Principal Investigator rDNA/Gene Transfer Training on NIH Guidelines

1. Principal Investigator rDNA/Gene Transfer Training on NIH Guidelines The University of Texas at Austin Office of Research Support and Compliance Pat Levin IBC Program Coordinator Environmental Health and Safety Rachel LeBansky Safety Specialist IV

2. Recombinant DNA Program • The office of Biotechnology Activities at the NIH oversees rDNA research, including human gene transfer • Manages the rDNA Advisory Committee (RAC) • Administers the NIH Guidelines for Research Involving Recombinant DNA Molecules • Partners with Institutional Biosafety Committees (IBC) in the oversight of rDNA research

3. Definitions • Recombinant DNA- (i) molecules that are constructed outside living cells by joining natural or synthetic DNA segments to DNA molecules that are replicated in a living cell or (ii) molecules that result from the replication of those described in (i) above. (www4.od.nih.gov/oba/Rdna.htm) • Select Agents – specific microbiological agents and toxins of biological origin that have the potential to be weaponized and are regulated by the CDC/USDA. (www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm)

4. Why is Training Required for PI’s and lab personnel? • NIH Guidelines requires training for all PI’s and their lab personnel conducting recombinant DNA (rDNA) research Regardless of source of funding. • Noncompliance can result in suspension, limitation, or termination of NIH funds for rDNA research for the entire institution.

5. NIH GuidelinesContent • Section I – Scope • Section II – Safety Considerations • Section III – Types of Experiments Covered • Section IV – Roles and Responsibilities • Appendices

6. NIH GuidelinesSection I Scope of the NIH Guidelines • Purpose • Definition of rDNA Molecules • General Applicability • Compliance with NIH Guidelines • General Definitions

7. The NIH Guidelines Apply to… • rDNA research that is • Funded by the NIH • Performed at or sponsored by an institution that receives any NIH funding for recombinant DNA research • Rationale: For biosafety to be meaningful, it has to be observed by all investigators at an institution

8. Are the NIH Guidelines optional? What are potential consequences of noncompliance with the NIH Guidelines? • Suspension, limitation, or termination of NIH funds for recombinant DNA research at the institution • Additional requirement for prior NIH approval of any or all recombinant DNA projects at the institution

9. The Advent of Recombinant DNA Technology • Emergence of recombinant DNA technology (mid- 1970’s) • Concerns among both scientific community and general public • Public health and safety • Environmental impact • Potential ethical and social implications

10. Published in July 1976 Established responsibilities of investigators and institutions The First NIH Guidelines

11. PUBLIC CONCERN Public concerns about the potential public health and environmental consequences of rDNA and gene transfer research such as: • Expanding programs of biodefense research • Emerging infectious disease threats (SARS, Avian flu) • Human gene transfer continues to raise many safety, ethical, and scientific issues in need of public discussion and analysis • Advances in technology which enable unprecedented research • Reverse engineering of 1918 flu virus • Synthesis of the polio virus

12. NIH Guidelines for Research Involving Recombinant DNA Molecules A scientifically-responsive document that will continue to evolve • Has undergone multiple revisions since 1976 • Latest version -April 2002 • http://www.utexas.edu/research/rsc/ibc/links.php

13. Specifics vs. Intent • “The NIH Guidelines will never be complete or final since all conceivable experiments involving recombinant DNA cannot be foreseen. Therefore, it is the responsibility of the institution and those associated with it to adhere to the intent of the NIH Guidelines as well as to the specifics.” • Good judgment is key • UT IBC can help

14. NIH Guidelines – Section II

15. NIH Guidelines Section II • Safety Considerations • Risk assessments: (Appendix B) RG 1 RG 2 RG 3 RG 4

16. BSL1 BSL2 BSL3 BSL4 NIH GuidelinesSection II • Containment • Physical (Appendix G Practices • Equipment/Facilities • Criteria of Biological Agents • Biological (Appendix I) • Survival/Transmission

17. It is the PI’s Responsibility to determine Risk Group Examples of the four Risk Groups: Risk Group 1: Klebsiella oxytoca, asporogenic B.subtilis Risk Group 2: Bortedella pertussis, Chlamydia trachomastis, E. coli, C. diphtheriae, Shigella, Vibrio cholerae, Yersinia entercolitica, all human adenovirus types, Grp A Arboviruses, hepatitis A-E virus, Herpes viruses, Influenza Types A, B,& C, all human papilloma viruses, retroviruses HIV, SIV, HTLV, VSV Risk Group 3: Brucella abortus, Coxiella burnetii, Francisella tularensis, M. tuberculosis, R. Typhi, Yersinia pestis, Coccidioides immitis, Histoplasma capsulatum, Lassa virus, Ebola & Marburg viruses Risk Group 4: There is no RG4 work done at UT

18. IBC, RAC, NIH Director IBC, OBA (in consult with experts) IBC, IRB, RAC IBC IBC (notification) Exempt NIH Guidelines – Section III

19. NIH GuidelinesSection III - Levels of Review

20. Key Portions of the NIH Guidelines for Animal Research • Section III-D-4 Experiments Involving Whole Animals – IBC Approval Before Initiation • Experiments in which: • the animal’s genome has been altered by stable introduction of rDNA into germline, or • rDNA modified microorganisms are tested on whole animals • BL2 or BL2-N or greater containment

21. Key Portions of the NIH Guidelines for Animal Research • Section III-D-5 Experiments Involving Whole Plants – IBC Approval Before Initiation • Experiments in which: • Plants genetically engineered by rDNA methods, or • Plants are used with recombinant DNA-modified insects • Generally BL2-P through BL4-P, depending on risk

22. Key Portions of the NIH Guidelines for Animal Research • Section III-E-3 Experiments Involving the Generation of Transgenic Rodents – IBC Notice at Initiation • Experiments in which: • Rodent’s genome has been altered by stable introduction of rDNA into germline • BL1 containment is appropriate

23. Key Portions of the NIH Guidelines for Animal Research • Section III-F (and Appendix C-VI) - Exempt Experiments • The purchase or transfer of rodents for experiments that require BL-1 containment • Further manipulations of these animals with recombinant DNA are not necessarily exempt from the NIH Guidelines

24. NIH GuidelinesSection IV Roles and Responsibilities • Principal Investigator (PI) • Biological Safety Officer (BSO) • Institutional Biosafety Committee (IBC) • Institutional Officer (IO) • NIH • This is the reporting order for incidents, accidents, or spills

25. PI Responsibilities NIH Guidelines • IBC approval approval must be granted prior to initiating or modifying any recombinant DNA research covered under sections III-A to III-D. • Determine whether experiments are covered under NIH Guidelines and notify the IBC as appropriate. • *Be adequately trained in good microbiological techniques*. • Adhere to IBC emergency plans for spills and personnel contamination. • Report any incidents, accidents, or spills to BSO and IBC..

26. NIH GuidelinesUT- Austin Responsibilities • Establish and implement policies for the safe conduct of recombinant DNA research. • Establish an Institutional Biosafety Committee. • Assist and ensure compliance with the NIH Guidelines by investigators. • Ensure appropriate training for IBC Members and PI’s • PI’s are responsible for the actions of their lab and compliance with the guidelines • Determine necessity for health surveillance of personnel. • PI’s work with IBC in reporting any significant incidents to NIH-OBA.

27. NIH GuidelinesAppendices • Appendix A – Exemptions: Natural Exchangers • Appendix B – Classification of Etiologic Agents • Appendix C – Exemptions under III-F • Appendix D – Major Actions • Appendix E – Certified Host-Vector Systems • Appendix F – Biosynthesis of Toxic Molecules • Appendix G – Physical Containment • Appendix H – Shipment • Appendix I – Biological Containment

28. NIH Guidelines Appendices • Appendix J – Biotechnology Research Subcommittee • Appendix K – Large Scale Physical Containment • Appendix L – Gene Therapy Policy Conferences • Appendix M– Points to Consider in Human Gene Transfer Research • Appendix P– Physical and Biological Containment: Plants • Appendix Q– Physical and Biological Containment: Animals

29. Key Portions of the NIH GuidelinesAppendix B Appendix B • Classification of human etiologic agents on the basis of hazard • Bacterial • Fungal • Virus • Prion • Parasites Microsporum RG2 Fasciola hepatica RG2 Brucella abortus RG3 Epstein Barr RG2 Ebola virus RG4

30. Key Portions of the NIH GuidelinesAppendix G Appendix G • Specifies details of containment and confinement for standard laboratory practices • Defines Biosafety Level 1 through Biosafety Level 4 • Appropriate for animals that are worked with in a laboratory setting

31. Key Portions of the NIH GuidelinesAppendix I Appendix I • Biological containment barriers • Limit the infectivity of a vector or vehicle (plasmid or virus) for specific hosts • Limit dissemination and survival of a vector in the environment • Vectors can be genetically designed to decrease, by many orders of magnitude, the probability of dissemination of recombinant DNA outside the laboratory

32. Key Portions of the NIH GuidelinesAppendix Q Appendix Q • Applies when research animals are of a size or have growth requirements that preclude laboratory containment • For example, cattle, swine, sheep, goats, horses, poultry, etc. • Addresses containment and confinement practices in animal facilities (BL1-N to BL4-N)

33. Key Portions of the NIH Guidelines for Animal Research • Primates - Appendix G or Q? • Depends on the conditions under which the primates are being housed and used in experimentation • Primates used in high-level, laboratory containment conditions; Appendix G applies • In other settings, primates may be worked with in settings akin to those described in Appendix Q • Professional judgment is key - IBC/IACUC can help!

34. Key Portions of the NIH GuidelinesAppendix M 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. • Requirements for Protocol Submission, Review, and Reporting

35. Key Portions of the NIH Guidelines for Animal Research • Appendix M • Applies to human gene transfer experiments • Includes many considerations related to preclinical studies with animals • Expedited safety reporting requirements amended to include specifically the reporting of animal data “that suggest a significant risk for human research participants.”

36. Questions?

37. Institutional Biosafety Committees: The Linchpin of Local Oversight

38. InstitutionalBiosafety Committee • IBCs were originally established specifically for the review of rDNA research, but includes more oversight now • Reviews research with biohazardous risks • Infectious agents • Highly toxic chemicals • Biological toxins

39. IBC Membership • No fewer than 5 individuals • Appropriate rDNA expertise collectively • Plant and animal experts, biosafety officer as appropriate • Expertise in assessment of risk to environment and public health • At least two public/community members not affiliated with the institution • Appointed by Institutional Official (VP for Research)

40. Biological Safety Officer • BSO must be appointed and made a member of the IBC if research is: • Large scale (>10 L) • BL-3 • The BSO’s duties include: • Periodic inspection of labs • Reporting to the IBC and institution of any problems, violations, research-related accidents or illnesses • Developing emergency plans for handling accidental spills and personnel contamination • Advice on lab security • Technical advice to PIs and IBCs on research safety procedures

41. IBC Responsibilities • Recombinant DNA research for conformity with the NIH Guidelines • Minimize potential risk to environment and public health • Containment levels per NIH Guidelines • Adequacy of facilities, SOPs, PI and lab personnel training • Institutional and investigator compliance; e.g., adverse event reports

42. IBC Responsibilities In basic and preclinical research, IBCs have the authority to: • Lower containment levels for certain experiments in which DNA from Risk Group 2-4 is cloned in non-pathogenic organisms • Set containment levels for experiments involving whole plants and animals • Periodically review institutional compliance with NIH Guidelines • Adopt emergency plans covering spills, contamination, other accidents

43. The Importance of IBCs and BSOs to Investigators • The IBC and BSO can help you to: • Ensure that you are working with recombinant DNA safely • Meet all compliance requirements associated with NIH funding for research involving recombinant DNA • Avoid preventable accidents and incidents that might cause harm or undermine public confidence in your research activities • Obtain biosafety advice on an ongoing basis

44. IBC IRB IACUC IBCs and Other Institutional Oversight Committees

45. IBC and IACUC Review of Animal Research Utilizing Recombinant DNA

46. IBCs and IACUCsAnimal Research • Joint purview, and ideally collaborative review, over certain types of research • Transgenic or cloned animals • Use of recombinant DNA molecules in animals • Pre-clinical studies and data assessment for human gene transfer protocols

47. Animal Research with rDNA: Points to Consider • Containment procedures (SOP’s) • Physical and biological • Plans for recapture of escapees • Consequences should containment fail • Procedures for transfer of animals • Transportation procedures • Disposal and destruction methods • Breeding SOP’s • Occupational biosafety concerns • Personal protective equipment • Decontamination

48. IRB and IBC Human Gene Transfer Research

49. IBC Meetings • Frequency Required to meet least quarterly, currently scheduled as bi-monthly. IBC meeting dates are posted on the web • Open Meetings IBC meetings are open to the public

50. Heightened Institutional Responsibility forSecurity Select Agent Rule Agents and toxins as listed by CDC/USDA/HHS • Must be registered with agency • Must develop written security plans • Federal agencies may not release information about the location of Select Agents.