1. Institutional Biosafety Committee Trends and Human Gene Transfer Considerations
Arizona Biosafety Alliance
January 24, 2014
Chris Jenkins, PhD, MPH, RBP, CHMM
Senior Director of IBC Services and Biosafety Consulting

2. Overview
Introduction and Research Context
Specific Aims
Study Design and Methods
Results
Discussion and Future Research
Human Gene Therapy Considerations

3. Feature Presentation 2:35 of Outbreak (1995) Star-studded cast:
Morgan Freeman
Dustin Hoffman
Rene Russo
Kevin Spacey
Patrick Dempsey
Cuba Gooding Jr.
Donald Sutherland (underrated)

4. Biosafety Levels 1 & 2
BSL-1
BSL-2

5. Introduction and Research Context

6. Research Problem
Current oversight system intended to provide institutional flexibility, with no “one-size-fits-all” solution.
Frederickson, 1982
No uniform standard exists for oversight of biological materials used in research as there are for animals, radioactive materials, or human subjects.
In practice, with no baseline, institutional research compliance leadership lack understanding on biological materials oversight.

7. Research Questions
What is the regulatory extent of biological materials oversight in the United States for life sciences research entities?
What insights can a cross-sectional survey provide into the current state of local biological materials oversight at life sciences research entities?

8. What are Biological Materials?
Biological Materials or Biohazards are infectious agents or hazardous biological materials that present a risk or potential risk to the health of humans, animals or the environment.
CDC BMBL 5th ed., 2010
Examples:
Whole or “Wild-type” Microorganisms
Biological toxins
Blood or other potentially infectious materials
Recombinant or synthetic DNA resulting in organisms
Image courtesy of the CDC

9. Recombinant and Synthetic Nucleic Acids
“Molecules that are constructed outside living cells by joining natural or synthetic DNA segments to DNA molecules that can replicate in a living cell, or molecules that result from the replication of those described above.”
National Institutes of Health, 2013
NIH funded research involving recombinant DNA (and in March, 2013, recombinant or synthetic nucleic acids, rsNA) requires a risk assessment by a local Institutional Biosafety Committee.
Image courtesy of the Scientific American

10. Recombinant and Synthetic Nucleic Acids
“Molecules that are constructed outside living cells by joining natural or synthetic DNA segments to DNA molecules that can replicate in a living cell, or molecules that result from the replication of those described above.”
National Institutes of Health, 2013
NIH funded research involving recombinant DNA (and in March, 2013, recombinant or synthetic nucleic acids, rsNA) requires a risk assessment by a local Institutional Biosafety Committee.
Image courtesy of the Scientific American

11. History of Recombinant DNA Technology and Oversight
Dr. Kornberg, 1968 U.S. Senate Subcommittee
Vettel, 2006
Moratorium on rDNA experimentation after Paul Berg generated first genetically modified replication competent E. coli in 1973
Jackson, Symons, and Berg, 1972
Gordon Conference Session request to National Academy of Sciences
Singer and Sol, 1973
Assessment of recombinant DNA risks to be handled at Asilomar State Beach
Berg, Baltimore et al, 1974
Above, Dr. Frederickson, Director of the NIH, Below, Dr. Paul Berg. Images courtesy of NIH and Nobel Institute
Q1. Dr. Arthur Kornberg directed the subcommittee's attention toward the rapidly progressing field of molecular biology (Vettel 2006), (1968). Dr. Kornberg noted important developments were near fruition and that the potential social impact of these advances could be far-reaching (Vettel 2006). Dr. Kornberg was referring to discoveries which would provide the technical framework for the specialty commonly known as genetic engineering (1968). New techniques developing in this field would enable a researcher to recombine DNA, the hereditary material of the cell, in a very precise manner (1968). The rDNA introduced could provide a cell with the ability to manufacture products (for example, insulin) which were previously not part of the cell's make-up (Johnson).
Q2. Singer and Soll

12. Asilomar Conference
The primary goal of the meeting was whether to lift the recombinant DNA moratorium and under what set of prescribed conditions.
Berg, Baltimore et al., 1975
While little data beyond Berg’s experiment existed at the time, despite opposition, the Conference ended with the understanding rDNA research should proceed but under strict guidelines.
Berg and Singer, 1995
1976, NIH Guidelines for Research Involving Recombinant DNA Molecules issued
Frequent revisions through 2013
Images courtesy of NIH

13. The Role of the Institutional Biosafety Committee and Risk Assessment
Institutional Biosafety Committee (IBC)
Capability to assess the safety of rDNA research
Be able to identify any potential risk to public health or the environment (NIH Guidelines, 2013)
Risk Assessment
Identify hazardous characteristics
Evaluate exposure and consequences
Determination BSL, work practices, safety equipment, and facility design to prevent exposure (CDC BMBL 5th ed., 2010)
Microsoft Office open-source images

14. United States Regulatory Oversight of Biological Materials in Research
NIH
rDNA
Dual Use Research of Concern
Gene Therapy
CDC/USDA
Select Agent Program
Importation
Federal OSHA
Bloodborne Pathogens
General Duty Clause
Others Peripherally Associated
Images courtesy of respective government entities

15. Figure 1. Biosafety & Biocontainment Regulations, Standards, and Guidelines Pertinent to High Containment and Maximum Containment Research (USDA Federal Task Force, 2009)

16. Figure 2. Biosafety & Biocontainment Oversight (USDA Federal Task Force, 2009)

17. Figure 2. Biosafety & Biocontainment Oversight (USDA Federal Task Force, 2009)

18. Specific Aims

19. Specific Aims
Specific Aim #1: To investigate United States life sciences regulation for research involving biological materials to assess the adequacy of biosafety and biosecurity oversight.
Specific Aim #2: To evaluate IBCs charged to oversee research with biological materials to determine whether additional guidance and regulation is needed to protect staff, biological materials, and public health.

20. Study Design and Methods

21. Survey Methodology Cross-Sectional Survey of NIH-OBA Registered IBCs
FOIA #40395 (August, 2012), 857
FOIA #41293 (May, 2013), 866
FOIA #42013 (December, 2013), 868
Survey Design
22 Questions
Institutional Type and Constituency of the IBC
Biological Materials Review
Protocol Review Determinations

22. Full List of Survey Questions
Institution Type (small, medium large University, research institute, private industry, government, non-profit, other)
Type of Biological Materials Research Reviewed by the IBC (rDNA, non-rDNA whole microorganisms, BBP, toxins, Select Agents, Dual Use)
Is your IBC administered internally (support staff on location) or externally (third-party)
Number of protocols reviewed over IBC active date range (1976 to 2012) at BSL-1, BSL-2, BSL-3, and BSL-4)
Estimated Time for Review per Protocol (from receipt, processing, inspection, IBC meeting, and follow-up items, approval) (0 hours to indefinite)
What type of renewal strategy is employed upon approval?
Number of Investigators with Approved Protocols
By year from the inception of the IBC, how many initial protocols are approved each year (1976 to 2012)
If yes to Recombinant DNA Research, what categories of rDNA research have been reviewed? How often are IBC meetings held?
How many Full-Time Employees support the Institutional Biosafety Committee (0 to indefinite)
How much financial support does the IBC receive from the Office of Research ($0 to indefinite)?
If yes, is the IBC Chair Compensated for time spent (Yes/No/N/A)
If yes, are other Committee members compensated for time spent serving on the IBC?
How much financial support does the IRB receive from the Office of Research ($0 to indefinite)
How much financial support does the IACUC receive from the Office of Research ($0 to indefinite)
Committee Composition (NIH Roster and Function Support)
Number of scientific members
Number of biological safety professionals
Number of external (public) members
Number of administrative staff
Other members (include title and number)
If known, the number of NIH reported laboratory accidents involving recombinant DNA at the entity from

23. NIH-OBA Registered IBCs
9/11 and Anthrax Letters
: 12 IBCs added per year
: 43 IBCs added per year
From 1976 to 2000, an average increase of 12 IBCs per year.
From 2001 to 2013, average jumps to 43 new IBCs per year, with the latest total in May, 2013 at 866.

24. Image courtesy of Qualtrics®
Survey Dissemination
Qualtrics®
Pilot Study
12/1/2012 to 12/15/2012
Full Study
1/15/2013 to 2/15/2013
Reminders sent at 2 week and 4 week intervals
Image courtesy of Qualtrics®

25. Results

26. Response Rates and Institutional Demographics
181/817 Responses in Main Survey (21.2%)
13/40 in Pilot Survey (32.5%) – not included
Institution Type
Number
Small University 19
Medium University 35
Large University 60
Research Institute 18
Private Industry 11
Government 9
Non-Profit 14
Other
Total
177

27. Institution Type by Biological Materials Research
Small U.
Medium U.
Large U.
Research I.
Private I.
Government
Non-Profit
Other
Total
rDNA 18 35 59 17 11 8 13
172
Microorgs. 24 43 2 5
109
OSHA BBP 10 22 37 3
103
Bio. Toxins 41 9 6 7 94
Select Agents 20 40 93
DURC 30 56 1 4
173
X2: 53.18
Df: 28
p-value: 0.00
Focus on the total category, and describe other. Other includes field releases.

28. Institution Type by Biological Materials Research
Count
Institution Type

29. Type of Recombinant DNA Research Reviewed
172 Responses
Institution Type
Number
Laboratory Benchtop
156/172
Animal Research
124/172
Plant Research
67/172
Gene Therapy
54/172
Other
10/172
*Other = field releases

30. Institution Type by Recombinant DNA Research Reviewed by the IBC
Small U.
Medium U.
Large U.
Research I.
Private I.
Government
Non-Profit
Other
Total
Lab Benchtop 18 31 60 15 7 10 8
156
Animal 13 26 51 14 3 6
124
Plant 11 42 2 1 67
Gene Therapy 12 9 54 4 34 17
172
X2: 53.18
Df: 28
p-value: 0.00

31. Institution Type by Recombinant DNA Research Reviewed by the IBC
Count

32. IBC Review Process and Institutional Support
Range of Research Investigators at an institution
1 to 800 (172 responses)
Protocol Review Process
Average time spent preparing protocol for IBC review
5.2 hours, with a range from 1 hour to 40 hours (110 responses)
Average time from PI submission to IBC approval
~29 days, with a range from 7-90 days (62 responses)
Compensation
IBC Chair: 33/139 (23.7%)
IBC Members 7/128 (5.5%)

33. IBC Protocols By Year
R² =
Count
Year

34. Institutional Case Study

35. Survey Response Comments
IBCs are under-supported compared to peer Research Compliance Committees (IRB and IACUC)
Low to no institutional interest in IBC support except for Select Agent Programs or following exposure incidents
Recombinant DNA not as risky as wild-type and bloodborne pathogens research
Review of IBC protocols differs by Biosafety Level (BSL) and type of research
Further quantitative data on trends over time is needed
IRB and IACUC
Little institutional interest except for Select Agents or incidents.

36. Discussion and Future Research

37. Observed Trends in Biological Materials Oversight and IBCs
Research involving biological materials has increased over time
Protocol review data
Expansion of IBC review beyond NIH Guidelines and Select Agent requirements
Institutional support minimal beyond staffing
This is evidenced by the increasing number of registered IBCs from the Hackney surveys to this cross-sectional survey, and the case-studies of protocols reviewed by year.
IBC also include review of whole microorganisms, blood borne pathogens, and biological toxins. In addition, IBCs are tasked with fulfilling federal requirements for the review of new DURC requirements, essentially asking institutions to conduct an analysis of whether a research project should ethically be allowed to proceed.

38. Observed Trends in Biological Materials Oversight and IBCs
Institutions registering multiple IBCs to handle different aspects of review
Inconsistent or irregular data on laboratory incidents with biological materials
172/173 reviewed recombinant DNA, as required by NIH funding
One example is a mid-size academic entity with a hospital system employing one IBC to conduct biosafety assessments of rsNA laboratory, animal, and plant research, and the other IBC to conduct research of rsNA into human subjects.
Only one institution has an IBC without recombinant DNA, indicating review covering non-NIH required work

39. Directions for Future Research
Annual survey of institutions through FOIA requests to NIH- OBA to build data set
Explore case study analysis of individual institutional trends
Research human gene therapy biosafety and participants safety through FDA FOIA on severe adverse events related to recombinant products
Evaluate implications of Dual Use Research of Concern regulation on IBCs
Highlight policy gaps and risks in United States biological materials oversight

40. Upcoming Publications
Journal of Research Administration, Society of Research Administrators, in review, Spring 2014.
Biosecurity and Bioterrorism: Biodefense Strategy, Practice, and Science. UPMC Center for Health Security (formerly Center for Biosecurity at UPMC), in draft, Summer 2014.

41. Selected References
(1968). Hearings on the Human Impact of Advances in Biological Science. Senate Subcommittee on Government Research, National Institutes of Health: 6.
Berg, P., et al. (1974). "Potential Biohazards of Recombinant DNA Molecules." Science 185(4148): 303.
Berg, P., et al. (1974). "Letter: Potential biohazards of recombinant DNA molecules." Science 185(4148): 303.
Berg, P., et al. (1975). "Asilomar Conference on Recombinant DNA Molecules." Science 188(4192):
Berg, P. and M. F. Singer (1995). "The recombinant DNA controversy: twenty years later." Proc Natl Acad Sci U S A 92(20):
Frederickson, D. S. The recombinant DNA controversy : a memoir : science, politics, and the public interest Washington, D.C., ASM Press.xix, 388 p., [8] p. of plates : ill. ; 23 cm.
Harris, K. (March, 2005). The NIH Guidelines and IBC Responsibilities. IBC Basics, Town and Country Resort Hotel, San Diego, California.
Jackson, D. D., R. H. Symons, and Paul Berg. (October 1972). ""Biochemical Method for Inserting New Genetic Information into DNA of Simian Virus 40: Circular SV40 DNA Molecules Containing Lambda Phage Genes and the Galactose Operon of Escherichia coli."." Proceedings of the National Academy of Sciences of the United States of America 69(10):
Johnson, J. A. The NIH Recombinant DNA Guidelines: Brief History and Current Status. . Washington D.C., USA UNT Digital Library.
National Institutes of Health, O. o. B. A. (2013). "NIH Guidelines for Research Involving Recombinant and Synthetic Nucleic Acid Molecules."
Paul Berg, D. B., Sydney Brenner, Richard Roblin III, Maxine Singer (June 1975). "Summary Statement of the Asilomar Conference on Recombinant DNA Molecules." Proceedings of the National Academies of Sciences, United States of America 72(6):
Pike, R. M. (1979). "Laboratory-associated infections: incidence, fatalities, causes and prevention." Annual Review of Microbiology 33:
Resnik, D. B. (2010). "Dual-Use Review and the IRB." J Clin Res Best Pract 6(1).
Raymond W. Hackney, J., Theodore A. Myatt, Kathleen M. Gilbert, Rebecca R. Caruso, and Susanne L. Simon (2011). "Current Trends in Institutional Biosafety Committee Practices." Applied Biosafety 17(1):
Singer, M. and D. Soll (1973). "Guidelines for DNA hybrid molecules." Science 181(4105): 1114.
U.S. Department of Health and Human Services, C. f. D. C. a. P., and the National Institutes of Health (2009). Biosafety in Microbiological and Biomedical Laboratories. Washington, D.C., U.S. Government Printing Office.
Vettel, E. J. (2006). Biotech: The Countercultural Origins of an Industry, University of Pennsylvania Press.
Zilinskas, R. A. and B. K. Zimmerman (1986). The gene-splicing wars: Reflections on the recombinant DNA controversy.

42. Human Gene Therapy Considerations

44. Ex vivo Gene Therapy Cells from diseased person are removed
Then, they are treated in lab (using techniques similar to bacterial transformation)
Finally, they are reintroduced to the patient
More effective than in vivo
Transfection is the introduction of DNA into animal or plant cells

45. In vivo Gene Therapy
Introducing gene directly into tissues or organs without removing body cells
Challenge is delivery only to intended tissues
Viruses act as vectors for gene delivery, but some injected directly into tissue

46. Delivery of Therapeutic Genes
Therapeutic genes often called payload
May require long-term expression of corrective gene
Others require rapid expression for short periods of times

47. Viral Vectors
Viral vectors use viral genome to carry therapeutic gene(s) and to infect human body cells
Adenovirus (common cold)
Adeno-Associated Virus
Retrovirus (HIV)
Herpes Simplex Virus (cold sores)
Vaccinia Virus
Viruses must be engineered so that they can neither produce disease nor spread (extremely effective at infecting human cells)

48. Vector Transfection
Targeted gene therapy may result since some viruses infect certain body cells
Adenoviruses infect both dividing and non-dividing cells effectively
Adeno-Associated viruses do not cause illness in humans, can infect a wide variety of cells, & integrate 95% of time in same location
Retroviruses are of interest because they insert DNA in to the genome of host where it remains permanently (integration), but often, randomly
Herpes simplex viruses (HSV-1) strain primarily affects central nervous system (CNS)
May help develop treatments for Alzheimer’s, Parkinson’s, etc.
Although viral vectors may help, most human cells are not easily transfected

49. Curing Genetic Disease
More than 3,000 human genetic diseases are caused by single gene mutations
These are strong candidates for treatment by gene therapy
Cystic Fibrosis
Huntington’s disease
Tay-Sachs
Hemophilia
Sickle cell disease
Phenylketonuria

50. First Human Gene Therapy
Ashanti de Silva (4 years old) with severe combined immunodeficiency (SCID) treated in 1990 at NIH in Maryland
Lacked functioning immune system because of a defect in gene called adenosine deaminase (ADA), which is involved in metabolism of dATP (nucleotide precurosor used for DNA synthesis)
Accumulations of dATP are toxic to T cells
Normal gene cloned into vector introduced into nonpathogenic retrovirus

51. First Human Gene Therapy Success
Ex vivo approach used
T cells isolated from blood
Multiple treatments required
Within a few months, T cell numbers increased
After 2 years, ADA enzyme activity was high
She is currently enjoying a healthy life

52. Risks of Gene Therapy
Discussions of safety intensified when 18- year old Jesse Gelsinger died during a clinical trial at the University of Pennsylvania in 1999.
Complications related to adenovirus that was used.
Ornithine transcarbamylase deficiency (affects ability to break down dietary amino acids)
1st person to die as a result of gene therapy.

53. Success of Gene Therapy
Success in Rhys Evans, a child born with X-linked Severe Combined Immunodeficiency Syndrome (SCIDS – aka bubble boy), in 2002
The team took stem cells that gave rise to immune cells from the boy’s bone marrow
They used a modified form of a retrovirus as a vector
The engineered stem cells were then returned to the boy’s body
Now, he has normal levels of T cells

54. Unresolved Questions Can gene expression be controlled in the patient?
What happens if normal gene is overexpressed?
How long will the therapy last?
What is the best vector to use?
What is the minimum number of cells needed to infect to achieve success?

56. Human Gene Transfer Biosafety Considerations
Consider the vector (replication competent, incompetent, attenuations)
Consider the transgene (oncogene, proto-oncogene, immune stimulator)
Consider mode of delivery (injection, cath lab)
Comprehension of risk by populations traditionally not serviced by biosafety professionals
Physicians/Clinicians
Pharmacists
Nursing Staff
Infection Control vs. Biosafety

57. Community IBCs Challenges Highly experienced Well connected
Interface between laboratory requirements (NIH Guidelines) with clinical practice
For multi-site, Sponsor trials, the PI is generally less engaged in the research. Education with the Sponsor and the PI is required.
Highly experienced
IBC reviews since 2000
Hundreds of sites – all sizes!
Nearly every state; Canada and 8 other countries
Wide range of technologies reviewed, including many first-in-human
Well connected
Extensive network of biosafety professionals
Close communication with NIH OBA (able to ask “hypotheticals”)
Proving ground for NIH-OBA

58. Thank You – Questions?