Oregon Health and Science University — Hazardous Materials Safety Interpretation

Page 1U.S. Department of Transportation Pipeline and Hazardous Materials Safety Administration SEP - 4 2009 1200 New Jersey Ave., SE Washington, DC 20590 Ms. Debra A. Brickey, PhD Responsible Official Select Agent Program, Central and Waterfront Campuses Oregon Health and Science University, PP- 1 70 3 18 1 SW Sam Jackson Pk Rd Portland, OR 97239-3098 Ref. No. 09-0 186 Dear Dr. Brickey: This responds to your June 3 1,2009 email requesting an interpretation of the applicability of the Hazardous Materials Regulations (HMR; 49 CFR Parts 17 1-1 80) to the transport of Division 6.2 materials. Specifically, you ask whether mice infected with a replication defective adeno-associated virus (AAV) are subject to the requirements of the HMR. The information attached to your email indicates that the mice are infected with a type of AAV that does not cause disease in humans or animals. The AAV injected into the mice is inactivated such that it does not replicate itself in the host; thus, the mice do not contain infectious AAV viral particles. Under tj 173.134, a Division 6.2 Infectious substance is defined as a material known to contain or reasonably expected to contain a pathogen, such as a virus, that can cause disease in humans or animals. Additionally, under exceptions provided in 173.1 34(b), a material containing pathogens that have been neutralized or inactivated such that they no longer pose a health risk is not subject to the requirements of the HMR as a Division 6.2 material. Based on the information provided in your email, it is the opinion of this Office that the mice do not meet the definition of a Division 6.2 material and are not subject to the HMR. I hope this information is helpful. If you have further questions, please contact this ofice. Sincerely, ief, Standards Development of Hazardous Materials Standards#

Page 2Der 6( r nde~eR age 1 of 4 Drakeford, Carolyn (PHMSA) From: Gorsky, Susan (PHMSA) Sent: Friday, August 07, 2009 3:00 PM To: Drakeford, Carolyn (PHMSA) Cc: Lavalle, Diane (PHMSA); Mazzullo, Ed (PHMSA) Subject: FW: OHSU Animal shipment to Germany From: Debra Brickey [mailto: brickeyd@ohsu.edu] Sent: Friday, August 07, 2009 2:35 PM To: Special Permits (PHMSA) Cc: Sally Finch; Kim Saunders; John Brigande; Gwynn Daniels Subject: RE: OHSU Animal shipment to Germany Hi Diane, Thanh you for J our assistance. 1 talked thi:, over nith the OSHIJ director of the Deparl~lle~ll oTCornparative blt.dlcine and we both agreed that at thesc animals would not havc an) non-integrated ~,cplication-~ncompctent adeno-associated virus. I'liese nlice would 1101 pose a harard to humails or othcr ailirnals duc to the procedure done to them as embrqos. If you could provide us with an email approval and a written approval so that we may ship as soon as animals are ready, it would be very appreciated by all of us here at OHSIJ. Debra I1cbr:i A. Hrickq. Phi) Laboratory Safety Ad\ isor Biosafety & Chemical Hygiene Ofiicer Kcsponsiblc Official Select Agent Program. CIentral and Waterfront Campuses Oregon Health and Science I_Jniversilj PP- 1 70 3 1 8 1 S&' Satn Sacl<son Pk lid T'ortland. OR 97239-3098 503-494-0655 hrickeydllu'cohsu.edu From: specialpermits@dot.gov [mailto:specialpermits@dot.gov] Sent: Friday, August 07, 2009 7:01 AM To: Debra Brickey Cc: Kenneth.Herzog@dot.gov; Darral.Relerford@dot.gov Subject: RE: OHSU Animal shipment to Germany Hi Debra, Your email indicates the mice are not pathogenic to other mice, rodents, or humans. We need to be sure the mice are not pathogenic to animals other than rodents. If you can assure us that the mice are not pathogenic to humans or animals, then we would agree they are not regulated as Division 6.2 materials under#

Page 3Page 2 of 4 the HMR. We can respond by email immediately, a written response will take a bit. Thanks, Diane LaValle Transportation Specialist From: Debra Brickey [mailto:brickeyd@ohsu.edu] Sent: Thursday, August 06, 2009 3:23 PM To: Special Permits (PHMSA) Subject: FW: OHSU Animal shipment to Germany Importance: High 1'0 the atteritiori of l)arrell Kelcford. I'his In reference to the permit for Oregon Ilealth & Scicncc Llniversity (OlISU) to ship mice from thc US to Germany on passenger aircraft. From: Debra Brickey Sent: Friday, July 31, 2009 10:45 AM To: 'kenneth.herzog@DOT.gov' Subject: OHSU Animal shipment to Germany Importance: High Kcnny, Here is Dr. Soh11 Rriga~ide'b explanation of his virul; work in these mice. What it boils down to is that the virus Is not in the mice at the tirllc of shipping. 'nit genes carried by the virus are in tlic mice but a11 of the virus has been eliminated by the mouse cells. The delays have lead to a lnillilnum cost oC$6000 dollars to the lab due to the age requireriierlt for the mice needed for the experiment an expedited assessment uould be greatly appreciated by all those involved in thc pcnriit process at 01 ISU and Gcnnatiy. If based on this you decide that a permit is not required, could you givc us a written dcclar ation of this assessment? Do you think we could have resolution 011 this by earlj next ~ c c k ' ? 'I'hanks. Debra Debra A. Hricl<ey. PhI) Laboratory Safety Advisor Biosafe'ety 8r. Chemical Hygiene Oflicer liesponsible Official Select Agent Program. Central C:tnlpus Oregon Health and Science University PP- 1 70 3 1 8 1 SW Sam Sackson Pk Kd Portland, OR 97239-3098 503-494-0655 brickcyd(~ohsu.edu , , ,,, From: John Brigande Sent: Thursday, July 30, 2009 8:34 PM To: Debra Brickey; Sally Finch Cc: Chris Bresee#

Page 4Page 3 of 4 Subject: RE: Animal shipment to Germany Deb, The nild type adeno-associated virus (AAV) is a member of the parvo\rirus famil! that displays 110 pathogenicity in hu~llans. 1 hc t irus is composcd of a single strandcd l>KA gellome that comes in hio versions: sense or coding and anti-sense or non-coding. l'he wild type virus in humans can integrate into a specific site called AAVSl in human chromosome 10. Because i: is not associated with ally pathogenic or disease slate in humaris and because it does integrate, AAV has bee~i detclopcd as a gcnc tl~erapy vcctor for usc i l l trcat~ng hu~na~l discases through genc replacement. Currently. there are cli~iical trials using AAV-based vectors for gene therapy for lipid storage disease: muscular d j strophy; and C'anava~i disease. Completed hulnan cli~lical trial> using AAV-based gene tllcrapy ~cctors ]lave focused on cystic fibrosis. al-tllritis. and l~ereditary empllyscnla alllong other d~seascs. The vector we are working with is a replication defective versio~~ of the non-pathogenic wild Qpe t irus. Our v i ~ us is also non-pathogenic, and it docs no1 lrlake more of itself in thc infected host. I'hc gellome of our virus has deletions of DNA seqilelice that encode proleiris essential for the virus to replicate. Hence. our virus infects cells but callnot replicate in them to produce more of itself. '["lie i~~fectior~ penl~lts our gene to bc esprcsscd in cells in the inner ear so \YC can learn if our pcrlc replacement restores hearing function. l'he infected cells cannot mahe tnore viral particles, and they therefore do not harbor infectious AAV viral particles. The animals we seek to ship to our colleague in Germanj Mere infected with the replication defective A4V virus lien they were en~brjos groni~lg in the uterus of the tkmale mouse. Tliey are born about a week later, mature to 31 days, and arc then tested for hca~ing function. 'The ncxt step is to have tlie scnsorq hair cells evaluated b) electrophysiology: our colleague in (;ermanj is one of tlie only people on earth who does the specialized hind of recordings ncccssary to validate hair cell function. 'I here arc no otller optio~ls for us to sccure the elcctrophysiological analyses. Wc are higllly confident that the adult tnicc that wcre cvposcd to 11 AV wllcn thcy bere cml~rjos g ~ o n 111g in the uterus do not shed virus: thcy carinot since they arc unable to iiiakc riew tirus. 'She .4AV virus wc usc is simplj a gene shuttle to bring our cargo. the genes of interest. into the target cells in the i~i~ier ear. very sinlilar to the AAV " tiral vectors used for human gene therapy. We bclie\le our mice pose 110 risk wllatsoe\.c~ to othcr nllce. ~odents. or humans. Wc have gcileratcd 30 adult rr~icc to datc that ucrc csposed to cxpcrirrlcr~tal arid coiitrol virus. Wc had 4 possible ship dates for these animals that have all passed. Estimating the cost of this extremely difficult. Each i11.iected mouse is worth about $200 US[> givc~l what it costs to generate the arlio~als. maintain them. and perform thc surgery and irijections. 'l'liis is tliereforc about $6.000 lost thus Car, and it's probabl~ a lot\ estimate. 1 have not tigured in the technician salary tirile to perform the preoperative and postoperative care. or my salarj as I do all of the in-jcctions. t3ut more importa~lt than the money lost is the corlipletc breakdown of basic science research that has occurred: we cannot adtance this project li~rlher without the electrophjsiological recording$. I hope that providing 1011 with tllcse details \+ill help the 110 1' and FAA criticallj evaluate our request I b r a passenger flight permit to get these mice to Germany. 1 atn happy lo fly to the DOT or the FAA home offices at rnq expcrlse and present a scssion on replication detective AAV V I J us ~f requested. 1 do appreciate your support of our efforts. John From: Debra Brickey Sent: Thursday, July 30, 2009 12:53 PM To: John Brigande; Sally Finch Subject: Animal shipment to Germany#

Page 5Page 4 of 4 Hey John, I spoke to Kenneth Herzog and he had heard from the FAA and their suggestions and then said it had to go to a technology advisor. I tried to explain more fully to Kenneth that the mice should not be categorized as Category B because there was no longer live virus in the animals. He asked me to write up an explanation to give to their technology representative but I first wanted to confirm my understanding of what happens with you John and also give an estimate of cost including time spent on the missed two previous shipments due to delays in getting this approval. So, my understanding is that the adenovirus construct is injected into embryos in the inner ear cells. The adenovirus expresses the prescribed proteins but since it cannot replicate it would be gradually degraded and expression lost as the animal develops (it does not integrate into the DNA correct?). By the age of shipment the animals would no longer have adenovirus in their bodies. So that the animals are essentially pathogen free. Do you want to add anything to that or clarify any part of that? Please let me know your costs and comments ASAP since I would like to email this to him later today or early tomorrow? I apologize for all the hassle this has caused for you. Debra#