal Care and Use Committee(IACUC) must participate in the review of any of the aforementioned experiments. While the IACUC traditionally examines pain and suffering, euthanasia and vivaria housing, the safe work practices described in Biosafety Levels 1-3 should be followed.

IBC-IACUC INTERACTIONS The IBC and IACUC should also ensure: biohazard vivaria areas are kept clean animal carcasses are properly disposed of infected animals are housed separately infected animals are transported safely infected animals do not infect humans

REPORTING ACCIDENTS AND INCIDENTS All laboratory accidents or incidents involving recombinant DNA molecules must be reported to NIH, OBA. All adverse events in gene transfer experiments must be reported to NIH, OBA, even if thought not to be in conjunction with the gene transfer intervention.

RAC APPROVAL The Institutional Biosafety Committee may not authorize initiation of experiments which are not explicitly covered by the NIH Guidelines until NIH (with the advice of the RAC when required) establishes the containment requirement. lV-B-2-b-(8)

IBC ADMINISTRATIVE REQUIREMENTS The University of Louisville shall file an annual report with NIH/OBA which includes: (i) a roster of all IBC members clearly indicating the Chair, contact person, Biological Safety Officer (if applicable), and identify the specific expertise of the committee members, e.g., animal expert, human gene therapy expert. (ii) biographical sketches of all IBC members (including community members).

IBC ADMINISTRATIVE REQUIREMENTS Upon request, the University of Louisville shall make available to the public all IBC meeting minutes and any documents submitted to or received from funding agencies.

IBC ADMINSITRATIVE REQUIREMENTS If public comments are made on IBC actions, the institution shall forward both the public comments and the IBC’s response to the Office of Biotechnology Activities, National Institutes of Health/MSC 7010, 6000 Executive Boulevard, Suite 302, Bethesda, Maryland , (301)

CONCLUSION This presentation was designed to inform the audience on the key provisions of the NIH Guidelines. The IBC must interact as necessary with other institutional committees and ensure that research involving recombinant DNA molecules is adequately reviewed.