BioSafety-’13-Outreach Changes to the NIH Guidelines - What They Mean to You - David W. Emery, PhD Research Associate Professor of Medicine & Chair, Institutional Biosafety Committee Eric Stefansson Manager EH&S Research and Occupational Safety

Refresher - Oversight of research with biohazardous agents at the University of Washington Changes to the NIH Guidelines What you need to do Resources for help Outline BioSafety-’13-Outreach

Infectious agents - Microorganisms (E. coli → Tuberculosis) - Viruses (AAV → HIV) - All human source materials (including cell lines) Recombinant DNA - Transfer DNA/RNA to or from microorganisms or any living cells - Transgenic mice - Transgenic plants - Clinical gene therapy trials - Synthetic DNA (PCR, sequencing, etc…) Select Agents (high security) Laboratory settings Live animals Clinical trails & Field tests Types of Biohazardous Research BioSafety-’13-Outreach

Oversight of Biohazardous Research - a team approach - EH&S Research & Occupational Safety (ROS) Institutional Biosafety Committee (IBC) Investigator BioSafety-’13-Outreach

Staffed by biosafety professionals Responsible for administrating biosafety programs and monitoring compliance with applicable regulations & policies. Key Functions: Facility design review Review research protocols and identify hazards Provide training, consultation, clearances and set OH requirements Site assessments/inspections UW Biohazard Safety Manual Support IBC EH&S / ROS BioSafety-’13-Outreach

Institutional Biosafety Committee Composed of faculty, staff, and community members Committee meetings open to public - public trust! Key Functions: Review & recommend institutional policies for research involving rDNA and biological agents. Review individual research proposals for compliance with Federal, State, local, and institutional regulations: - Biosafety containment levels (BSL1, 2, 3) - Adequacy of facilities, SOPs, training Independent approval authority for specific proposals. Monitor and assure compliance with the NIH Guidelines BioSafety-’13-Outreach

Federal Authority for IBC  Institutions receiving funding from the National Institutes of Health must assure that ALL research is carried out in compliance with the NIH Guidelines for Research Involving Recombinant DNA (& Infectious Agents). (  Assurance of compliance is accomplished through the Institutional Biosafety Committee (IBC) in collaboration with the Institutional Health and Safety Departments. ( BioSafety-’13-Outreach

Investigator Obligations (per NIH Guidelines) Wait to initiate or modify recombinant DNA research that requires IBC approval until approval is granted. Identify which section(s) of the NIH Guidelines apply to their research and notify the IBC as appropriate. Be adequately trained in good microbiological techniques (and adequately train lab personnel). Adhere to IBC determinations for containment and plans for spills and personnel contamination. Report any significant problems or violations to the IBC and NIH Office of Biotechnology Activities within 30 days (immediately in cases of exposure). BioSafety-’13-Outreach

Biosafety Standards Biosafety standards are defined and codified: Institution (Administrative Policy, EH&S/ROS, IBC) NIH Guidelines CDC/NIH BMBL Biosafety in Microbiological and Biomedical Laboratories US Department of Agriculture Select Agent Rule, 42 CFR Part 73 WISHA/WAC Occupational Health & Safety Standards WAC Bloodborne Pathogens Standard Seattle Municipal Code Infectious Waste Management BioSafety-’13-Outreach

Biosafety Standards Biosafety standards are defined and codified: Institution (Administrative Policy, EH&S/RBSO, IBC) NIH Guidelines CDC/NIH BMBL Biosafety in Microbiological and Biomedical Laboratories US Department of Agriculture Select Agent Rule, 42 CFR Part 73 WISHA/WAC Occupational Health & Safety Standards WAC Bloodborne Pathogens Standard Seattle Municipal Code Infectious Waste Management BioSafety-’13-Outreach

What Changed ?  In September 2012 the National Institutes of Health (NIH) Office of Biotechnology Activities (OBA) published final changes to the NIH Guidelines which it deemed necessary to reflect the emerging technologies involving "synthetic DNA"...  Prior to these changes, recombinant DNA was defined, in part, as "joining segments of DNA from different sources"; the new rules were necessary to capture the use of recDNA generated by artificial synthesis (synthetic DNA).  This precipitated several other changes to NIH Guidelines... BioSafety-’13-Outreach

What Changed ? (continued) Synthetic DNA is treated the same as Recombinant DNA Recombinant DNA outside of cells remains "exempt" from the NIH Guidelines, with some exceptions New rules for recDNA prepared in liposomes, other "vehicles" New definition and rules for "safe" recDNA inside cells New rules for chemically modified recDNA New rules for drug resistance genes in pathogenic agents New rules for synthetic DNA that exists in nature New rules for clinical gene therapy New rules for risk assessment Reconfirm concept that all recDNA is hazardous BioSafety-’13-Outreach

Synthetic DNA = Recombinant DNA (i) molecules that are constructed outside of living cells by joining natural or synthetic DNA segments to DNA molecules that can replicate in a living cell, or (ii) molecules that result from the replication of those described in (i) above. (i) molecules that a) are constructed by joining nucleic acid molecules and b) can replicate in a living cell, i.e., recombinant nucleic acids; (ii) nucleic acid molecules that are chemically or by other means synthesized or amplified, including those that are chemically or otherwise modified but can base pair with naturally occurring nucleic acid molecules, i.e., synthetic nucleic acids; or (iii) molecules that result from the replication of those described in (i) or (ii) above. Old Definition (Recombinant DNA) New Definition (Recombinant & Synthetic Nucleic Acid Molecules) BioSafety-’13-Outreach

recDNA Outside Cells Remains Exempt Synthetic or recombinant nucleic acids "that are not in organisms, cells, or viruses and that have not been modified or manipulated (e.g., encapsulated into synthetic or natural vehicles) to render them capable of penetrating cellular membranes" are exempt from the NIH Guidelines. New Section III-F-2 (exempt from the NIH Guidelines) BioSafety-’13-Outreach

recDNA Prepared in "Vehicles" Synthetic or recombinant nucleic acids that are "encapsulated into synthetic or natural vehicles" so as to render them "capable of penetrating cellular membranes" are no longer exempt from the NIH Guidelines, but instead fall under Section III-E (must be reported to the IBC). New Upgrade from Section III-F to III-E Examples: - Liposomes / polyplexes - Nanoparticles - Synthetic scaffolds A new section on the Biological Use Authorization (BUA) application specifically asks about this work BioSafety-’13-Outreach

"Safe" recDNA Oligonucleotides or other synthetic nucleic acids (<100 bases) that meet the following criteria: a) Do not contain an origin of replication or elements known to interact with either DNA or RNA polymerase, and b) Are not designed to integrate into DNA, and c) Do not produce a toxin that is lethal for vertebrates (LD50 < 100 ng/Kg) Examples: - siRNA, even when administered to animals - nucleic acid adjuvants for immunization - short morpholinos New Downgrade from Section III-E to III-F BioSafety-’13-Outreach

Chemically Modified recDNA Recombinant or synthetic nucleic acid molecules that are chemically modified but can still "base pair with naturally occurring nucleic acid molecules" now must be treated the same as their unmodified "functional equivalent". Examples: - morpholinos not fitting the category of "safe" - bioengineering studies with base analogs New Upgrade from Section III-F to III-E BioSafety-’13-Outreach

Drug Resistance Genes in Pathogens Section III-A - Review by the NIH RAC Old Definition New Definition The deliberate transfer of a drug resistance trait to microorganisms that are not known to acquire the trait naturally, if such acquisition could compromise the use of the drug to control disease agents in humans, veterinary medicine, or agriculture... The deliberate transfer of a drug resistance trait to microorganisms that are not known to acquire the trait naturally, if such acquisition could compromise the ability to control disease agents in humans, veterinary medicine, or agriculture... Consideration should be given as to whether the drug resistance trait to be used in the experiment would render that microorganism resistant to the primary drug available to and/or indicated for certain populations, for example children or pregnant women. Special provisions are provided for consulting the Office of Biotechnology Activities. BioSafety-’13-Outreach

DNA Sequences that Exist in Nature Old Definition (Exempt, Section III-F-2) New Definition (Exempt, Section III-F-3) Research with DNA molecules that are essentially copies of DNA found in nature (paraphrased). Rational: Assure that attempts to reanimate extinct pathogens (e.g influenza) are reviewed by the IBC prior to initiation of the research. Research with recombinant or synthetic nucleic acids "that consist solely of the exact recombinant or synthetic nucleic acid sequence from a single source that exists contemporaneously in nature." BioSafety-’13-Outreach

New Rules for Clinical Trials a) Less than 100 nucleotides b) Do not contain an origin of replication or elements known to interact with either DNA or RNA polymerase, and c) Are not designed to integrate into DNA Note: All clinical trials with recombinant and/or synthetic DNA, even if meeting the criteria for "safe", must still be reviewed and approved by the IBC prior to initiation. New rules exempt clinical trials with "safe" recombinant and/or synthetic DNA from review by the RAC BioSafety-’13-Outreach

New Rules for Risk Assessment 1) The Institution, IBC, and Principal Investigators must adhere to the intent of the NIH Guidelines as well as to its specifics. 2) For organisms containing recDNA from multiple sources, the risk assessment should include both the origin of those sources and function of the individual genes. 3) Both the Principal Investigator and the IBC should consider whether the "combination of certain sequences in a new biological context may result in an organism whose risk profile could be higher than that of the contributing organisms or sequences". In short: The PI and the IBC need to carefully assess the potential hazard of highly engineered organisms based on expected biology as well as by category. BioSafety-’13-Outreach

All recDNA Still Deemed Hazardous Even though much work with recombinant and/or synthetic nucleic acids remains exempt from the NIH Guidelines: 1) Contact EH&S Research and Occupational Safety (ROS) to help determine if your research is exempt. 2) ALL forms of recombinant and/or synthetic DNA, no matter how trivial or exempt from the NIH Guidelines, are considered a biohazard and must be handled and disposed of in an appropriate manner! BioSafety-’13-Outreach

What You Need To Do Review your research program to determine if these changes to the NIH Guidelines apply to your research. If you received Biological Use Authorization during the past three years and you believe your research approval would be impacted by these changes, please submit a Request for Change form no later than February 28. If you are currently working without an EH&S Biological Use Authorization and you believe your research falls under the amended NIH Guidelines, please submit a BUA application no later than February 28. Please keep these changes in mind as you plan your future research efforts. BioSafety-’13-Outreach

Examples You use liposomes or other forms of DNA "vehicles" (may raise containment level) You use siRNA or other forms of "safe" recDNA in animals (may lower containment level) You work with recombinant / synthetic nucleic acids and have never registered with EH&S. EH&S Research and Occupational Safety will be reviewing past approvals for these and other specific types of research and may contact you if needed. BioSafety-’13-Outreach

Deadlines All new applications and change forms must be submitted no later than February 28, If you miss this due-date, your application my not be reviewed and approved be the end of March 2013 (the due-date set by the NIH). Please remember that failure to comply with the NIH Guidelines, including properly registering your research, puts your funding, and that of your colleagues, at risk. BioSafety-’13-Outreach

References / Contacts EH&S Research and Occupational Safety: , Biological Use Authorization applications: NIH Guidelines: FAQs for changes to the NIH Guidelines: NIH: EH&S: BioSafety-’13-Outreach

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