Research Involving Recombinant DNA Molecules

1. Research Involving Recombinant DNA Molecules Training Drexel University August 2008

2. A Brief History of Relevant Scientific Advances A 20 year history between 1953 – 1973 characterized by unprecedented rates of advancement in the fields of genetics and molecular biology. Structure of DNA defined Restriction enzymes were discovered Successful recombination experiments conducted

3. Gordon Research Conference on Nucleic Acids - 1973 “The capability to perform DNA recombinations, and the potential hazards, had become apparent…”(Federal Register, Vol. 41, No. 131 July 7, 1976) As a result of this conference, scientists called for a worldwide moratorium on certain kinds of experiments.

4. Letter from National Academy of Sciences (NAS) “First, and most important, that until the potential hazards of such recombinant DNA molecules have been better evaluated or until adequate methods are developed for preventing their spread, scientists throughout the world join with the members of this committee in voluntarily deferring [certain] experiments” Letter From NAS published in Science 185, 303, (1974)

5. Asilomar Conference on Recombinant DNA Recommendations of Asilomar Conference on Recombinant DNA (February 1975): Recombinant DNA work should proceed Containment levels of possible hazard for various types of experiments. Certain experiments were judged to pose such serious potential dangers that the conference recommended against their being conducted at the present time. Guidelines of the Conference were published in Science, Nature, and Proceedings of the National Academy of Sciences in 1975.

6. Initiation of RAC and the NIH Guidelines NIH appointed Recombinant Advisory Committee (RAC) Using Asilomar report, drafted NIH Guidelines (1975-1976) NIH Guidelines were more conservative and more detailed than Asilomar Guidelines. NIH Guidelines published in Federal Register on July 7, 1976 Required Institutional Biohazard Committees and outlined responsibilities. NIH Guidelines have been changed over 50 times since 1976, and continue to be a living document.

7. Scope of NIH Guidelines The purpose of the NIH Guidelines is to specify practices for constructing and handling: recombinant deoxyribonucleic acid (DNA) molecules, and organisms and viruses containing recombinant DNA molecules. Guidelines apply to all recombinant DNA research that: is sponsored, funded, or in any way supported by NIH in U.S. is conducted abroad and is funded by NIH. involves testing in humans of materials containing recombinant DNA developed with NIH funds.

8. Compliance Required Non-compliance may result in: (i) suspension, limitation, or termination of financial assistance for the non-compliant NIH-funded project and of 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. “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.” - NIH Guidelines

9. Committee Membership per NIH No fewer than 5 individuals Plant and animal experts Biosafety Officer (BSO) Appropriate recombinant DNA expertise collectively Expertise in assessment of risk to environment and public health At least two members not affiliated with the institution Section IV-B-2-a Assembling an Institutional Biosafety Committee (IBC)

10. Drexel’s Institutional Biosafety Committee (IBC) Composition: 12 members and two community members. Meetings and Location: Meets 4th Tuesday of each month at the Office of Regulatory Research Compliance, 10th floor, 3-Parkway Building, 1601 Cherry Street, Philadelphia, PA 19102 Guidelines and Forms can be found at: http://www.research.drexel.edu/compliance/biosafety/guidelines.aspx What’s the main goal? Provide an independent, peer review of recombinant DNA research for conformity with the NIH Guidelines Identify and mitigate potential risk to environment and public health Serve as a technical resource for both administration and researchers

11. IBC Responsibilities and Limitations Responsibilities Prescribe containment levels per NIH Guidelines(IBC can raise or lower the level) Ensure adequacy of facilities, SOPs, PI and lab personnel training Ensure institutional and investigator compliance (e.g., adverse event reports) Limitations The IBC may NOT authorize initiation of recombinant DNA experiments not explicitly covered by the NIH Guidelines until NIH (with the advice of the RAC when required) establishes the containment requirement.

12. IBCs and Exempt Research Does the IBC determine what research is exempt? Yes This is a matter of institutional policy. See the IBC Guidelines at: http://www.research.drexel.edu/compliance/biosafety/guidelines.aspx IBC may wish to designate member, chair, or BSO as first line of review to make determinations about what is exempt and what requires full review NIH Office of Biotechnology Activities (OBA) can help with determinations

13. IBCs and Risk Assessment Primary function of IBC Ensure safety of laboratory workers, the community, and the environment. Recombinant DNA research poses ethical as well as safety concerns. Nature of research lends itself to unique hazards that need peer and community review.

14. Risk Assessment Guidelines also require comprehensive risk assessments to be conducted Comprehensive risk assessment should consider: Virulence Pathogenicity Infectious Dose Environmental Stability Route of Spread Communicability Operations Quantity Availability of vaccine or treatment

15. Biological Hazard Assessment and Biosafety Levels Hazards The hazard associated with biological agents varies depending on many factors. In general, agents are classified into four Risk Groups (RG) established by WHO and used by CDC This Assessment is used to set appropriate containment conditions for the experiment. Biosafety Levels NIH describes 4 Biosafety Levels, designated BL1, BL2, BL3, and BL4 Each level requires a different scale of containment and/or combinations of laboratory practices, containment equipment, ventilation, and laboratory design

16. Vertebrate Animal Biosafety Levels and Hazard Assessment Guidance for the use of experimentally infected animals housed in indoor research facilities Institutional management must provide facilities, staff and established practices designed to ensure appropriate levels of environmental quality, safety, security and care. In general, the biosafety level recommended for work with infectious agents in vivo is comparable to those recommended for in vitro activity. The are, however, additional concerns that are not present in a standard micro lab. Animals may: - Create Aerosols - Bite or Scratch - Be infected with a zoonotic agent Animal Biosafety Levels NIH describes 4 Animal Biosafety Levels, designated ABL1, ABL2, ABL3, and ABL4 As with previously mentioned biosafety levels, each animal biosafety level requires a different scale of containment and/or combinations of laboratory practices, containment equipment, ventilation, and laboratory design

17. Biohazard Containment Biological Containment Contained in NIH Guidelines, Appendix I (http://www4.od.nih.gov/oba/) The application of highly specific biological barriers Barriers that limit either the infectivity of a vector, or Barriers that limit its dissemination and survival in the environment.

18. Virulence Will the experiment somehow give the organism specific determinants (virulence factors) that permit its establishment within a host and its ability to be transmitted to new hosts? Ability to attach to host cells (pili, adhesins) Serum resistance Capsules (Cryptococcus neoformans) Enzymes that destroy host cells

19. Toxicity Campylocacter jejuni (enterotoxin) Vibrio cholerae (Cholera toxin) Staphylococcus aureus (α-toxin) NIH OBA and IBC approval required before initiation of experiments involving the cloning of toxin molecules with LD50 of less than 100 nanograms per kilogram body weight.

20. Pathogenicity Normal flora of literally thousands of species of microorganisms endogenous to humans Establishment of disease relies on the ability of a microorganism to enter the host and initiate infection Will genetic manipulation give organism a competitive edge?

21. Human Gene Transfer An experiment involving the deliberate transfer of recombinant DNA into human research participants. Clinical Trials involving gene transfer are reviewed by our external Western IRB Contact Office for Regulatory Research Compliance (215-255-7857) for additional information.

22. Expanding Role of IBCs In addition to recombinant DNA and gene therapy protocols, IBCs have roles in: Select Agent Registration New Defense Initiatives

23. Drexel University Contact Information DEPARTMENT OF SAFETY AND HEALTH 400 N. 31st Street Philadelphia, PA 19104 Telephone: 215-895-5891 Fax: 215-895-5926 Web site: http://www.drexelsafetyandhealth.com/ OFFICE FOR REGULATORY RESEARCH COMPLIANCE Suite 10100, 3 Parkway Building 1601 Cherry Street Philadelphia, PA 19102 Telephone: 215-255-7857 Fax: 215-255-7874 Web site: http://www.research.drexel.edu/compliance/

24. NIH OBA Contact Information OFFICE OF BIOTECHNOLOGY ACTIVITIES Office of Biotechnology Activities National Institutes of Health 6705 Rockledge Drive, Suite 750, MSC 7985 Bethesda, MD  20892-7985 Phone: 301-496-9838 Fax: 301-496-9839 Email: oba@od.nih.gov Website: http://www4.od.nih.gov/oba/