Implementation of the NIH Guidelines 

Robert J. Hashimoto, CBSPUniversity of MinnesotaMinneapolis, Minnesota

November 15, 2007

INTRODUCTION

The National Institutes of Health (NIH) requires that any institution that receives funding from the NIH must comply with the NIH Recombinant DNA Guidelines. This presentation will summarize the NIH Guidelines and emphasize how the University of Minnesota is obligated to follow the contents.

REGULATORY OVERVIEW

The institutional review of recombinant DNA experiments is based on the five sections and appendices of the following guideline:

The NIH Guidelines for the Use of Recombinant DNA Molecules,Published in FR, July 5, 1994 (59 FR 34496), Amendment Effective April 1, 2002, FR, April 1, 2002 (65 FR 60328)

Section I of the NIH Guidelines

Applicability

Compliance

SECTION I: APPLICABILITY

The NIH Guidelines are applicable for:Research that is conducted at or sponsored by an institution that receives any support for recombinant DNA research from NIH, including research performed directly by NIH. An individual who receives support for research involving recombinant DNA must be associated with or sponsored by an institution that assumes the responsibilities assigned in the NIH Guidelines. (Section I-C-1-a-(1))

(e.g., PI with NIH Grant)

SECTION I: APPLICABILITY OF THE GUIDELINESThe NIH Guidelines are applicable for:

Research that involves testing in humans of materials containing recombinant DNA developed with NIH funds, if the institution that developed those materials sponsors or participates in those projects. Participation includes research collaboration or contractual agreements, not mere provision of research materials. (Section I-C-1-a-(2). (e.g., PI with NIH Grant conducting a Section III-C experiment must receive both IRB and IBC approval with NIH review)

SECTION I: Compliance with the NIH Guidelines

As a condition for NIH funding of recombinant DNA research, institutions shall ensure that such research conducted at or sponsored by the institution, irrespective of the source of funding, shall comply with the NIH Guidelines. (Section I-D-1)

(If you receive NIH money, you must follow the NIH Guidelines)

SECTION I: Compliance with the NIH Guidelines

All NIH-funded projects involving recombinant DNA techniques must comply with the NIH Guidelines. Non-compliance may result in: (i) suspension, limitation, or termination of financial assistance for the noncompliant NIH-funded research project and of NIH funds for other recombinant DNA research at the institution, or (ii) a requirement for prior NIH approval of any or all recombinant DNA projects at the institution. (Section I-D-1) (Failure to follow NIH Guidelines may result in forfeiture of the NIH grant from not just the PI who violates the Guidelines but all NIH funds at the institution.)

SECTION I: Compliance with the NIH Guidelines

All non-NIH funded projects involving recombinant DNA techniques conducted at or sponsored by an institution that receives NIH funds for projects involving such techniques must comply with the NIH Guidelines. Noncompliance may result in: (i) suspension, limitation, or termination of NIH funds for recombinant DNA research at the institution, or (ii) a requirement for prior NIH approval of any or all recombinant DNA projects at the institution.(Section I-D-2)

(If a non NIH funded PI violates the NIH Guidelines, it may result in the forfeiture of NIH funding from all other NIH grants.)

Section II of the NIH Guidelines

Safety

Risk Assessment

Containment

SECTION II: SAFETYRisk Assessment and Risk Groups (RG)

The Guidelines require that the Principal investigator make an initial assessment of risk based on the Risk Group (RG) of an agent (see Appendix B of the Guidelines, Classification of Human Etiologic Agents on the Basis of Hazard). This is based on the World Health Organization (WHO) Risk Classification.

SECTION II: SAFETYDefinition of Risk Groups (RG)

Risk Group 1: not associated with disease in healthy adult humans (e.g. E-coli K-12).

Risk Group 2: associated with human disease which is rarely serious and preventive or therapeutic interventions are often available. (e.g., HBV)

Risk Group 3: associated with serious or lethal human disease for which preventive or therapeutic interventions may be available.(e.g., M. tuberculosis)

Risk Group 4: likely to cause serious or lethal human disease for which preventive or therapeutic interventions are not usually available. (e.g., Ebola)

SECTION II: SAFETY Risk Assessment

The Biosafety Officer and Institutional Biosafety Committee will review the agent itself and the proposed experiment procedures to determine the appropriate level of containment.

The levels of containment or Biosafety Levels are described in Appendix G of the Guidelines and are also found in the Fifth Edition of the Centers for Disease Control and Prevention (CDC) guideline, Biosafety in Microbiological and Biomedical Laboratories.

Section III of the NIH Guidelines

Experiments That Require IBC Approval

Section III-A through Section III-E-3

Recombinant DNA ExperimentsSection III-A of the NIH Guidelines

Deliberate transfer of a drug resistant trait to microorganisms that could compromise medicine or

agriculture in its control.

Creation of a multi-drug resistant virus or bacteria that would compromise medicine, agriculture or public health, e.g., a recombinant HBV virus that was would render the current HBV vaccine series as ineffective.

These Experiments require Institutional Biosafety Committee Approval, RAC Review, and NIH Director Approval Before Initiation and is considered a Major Action under the NIH Guidelines.

Recombinant DNA ExperimentsSection III-B of the NIH Guidelines

The deliberate formation of recombinant DNA containing genes for the biosynthesis of toxin molecules lethal at an LD50 of less

than 100 nanograms per kilogram of body weight.

These experiments involve the potential manipulation of sequences from microbial toxins such as the botulinum toxins, tetanus toxin, diphtheria toxin, and Shigella dysenteriae neurotoxin, e.g., creation of recombinant organism, such as E-coli K12 with a sequence to synthesize Clostridium botulinum toxin

These experiments require NIH/OBA and Institutional Biosafety Committee Approval Before Initiation (and may require CDC Select Agent approval if non-exempt quantities are used).

Recombinant DNA ExperimentsSection III-C of the NIH Guidelines

The deliberate transfer of recombinant DNA or RNA derived from recombinant DNA into one or more human research participants (e.g., in vivo human

gene therapy)

These experiments often involve in-vivo human gene therapy but may also include vaccine studies, e.g.,an adenoviral mediated gene transfer experiment

These experiments require Institutional Biosafety Committee and Institutional Review Board Approvals and RAC Review Before Research Participant Enrollment

Recombinant DNA ExperimentsSection III-D of the NIH Guidelines

Section III-D of the NIH Guidelines

Experiments that Require Institutional Biosafety Committee Approval Before Initiation

The Principal Investigator must submit a registration document to the Institutional Biosafety Committee which contains the following information:

the source(s) of DNA the nature of the inserted DNA sequences the host(s) and vector(s) to be used if an attempt will be made to obtain expression of a foreign

gene, and if so, indicate the protein that will be produced the containment conditions that will be implemented as

specified in the NIH Guidelines.

Recombinant DNA ExperimentsSection III-D of the NIH Guidelines

Section III-D-1 of the NIH Guidelines

The use of Risk Group 2-4 human or animal pathogens as part of a host-vector system at a containment level determined by the IBC.

Section III-D-2 of the NIH Guidelines

The use of DNA from Risk Group 2-4 human or animal pathogens being cloned into non-pathogenic prokaryotic or lower eukaryotic host-vector systems at a containment level determined by the IBC.

Recombinant DNA ExperimentsSection III-D of the NIH Guidelines

Section III-D-3 of the NIH Guidelines

The Use of infectious DNA or RNA viruses or RNA viruses in the presence of helper virus in Tissue Culture Systems.

Section III-D-4 of the NIH Guidelines

Whole animals in which the animal's genome has been altered by the stable introduction of recombinant DNA, or RNA derived therefrom, into the germ-line (transgenic animals).

Recombinant DNA ExperimentsSection III-D of the NIH Guidelines

Section III-D-5 of the NIH GuidelinesExperiments to genetically engineer plants by recombinant DNA methods, to use such plants for other experimental purposes,(e.g., response to stress), to propagate such plants, or to use plants together with microorganisms or insects containing recombinant DNA.

Section III-D-6 of the NIH GuidelinesExperiments involving more than 10 liters of culture. The appropriate containment will be decided by the IBC. Where appropriate, Appendix K, Physical Containment for Large Scale Uses of Organisms Containing Recombinant DNA Molecules, shall be used.

Recombinant DNA ExperimentsSection III-E of the NIH Guidelines

Section III-E of the NIH GuidelinesExperiments that Require Institutional Biosafety Committee Notice Simultaneous with Initiation

Experiments not included in Sections III-A through III-D are considered in Section III-E. All such experiments may be conducted at BL1 containment.

The Institutional Biosafety Committee reviews and approves all such proposals, but Institutional Biosafety Committee review and approval prior to initiation of the experiment is not required.

Recombinant DNA ExperimentsSection III-E of the NIH Guidelines

Section III-E-1 of the NIH Guidelines

Experiments involving the formation of recombinant DNA molecules containing no more than two-thirds of the genome of any eukaryotic virus.

Section III-E-2 of the NIH Guidelines

  This section covers experiments involving recombinant DNA-modified whole plants, and/or experiments involving recombinant DNA-modified organisms associated with whole plants, except those experiments that fall under Section III-A through III-D. BL-1 containment is adequate.

Recombinant DNA ExperimentsSection III-E of the NIH Guidelines

Section III-E-3 of the NIH Guidelines

Experiments Involving Transgenic Rodents

  This section covers experiments involving the generation of rodents in which the animal's genome has been altered by stable introduction of recombinant DNA, or DNA derived therefrom, into the germ-line (transgenic rodents). Only experiments that require BL1 containment are covered under this section; experiments that require BL2, BL3, or BL4 containment are covered under Section III-D-4, Experiments Involving Whole Animals.

Section IV of the NIH Guidelines

Institutional ResponsibilitiesDuties of IBC

Duties of Biosafety OfficerComposition of IBC

SECTION IV: RESPONSIBILITIES OF THE IBCSection IV-B-2. The University of Minnesota shall establish an

Institutional Biosafety Committee which must be comprised 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 and environment.

At least 2 members are non-affiliated The duties of the IBC is not limited to reviewing

recombinant DNA research

SECTION IV: RESPONSIBILITIES OF THE IBC

Requirement Reference

Annual Report To NIH Section IV-B-2-a-(3) Write Meeting Minutes Section IV-B-2-a-(7) Conflict of Interest Section IV-B-2-a-

(4) Reporting Incidents Section IV-B-2-b-

(7) Adequate IBC Training Section IV-B-1-h

SECTION IV:KEY DUTIES OF THE IBC

Assignment Reference

Set Containment Level Section IV-B-2-b-(2-4) Notify PI of Approval Section IV-B-2-b-(2-3) Assess Lab Facilities Section IV-B-2-B-(1) Monitoring Projects Section IV-B-2-B-(5) Establish Emergency Plans Section IV-B-2-B-(6) Ensure NIH review Section IV-B-2-B-(8)

THE BIOSAFETY OFFICERSection IV-B-3-a, b: ROLE OF THE BIOSAFETY OFFICER

The institution shall appoint a Biological Safety Officer if it engages in large-scale research or production activities involving viable organisms containing recombinant DNA molecules. 

The institution shall appoint a Biological Safety Officer if it engages in recombinant DNA research at BL3 or BL4. The Biological Safety Officer shall be a member of the Institutional Biosafety Committee.

 

THE BIOSAFETY OFFICERDuties of the Biosafety Officer

Assignment Reference

Conduct Inspections Section IV-B-3-c-(1) Notify IBC, Chair Section IV-B-3-c-(2) Investigate Accidents Section IV-B-3-c-(3) Establish Emergency Plans Section IV-B-3-c-(3) Advise on Lab Security Section IV-B-3-c-(4) Provide Technical Advice Section IV-B-3-c-(5)

Section IV-B-7: ROLE OF THE PI

On behalf of the institution, the Principal Investigator is responsible for full compliance with the NIH Guidelines for any recombinant DNA research. The PI shall:

Initiate or modify no recombinant DNA research which requires IBC approval until that research has been approved by the IBC and has met all other requirements of the Guidelines

Report any significant problems, violations of the NIH Guidelines, or any significant research-related accidents and illnesses to the Biological Safety Officer and IBC.

Section IV-B-7-c: PI SUBMISSION TO THE IBC

The Principal Investigator shall: Make an initial determination of the required

levels of physical and biological containment in accordance with the NIH Guidelines; and select appropriate microbiological practices and laboratory techniques for the research;

Section IV-B-7-c: PI SUBMISSION TO THE IBC

The Principal Investigator shall: Submit the initial research protocol and any

subsequent changes if the experiment is covered under Sections III-A through III-E-3 to the IBC for approval. The PI must determine which section of the Guidelines that the experiment falls under. (e.g, III-D-1, Experiments involving RG2 agents as a host vector system) and indicate this on the IBC Application Form.

This is now an inspectable item on an NIH Site Visit.

Section IV: PI DUTIES AND RESPONSIBILITIES

Responsibility Reference

Train Lab Staff (Micro) Section IV-B-7-a, d Ship Agents Safely Section IV-B-7-a Follow Emergency Plans Section IV-B-7-a Notify NIH (e.g., HGT) Section IV-B-7-b Update Protocol Section IV-B-7-c Notify IBC, Chair Section IV-B-7-e Make Protocol Available Section IV-B-7-d Offer Medical Surveillance Section IV-B-7-d Supervise Lab Members Section IV-B-7-e Maintain Safety EquipmentSection IV-B-7-e

Section V of the NIH Guidelines

Footnotes and ReferencesInclusion of key references such as Restrictions for Select Pathogens (Variola major)USDA permit requirements

The Key Appendices

Appendix BAppendix GAppendix KAppendix MAppendix PAppendix Q

APPENDIX B.CLASSIFICATION OF HUMAN ETIOLOGIC AGENTS ON THE BASIS OF HAZARD

This appendix includes those biological agents known to infect humans as well as selected animal agents that may pose theoretical risks if inoculated into humans. Included are lists of representative genera and species known to be pathogenic; mutated, recombined, and non-pathogenic species and strains are not considered. Non-infectious life cycle stages of parasites are excluded. The WHO Risk Group classification is used in the guidelines.

APPENDIX G.PHYSICAL CONTAINMENT

Appendix G specifies physical containment for standard laboratory experiments and defines Biosafety Level 1 - Biosafety Level 4, consistent with the CDC Guideline, Biosafety in Microbiological and Biomedical Laboratories. The following appendices shall be used to determine containment in the following instances:

For large-scale (over 10 liters) research or production, Appendix K shall supersede Appendix G.

For certain work with plants, Appendix P shall supersede Appendix G.

For certain work with animals, Appendix Q supersedes Appendix G.

APPENDIX KLARGE SCALE CONSIDERATIONS

Appendix K specifies physical containment guidelines for large-scale (greater than 10 liters of culture) research or production involving viable organisms containing recombinant DNA molecules.

It shall apply to large-scale research or production activities as specified in Section III-D-6, Experiments Involving More than 10 Liters of Culture.

For the purpose of large scale research or production, four levels of containment are established. These are referred to as BL1-Large Scale (LS), BL2-LS, BL3-LS, and Good Large Scale Practice (GLSP)

APPENDIX M. POINTS TO CONSIDER IN THE DESIGN AND SUBMISSION OF PROTOCOLS FOR THE TRANSFER OF RECOMBINANT DNA MOLECULES INTO ONE OR MORE HUMAN RESEARCH PARTICIPANTS

Appendix M describes the process for review & submission of research proposals involving the deliberate transfer of recombinant DNA, or DNA or RNA derived from recombinant DNA, into human subjects (human gene transfer).

Concurrent Institutional Review Board (IRB) approval is required.

APPENDIX M. POINTS TO CONSIDER IN THE DESIGN AND SUBMISSION OF PROTOCOLS FOR THE TRANSFER OF RECOMBINANT DNA MOLECULES INTO ONE OR MORE HUMAN RESEARCH PARTICIPANTS

Much of Appendix M summarizes information that is found in the Clinical Protocol, Investigational Brochure and Human Subject Consent Form.

This appendix must be completed by the PI and include local considerations, such as the phone number to report adverse events, not by the sponsor.

APPENDIX P. PHYSICAL AND BIOLOGICAL CONTAINMENT FOR RECOMBINANT DNA RESEARCH INVOLVING PLANTS Appendix P specifies physical and biological containment

conditions and practices suitable to the greenhouse conduct of experiments involving recombinant DNA-containing plants, plant-associated microorganisms, and small animals and will supersede Appendix G when the research plants are of a size, number, or have growth requirements that preclude the use of containment conditions described in Appendix G.

Four biosafety levels (referred to as Biosafety Level 1 – Plants or BL1-P, BL2-P, BL3-P, and BL4-P) are used for research involving recombinant DNA molecules in plants requiring special containment.

APPENDIX Q.PHYSICAL AND BIOLOGICAL CONTAINMENT FOR RECOMBINANT DNA RESEARCH INVOLVING ANIMALS

Appendix Q specifies containment and confinement practices for research involving whole animals, both those in which the animal's genome has been altered by stable introduction of recombinant DNA, or DNA derived therefrom, into the germ-line (transgenic animals) and experiments involving viable recombinant DNA-modified microorganisms tested on whole animals.

For the purpose of animal research, four levels of containment are established in Appendix Q. These are referred to as BL1-Animals (N), BL2-N, BL3-N, and BL4-N.

How does this affect me?

Apply to the IBC

If you are conducting any work with recombinant DNA molecules, then you must complete an IBC application, regardless of your source of funding.

The experiment may or may not require full committee review but the University reviews alI experiments.

Apply to the IBC-Which Section of the Guidelines? Based on the agent used, e.g. adenovirus 5,

determine what risk group it is by referring to Appendix B of the Guidelines. In our example of adenovirus, adenovirus is a Risk Group 2 organism.

Once the Risk Group of the agent is known, determine the type of experiment described in Section III of the NIH Guidelines (e.g. Section III-D-1, the use of Risk Group 2 agents as part of a host vector system).

Apply to the IBC-Which Level of Containment? Based on the agent used, e.g. adenovirus 5, and the

experiment type (in our example, Section III-D-1), determine the level of containment or biosafety level you propose for this experiment by reviewing Appendix G.

Usually a RG2 agent will be used at Biosafety Level 2 but depending on the procedures, the biosafety level may be lower or higher.

The IBC will review the Scope of Work narrative to review the procedures performed and make the final determination of the Biosafety Level.

THE IBC APPLICATION FORM BIOSAFETY LEVELS

The biosafety level may increase for: large volumes of culture aerosol generating procedures high concentrations the use of transgenic animals the use of vectors and vulnerable animals inserted genetic sequences that increase

virulence the use of drug/antibiotic resistant strains

THE IBC APPLICATION FORM -BIOSAFETY LEVELS

The biosafety level of the experiment may decrease if:

the organism is attenuated, inactivated, or replication deficient;

the strain is not infectious to humans and a vulnerable species of animal is not used

sequences that do not code for virulence are used and are not being isolated prior to use

the NIH has approved that identical work is performed at a lower level of containment

THE IBC APPLICATION FORM –SCOPE OF WORK

The previous two slides illustrate why the Scope of Work section is so important. Without the information describing the experiment, the IBC may not accurately assess the level of containment needed to safely work with the agent.

Review the Scope of Work narrative to get an idea of what the IBC needs to review.

Note: the IBC may use Appendix K, M, P or Q based on the type of experiment conducted!

CONCLUSIONThe NIH Recombinant DNA Guidelines describe the safe use, handling and administrative requirements for an institution to work with recombinant DNA. It also describes the requirements for the IBC and Biosafety Officer to manage the use of these materials. The University of Minnesota is obligated to follow the guidelines or potentially forfeit NIH funding.