Discussion paper 2003

Regulation of DNA vaccines

The Norwegian Biotechnology Advisory Board and gene therapy on animals The Norwegian Biotechnology Advisory Board

Editor-in-chief: Sissel Rogne Editor: Grethe S. Foss Publisher: The Norwegian Biotechnology Advisory Board Printed in Norwegian 24 March 2003 Printed in English 10 November 2004 (500 copies) Available online in English since November 2003 ISBN: 82-91683-20-4 Postal address: P.O.Box 522 Sentrum NO-0105 Oslo, Norway Internet: www.bion.no E-mail: [email protected] Layout: The Norwegian Biotechnology Advisory Board

The Norwegian Biotechnology Advisory Board is an independent body appointed by the Norwegian government and was established in 1991. The Board is founded in the Act relating to the application of biotechnology in medicine and the Act relating to the production and use of genetically modifi ed organisms.

The main tasks of the Norwegian Biotechnology Advisory Board are to identify and examine the ethical questions raised by applications of modern biotechnology on humans, animals, plants and microorganisms, provide advice that can assist policy-making and stimulate public debates on the issues.

The Board consists of 24 members and has observers from six ministries. The Board’s secretariat has fi ve to eight employees. For 2003 the budget of the Biotechnology Advisory Board is 6.3 million NOK (appr. 750.000 €).

2 Regulation of DNA vaccines and gene therapy on animals

Preface

At present the Norwegian Gene Technology The Biotechnology Advisory Board has Act provides no clear answer as to how discussed regulatory alternatives for DNA animals receiving DNA vaccines and gene vaccines and gene therapy on animals therapy are to be regulated and whether in the light of the internal seminar and or not they are to be termed as genetically the discussion paper. The Biotechnology modifi ed. Advisory Board’s recommendations are set out in its reply letter to the Ministry of the The Norwegian Biotechnology Advisory Environment dated 26 February 2003, which Board raised this problem for the fi rst time is an enclosure to this discussion paper. at an internal seminar in Namsos on 5 September 2001. In the light of the seminar, Dr Grethe S. Foss from the Biotechnology the Ministry of the Environment asked the Advisory Board’s secretariat has been Biotechnology Advisory Board to discuss how responsible for preparing the discussion paper, DNA vaccines and gene therapy on animals and members of the Biotechnology Advisory should be regulated and what status should Board and the secretariat, specialists and be given to DNA-treated animals. others have provided valuable contributions and comments. Since the problem is highly complex, the Biotechnology Advisory Board’s secretariat The discussion paper has been translated has drafted this discussion paper setting out with the assistance of Ms Anne Bryn and Dr the various aspects of the issue. Graham Harrod.

Werner Christie Sissel Rogne Chairman Director

3 The Norwegian Biotechnology Advisory Board

4 Regulation of DNA vaccines and gene therapy on animals

Contents

1. Background for the discussion paper 6

2. Introduction 8 3. Biology: DNA vaccines and gene therapy 10 3.1 Defi nitions 10 3.2 DNA vaccines 10 3.3 Gene therapy 12 3.4 Stability of the added genetic material in the animal 13 3.5 Possible integration into chromosomes 13 3.6 New properties of DNA-treated animals 14

4. Current regulatory framework – national and international 16 4.1 GMO and DNA under current Norwegian legislation 16 4.2 Defi nitions and intentions of the Gene Technology Act 18 4.3 New EU Directive on the deliberate release of GMOs 21 4.4 Cartagena Protocol on Biosafety 23

5. Possible regulatory methods 25 5.1 Regulating the animal 25 5.2 Regulating treatment 26

6. Considerations to be made when selecting regulatory method 27 6.1 Environment 27 6.2 Animal welfare 29 6.3 Health 32 6.4 Consumers 34 6.5 Business interests 35 6.6 International cooperation 37

7. Summary of regulatory alternatives 40 7.1 Regulating the animal 40 7.2 Regulating treatment 42

8. References 44

Enclosure: The Norwegian Biotechnology Advisory Board’s recommendations for a regulatory method

5 The Norwegian Biotechnology Advisory Board

1. Background for the discussion paper

New and promising methods for preventing therapy are to be regulated. The ambiguity and combating disease are now being lies in the Act’s defi nition of GMOs as developed for humans and animals in the “microorganisms, plants and animals in form of DNA vaccines and gene therapy. which the genetic material has been altered Both methods are based on the transfer of by means of gene or cell technology” and more genetic material to cells in the body. particularly what an alteration of “genetic material” implies. Would there have to be a Besides good nutrition and clean water, heritable alteration, or would it suffi ce if one of the animal’s cells had received a foreign vaccines have revolutionized public health. DNA fragment? No biological answer to Traditionally, weakened or dead disease this question is evident and, in this respect, organisms have been used as vaccines. Most it remains up to the public authorities to vaccines provide such good protection that defi ne an appropriate regulatory system for the benefi ts more than outweigh the rare human-created technologies. cases of disease that may occur as a result of the vaccine. However, for many disease organisms, especially viruses, it has proven The problem has become of current diffi cult to develop vaccines. DNA vaccines interest following specifi c enquiries made are, moreover, cheap to produce and stable to the Ministry of the Environment and under transportation. Trials are currently the Norwegian Directorate for Nature in progress to develop DNA vaccines against Management by two different fi rms – one malaria and AIDS. wishing to develop DNA vaccines for fi sh, the other to enhance fi sh growth through gene therapy by introducing a growth In order to gain public acceptance for the hormone gene into somatic cells in the fi sh. use of genes for medical and veterinary These fi rms asked whether vaccinated fi sh purposes, there is a need for information or fi sh treated by means of gene therapy and an appropriate regulatory framework, of would be subject to the provisions of which risk assessment must be an integral the Gene Technology Act. In view of the part. methods applied, interpretations of the Gene Technology Act and the preparatory The methods used in DNA vaccines and work of the Act, the Directorate held that gene therapy are similar to those applied in DNA vaccines and gene therapy would be the genetic modifi cation of organisms. This regulated by the Gene Technology Act, which raises the question of whether an animal would mean that animals receiving such is to be considered a genetically modifi ed treatment would be classifi ed as genetically organism (GMO) when it has received a modifi ed. DNA vaccine or gene therapy. If this is the case, the Act relating to the production and The Biotechnology Advisory Board raised use of GMOs (the Gene Technology Act) this issue at a seminar held in Namsos on becomes relevant and regulates the animal’s 5 September 2001, where invited lecturers movements in nature as a deliberate release from industry and public administration of a GMO. Human beings are not covered by discussed DNA vaccines, recombinant living the Gene Technology Act. Hence, under no viral vaccines, various risk elements involved circumstances, will humans receiving gene and the environmental administration’s therapy be classifi ed as GMOs under today’s assessment of the situation to date. On the legislation. basis of the different views expressed, the Ministry of the Environment wanted the At present, the Gene Technology Act regulation of DNA vaccines and gene therapy provides no clear-cut answer as to how of animals to be assessed on grounds of animals receiving DNA vaccines and gene general principles, pinpointing the various

6 Regulation of DNA vaccines and gene therapy on animals

consequences and considerations that are should be regulated. The fundamental and involved. practical aspects of the issue have been set out and possible alternative regulatory The Gene Technology Act is currently due methods have been outlined. The various for revision as a result of the new EU considerations that have been stressed in the GMO debate have been assessed in Directive (2001/18/EC) on the release of relation to DNA vaccines and gene therapy GMOs. In conjunction with such a revision, on animals, and the major consequences the authorities wish to defi ne precisely how and challenges entailed by the various DNA vaccines and gene therapy on animals regulatory alternatives have been set out. are to be regulated. Through the work on this discussion paper, members of the Biotechnology Advisory The Ministry of the Environment asked Board and the secretariat, specialists and for the assistance of the Biotechnology others have made valuable contributions Advisory Board in these endeavours. The and comments. The Biotechnology Advisory present paper has been drawn up as an Board’s recommendations for a regulatory underlying document for discussions on how system have been set out in a separate DNA vaccines and gene therapy on animals document (see enclosure).

Photo: Tom A. Kolstad, Aftenposten / Scanpix

7 The Norwegian Biotechnology Advisory Board

2. Introduction

Throughout the ages, humans have of animals raises a question of values that infl uenced the species and ecosystems of engages many people. Staking out the their surroundings. Modern agriculture borderline between what might constitute is the result of millennia of selection and the medical treatment of an animal and deliberate breeding. The last decade has what might entail the genetic modifi cation of seen the development of mutants and new that animal will have serious consequences varieties by means of radiation and chemical for researchers and manufacturers. Few treatment. However, with the advent of people would like to see genetically modifi ed gene technology, the power of humans over salmon in our shops today. If DNA-vaccinated nature has signifi cantly increased. We are animals are defi ned as genetically modifi ed, now capable of deliberately changing the this would, in practice, prevent the use of DNA heritable material of animals and plants, vaccines on animals, even if such vaccines combining genes from entirely different might be better than today’s vaccines for species and thereby infl uencing evolution consumers, the animals, the environment and development to a greater degree and the manufacturer. On the other hand, and at a swifter pace than ever before. if we opt for a liberal regulation of DNA The consequences may be unpredictable vaccines and gene therapy, the intentions of and unmanageable. Hence, as far back the Gene Technology Act might conceivably as 1975, the researchers who developed be circumvented, for example, by means of gene technology also drew up guidelines gene therapy with genes that might give for the use of this technology. Legislation the animal new traits and that might also subsequently developed is based on these become heritable. guidelines. New gene technology applications give rise The Norwegian Gene Technology Act of to new grey areas that the 1993 defi nition of 1993 covers living organisms whose genetic GMOs has failed to take into account. DNA composition has been altered by means vaccination and gene therapy may be able to of gene or cell technology. The deliberate prevent and treat diseases in animals and release of GMOs requires an authorisation humans for which there is no treatment that is based on an impact study taking today. But the concepts of vaccine and into account the environment, health, therapy can be stretched. Vaccine trials have benefi t to society, ethics and sustainable been conducted producing immune responses development. Humans are exempted aimed at eliminating the boar taint in pigs from the Gene Technology Act and are and giving ewes more lambs (1). In trials with never classifi ed as “genetically modifi ed”. gene therapy on fi sh, the addition of a growth The use of gene technology on humans is hormone gene results in signifi cantly faster instead regulated by the Act relating to the growth than in untreated fi sh. The same application of biotechnology in medicine (the effect could be imagined without the use of Biotechnology Act). This Act prohibits any DNA, as with the direct administration of heritable alteration of humans, and the use hormones. But the picture for DNA vaccines of gene therapy is restricted to the treatment and gene therapy is further complicated by of somatic cells in persons who are seriously the fact that there is the possibility that the ill and then only subject to special approval, genetic material added may move around after the Biotechnology Advisory Board and inside the animal as well as outside in the other bodies have had the opportunity of surrounding environment. Furthermore, considering the case at hand. there is also the slight possibility that it might become integrated into the hereditary At present, the deliberate release of GMOs material of the reproductive cells, causing is a topic of intense debate and subject to the alteration to be passed to the offspring. extensive regulation. The genetic modifi cation The problem becomes all the more diffi cult

8 Regulation of DNA vaccines and gene therapy on animals

when merely classifying the treated animal one might then be more concerned about as genetically modifi ed, irrespective of added, free DNA molecules than about any whether it has been approved for deliberate alterations in the animal’s chromosomes. If release or not, complicates the marketing economics and trade are deemed the most of the product. Caution must, therefore, important, it might be appropriate to focus be exercised when manoeuvring in such a on applying the same regulatory system as minefi eld, and the regulation of DNA-treated our most important trading partners. In animals must be thoroughly assessed and addition to all these considerations, it is vital well founded – for environmental, ethical as that any regulatory method is biologically well as socio-economic reasons. well-founded and possible to implement legislatively and in practice. A variety of arguments have been put forward in the debate on GMO releases in defence of Since both practical aspects and matters a restrictive approach. Some place emphasis of principle must be considered, there are on safe food, others on preventing threats to many ways of looking at the regulation of ecosystems, whereas yet others are concerned DNA vaccines and gene therapy on animals. about the power balance between developing Biologically speaking, one could look at how countries and large multinational companies. the genetic material enters, what becomes DNA vaccination and gene therapy of of it, its function, the techniques applied or animals is an area where the benefi t to what the purpose of the treatment is. In the society may be high and where a balancing of eyes of public authorities, it might be useful to different considerations becomes apparent. look at the laws that must be complied with, Considerations that are given the greatest which mechanisms that are available and emphasis will underpin the preferred what is achievable in practical terms. From defi nition of GMOs. If emphasis is placed on preserving the species, the borderline might a societal perspective, it might be expedient be determined by whether the alteration is to consider what the consequences of the heritable or not. If this is a vital matter of different regulatory alternatives might be principle, signifi cant importance might also and what is useful and ethically justifi able. be attached to the small possibility that exists of a naked DNA being spread throughout the In this document, the biological and body and integrated into the chromosomes legislative framework of the problem have of reproductive cells. If the animal’s intrinsic been described fi rst - in Chapters 3 and value is what counts, any alterations that 4 respectively - followed by a review of the introduced gene entails will be of crucial possible regulatory methods in Chapter 5. importance, whether it be heritable or not. Important considerations that should be If the spread of genetic material to other assessed and how these are addressed by the organisms is considered the most important different regulatory methods are described aspect, the deciding factor might then be in Chapter 6. Chapter 7 provides a summary whether a foreign DNA is still present of the different regulatory methods, with when the animal dies/is eaten, and perhaps the consequences and challenges involved.

9 The Norwegian Biotechnology Advisory Board

3. Biology: DNA vaccines and gene therapy

3.1 Defi nitions heritable genetic modifi cation. The objective, however, varies, and this is refl ected in The defi nitions of DNA vaccines and gene the choice of elements built into the new therapy vary in literature. Often, gene genetic material, where and how the genetic therapy also encompasses the use of DNA material is added and what is focussed on vaccines. The Norwegian Biotechnology Act, when results are measured. which regulates gene therapy on humans, applies this type of broad defi nition of gene 3.2 DNA vaccines therapy. When discussing DNA vaccination and gene therapy on animals, it might be For some time, it was thought that naked useful, however, to distinguish vaccine DNA outside living cells would rapidly be purposes from other purposes. It might degraded. But surprisingly enough, it has also be appropriate to use a specifi c term been seen that cells in the body are capable of to describe the transfer of genetic material taking up naked DNA and expressing genes not involving the use of genetically modifi ed encoded by this DNA (2). This discovery has microorganisms. In this document, therefore, encouraged the development of vaccines the terms have the following meanings: based on naked DNA.

DNA vaccine: The intentional transfer of genetic material Properties of DNA vaccines (DNA or RNA) to somatic cells for the DNA vaccines consist of a DNA molecule, purpose of infl uencing the immune system. generally a circular plasmid, with a gene that codes for the protein against which an immune response is desired. The Gene therapy: protein-coding entity is surrounded by The intentional transfer of genetic material regulatory sequences that ensure good to somatic cells for purposes other than protein production. Plasmids will also have infl uencing the immune system. a sequence allowing replication in bacteria and often a gene for a selectable marker. The DNA treatment: future may see the replacement of antibiotic The intentional transfer of genetic material resistance genes by other less controversial to somatic cells, either in the form of DNA selectable marker genes. DNA vaccines may vaccines or gene therapy, in ways that do be administered by a variety of methods. not entail the use of genetically modifi ed In present day trials, injection is the most microorganisms. effi cient method, but other means of delivery are in the process of being developed based In gene therapy as with DNA vaccines, on, for example, immersion, spraying, gene genetic material is transferred to some of gun and electroporation. the animal’s cells for the purpose of carrying out a specifi c function. This function may DNA vaccines possess new, promising be fulfi lled by the cells producing proteins properties compared to earlier vaccines. from the added genes, or by the added They are capable of providing a broad, long- genetic material directly infl uencing genetic lasting immune response, they do not require material or other molecules in the cell. complete knowledge of the pathogenic Genetic material may be transferred either organism, they are relatively simple, cheap as naked DNA, as encapsulated DNA or via and quick to produce and they are stable genetically modifi ed viruses, bacteria etc. at room temperature and therefore easy to transport and store. To achieve a good In general, the same molecular tools are vaccinating effect, the DNA vaccine should used in DNA vaccination, gene therapy and be present and active for only a short period

10 Regulation of DNA vaccines and gene therapy on animals

of time. During this period, a large amount uncharted in detail, and the microorganisms of protein will be formed, giving rise to an are classifi ed as not genetically modifi ed. The immune response. genetic alterations involved may, however, be fairly extensive. Recently, a comparison was made of the heritable material of the Vaccine principles BCG-vaccine with that of the tuberculosis DNA vaccines are a type of subunit vaccine, bacterium, and an area of nine genes was in the sense that they produce only a identifi ed as missing in the vaccine (3). selected protein from a microorganism. In With the advent of gene technology it is other forms of subunit vaccines, the protein now possible to deliberately alter or remove may be purifi ed from the microorganism pathogenic elements in the heritable material itself, or it may be produced by means of of microorganisms. This method is much gene technology by cloning the gene for used for viruses in particular. Genetically the subunit from the heritable material of modifi ed viruses may be used as vaccines. the pathogenic microorganism. Selecting These are termed homologous viral vaccines only one component of a microorganism when the pathogenic virus itself has been as a vaccine, involves certain limitations. modifi ed. Genetically modifi ed viruses may, The immune response generated by DNA however, also be used as vectors to produce an vaccines is limited to targeting proteins and immune reaction against proteins from other not other types of complex molecules on the microorganisms. In this instance, the gene surface of the microorganism. On the other for the desired protein is introduced into the hand, there is little risk that the vaccine itself heritable material of the vector virus. These will cause disease, as it does not contain the are then called heterologous viral vaccines or other components of the microorganism. viral vectors.

Traditionally, vaccines have been developed in two different ways: either How vaccines work by using dead microorganisms or with Live vaccines will often provide better the help of attenuated (weakened), living protection, as they activate a broader part of microorganisms. A vaccine consisting of the immune system than dead or inactivated dead or inactivated microorganisms will microorganisms do. Although DNA vaccines normally generate an immune reaction are not living, they are more closely related against structures on the surface of to genetically modifi ed viral vaccines than the microorganism, which in turn will their name indicates. This is an important provide protection against later infections. aspect to be borne in mind when considering Alternatively, vaccines can be based on how such vaccines should be regulated. variants of living bacteria and viruses that DNA vaccines and viral vaccines both are less pathogenic than those against possess certain traits of an ordinary viral which protection is required. Variants can be infection in the way in which they work. closely related pathological organisms that They provide better and more long-lasting prefer other host animals (e.g. cow pox) or protection than vaccines consisting of the pathogenic organism may be attenuated inactivated microorganisms, since DNA through breeding and selection. For many vaccines and viruses both result in genetic decades, microorganism attenuation has material entering into the cells, thereby been used to develop new vaccines in the triggering the new production of proteins. form of mutant strains. One example is the Such proteins retain their natural form and BCG-vaccine against tuberculosis. There is, are not deformed, as is often the case when however, a certain risk that living vaccines microorganisms are inactivated. This leads may revert to their pathogenic variant. to the activation of B cells and an antibody Today’s incidences of polio in the western response (“humoral immune response”) that world are caused by reversion of a living, more readily recognizes naturally existing attenuated vaccine. microorganisms. Moreover, DNA vaccines and viral vaccines activate another branch Genetic alterations in deliberately of the immune system, i.e. the T cells, also attenuated strains are normally unknown or called the “cellular immune response”.

11 The Norwegian Biotechnology Advisory Board

T cells keep track of what is going on forms of gene therapy, the aim is even to inside the cells of the body by means of a integrate the genetic material into the mechanism where fragments of the cell’s chromosomes. contents are transported with the major histocompatibility complex (MHC) to the Gene therapy often requires a more targeted cell surface and presented to the immune and fi nely tuned technology than is the cells. In this way, virus-infected cells can be case for DNA vaccines. In gene therapy, the detected and the infection stopped by the T genetic material must reach the right cells, cells killing off the infected cells. as well as generate the desired degree of activity. For this reason, the development of Similarities between DNA vaccines and gene therapy has not advanced as far as the viruses development of DNA vaccines. Although viruses are, in many instances, called microorganisms, their structure is Gene therapy may aim at different objectives. exceedingly simple and they are dependent One objective might be to add genes coding on the machinery of host cells to reproduce. for desired proteins, which can either Generally, they only consist of heritable contribute to a new function in the cell or material encapsulated by proteins and in replace proteins that are no longer effective some cases a membrane. In the heritable due to harmful mutations. It is also possible material, many viruses have genes for to add genetic material that specifi cally enzymes that are necessary for the affects the expression of a protein. In this way, multiplication of the virus. With certain viral genes that code for undesirable or harmful infections, therefore, the virus’ heritable products may be neutralized by inhibiting material alone is suffi cient to produce new or preventing the production of the protein. viral particles. Hence, this type of viral When adding small DNA or RNA molecules infection can also be formed by the cell that correspond to the desired gene and taking up naked viral heritable material. therefore attach themselves to the gene as A DNA vaccine consisting of complete antisense, the gene’s transcription may be heritable material from such viruses could, directly inhibited or the gene’s mRNA may therefore, trigger a new viral infection. be prevented from serving as a template for Recently, researchers have succeeded in protein production. Some RNA molecules manufacturing infectious poliovirus by (ribozymes and siRNA) may even be designed means of chemically synthesing the genetic so that the gene’s mRNA molecules are material (4). This has subsequently raised specifi cally fragmented. the question of when naked genetic material is to be classifi ed as an organism. Inversely, In order to introduce new genetic material it is conceivable that DNA vaccines could be into cells, a series of vector systems have been given a viral form by encapsulating the DNA developed. As for vaccines, the new genes may in a viral particle. Since there is the chance be delivered with plasmids or viral vectors. of an indistinct borderline between virus Small DNA molecules (oligonucleotides) may and DNA, it is important, when considering be introduced in their existing form, or they appropriate regulatory mechanisms, that may be wrapped into appropriate molecular DNA vaccines and genetically modifi ed viral packages, e.g. liposomes or other vesicles. vaccines are viewed in the same context. RNA molecules as well may be delivered directly, but they may also be introduced in 3.3 Gene therapy the form of a gene that will be transcribed to RNA inside the cell. Contrary to DNA vaccination, which aims to produce large amounts of protein in a short The challenges facing gene therapy today span of time so as to generate an immune reside in the need to achieve good biological response, gene therapy is often aimed at and therapeutic effects. To do so, the genetic achieving a long-lasting, physiologically material must be able to reach the right cells, matched expression of the gene, without they must remain there and be precisely as activating the immune system. In certain active as required. Systems for the delivery

12 Regulation of DNA vaccines and gene therapy on animals

and targeting of genetic material are being and long-term expression of the genetic continuously developed and, so far, there are material. few good strategies for gene therapy. If retention of the genetic material in 3.4 Stability of the added genetic the cells is desired, elements allowing material in the animal replication in the host cell may be built into the genetic construct. Most plasmids contain an element enabling replication in bacteria, NA vaccines are distinct from gene D but this element does not normally work in therapy in that a short-term presence of animal cells. Many viral vectors are capable DNA in the animal is the desired result, of amplifi cation in animal cells. Some will whereas gene therapy often aims at ensuring cause an infection, which will transfer the the presence of the added genetic material added genetic material into nearby cells, as over a longer period of time. well as into the surroundings in the form of viral particles. The most common method When DNA vaccines have been added to the for achieving long-term presence in the body, either by intramuscular injection or cell is, however, to use a mechanism that other means, most of the added DNA will integrates the genetic material into the cell’s normally be degraded in a short space of time, chromosomes. while a portion will be absorbed by the cells. Some of the DNA may, however, transfer into lymph and blood and thereby spread to the 3.5 Possible integration into rest of the body. Spermatozoa are capable chromosomes of taking up foreign DNA, a phenomenon that is currently being used to develop new In gene therapy, viral vectors are occasionally methods for the genetic modifi cation of used to ensure the incorporation and lasting animals by treating spermatozoa with DNA presence of genetic material in the cell. One (5) and by injecting DNA into animal testes viral vector used (adeno-associated virus) (6). even possesses a mechanism enabling its relatively precise integration into a specifi c In gene therapy, genetic material is often area of the human chromosome 19. However, meant to reach a specifi c type of cell in it has been seen that it is also capable of the body and it may need to move to its integrating into other areas (7). destination via the blood. Work is currently ongoing to develop specifi c molecular tags There is, however, always a slight probability so as to ensure that the DNA is taken up that added DNA will be integrated into the only by the target cells. Until this problem chromosomes of the cell, independently of has been resolved, methods are being used whether such integration is intended or not. where target cells are more easily accessible. The probability may vary according to the One such example is the trial gene therapy type of tissue, means of delivery and amount in cancer treatment, where immune cells are of DNA introduced. The traditional method extracted from the body, given gene therapy, for producing heritably genetically modifi ed selected for DNA uptake and thereafter (transgenic) animals makes use of this reintroduced into the body. probability. In such instances, many plasmid copies are injected into a series of fertilized Once inside the cell, many partly unknown eggs and the DNA will be integrated into only factors will affect the DNA’s stability. a small proportion of the eggs and thereby Without elements enabling DNA replication, produce transgenic offspring. it will remain active for a certain period of time and subsequently degrade. How long It is impossible to predict the effects of this will take will depend on both DNA incidental incorporation. The added gene may sequence, DNA structure and cell type. be introduced into an area of no consequence To achieve the best effect of gene therapy, or it may end up in the middle of another work is in progress to optimise the stability gene and disrupt it. Hence, receiving gene

13 The Norwegian Biotechnology Advisory Board

therapy and DNA vaccines will involve a functions as heritable material in other certain risk of developing cancer or other contexts. Many viruses have RNA as their diseases, as a result of the effect of the genes heritable material instead of DNA. Some of at the site of integration. Cases of cancer due these (retroviruses) possess a gene for the to integration have recently led to the halt of enzyme reverse transcriptase that converts several gene therapy trials on humans (8). RNA into DNA following infection. In a Such a possibility will exist, irrespective of subsequent phase, the virus may ensure that the type of DNA added. Many integrative the DNA version of the heritable material is mechanisms and transposable elements integrated into the host cell’s chromosomes. are based on relatively short sequences, so However, retroviruses require quite specifi c the use of short DNA molecules provides sequences for conversion from RNA to DNA, no guarantee against integration. The as well as for integration. The likelihood of likelihood of integration may be reduced by RNA vaccines being integrated while a cell avoiding elements that are known to promote is simultaneously infected with a retrovirus integration and by conducting further is, therefore, exceedingly small, as long as research on the stability and properties of integrative sequences are avoided. genetic materials in the target organism. 3.6 New properties of DNA- If the added genetic material circulates in the treated animals body, is taken up by reproductive cells and also is integrated into the chromosomes there, Even if DNA vaccination and gene therapy the DNA-treated animal could theoretically results in DNA being taken up by only a produce heritably genetically modifi ed few cells, it might still be enough to give the offspring. Gene therapy could conceivably animal new physiological properties, in the allow the administration of DNA capable of same way as for genetically modifi ed animals. translocating to the reproductive cells and the One such example is the introduction of a subsequent selection of offspring possessing growth hormone gene into fi sh, where the properties attributable to integration and cells taking up the gene secrete hormones heritable genetic modifi cation. Furthermore, that affect the entire animal, producing such a method might possibly be resorted fi sh that grow signifi cantly faster. A similar to if someone wished to misuse a regulatory effect could also be achieved without the system that is more liberal where gene use of DNA, by giving the fi sh hormone therapy is concerned, compared to heritable injections. The difference is that the use of genetic modifi cation. DNA moves the hormone production into the animal itself. Using RNA instead of DNA might possibly avoid the problem of unwanted integration The concept of vaccine may, moreover, be into chromosomes. In the cell, mRNA extended to have effects above and beyond serves as the link between genes and the that of disease prevention. Immune reactions machinery for protein production. By may be used to alter the hormone balance of adding RNA that functions as mRNA, it animals and change the animal’s natural would be possible to enter at the post-DNA characteristics. But the same effects could step of the information ladder and still also be achieved by means of traditional achieve the required protein production. vaccine methods and without the use of After DNA vaccines, RNA vaccines appear DNA. to be the next step in the fi eld of vaccine development. RNA is already in use in the Some of the novel properties that could fi eld of gene therapy. One disadvantage of be introduced by means of DNA vaccines using RNA is that it is more complicated and gene therapy might conceivably lead to produce and less stable than DNA. RNA to a selective advantage in the animal’s molecules for use in gene therapy are ecosystem, if such properties are inherited. now being chemically modifi ed to improve The fact that there is a slight possibility their stability. Although RNA is not the that the added DNA could be integrated heritable material in the animal cell, it still and passed on to the next generation could,

14 Regulation of DNA vaccines and gene therapy on animals

therefore, play a role when choosing the type be the same as those of heritable genetic of genes that should be permitted for the DNA modifi cation, even though their likely treatment of animals. The consequences of occurrence is signifi cantly smaller. With DNA treatment using genes that introduce repeated releases, non-heritable properties a selective advantage for the animal could could also have an ecological effect.

Photo: Stuart McEvoy, EPA / Scanpix

15 The Norwegian Biotechnology Advisory Board

4. Current regulatory framework – national and international

4.1 GMO and DNA under When naked recombinant DNA is used as a medicinal product, it is governed by the current Norwegian legislation Medicinal Products Act. When defi ning more precisely the regulation of DNA vaccines and gene The Gene Technology Act is restricted therapy on animals, it is important to have to genetic modifi cations using gene and a clear picture of what the Gene Technology cell technology. Radiation and chemical Act provides for in these areas today. But the mutagens have long been used to produce Gene Technology Act is not the only piece of new, improved strains of species, in research legislation dealing with genetically modifi ed as well as in production, and this has given organisms. rise to heritable alterations in the form of genetic mutations. Some of these mutations present altered properties – in a positive Choices made in current legislation or negative way – whereas the majority of The Act relating to the production and such mutations are not apparent. The use use of genetically modifi ed organisms of mutagens or radiation is not regarded as genetic modifi cation, even though the (Gene Technology Act) regulates living, purpose, in these instances as well, has genetically modifi ed organisms in Norway. been to achieve alterations in heritable In this context, GMOs are defi ned as ”micro- properties. organisms, plants and animals in which the genetic material has been altered by means of gene or cell technology”. Human beings Below follows a review of other legislation are not covered by the Gene Technology Act. that is particularly relevant to the regulation of DNA vaccines and gene therapy on animals. The Gene Technology Act itself is As long as a GMO is alive, the Gene dealt with in sub-chapter 4.2. Technology Act applies. For dead GMOs or for products produced from GMOs, other legislation applies, depending on Food Act the purpose involved. Once a dead GMO When mentioning the regulation of becomes food, it is regulated by the Food Act. genetically modifi ed foodstuffs, the Food If used for animal feed, the Animal Feed Act Act refers to the Gene Technology Act’s applies. Medical and veterinary medicinal defi nition of a GMO. It seems logical, products manufactured with the help of gene therefore, that the administration of the technology are subject to the Norwegian Food Act should comply with the regulatory Medicinal Products Act. Furthermore, the system applying to DNA-treated animals Animal Welfare Act restricts the type of as stipulated in the Gene Technology Act. genetic modifi cations that may be performed In consequence, an interpretation of DNA- on animals. treated animals as GMOs would require the approval and labelling of products from such Naked recombinant DNA is regulated by the animals in Norway. When importing similar Gene Technology Act solely when the genetic foodstuffs from abroad, these too would material is included in the production in principle have to be labelled as GMOs, of a GMO. Furthermore, the health and but, in that case, only after the product environmental risks of spreading genetic has been approved by the Norwegian Food material from the dead organism must be Control Authority (SNT). Such approval considered when applying for the deliberate may, however, be granted without the release of a living GMO. The Animal Feed same comprehensive impact study that is Act has introduced a ban on the presence of required for the deliberate release of living antibiotic resistance genes in animal feed. GMOs. When applying for approval, health

16 Regulation of DNA vaccines and gene therapy on animals

effects will be examined, in addition to other Animal Health Act societal considerations. Section 9 of the Animal Health Act states that ”vaccines for the vaccination of animals The Norwegian Food Control Authority also (livestock, game) may be used only when administers the Regulation on maximum the vaccines and vaccinations have been residue limits for veterinary medicinal approved by the Ministry. The Ministry products in foodstuffs of animal origin, which may issue regulations on the use of vaccines includes “provisions on the determination for the vaccination of animals (livestock, of maximum residue limits for veterinary game).” All DNA vaccines for livestock and meidicinal products in all foodstuffs of game animals are, therefore, subject to animal origin, including meat, fi sh, milk, the approval of the Norwegian Medicines eggs and honey. The Regulation does not Agency and the Norwegian Animal Health apply to active substances of biological origin Authority before use. that are intended to produce active or passive immunity or to diagnose a state of immunity, and that are used in immunological Animal Welfare Act veterinary medicinal products”. Here, an The Animal Welfare Act was amended when exemption has been made for vaccines, and the Gene Technology Act entered into force. hence the Regulation applies to gene therapy Section 5 of the Act provides for restrictions products, but not to DNA vaccines. on the genetic modifi cation of animals for breeding purposes: “It is prohibited to alter the heritable material Animal Feed Act of animals by means of gene technology Genetically modifi ed animal feed products methods or by means of traditional breeding must be labelled for sales purposes, as well methods if: as when they are ingredients of feed mixes. 1. it makes the animal unfi t for carrying Here too, it would be logical to rely on the out normal behaviour or if it adversely defi nitions set out in the Gene Technology affects its physiological functions; Act. Furthermore, it has been established 2. the animal is caused unnecessary that the use of genetically modifi ed feed in suffering; production does not require the labelling of 3. the alterations arouse general ethical fi sh or meat products, as long as the genetic concerns. composition of the end product remains It is prohibited to breed animals covered by unchanged (9). the fi rst paragraph.”

Biotechnology Act The animal’s natural characteristics are Gene therapy on humans is regulated thereby safeguarded by the Act as far as by the Act relating to the application of heritable alterations are concerned. The biotechnology in medicine. The conditions same does not apply, however, when altering thereof are set out in Section 7-1: the individual animal without affecting “The human heritable material may only be its genome. However, if the DNA in the altered by means of somatic gene therapy treatment should prove to be heritable and the alteration affected the animal’s natural for the purpose of treating serious disease or characteristics, the fi nal paragraph of this preventing serious disease from occurring. Section would then prohibit any further Germline gene therapy is prohibited.” breeding of the animal. Although humans who have undergone gene therapy treatment cannot be classifi ed as genetically modifi ed organisms, The use of animals for research purposes is environmental, as well as other concerns will regulated under Section 21 of the Act, Use of be assessed for every individual application animals for research purposes: for gene therapy. The defi nition of gene “It is prohibited to perform biological therapy set out in the Biotechnology Act also experiments on animals without specifi c covers any use of DNA vaccines on humans. authorisation. Such authorisation may be

17 The Norwegian Biotechnology Advisory Board

granted if the purpose is to determine the Most DNA vaccines would be defi ned as type of disease that animals or humans medicinal products, since they prevent are suffering from, or if the purpose is to disease in animals. Also gene therapy, prevent or eradicate disease. Authorisation which is not disease-related, but which may also be granted when the purpose is affects physiological functions, would fall related to research, manufacture or testing of within the defi nition of a medicinal product. medicines, medications, preparations, toxins, Furthermore, the wording allows the etc. for use on humans, animals or plants. Norwegian Medicines Agency to determine Such trials must be conducted in such a way which products are to be classifi ed as medicinal as to prevent any risk of the animal suffering products and which are not. Section 4-6 of the more than is strictly necessary for the purpose Medicinal Products Regulation stipulates in question”. More detailed guidelines and that any application for authorisation to authorisation procedures are available in the place on the market a medicinal product for Animal Welfare Act’s Regulation on animal animals should include “an assessment of the trials. Here, however, an exemption has possible risk to the environment and to the been made for “treatment and interventions health of humans and animals with normal conducted in clinical veterinary activities use of the medicinal product, as well as all in accordance with recognized methods and documentation on which said assessment trials relating to breeding/farming, feeding has been based”. Medicinal products based and the environment (livestock and aquatic organisms), unless there is reason to believe on gene technology are granted market that the trials will result in an abnormal authorisation by way of a common European physiological condition for the experimental application procedure. animal.” Gene therapy trials causing such conditions for the animal require, therefore, The Norwegian Medicines Agency is also permission from the Experimental Animals responsible for assessing the clinical testing Committee, whereas trials with vaccines of medicinal products on animals. Specifi c according to recognized methods require no guidelines have, for example, been drawn such permission under the Animal Welfare up for the clinical testing of vaccines on fi sh. Act. On the other hand, animal trials involved in the development of medicinal products for Medicinal Products Act humans are not subject to prior assessment by the Norwegian Medicines Agency. Section 2 of the Medicinal Products Act defi nes what is considered to be a medicinal product: “For the purpose of the present Act, medicinal 4.2 Defi nitions and intentions of products are defi ned as substances, drugs the Gene Technology Act and preparations that are intended for or presented for use to prevent, heal, cure Today, the defi nition of a GMO under or alleviate disease, disease symptoms or the Gene Technology Act is as follows: pain, to infl uence physiological functions “Microorganisms, plants and animals in in human beings or animals, or, through which the genetic material has been altered internal or external use, to detect disease. The by means of gene or cell technology.” King may issue more detailed regulations The individual concepts are further defi ned defi ning what is to be considered a medicinal as follows: product. Such regulations may stipulate that “Microorganisms: any cellular or non- certain substances, drugs or preparations are cellular microbiological entity that is able to always to be considered medicinal products, reproduce or transfer genetic material” regardless of whether they might have other applications, and that certain other “Gene technology: techniques that involve substances, drugs or preparations, falling heritable material being isolated, charac- within the scope of the provision set out in the terized, modifi ed, and introduced into living fi rst paragraph above, may still be considered cells or viruses” to be non-medicinal products.” “Cell technology: techniques for the production

18 Regulation of DNA vaccines and gene therapy on animals

of living cells with new combinations of What is meant by the term “genetic material” genetic material by the fusion of two or more is not further developed in the Proposition to cells.” the Odelsting. It is not entirely clear whether this applies to all addition of new genetic material or only when the added genetic Expanding on the defi nitions material is heritable. Proposition No. 8 to the Odelsting* (1992-93), in which the draft of the Gene Technology However, reference is made to examples that Act was submitted, provides further details fall outside the scope of the Act (page 68 of on how legislators have worked their way to Proposition No. 8): a defi nition of GMOs. This might shed some This means that work with genetic material light on the possible interpretations of how that has been extracted from an organism DNA-treated animals are to be regulated. or is synthesized, as well as biochemical research on polynucleotides, fall outside the Could naked genetic material per se be scope of the Act when the direct aim is not to regarded as a GMO? introduce heritable material into a living cell One possible interpretation of the defi nition or virus.” of microorganisms might be that naked DNA, when self-replicating, is a “non- This statement could indicate that it cellular, microbiological entity that is able to would suffi ce that the genetic material was reproduce”, and consequently might per se be introduced and that there is no requirement defi ned as a GMO. However, the preparatory as to the material being inherited. work provides no support for such an interpretation. Here, endorsement was Moreover, the meaning of the concept of instead given to the then EC’s interpretation “genetically modifi ed organism” is further of what the concept of “non-cellular expanded (page 70): microbiological entity” meant (Proposition “A genetically modifi ed animal or a so-called No. 8 to the Odelsting, page 70): “transgenic animal” also refers to the progeny “An interpretation was reached which of a crossing of a transgenic animal with meant that the defi nition covered viruses, a non-modifi ed animal, as well as animals including bacteriophages, but not plasmids with transplanted transgenic tissue.” or other naked genetic material. The Ministry endorses this delimitation. Hence, Transplanted transgenic tissue would not the defi nition of microorganisms includes normally lead to a heritable alteration, viruses, bacteria, unicellular plants and which might indicate that the idea was animals, plant and animal cells (including that genetic modifi cation did not have to be human cells) in culture and microscopic heritable to be covered by the current Gene yeast and mould fungi.” Technology Act.

In consequence, naked genetic material per se falls outside today’s defi nition of a Is the method used for adding naked GMO. (Entire viral genomes are defi ned as genetic material important? viruses.) What the Act’s defi nition of “gene technology” (see above) actually implies may be somewhat unclear. The defi nition could be Would an organism to which naked read to mean that genes are either isolated, genetic material has been added be or characterized, or modifi ed, or introduced classifi ed as a GMO? into living cells or viruses. Alternatively, The Gene Technology Act applies to living the defi nition of gene technology could organisms whose genetic material has been mean that genes are isolated, as well as altered by means of gene or cell technology. characterized, as well as modifi ed, as well as introduced into living cells or viruses. These *Note: Odelsting: one of two divisions of the different meanings could have an impact on Norwegian Parliament which vaccine technologies might today be

19 The Norwegian Biotechnology Advisory Board

considered as gene technology. DNA vaccines and gene therapy on animals should be defi ned as genetic modifi cation. It On page 70 of the Proposition to the is possible, by relying on various sentences in Odelsting, there is further emphasis on the wording of the Act and of the Proposition the importance that the uptake of genes to the Odelsting, to fi nd support for the has for the defi nition of “gene technology”: interpretation that DNA-treated animals are “One absolute condition is that the aim is the GMOs and as well to interpret the wording uptake of heritable material in a living cell or to mean that they are not to be considered in a virus. There is no condition that the gene as GMOs. The picture might possibly be must be isolated, as well as characterized, clarifi ed by examining the intentions behind as well as modifi ed. One of these work the Gene Technology Act. operations, combined with the introduction of the heritable material into a living cell or Intentions of the Gene Technology virus (see above), would suffi ce.” Act

However, it is still not entirely clear what In the context of a regulatory system is meant by the terms “introduction” for GMOs, many considerations have and “uptake” and whether this entails been underscored. These are refl ected in incorporation into chromosomes or not. the recommendation that the Standing Committee on Local Government and the Environment of the Norwegian Storting In gene therapy, there are several possible (parliament) tabled in May 1991: delivery methods for the genetic material. “When deliberately releasing genetically When considering how relevant the method modifi ed organisms into nature, the majority of introduction is, it is important to bear of the Committee, like the Government, in mind that the Act’s technical area of believes that the regulatory framework application covers production, in addition to should be restrictive […] A prerequisite use: “The Act applies to the production and for permission for deliberate release must use of genetically modifi ed organisms. The be that there is no danger of unwanted provisions of the Act relating to genetically modifi ed organisms also apply to substances ecological or health effects and of the and products that consist of or contain unwanted spread of the organism or its genetically modifi ed organisms.” genetic material. The majority would also stress that permission must be contingent on the utility value involved and the ethical, This is further expanded on page 68 of the health and ecological issues that deliberate Proposition to the Odelsting: release raises following controlled trials and “The term “production” means all the steps impact and risk assessments.” in a process that specifi cally lead to an organism being genetically modifi ed”. In the interests of the environment, any detrimental spread of either GMOs or their Hence, there is no requirement as to how genetic material is unwanted. An important the genetic material is introduced. For factor to be borne in mind when considering substances or products consisting of or DNA vaccines and gene therapy is, therefore, containing genetically modifi ed organisms, how the added genetic material is treated by the requirement is that the GMOs must be the body and any possible environmental living: “The Act does not apply to products effects the naked genetic material might that are produced by means of genetically have when liberated from the animal. The modifi ed organisms when the end product intention of the Act provides no clarifi cation does not contain living organisms.” on the status awarded to the treated animal. It might still be considered either as a GMO This review shows that even when the per se, or it may be regarded and evaluated wording of the Act is examined in the light of as a part of the environment into which Proposition No. 8 to the Odelsting (1992-93), the genetic material has been deliberately it is still not clear whether or not the use of released.

20 Regulation of DNA vaccines and gene therapy on animals

4.3 New EU Directive on the for genetic modifi cation are mentioned: deliberate release of GMOs Annex 1A, Part 1: Techniques of genetic modifi cation referred to in Article 2(2)(a) are inter alia: The EU Parliament has approved a new directive (2001/18/EC) for the deliberate 1) recombinant nucleic acid techniques release of genetically modifi ed organisms. involving the formation of new combinations The Directive states that the deliberate of genetic material by the insertion of nucleic release of GMOs must be based on the acid molecules produced by whatever means precautionary principle. DNA vaccines and outside an organism, into any virus, bacterial gene therapy are not specifi cally mentioned plasmid or other vector system and their in the new Directive, but its defi nitions incorporation into a host organism in which do provide indications of how the EU will they do not naturally occur but in which they regulate DNA treatment and DNA-treated are capable of continued propagation; animals. 2) techniques involving the direct introduction into an organism of heritable material prepared outside the organism Defi nition of a GMO in EU Directive including micro-injection, macro-injection 2001/18/EC and micro-encapsulation; In the English version of the Directive, 3) […] organisms and GMOs are defi ned as follows: It is conceivable that DNA treatment could 1) “organism” means any biological entity be covered by item 1 in those instances where capable of replication or of transferring the inserted genetic material could propagate genetic material; in the organism. In that case, it would apply 2) ”genetically modifi ed organism (GMO)” to a greater degree to gene therapy than to means an organism, with the exception of DNA vaccines. However, there is still room human beings, in which the genetic material for other interpretations based on what the has been altered in a way that does not term ”propagation” entails. occur naturally by mating and/or natural recombination; DNA treatment appears, moreover, to fall Within the terms of this defi nition: under item 2, which mentions the direct introduction of heritable material into an (a) genetic modifi cation occurs at least organism. However, it is not clear whether through the use of the techniques listed in ”heritable material”, in this context, means Annex 1A, Part 1; nucleic acids in general or whether it means (b) […] that the material indeed must be heritable. Under item 1 of the Annex, the expressions The defi nition of organism is very similar ”nucleic acid molecules” and ”genetic to the one used in the Norwegian Gene material” are used instead of ”heritable Technology Act and it is not specifi ed material”. The same distinction is also to whether replicating plasmids are covered by be found in the Danish and French texts. the defi nition. In the preparatory work of the This could indicate that the EU’s defi nition Norwegian Act, the then EC defi nition was of genetic modifi cation does not include the used as a basis when excluding plasmids direct introduction of nucleic acid molecules from the defi nition of an organism. During that are not heritable, but this is not the negotiation of the Cartagena Protocol, explicitly stated. An alternative explanation the EU Commission stated that it did not might be that the term is also meant to cover consider plasmids and other naked genetic other types of molecules that are capable of material to be organisms. transferring properties, such as prions.

However, treating an animal with naked Hence, the new EU Directive does not appear DNA could conceivably lead to the animal to be entirely unambiguous as to whether becoming a GMO. Attached to the defi nition DNA-treated animals in general are to be of a GMO is an Annex in which techniques considered as genetically modifi ed. Here,

21 The Norwegian Biotechnology Advisory Board

there is room for discretion and different should apply to both deliberate release and interpretations. product approval, the Directive with its related Regulations could be interpreted to There are few international statements mean that the treatment of animals with that explicitly take a position on the genetically modifi ed medicinal products not status to be given to DNA-treated animals. even makes the animals genetically modifi ed However, the Agriculture and Environment while alive, even though the genetically Biotechnology Commission (AEBC) of Great modifi ed product is present in the animal for Britain published the report ”Animals a certain period of time. and Biotechnology” on 3 September 2002. On page 13, DNA-vaccinated animals are A further exception has been made for mentioned: ”Importantly, the foreign DNA genetically modifi ed medicinal products, is not expected to integrate into the host’s and this applies to the product itself. In genome and so the vaccinated animal is not recital 31 of EU Directive 2001/18/EC, it is genetically modifi ed.” stated that genetically modifi ed medicinal products that are to be placed on the market Applying this criterion, the majority of DNA- are not covered by the Directive, but that vaccinated animals would not be genetically an environmental risk assessment should, modifi ed, whereas gene therapy could imply nevertheless, be carried out: genetic modifi cation. ”(31) Part C of this Directive does not apply to products covered by Council Regulation (EEC) No. 2309/93 of 22 July 1993 laying Special rules for genetically down Community procedures for the modifi ed medicinal products authorisation and supervision of medicinal In the EU, the aim is that a single application products for human and veterinary use should suffi ce in order to assess the and establishing a European Agency for the deliberate release of a GMO and to approve Evaluation of Medicinal Products, provided products produced from this GMO as food that it includes an environmental risk or animal feed. In conjunction with the assessment equivalent to that provided for by Directive, therefore, new regulations have this Directive.” been issued on the traceability and labelling (Part C: Placing on the market of GMOs as of genetically modifi ed food and feed. This or in products; Part B: Deliberate release of is formulated in Regulations 2001/0173 and GMOs for any other purpose than for placing 2001/0180. In these documents, approval and on the market.) labelling requirements are linked to the food or feed, or the ingredients of these, that have A GMO that is to be placed on the market as been produced from a GMO, but not to those a veterinary medicinal product is, therefore, that have been produced with a GMO. The not directly covered by the deliberate release criterion is whether the material from the Directive. The exemption presupposes, GMO is present in the product. It is specifi ed however, that an equivalent environmental that products from animals having received risk assessment is evaluated by the same genetically modifi ed feed and genetically body that considers other deliberate releases modifi ed medical products are not covered of GMOs. This type of environmental risk by the Regulation: assessment includes the species on which ”Thus, cheese produced with a genetically the medicinal product is to be used. As a modifi ed enzyme that does not remain in result, the animals will be covered by a the fi nal product and products obtained risk assessment, by being a part of the from animals fed with genetically modifi ed environment into which the medicinal GMO feed or treated with genetically modifi ed is to be released. medicinal products would be subject neither to the authorisation requirements, nor to The exemption set out in recital 31, entailing the labelling requirements laid down in the the requirement of an environmental risk proposed Regulation.” assessment, applies to medicinal products Since the aim is that a single application where a GMO is the product or is an

22 Regulation of DNA vaccines and gene therapy on animals

ingredient of the product. This applies, color additives, and vaccines.” therefore, to genetically modifi ed viruses for use in vaccines and gene therapy. However, The Pew Initiative on Food and Biotechnology it is unclear whether DNA treatment in the US pointed out in a recent report products are to be considered as genetically that the regulatory system in the US was modifi ed medicinal products, as no organism ambiguous as far as genetically modifi ed constitutes the product or is an ingredient of animals for food purposes were concerned, the product. and that genetically modifi ed salmon, for example, could not be regulated on the basis If products for DNA treatment are not of environmental considerations alone (10). to be considered as GMOs or genetically modifi ed medicinal products, there is no 4.4 Cartagena Protocol on requirement that they be assessed in terms of their environmental risks by the same Biosafety body that evaluates GMO releases. Such an environmental risk assessment could, When drawing up a Norwegian regulatory on the other hand, be relevant at the next system for DNA vaccines and gene therapy stage, when assessing whether the animal on animals, consideration should be given to treated with such products is to be regarded international agreements that Norway has as genetically modifi ed. signed.

The European Agency for the Evaluation The Cartagena Protocol applies to the of Medicinal Products (EMEA) has drawn biosafety of GMOs in the environment and up guidelines for medicinal product it is part of the Convention on Biological development, where DNA vaccines for Diversity. The Protocol has been ratifi ed by animals are covered by the following: Norway, the EU Commission and several -“DNA vaccines non-amplifi able in eukaryotic EU member states. It entered into force 11 cells for veterinary use” September 2003. -“Environmental risk assessment for immunological veterinary medicinal The objective of the Protocol is to take products” account of biological diversity in the context Here, there are requirements as to safety of the trade in and use of living, genetically and documentation on the spread of the DNA modifi ed organisms. It is worded as follows: in the animal, integration into chromosomes ”In accordance with the precautionary and toxicity for the animal itself and for the approach contained in Principle 15 of animal’s reproduction. the Rio Declaration on Environment and Development, the objective of this Protocol is to contribute to ensuring an adequate In the US, gene therapy and the heritable level of protection in the fi eld of the safe genetic modifi cation of animals are both transfer, handling and use of living covered by the concept of drug. The area modifi ed organisms resulting from modern is controlled by the Center for Veterinary biotechnology that may have adverse effects Medicine (CVM) under the Food and Drug on the conservation and sustainable use of Administration (FDA). On the FDA’s home biological diversity, taking also into account page, this is expanded under ”Q&A”: risks to human health, and specifi cally ”Most, but probably not all, gene-based focusing on transboundary movements.” modifi cations of animals for production or therapeutic claims fall under CVM regulation as new animal drugs.” ”The In the Cartagena Protocol, the term ”LMO”, animal drug provisions of the Federal Food, living modifi ed organism, is used for GMOs: Drug, and Cosmetic Act best fi t the transgenic ””Living modifi ed organism” means any animals that have agronomic traits now living organism that possesses a novel being investigated and developed. Other combination of genetic material obtained transgenics will no doubt come along that through the use of modern biotechnology; could be viewed as containing food additives, ”Living organism” means any biological

23 The Norwegian Biotechnology Advisory Board

entity capable of transferring or replicating medicinal products are not (Article 5): genetic material, including sterile organisms, ”Notwithstanding Article 4 and without viruses and viroids; prejudice to any right of a Party to ”Modern biotechnology” means the subject all living modifi ed organisms to application of: risk assessment prior to the making of a. In vitro nucleic acid techniques, decisions on import, this Protocol shall including recombinant deoxyribonucleic not apply to the transboundary movement acid (DNA) and direct injection of nucleic of living modifi ed organisms which are acid into cells or organelles, or pharmaceuticals for humans that are b. Fusion of cells beyond the taxonomic addressed by other relevant international family, agreements and organisations.” that overcome natural physiological reproductive or recombination barriers and The Protocol’s defi nition of an organism that are not techniques used in traditional includes biological entities that are capable breeding and selection.” of transferring or replicating genetic material. This defi nition is similar to The Protocol uses the term ”novel that used in the new EU Directive, but it combination of genetic material”, but it is mentions, in addition, sterile organisms, not specifi cally stated whether the novel viruses and viroids in particular. In the New combination has to be heritable. Hence, Penguin Dictionary of Biology, viroids are it is possible to interpret DNA treatment explained as follows: as a method of genetically modifying an ”Small naked RNA loops, 300-400 organism, and the animal as an LMO, for nucleotides long. Replicated by host enzymes as long as the novel combination is present. as their genomes do not encode any. Lacking However, the Protocol is only applicable as capsids, only able to pass from one damaged long as the animal is alive, and separate cell to another. Analagous to plasmids.” rules apply for organisms that are to be used directly as food or animal feed. The fundamental difference between viroids and self-replicating plasmids is not that The Cartagena Protocol, too, provides signifi cant, but plasmids are not explicitly exemptions for medicinal products, but as included in the defi nition of a living organism. far as veterinary medicinal products are Living, genetically modifi ed viral vaccines concerned, there is a difference compared for animals are, therefore, included in the with the EU Directive. In the Cartagena defi nition of an LMO, whereas DNA-treatment Protocol, medicinal products intended for products based on plasmids do not, according human use are exempted, whereas veterinary to the Protocol, appear to be LMOs per se.

24 Regulation of DNA vaccines and gene therapy on animals

5. Possible regulatory methods

There are two principal methods for 1c. GMO when the added genetic material regulating DNA vaccines and gene therapy is likely to become heritable on animals: regulating the animal that has Some genetic constructs might entail a undergone treatment or regulating the genetic alteration of reproductive cells. The treatment itself. A combination of both is, of animal would be classifi ed as genetically course, a third option. modifi ed only when it is likely that the genetic material added could be inherited. 5.1 Regulating the animal Here, an assessment would have to be made of what is an acceptable risk for incidental integration in the chromosomes egulating DNA-treated animals under R of reproductive cells. the Gene Technology Act could mean that the animal would, under certain circumstances, be classifi ed as genetically modifi ed. The 1d. GMO when the new genetic material key question would then be what the term has particular characteristics “genetically modifi ed” implies and, more specifi cally, the concept of “genetic material”. This alternative would provide guidelines Alternatively, a new category could be specifying when the genetic material is of introduced or DNA treatment could be such a nature that it would render the animal regulated under other legislation. The term genetically modifi ed, and when this is not “genetically modifi ed animal” has traditionally the case. It might apply to combinations of been used to designate animals that have the criteria set out in the alternatives above been produced by altering the chromosomal or if the animal acquires new properties DNA of a fertilized egg or embryo by means that alter its natural characteristics. In of gene technology. But what should be the cases of doubt, the considerations should be criteria to apply when determining whether made on a case-by-case basis. DNA treatment of a developed animal makes it genetically modifi ed? In this respect, there may be several alternative approaches: 1e. DNA-treated animals are given an entirely new designation 1a. GMO if genetic material has at any Rather than establishing a sharp distinction time been added to the animal between genetically modifi ed and non- genetically modifi ed, it might be possible to Animals that have received genetic material, either in the form of a vaccine or for other grade genetic modifi cation by introducing purposes, would be classifi ed as genetically one or more new categories, e.g. “DNA- modifi ed forever. A relevant consideration is treated” as a term to describe animals then whether this should also apply to the that have received DNA vaccines or gene offspring of such animals. therapy.

1b. GMO as long as the added genetic 1f. The term “GMO” is reserved for material is present deliberate, heritable modifi cations The animal will be classifi ed as genetically modifi ed only for as long as the genetic Another option might be to refrain from material added remains present in the regulating DNA-treated animals under the animal. In this case, it would have to Gene Technology Act, but instead reserve be shown probable that the DNA is no the term “GMO” for organisms that have longer present before the animal could be undergone deliberate, heritable genetic declassifi ed as genetically modifi ed. modifi cation.

25 The Norwegian Biotechnology Advisory Board

5.2 Regulating treatment values on which they have been founded, the considerations that regulators wish to DNA vaccines and gene therapy on animals make and how opposing considerations are might conceivably be controlled by regulating balanced. treatment instead of, or in addition to the DNA-treated animal itself. Since DNA treatment of animals moves in a grey area of the Gene Technology Act and since 2a. Genetic material for DNA treatment is a restrictive regulation of GMOs is widely regulated under the Gene Technology Act accepted in today’s society, it is important to determine whether the considerations on The Gene Technology Act’s current defi nition which this Act is based should also apply of organisms includes viruses, but not naked to DNA-treated animals. According to the DNA. The defi nition could be extended to intentions of the Gene Technology Act, the apply also to naked genetic material when production and use of GMOs should be such material is a replicating entity or fulfi ls without detrimental effects on health and the other selected criteria. environment. Furthermore, they should take place in an ethically and socially justifi able 2b. Specifi c legal provisions regulating the way, in accordance with the principle of DNA treatment of animals sustainable development. Any evaluation DNA treatment of animals could be of these overriding considerations depends, regulated in a separate act or in a separate however, on the values that underpin them chapter of the Gene Technology Act, where and how these are expressed. Hence, focus no defi nite position on the defi nition of GMO should be placed on considerations that would necessarily have to be adopted. might be important in such an evaluation, such as animal welfare, consumer interests, business interests and international 2c. DNA treatment of animals is covered cooperation. Furthermore, it is important by other legislation that any regulation of DNA-treated animals DNA vaccines and gene therapy on animals is biologically well-founded and that it is will, in most instances, fall within the scope possible to implement in legislation as well of other acts. Veterinary medicinal products as in practice. are, for instance, covered by the Medicinal Products Act and their use by the Animal The next chapter presents a discussion of the Health Act. Any amplifi cation of the Gene most important considerations that must be Technology Act in these areas would, made. The considerations mentioned have therefore, be superfl uous, but additional been selected in the light of the current provisions could be introduced under other situation, but, to the extent possible, legislation. also with a view to the consequences of future developments. For each specifi c Which of these regulatory systems would consideration, it is discussed whether the provide for appropriate regulation of DNA- different regulatory alternatives provide treated animals? It would depend on the appropriate safeguards.

26 Regulation of DNA vaccines and gene therapy on animals

6. Considerations to be made when selecting regulatory method

6.1 Environment restricted, but incidental integration can occur independently of the sequence. The development of transgenic animals mostly Environmental considerations depends upon incidental integration. Even Environmental considerations represent a if genetic sequences are as “innocuous” as core element of any regulation of genetically possible and do not contain any known modifi ed organisms, and they are, therefore, integrative sequences, it is impossible to important to any evaluation of how DNA- control where in the genome such incidental treated animals should be regulated. integrations may occur. They can occur in various locations in the genome and give rise to different effects. In this respect, there will Ethical principles related to the always be an element of uncertainty. What environment kind of effects could this produce? In many The value of the environment will, for many cases, it will affect the animal itself, in the people, mean the value that the environment form of cancer development or other diseases. holds for human beings – for people alive In rare instances, an alteration may occur today, as well as for future generations. giving rise to a selective advantage for the The interests of future generations are animal in the ecosystem, something that will central to the concept of “sustainable often be unwanted. development”. When assessing benefi t to the society, environmental considerations will be weighed against other considerations, Spread of genetic material and in such a way that we feel able to answer recombination for any balancing of the interests of today’s Genetic material may spread accidentally, e.g. society against the importance of the by the transfer of plasmids to other species interests of coming generations. However, when the animal dies or via body fl uids the environment may also represent a value secreted by the animal (11). A recombination that goes beyond that of its worth to human with natural viruses could, moreover, occur beings. In this instance, the environment inside the animal’s cells, especially with the is said to hold an inherent value. Thinking use of viral vectors (12). With free plasmids along these lines, many could argue that we there is a greater likelihood of mobility have no right to misuse nature, regardless than with a genetic alteration in the of how useful it might be to us. Both these chromosomes. It is conceivable, therefore, ethical principles could form the basis of an that added genetic material might be taken evaluation of the different environmental up by microorganisms living in the animal. considerations involved, and they will often lead to the same conclusions. Risk assessment and the precautionary principle Integration and ecological balance According to the Gene Technology Act, Long-term effects of DNA treatment might deliberate releases should have no occur when humans repeat the same detrimental effects on the environment. actions over a long period of time, like All applications for the deliberate release when vaccinating farmed fi sh, or when new of a GMO must, therefore, include a risk characteristics become heritable. For DNA- assessment accounting for its possible treated animals, heritable characteristics effects on the environment. The harmful could arise as a result of the intentional environmental effects that are feared integration of genetic constructs into include the disproportionate and irreversible chromosomal DNA, or they could be caused effect on non-target organisms, gene fl ow to by unintentional integration. The use wild congeners or closely related species, of integrative genetic elements may be recombination and the creation of, for

27 The Norwegian Biotechnology Advisory Board

example, new viral strains, and the spread animal species that is to be treated. In that of properties such as antibiotic resistance case, documentation for every new animal via horizontal gene transfer. Many of species and every new type of treatment will these aspects are diffi cult to assess, due be required. Depending on the environment to complex systems and unknown risks. in which the animal lives, it might also be Especially diffi cult is any estimation of the relevant to analyse possible impacts on long-term effects, since small and perhaps the environment surrounding the treated unknown selective advantages can have animal and to consider limiting its freedom major consequences in the long term. The of movement. precautionary principle should, therefore, form the basis of any deliberate release of GMOs into the environment. Safeguarding environmental interests by regulating the animal The probabilities of DNA-treated animals If DNA-treated animals are to be regulated spreading the genetic material and having as GMOs under the Gene Technology Act, effects on the environment will differ from animals that are not meant for contained those for heritably genetically modifi ed use will be considered in the same way as animals. Since risk is defi ned as probability deliberate release of GMOs, involving a times consequence, differing probabilities comprehensive study of the environmental will alone affect the risk. There might possibly impact in each individual case. Such a study be a greater likelihood of spread of genetic would also include the possible consequences material to microorganisms following DNA of any spreading outside the release area, injection than when the DNA is integrated the possibility of transfer of genetic material into the chromosomes. For DNA treatment, to other organisms, the likelihood of other there are four circumstances that all have interactions with the environment and the to occur for the genetic material to become possibility of pathogenic or harmful effects to heritable; the DNA added has to translocate health by living or dead GMOs. Furthermore, inside the body, it must be taken up by safety measures would be required to avoid reproductive cells, it must be integrated spreading, as would methods for traceability into the chromosomes of reproductive cells and internal control. Environmental without these cells dying, and the cells interests would, therefore, be safeguarded must subsequently lead to live offspring. under the rules applying to the deliberate Furthermore, added genes may – perhaps release of GMOs. to a lesser degree than for intentional, heritable modifi cation – give the animal Defi ning all animals that have ever received a selective advantage. But even though DNA treatment as GMOs would ensure that the probabilities of such events differ, the the animal’s local, ecological environment is unwanted environmental consequences may taken into account, and it would be possible still be the same, and the risk correspondingly to say “no” when required. This approach high. What an acceptable risk limit is, or would also presuppose a precautionary whether the precautionary principle should attitude, based on the assumption that it be applied, will have to be evaluated on a is impossible to rule out that some of the case-by-case basis. added genetic material may still be present in the animal and that it might have been In order to ensure that the possible effects integrated into its reproductive cells and on the environment following the use of lead to offspring with a heritable genetic DNA vaccines and gene therapy are taken modifi cation. into consideration, public authorities need to be able to assess potential environmental The precautionary principle would be effects. A risk assessment that also includes somewhat toned down if the possibility of environmental impacts will therefore be a temporary genetic modifi cation of the required. Since different animal species have animal is allowed for. In that case, trial different physiologies and genetics, trials are results would have to be produced showing needed to show the effects on the specifi c that the genetic material is degraded and no

28 Regulation of DNA vaccines and gene therapy on animals

longer present in the tissue. Furthermore, a Safeguarding environmental time margin would have to be set before an interests by regulating treatment animal would no longer be termed a GMO. By including plasmids and perhaps even other genetic constructs, in the defi nition of a In some gene therapies, the aim might be GMO, the spread of genetic material and its to integrate the genetic material into the effect on the environment could be assessed chromosomes of target cells. In such cases, before proceeding with DNA treatment. This there is a signifi cant possibility that the type of assessment could also include the gene construct might also be integrated into the chromosomes of reproductive cells. animals that receive DNA treatment, and the In a regulatory context, therefore, primary effects that the genetic material might have emphasis might be placed on the likelihood on them. On the other hand, an assessment of genetic material giving rise to a heritable of the local environment is diffi cult, as genetic modifi cation and less emphasis might authorisation will have been granted before be given to the presence of genetic material. having knowledge of the local environment Since integration rarely can be completely of application. ruled out, documentation must be required showing that the method has minimized the It is also possible to require an environmental probability of such an event. risk assessment of DNA treatment without extending the concept of GMO, regardless of A more detailed specifi cation of the criteria whether the regulatory system is formulated for classifi cation as a GMO, with the as a separate act (or a separate chapter of an subsequent requirement of prior approval act) or whether DNA treatment is merely to be for each individual release, might be an covered by existing legislation. In the latter alternative approach. Here, factors other alternative, however, it is important for the than environmental considerations could be safeguarding of environmental concerns that included, such as the genetic material’s effect DNA treatment, for all purposes whatsoever, on the animal’s natural characteristics. is subject to regulation, and not only when used for medicinal product purposes. The difference in environmental risk between DNA-treated animals and heritably genetically modifi ed (transgenic) animals would be made more apparent if the term 6.2 Animal welfare “DNA-treated” (or the like) was introduced as a separate category, in addition to Vaccines and disease treatment of animals “genetically modifi ed”. As is the case for are important for the welfare of the animal, GMOs, an impact study and approval can as well as to prevent the loss of production be required before DNA-treated animals effi ciency when the animal is a production could be released into the environment. In animal. At the same time, a key argument this situation, it is also possible to evaluate, often used against the genetic modifi cation on a case-by-case basis, whether or not the of animals is that modifi cation might be animal should be termed GMO and relate at the expense of the animal’s natural the environmental risk assessment to that characteristics and health. The use of DNA evaluation. vaccines and gene therapy on animals entail methods that are similar to genetic If a DNA-treated animal already has been modifi cation, but their purpose is often to defi ned as a non-GMO, an environmental improve the animal’s health and welfare. risk assessment of the treatment given to the Hence, it is important, when drawing up a animal could still be introduced. The animal regulatory system for DNA vaccines and gene could be regarded as part of the environment therapy, to consider possible animal welfare in which the genetic material is released (see interests, as well as the consequences that below). alternative regulatory regimes may lead to.

29 The Norwegian Biotechnology Advisory Board

Animal welfare interests has been found in the spermatozoa of pigs vaccinated with DNA (13). Some types of As in the case of assessing environmental added genes might be capable of altering the interests, different values may underpin the individual animal’s natural characteristics, considerations that one might wish to make whereas vaccination against disease is not in respect of animal welfare. Throughout perceived as a means of infl uencing the the ages, human beings have made use of animal’s natural characteristics. animals, and different animals could be said to represent greater or lesser value to humans. However, there is a difference Animal health following genetic between the purely utilitarian view of modifi cation or DNA treatment an animal, where the animal is regarded With many forms of genetic modifi cation exclusively as a creature for human use, of animals, the animal will notice little and attributing to the animal a value that or nothing of the genetic modifi cation. In represents an end in itself. Our wish to other instances, for example when animals preserve an animal’s natural characteristics are used for research on human genetic may be based either on the view that it is diseases, the animal may suffer as a result. of value to us that the animal preserves its The aim of animal vaccination, including natural traits and characteristics, or because DNA vaccination, will generally be to avoid we attribute an inherent value to the animal. disease, thereby contributing to improved If the animal is attributed an inherent value, animal health. With the gene therapy not it should be our duty to take account of this aimed at preventing disease, an evaluation in our treatment of the animal, above and would depend on the characteristics that beyond any value that we as humans might the animal acquires from the added genetic draw from such treatment. material (see below). Furthermore, DNA vaccination and gene therapy will always Infl uencing the natural characteristics of present a slight possibility of integration of animals the added genetic material into an adverse site in the chromosomes, leading to the Wild animals have developed through development of cancer or metabolic disorder evolution into the animals we see today. in the animal. Over a long period of time, humans have performed deliberate breeding of livestock, and, as a result, these animals are today Choice of method for disease prevention vastly different from their wild ancestors. and treatment The genetic modifi cation of animals could Many considerations come into play when be viewed as a continuation of this process, selecting the type of treatment for an but there are certain differences. The time animal, and much will depend on the role scale is different – with genetic modifi cation, played by the animals for human beings. major alterations may be achieved from one generation to the next. Furthermore, For farmed fi sh, a certain level of mortality entirely novel characteristics may be is calculated into production costs. Quite introduced from other species, so the range of different considerations may prevail when a possible alterations is signifi cantly broader. pet is taken to the vet. Gene technology offers, moreover, greater opportunity for carrying out intentional Many of today’s vaccines have side effects alterations and the control that humans for the animal. Fish may, for example, suffer have gained over the natural characteristics adhesions at the injection site when an oil of animals is, therefore, much greater. In adjuvant is used in the vaccine mixture. contrast to heritable genetic modifi cation, An adjuvant is necessary to achieve high gene therapy and DNA vaccination will, in effi cacy of vaccines based on inactivated principle, have an impact on the individual microorganisms. Viral vaccines and DNA animal and not on its offspring. However, vaccines may be administered without the there is the slight probability of a transfer of use of an oil adjuvant. DNA vaccines and added genetic material to offspring through other subunit vaccines are also safer than reproductive cells. For example, vaccine DNA attenuated viral vaccines, because disease

30 Regulation of DNA vaccines and gene therapy on animals

breakout caused by incomplete attenuation capacity and metabolism. Vaccine trials have or reversion can be avioded. already been conducted to eliminate the boar taint in pigs and to increase the number of Vaccines administered in the form of an lambs in ewes. By using vaccines against injection entail stress for the animal. certain molecules on fat cells, the fat deposit Immersion as a delivery method for fi sh in slaughter animals may be reduced, and and nose spray or edible vaccines for other the colour of the fat may also be infl uenced animals would represent better vaccination through the immune system (14). methods for the animals. Work is currently in progress to develop such delivery methods Many of these objectives could also be for DNA vaccines. achieved by means other than DNA treatment, for example with the use of When treating disease in animals, as in protein-based medicinal products. Instead of humans, it is always an advantage to have injecting fi sh with a growth hormone gene the opportunity of choosing the treatment to increase its size, the fi sh could receive the that is best for the animal in terms of good growth hormone directly, in the same way effect and few side-effects. In all likelihood, as cattle do in the US. Immune reactions owners of pets or production animals alike against the animal’s own proteins may also would prefer to use the best vaccines and be achieved by using other types of vaccines medicinal products available. With a more than DNA vaccines. At present, vaccines and restrictive regulation of DNA treatment in the use of these are regulated by the Animal Norway than in other countries, there is the Health Act, the Animal Welfare Act and the possibility that the best treatment may not Medicinal Products Act, and DNA treatment be available here in Norway. would also fall within the scope of this legislation. Disease in animals may also be limited by means other than vaccination. The animal’s In several contexts, proposals are currently resistance to attack may be improved with being made to genetically modify animals correct nutrition and good living conditions, for useful purposes. In Australia, there have and it may be better protected from sources been proposals to use genetically modifi ed of contagion if such sources are identifi ed and viral vaccines to inhibit the fertility of avoided. It is important, therefore, to look at rabbits by means of an immune response all the different aspects of vaccination. It is against the rabbit’s own egg cells (15) and conceivable that an animal’s conditions of to genetically modify carp so as to produce life might, in certain instances, be worsened male offspring only (16). The aim of both as a result of good vaccines, as the animal these proposals is to limit the spread of might then be pushed even further without these species in Australia. The future may falling ill. This aspect might be particularly see many more similar proposals involving relevant in the fi sh-farming industry. gene technology, and it might be easier to gain acceptance for treatment with only a temporary effect than for heritable, lasting DNA treatment of animals for other alterations. In certain cases, therefore, purposes DNA treatment could gain relevance as an When choosing a regulatory system for DNA alternative to heritable genetic modifi cation. vaccines and gene therapy on animals, it DNA treatment may be simple to develop is important to consider the fact that DNA and cheap to use and may thus contribute treatment may have applications beyond to a further instrumentalisation of animals. those of disease prevention and treatment. Any selective advantage resulting from Here, there may be areas that might confl ict such treatment might conceivably manifest with the intentions on which the Gene itself in the species in the long-term, should Technology Act is founded. the added DNA become integrated and be inherited by offspring. Any regulation of DNA Some of the vaccines of the future may aim treatment must also take these perspectives at controlling the animal’s reproductive into account.

31 The Norwegian Biotechnology Advisory Board

Safeguarding animal welfare would represent an arena for the deliberate interests by regulating the animal release of GMOs. For a certain period of time after treatment, the animal may contain Classifying all DNA-treated animals as the modifi ed DNA, and restrictions on the GMOs will require that an application is animal’s freedom of movement might become made for deliberate release both of production one of the criteria for releasing the DNA. animals and of pets following treatment. The use of DNA treatment would then be signifi cantly restricted, including those With specifi c legislation regulating DNA instances where such treatment would be in vaccines and gene therapy, animals treated the animal’s best interests. Even introducing with such methods could be subject to the concept of “temporary GMO”, would requirements limiting the possible spread require the approval of a deliberate release, of genetic material, for example, that such unless the animal’s freedom of movement animals might be banned from being used was so strictly limited that it could be for breeding purposes. This might infl uence termed “contained use” during the period the treatment given to the animal and when the animal is considered a GMO. It is perhaps even prevent it from receiving conceivable that a period of contained use the best treatment available. On the other could be implemented for farmed fi sh, as hand, it might also pave the way for the well as for production animals, but hardly development of good DNA vaccines for for all pets. animals. At the same time, special criteria could be formulated ensuring that DNA If an animal is termed a GMO only when and treatment does not adversely affect the if it has been shown probable that the added animal’s natural characteristics. genetic material will become heritable, the use of DNA vaccines might, to a greater The treatment of animals and veterinary degree, become practically viable, whereas medicinal products are areas regulated by many forms of gene therapy will be restricted other legislation. From an animal health by the approval requirement for deliberate perspective, the pros and cons of DNA release. The interests of preserving the treatment could be taken into account in animal’s natural characteristics will be such legislation, in the way in which the safeguarded if the regulatory system sets genetic modifi cation of animals is currently up criteria for the type of alterations that restricted under Section 5 of the Animal the added genetic material is permitted to Welfare Act related to breeding. produce.

If DNA-treated animals are not termed 6.3 Health GMOs as a result of their treatment, they may be freely treated with DNA vaccines Health considerations and gene therapy, provided that treatment is approved according to regulations in force. Safe food This would give producers of animal food The food we eat must be safe. This is an greater freedom in their choice of veterinary essential principle of food control. The health medicinal treatment for their animals, since hazard of novel foods is assessed in terms of they would neither be required to apply for whether the food is substantially equivalent approval of deliberate releases, nor to label to existing foodstuffs. The Norwegian Food their products as GMOs. Control Authority has issued rules that apply in this area. Genetically modifi ed food is to be assessed with regard to new Safeguarding animal welfare components, possible antibiotic resistance interests by regulating treatment genes and allergenic substances. Analyses With a concept of GMO that includes of nutritional and physiological effects may many of the DNA constructs of vaccines also be required, which mostly are carried and gene therapy, the animals themselves out in the form of animal feed trials.

32 Regulation of DNA vaccines and gene therapy on animals

If DNA-treated animals are used for food is progressing to develop gene therapies. production purposes, new components and So as to ensure the best possible medical foreign genes might also be present in the treatment of humans, it is important that food. There might conceivably be a greater the development of DNA vaccines and gene likelihood of intestinal uptake of plasmids therapy for humans is not hampered by an than uptake of chromosomal fragments excessively restrictive regulation of DNA- from transgenic animals. Also products from treated animals. This might become the case DNA-treated animals should, therefore, be if potential DNA vaccines and gene therapy assessed with regard to the possible health methods for humans were diffi cult fi rst hazard of the added genetic material and of to test on animals. A severe restriction of direct and indirect alterations caused by the DNA treatment on experimental animals in genes. Norway would mean that the development of DNA vaccines and gene therapy for In line with EU regulations, Norway also humans would be dependent on animal applies rules on the residual quantity of trials conducted abroad. The bulk of animal veterinary medicinal products in meat and trials, however, would be contained use and fi sh. Instead of zero tolerance, limit values would not entail a deliberate release of the have been set for the various medicinal animal into the environment. Distinguishing products, based on the principle that they between contained use and deliberate release must entail no hazard to health. This also in the context of DNA treatment could, applies to products from animals that may therefore, provide the appropriate safeguards become food, but that are not production in the interests of research. Moreover, the animals themselves, such as race horses, development of animal vaccines would lead for example. These rules would also apply to to increased research on vaccines in general veterinary medicinal gene therapy, but they and, in consequence, enhanced competence do not apply to vaccines. in this fi eld.

When assessing DNA vaccines and gene Protection against infections therapy for the medical treatment of humans, Animals may also involve a hazard to human the interests of human health and benefi t to health beyond their role as food. Interaction the community would be weighed against with animals may lead to infections and environmental considerations. Hence, it disease (zoonoses). Hence, any treatment of might be possible to defend a policy where animals must also be considered in the light humans are allowed DNA treatment, whereas of how such treatment may affect humans. a restrictive approach is chosen for the DNA For example, the infl uence of host specifi city treatment of animals in the environment. It in the development of viral vaccines for is, of course, possible for patients to choose animals may have an impact. Possible not to receive DNA treatment if they so health effects on humans in general, and wish. for veterinarians and animal producers in their working environment in particular, Safeguarding health aspects by are assessed before vaccines for animals regulating the animal are approved. However, the use of vaccines may also lead to the development of mutated Since food is assessed from a health point pathogenic organisms that might be of view with regard to its components and transferred to humans. On the other hand, residual medicinal products present, the less disease in animals in general might designation ”GMO” or ”non-GMO” should, also lead to fewer cases of animal diseases in principle, have no decisive impact on in humans. the health aspect of the food product. Nevertheless, it is conceivable that more work would be devoted to demonstrating Prospect of good medical treatment that a food product produced from a DNA- Several DNA vaccines are currently being treated animal is non-hazardous to human developed for humans, including vaccines health, if specifi c labelling was required for against malaria and HIV. In addition, work such a product.

33 The Norwegian Biotechnology Advisory Board

Any regulation of the DNA treatment of The introduction of a new labelling category, animals that is so restrictive as to obstruct e.g. ”DNA-treated” would provide more the build-up of research and competence balanced information, but also make it more could have an adverse effect on medical diffi cult for consumers to react to labelling. competence in the fi eld of DNA vaccines Furthermore, for many consumers it would and gene therapy for humans. A more subtle be important to have access to information regulatory system, setting up requirements about production methods; this is the reason for the characteristics of the genetic for the introduction of process labelling of material involved, could, on the other hand, GMOs in Norway and the EU. An alternative help in the development of DNA treatment could be to make information available to for medical use in humans. consumers in other ways than by labelling.

Safeguarding health aspects by Wide selection, good quality and low regulating treatment prices New veterinary medicinal products based Consumers are also interested in having a on biotechnology and gene technology wide selection in the supermarkets, good must all be assessed for health risk – for quality products from healthy animals and the veterinarian using them, but also for low prices. Effectively combating disease possible effects on others. This holds true in animal production for food purposes is irrespective of whether the medicinal product an important factor in this respect. Hence, is classifi ed as a GMO or not. However, if the it is also in the consumer’s interests that GMO concept is extended to also apply to production runs as smoothly as possible and genetic constructs used in DNA treatment, that producers are allowed a choice of the gene therapy products and DNA vaccines medicinal products they believe to be the best for human use would have to be approved for their animals. On the other hand, highly as deliberate releases of GMOs prior to intensive animal production may lead to use. Human beings themselves would not, poor conditions of life for the animals and to however, be regarded as GMOs. poorer quality of the products. Good business development may contribute to added value in society and improved purchasing power 6.4 Consumers for the individual consumer.

Consumer interests Confi dence in the authorities It is natural to feel scepticism towards new Good information and freedom of choice and unfamiliar technologies, and perceived The introduction of labelling requirements risk may often deviate from actual risk. for genetically modifi ed food is a consequence It is important that a regulatory system of the fact that people have different views creates confi dence among consumers and on such products. The same might also be that they feel that their interests are relevant for products produced from DNA- being safeguarded. This applies not only treated animals. Consumers are largely able to objective information, but also to control to infl uence the market, and a labelling of measures and risk research and is helped salmon as genetically modifi ed as a result of by ensuring that the regulatory system DNA vaccination, could lead to a decline in coincides with the ethical principles of the the sale of fi sh. If the labelling requirement general public. If the principle objection to is also to cover imported products produced GMOs is founded on resistance to intentional from animals that have received DNA heritable alterations, a concept of GMOs vaccines, the number of GMO products may that also includes DNA-vaccinated animals become so vast that many consumers could might be perceived as an unfortunate gradually resign and end up thinking that watering-down of the concept, so that it loses ”everything” is genetically modifi ed. much of its force.

34 Regulation of DNA vaccines and gene therapy on animals

Safeguarding consumer interests 6.5 Business interests by regulating the animal With a regulatory system where DNA- Taking account of business treated animals are considered as GMOs at interests their time of slaughter, the products would be In the interests of business development, labelled ”genetically modifi ed”. Consumers it is generally important to have clear-cut, may either feel that their confi dence in the precise guidelines so that companies can public authorities is enhanced as a result, or avoid making unsound investments. At there might be an unworkable watering-down the same time, there is a case for avoiding of the concept if too many products would an excessively complicated and laborious be given this labelling. Reduced confi dence bureaucracy that might give rise to serious with certain consumers can be the result if practical diffi culties. the regulatory system runs contrary to their arguments for wanting GMO products to be Today’s markets strive to achieve a level labelled. The supply of food products could playing fi eld in order to promote competition also decrease and prices rise, as a result of between companies. In this respect, Norway import restrictions. is part of the EEA and free competition applies across national borders. For If choosing the term “DNA-treated” to Norwegian producers wishing to be part designate animals that have received DNA of this market, it will be important that vaccines or gene therapy, and by labelling Norway’s rules are no more restrictive than those of our trading partners in the fi elds of the products accordingly, consumers would natural resource use and the development have to deal with a new category of products. and sale of products. Sound public information would be required, and it is not entirely evident how consumers would react. Furthermore, the supply of food Development of veterinary medicinal products might be limited, due to import products restrictions, depending on the rules that Norway has approximately half of the apply to the import of products from DNA- world’s market for fi sh vaccines, and treated animals and on whether these rules there is a large potential market in the deviate from those of our trading partners. aquaculture industries. Firms and research environments in Norway are in the forefront If DNA-treated animals are not regarded as of developments of medicinal products for fi sh. This has given rise to high-competence GMOs and are not labelled in any particular environments that, in turn, provide good way, it will be up to the individual consumer breeding grounds for further growth in to obtain information about any medicinal the industry. Most fi rms have been bought treatment of the animals and otherwise up by large, international pharmaceutical trust that residual medicinal products are companies that have seen the potential well below the level involving health risks. of the fi sh-farming industry. These are This alternative would entail few import companies capable of contributing with their restrictions on animal products. experience and fi nancial power to ensure that new medicinal products are approved through centralised procedures. Safeguarding consumer interests by regulating treatment DNA vaccines are relatively simple to By regulating treatment and not the animal, develop, since fi nding the right culture consumers would be allowed no choice from conditions for new pathogenic organisms shop shelves based on personal opinion. is not needed. The vaccines are cheap However, an appropriate regulation of to produce, since standard methods for treatment could contribute to good quality purifi cation may be used. Furthermore, DNA of the products. is stable at a wide range of temperatures and

35 The Norwegian Biotechnology Advisory Board

does not require refrigeration In addition, if Nonetheless, fi sh-farmers in Norway today the vaccine can be administered orally or by want to offer “clean” products. This is in line means of immersion instead of injection, the with rising consumer awareness, which has vaccination process becomes signifi cantly also led to increased sales of organic food. cheaper. DNA vaccines seem, therefore, Fish-farmers depend on the confi dence of to hold a great potential as medicinal consumers and, at the present time, are, products. therefore, not interested in exploring the possibility of genetically modifi ed products. Pharmaceutical manufacturers would If DNA treatment is regulated by means like to see a liberal regime allowing for of a liberal regime, Norwegian farmed fi sh the development of the best vaccines or may face unwanted competition from, for therapies, irrespective of the method used. example, imported fi sh that has received On the international market, good effects gene therapy in the form of growth and good prices would sell best. Specifi c hormone genes. A regulatory system for medicinal products could replace the more genetic modifi cation and DNA treatment general drugs that have side effects, in in Norway that takes due account of the the same way as vaccines against specifi c environment could, on the other hand, be diseases have replaced the use of antibiotics used in the marketing of Norwegian food in the fi sh-farming industry. This is a products as more environmentally-friendly trend that would benefi t society and that than products from many other countries. might, therefore, pay off to focus on. Having national cooperating partners and users will encourage the development of medicinal Agrobusiness products here in Norway. A more restrictive For the agricultural sector, as for regulatory system in Norway compared to aquaculture, it is a matter of having a other countries might, on the other hand, wide range of effective medicinal products hamper the development of such products capable of combating disease in livestock in this country. But a strict regulatory more effi ciently and lower the production system could also contribute to putting costs. New properties may contribute pressure on industry to produce even to the further development of products. more environmentally-friendly medicinal Genetic modifi cation, gene therapy and products. DNA vaccines could be used to achieve such aims. On the other hand, producers are best served if confi dence among Aquaculture industry consumers is retained, and as long as there A wide choice in the types of vaccine is resistance to products labelled as GMOs, technologies offered would make it easier many producers will seek to avoid this to combat disease effi ciently in fi sh- label on their products. farming facilities. Furthermore, if injections can be avoided, and the vaccines can be administered either orally or by means of Wilderness sector immersion, the vaccination process would The development of medicinal products to become cheaper to perform. combat disease in wild animal populations has also begun. One example is a genetically Medicinal products may also contribute modifi ed vaccine that was introduced into to increasing production effi ciency beyond fox bait in Belgium to combat rabies. that of combating disease, for example, Recently, the fi rst DNA vaccine in practical by promoting rapid growth and other use was given to the endangered Californian properties. In this way, a free choice of the condor to protect it against the fatal West best medicinal products, including DNA Nile virus (17). Medicinal products of this vaccines, living genetically modifi ed viral type could also replace more unspecifi c vaccines and gene therapy, may contribute measures, such as rotenone treatment to increasing the production in fi sh-farming of rivers aimed at eradicating the salmon facilities. parasite Gyrodactylus salaris.

36 Regulation of DNA vaccines and gene therapy on animals

Safeguarding business interests by in Norway. This depends, however, on regulating the animal whether the sales potential is considered signifi cant enough, something that may, in If DNA-treated animals are regulated as turn, adversely affect price and supply, seen GMOs, developing and selling DNA vaccines from a user perspective. Hence, users should will become a complicated matter. Many also have the possibility of applying for the animal producers will refrain from using authorisation of a product. DNA treatment if they risk having to label the animal as a GMO, and their use will require authorisation for every new deliberate When regulating DNA treatment in release. Restricted freedom of choice might ways other than as a method for genetic conceivably hamper the fi ght against animal modifi cation, guidelines should be clear diseases and keep production costs at a and should avoid imposing an unnecessary higher level. By regulating DNA-treated workload on producers and users. The animals as temporary GMOs, many DNA import and use of medicinal products and vaccines could be used, but there might be the import and export of animals would be additional costs involved in keeping animals simplest if Norway and the EU practised the contained for a certain period of time. Also a regulations in the same way. labelling as ”DNA-treated” would go against the wishes of many animal producers. 6.6 International cooperation If DNA-treated animals do not need to be specifi cally labelled, and authorisation does not have to be applied for prior to each Interests of international individual treatment, this will enable a cooperation much more liberal use of DNA vaccines and Import and export agreements gene therapy in animal production. It might also contribute to growth in the development If the defi nition of a GMO is interpreted of veterinary medicinal products in Norway. differently in Norway compared to its If the criteria determining which treatments surrounding countries, including the EU make an animal a GMO, are not clear, and area, this may lead to diffi culties for import applications are assessed on a case-by-case and export activities. DNA treatment basis, pharmaceutical companies and users that in Norway would render the animal will need clear-cut guidelines and reasonably genetically modifi ed, would perhaps, for predictable approval procedures. this reason, not be practised here. In that case, some products might be outpriced by cheaper products from other countries. Irrespective of regulatory method, it will be When importing animal products for food possible for individual producers to continue and animal feed, labelling of a product as to engage in the types of production that do genetically modifi ed (or as DNA-treated) not involve the use of DNA treatment, if they may be required, and if products are to be should so wish. labelled, they must fi rst be approved. If the products are not classifi ed as GMOs in their Safeguarding business interests by country of origin, it might be diffi cult to regulating treatment obtain suffi cient information to carry out a thorough evaluation here in Norway of every By extending the GMO concept to include product prior to its import into the country. forms of naked genetic material, or if DNA If the animal itself is classifi ed as a GMO, treatment is subject to the same regulatory the import of living animals will have to be system, the authorisation procedures considered as a deliberate release of a GMO. applying to DNA treatment will be the In this context, the necessary information same as those applying to the use of living, might be more readily available in the form genetically modifi ed viral vaccines. Many of vaccination certifi cates, etc. However, a major pharmaceutical companies will have regulatory system of this type would place the necessary funds and experience to strong demands on public authorities. carry through an authorisation procedure Furthermore, in order to contribute to a

37 The Norwegian Biotechnology Advisory Board

good climate of cooperation, unnecessary basis of sound justifi cation, opts for a regime accusations of setting up trade barriers of clear regulation and argues well in favour should be limited as far as possible. of such a system, it might contribute to a debate on regulatory systems within the EU Since the legal basis of the new EU and in other countries as well. Directive on the deliberate release of GMOs is founded on the Amsterdam Treaty and Developing countries since it does not allow for stricter rules in The development of DNA vaccines may the individual states, it will be up to Norway give rise to new vaccines for both human to negotiate, through the EEA Agreement, if and animal use in developing countries, the Norwegian authorities wish to apply a where thermal stability and reasonable different regulatory system in this country. If Norway chooses to regulate DNA-treated production costs constitute essential animals as GMOs and the EU does not do so, aspects. A regulatory system that promotes the key question at the next juncture may the development of DNA vaccines providing be whether many animal products from EU a high level of security for health and the states will have to be stopped at the border environment could, therefore, contribute on the grounds of a lack of authorisation and to more effi ciently combating disease in labelling of the products as GMO products. developing countries. Since DNA vaccines are so far not in use for production animals in the EU, it is unclear, Safeguarding the interests of however, whether the various EU member states will interpret and apply EU rules international cooperation by differently. regulating the animal The EU’s deliberate release directive is open to differing interpretations of the concept The sea as an arena of deliberate release of GMOs, and by linking the regulation of Norway’s regulatory practice on issues of DNA-treated animals to an interpretation genetic modifi cation and the DNA treatment that agrees with the EU Directive, Norway of animals in nature may entail consequences could work to establish a constructive for other countries beyond the regulatory dialogue with its trading partners in the EU. scope of current trade agreements. This If Norway chooses to retain the possibility of applies in particular to fi sh and other marine exercising discretion in the interpretation animals, but also to birds and insects. of the GMO concept, any divergent practice could be adjusted if trade confl icts arose. Similarly, Norway will be affected by the practices of other countries. A regulatory system applying to this fi eld should, Safeguarding the interests of therefore, be regarded as an international international cooperation by matter. regulating treatment If the defi nition of GMO in Norway is extended Possible means of infl uence to include plasmids, the import of many It is unclear whether authorised products medicinal products could get complicated, for DNA treatment purposes are to be because they would then be considered as considered as genetically modifi ed medicinal GMOs. Viral vaccines from the EU will products in the EU and also whether DNA have undergone an environmental risk treatment that is non-medical will render the assessment as GMOs, but perhaps not DNA animal genetically modifi ed. Different EU vaccines to the same extent. It also would member states may interpret EU Directive infl uence relations between Norwegian and 2001/18/EC differently. In this respect, international researchers working with Norway could contribute to clarifying the DNA vaccines and gene therapy for humans, problem and work to achieve a regulatory since many of these genetic constructs would practice that meets the challenges involved also be defi ned as GMOs. Furthermore, it in a constructive manner. If Norway, on the would complicate Norwegian participation

38 Regulation of DNA vaccines and gene therapy on animals

in research projects in the fi eld of molecular product, this too might hamper trade with biology in general, where the exchange of countries that have a different regulatory plasmids constitutes an important element. system. However, there would also be the possibility of contributing to the setting If a regulatory system is linked to the up of a common set of international use of DNA treatment and not to the rules for DNA treatment of animals.

Photo: Karin Beate Nøsterud, VG / Scanpix

39 The Norwegian Biotechnology Advisory Board

7. Summary of regulatory alternatives

7.1 Regulating the animal • The development of DNA vaccines for use on humans might also be hampered.

1a. GMO if genetic material has at Challenges: any time been added to the animal • The concept of GMO might be watered down. Considerations made: • There might be the possibility of methods • It would be impossible to rule out that for the indirect DNA treatment of animals some of the DNA might have reached the not performed “by means of gene or cell reproductive cells and been incorporated technology” in the form of intentional there. Furthermore, some of these animals “introduction into living cells”, e.g. in the might escape and the genes might be passed form of animal feed additives. on. • Animals having received live, attenuated • Naked, modifi ed DNA might be transferred viral vaccines have also had DNA or RNA to other species without the consequences introduced into the cells. It could therefore being known in advance. A precautionary be considered whether these animals attitude would indicate that this should be also should be termed GMOs. A genetic avoided as far as possible. characterization of attenuated viruses • With gene therapy, it would be possible might indeed lead to a situation where to select (intentionally or unintentionally) the requirements for being termed “gene animals where the DNA has been technology” would gradually be met. incorporated into the genome. Alternatively, it might be possible to select the offspring 1b. GMO as long as the added that have inherited the added genetic genetic material is present material. In that case, the method would be almost identical to the one commonly used to Considerations made: produce transgenic animals. • Genetic material might be spread to the • It would be possible for consumers to environment and it would be important, avoid the products of DNA-treated animals, therefore, to keep such material under without having to deal with a specifi c controlled conditions until it was degraded. category for such products. Hence, the animal would be regulated as Consequences: a GMO until it could be assumed that the added DNA was degraded. Replicating and • All animals that had received DNA vaccines integrating genetic constructs will remain or gene therapy would have to be classifi ed present for longer periods of time than as GMOs, thereby creating a labelling short DNA molecules, so their use would be requirement for the products. limited. • It might lead to confl icts due to import • It would be important not to water down restrictions, because our trading partners the concept of GMO, and a distinction would, might have other labelling regulations. therefore, be made between the animals that It might also become diffi cult to obtain were known to have undergone heritable information about the DNA treatment alterations and the animals that had received of imported animals and products from DNA vaccine/gene therapy on somatic cells countries applying different labelling only. The likelihood of incorporation of added regulations. DNA in the chromosomes of reproductive • Different combinations of animals and cells would be considered so slight that it vaccinations would have to be assessed as could be ignored. separate releases and this might hamper the development and use of such medicinal Consequences: products. • The animal would be termed genetically

40 Regulation of DNA vaccines and gene therapy on animals

modifi ed when foreign genes are present in information on any DNA-treatment would the animal’s cells. Hence, it would only be be required. classifi ed as a GMO for a certain period of Challenges: time and would, following an estimated DNA degradation period, no longer be considered • It would be diffi cult to estimate whether a GMO. the added genetic material might be • It would be possible to use DNA vaccines heritable, and trials would be required in and at the same time take account of order to show that it would not be integrated environmental considerations. into reproductive cells. • In the event of a deliberate release, every vaccination case would have to be assessed 1d. GMO when the new genetic separately. If vaccination took place within material has particular charac- the context of contained use, involving teristics subsequent release or sale, the duration of the GMO period would still have to be Considerations made: determined. • Consideration could here be given to the • Imported animals and products would have possibility that certain types of genetic to be assessed, so as to determine whether or material might give the animal entirely not the products would need labelling. This new properties affecting its natural might lead to trade restrictions on certain characteristics. products. • It would also be possible to take account Challenges: of the risk of gene transfer, as well as of integration. • It would be diffi cult to know at what point in time the added DNA was degraded. Consequences: • It might be diffi cult to obtain information • This might enable the use of DNA about the DNA treatment of imported treatment in the fi ght against disease, while animals and products. restricting the use of gene therapy for other purposes. 1c. GMO when the added genetic • Detailed guidelines would need to be drawn material is likely to become up, taking into consideration the many different types of genetic constructs, and it heritable would be logical to process applications on a Considerations made: case-by-case basis. • • Emphasis would be placed on preserving It would be possible to exercise discretion, the species, by regulating possible heritable as well as to adjust practices en route, if so genetic modifi cation in the same way as desired. deliberate, heritable genetic modifi cation. At Challenges: the same time, emphasis would be put on the • The guidelines drawn up for manufacturers, likelyhood of integration occurring. producers and administrators would have to • The use of DNA vaccines in animal be clear-cut to avoid ambiguity. production might combat disease more effi ciently than traditional vaccines. 1e. DNA-treated animals are given Consequences: an entirely new designation • This would enable the use of DNA vaccines where there was only a very slight Considerations made: probability that the genetic material would • A distinction would be made between be inherited. heritable genetic modifi cation and DNA • Gene therapy on animals might, to a treatment, while simultaneously regulating greater degree than for DNA vaccination, DNA treatment. mean that the animal would be regulated • The animals would be given a designation as a GMO. that provided information about factual • When importing live animals, detailed circumstances.

41 The Norwegian Biotechnology Advisory Board

Consequences: 7.2 Regulating treatment • Food products could be labelled specifi cally with “DNA-treated” or the like. 2a. Genetic material for DNA • The concept of GMO would not be watered treatment is regulated under the down. Gene Technology Act • It would be possible to draw up a separate regulatory system for DNA treatment. Considerations made: • Imported food could be labelled in the same • Account would be taken of the fact that way, on the basis of information provided by certain genetic constructs may be replicated the producer. and spread in the surroundings, and the use • Labelling requirements might have a price- of more environmentally-friendly genetic raising effect. constructs would be encouraged. Challenges: Consequences: • It might be quite challenging for consumers • Certain DNA vaccines and gene therapy to have to deal with a separate category for products for human use might also be DNA-treated animals. classifi ed as GMOs. • There might be more complicated rules • If certain DNA constructs were defi ned as regulating the use of gene technology on GMOs in Norway only, the import of many animals. new medicinal products might become • A new labelling category might lead to diffi cult. trade barriers. • Norwegian participation in international research projects in the fi eld of molecular biology might be complicated. 1f. The term ”GMO” is reserved for deliberate, heritable modifi cations Challenges: • Naked DNA has not been regulated under Considerations made: the Gene Technology Act in the past, and if • The likelihood of heritable alterations any genetic constructs were included in the following DNA treatment is very slight, and GMO designation, this would mean that the the concept of GMO should not be watered rest of the act would have to be reconsidered down. in light of this development. • Animal producers should be given freedom to choose the treatment that is best for their animals and for production effi ciency. 2b. Specifi c legal provisions regulating the DNA treatment Consequences: • Animals that have been treated with Considerations made: DNA vaccines or gene therapy as medicinal • The difference between heritable genetic products would not be classifi ed as GMOs. modifi cation and DNA treatment would • Nor would other types of gene therapy on be clarifi ed in regulations and in terms of animals be covered by the Gene Technology possible required labelling. Act. • A separate requirement for an • Producers in Norway might experience environmental risk assessment might be favourable competitive conditions, but specifi ed. it might also reduce the possibility of Consequences: specifi cally marketing “clean” products. • The GMO concept would not be watered Challenges: down. • Treatment with DNA that is not a • The products could be developed and medicinal product might approach methods marketed, even though their use might be for deliberate genetic modifi cation, and clear- somewhat limited for Norwegian animal cut guidelines would have to be drawn up. producers.

42 Regulation of DNA vaccines and gene therapy on animals

Challenges: Products Act and other legislation, and • New grey areas might be created between unnecessary double regulation could be the statutory regulation of DNA treatment avoided. and the regulation of heritable genetic Consequences: modifi cation. • DNA-treated animals and the products of such animals would not be classifi ed as 2c. DNA treatment of animals is GMOs. covered by other legislation • There would be no additional regulation of the development and use of medicinal Considerations made: products based on DNA. • Genetic modifi cation would be considered to apply only to heritable properties, and the Challenges: preservation of species and the ecosystem • DNA treatment that, with a reasonable would not be considered threatened by DNA degree of probability, might result in heritable treatment that did not specifi cally aim at genetic modifi cation would not necessarily genetic modifi cation. be covered by the other legislation. The • DNA vaccines and gene therapy would, grey area of the Gene Technology Act would, in most cases, be covered by the Medicinal therefore, still need to be clearifi ed.

Photo: Heiko Junge / Scanpix

43 The Norwegian Biotechnology Advisory Board

8. References

1. Gudding, R. (2000) “Immunprofylakse 9. Circular 4039 R from Norwegian Food i veterinærmedisinen”, Scandinavian Control Authority, 1997. Veterinary Press, p. 130. 10. Gillis, J. (2003) “Old laws, new fi sh. 2. Wolff, J.A., Malone, R.W., Williams, P., Environmental regulation of gene-altered Chong, W., Acsadi, G., Jani, A. & Felgner, foods is a grey area”, Washington Post, 15 P.L. (1990) “Direct gene transfer into mouse January 2003. muscle cells in vivo”, Science 247, 1465-8. 11. Traavik, T. (1995) “Too early may be too 3. Lewis, K.N., Liao, R., Guinn, K.M., Hickey, late. Ecological risks associated with the M.J., Smith, S., Behr, M.A. & Sherman, D.R. use of naked DNA as a biological tool for (2003) “Deletion of RD1 from Mycobacterium research, production and therapy”, Report tuberculosis mimics bacille Calmette-Guerin for the Directorate for Nature Management, attenuation”, J. Infect. Dis. 187, 117-23. 1999-1.

4. Cello J., Paul, A.V. & Wimmer, E. (2002) “Chemical synthesis of poliovirus cDNA: 12. Traavik, T. (1999) ”An orphan in science: generation of infectious virus in the absence Environmental risks of genetically engineered of natural template”, Science 297, 1016-8. vaccines”, Report for the Directorate for Nature Management, 1999-6. 5. Lavitrano, M., Forni, M., Bacci, M.L., Di Stefano, C., Varzi, W., Wand, H. & Seren, E. 13. Gudding, R. (2000) “Immunprofylakse (2003) “Sperm mediated gene transfer in pig: i veterinærmedisinen”, Scandinavian Selection of donor boars and optimization of Veterinary Press, p. 121. DNA uptake”, Mol. Reprod. Dev. 64, 284-91. 14. Gudding, R. (2000) “Immunprofylakse 6. Sato, M., Ishikawa, A. & Kimura, M. (2002) i veterinærmedisinen”, Scandinavian “Direct injection of foreign DNA into mouse Veterinary Press, pp. 128-31. testis as a possible in vivo gene transfer system via epididymal spermatozoa”, Mol. Reprod. Dev. 61, 49-56. 15. O’Neill, G. (2002) “The contraceptive plague”, New Scientist 10 August 2002, 4-5. 7. Miller, D.G., Rutledge, E.A. & Russell, D.W. (2002) “Chromosomal effects of adeno- 16. Nowak, R. (2002) “Gene warfare”, New associated virus vector integration”, Nature Scientist 11 May 2002, p. 6. Genetics 30, 147-8. 17. Bouchie, A. (2003) Nature Biotechnology 8. Check, E. (2003) “Second cancer case halts 21, 11. gene-therapy trials”, Nature 421, 305.

44 Ministry of the Environment P.O. Box 8013 Dep. 0030 Oslo

Your ref.: 2001/2807 N/AMy Our ref.: 01/87 Date: 26 February 2003

Regulation of DNA vaccines and gene therapy on animals

In its letter of 02.10.01, the Ministry of the Environment asked the Biotechnology Advisory Board to discuss how DNA vaccines and gene therapy on animals should be regulated and what should be the status of DNA-treated animals under such a regulatory system.

New and promising methods for preventing and combating disease are currently being developed, in the form of DNA vaccines and gene therapy, which are based on the transfer of genetic material to cells of the body. These methods are, therefore, approaching the methods used to genetically modify organisms. This raises the question of whether an animal is to be considered a genetically modified organism (GMO) when it receives DNA vaccines or gene therapy. In that case, the animal would be regulated under the Gene Technology Act.

The Biotechnology Advisory Board raised this issue for the first time at an internal seminar in Namsos on 05.09.2001 and has, following the enquiry made by the Ministry of the Environment, drafted a separate document highlighting the various aspects of the problem (see enclosure). The Board has subsequently discussed possible regulatory methods for the DNA treatment of animals in the light of the internal seminar and the discussion paper and has arrived at a recommended regulatory method.

The concept of “DNA treatment” Vaccines and gene therapy may be administered in the form of genetically modified viruses and bacteria. However, the concept of “DNA treatment” is here used as the transfer of nucleic acids – either DNA or RNA – in ways that do not entail the use of genetically modified organisms. The term covers both DNA vaccines and gene therapy for purposes other than influencing the immune system. In the discussion on a regulatory system for the use of DNA vaccines and gene therapy on animals, it may be useful to apply a separate designation to define the transfer of nucleic acids that takes place without the use of genetically modified viruses or bacteria.

The Norwegian Biotechnology Act uses the term “gene therapy” to cover all intentional transfers of genetic material to cells in the human body. This use of the concept includes DNA vaccines and gene therapy in the form of naked or encapsulated DNA, as well as the transfer of genetic material by means of genetically modified viruses and bacteria. In the context of a statutory regulation of DNA vaccines and gene therapy on animals, the term “gene therapy” could be defined and used in the same manner, if conformity between the Gene Technology and Biotechnology Acts is required.

Regulating DNA treatment in a separate chapter of the Gene Technology Act Since DNA treatment may give rise to a wide range of possible consequences, the Biotechnology Advisory Board, with the exception of two of its members, would like to see DNA treatment regulated in accordance with the principles laid down in the Gene Technology Act. Board members Grethe Evensen and Christina Abildgaard, however, maintain that it would be natural

1 for Norway to follow the same regulatory practice as that adopted by the European Union in respect of DNA vaccines and gene therapy as medicinal products (see below).

DNA treatment may entail serious environmental consequences, also when the genetic material is not inherited. Furthermore, since DNA treatment involves a series of different methods and genetic constructs, it is important to apply a system of case-by-case evaluation, ensuring a thorough environmental impact assessment. The majority of the Board’s members recommends, therefore, that DNA treatment is regulated in the form of a separate chapter of the Gene Technology Act, specifying the rules that apply to DNA treatment. The Board believes that such a chapter could be drafted in such a way that it would apply to the DNA treatment of all types of multicellular organisms, including plants, and not only to animals. Board members Grethe Evensen and Christina Abildgaard support a system of case-by-case evaluation of applications, but argue that regulation through existing legislation would safeguard these aspects as far as medicinal products are concerned.

The production and use of genetically modified organisms is currently divided into contained use and deliberate release respectively, where the requirements applying to contained use must be fulfilled so as not to be considered a deliberate release. The Biotechnology Advisory Board suggests that this distinction is maintained. The rules and evaluations applying to contained use and deliberate release could, on the whole, be the same for DNA treatment as for the genetic modification of organisms, but the barriers against the spread of genetic material and the criteria for contained use could differ somewhat.

1) It might be useful to consider whether Section 5 of the Animal Welfare Act should be amended to also cover DNA treatment, and not only genetic modification related to breeding.

Definition of a GMO The Gene Technology Act’s definition of a “genetically modified organism” leaves room for interpretation as far as DNA-treated animals are concerned. A similar room for interpretation is also to be found in EU Directive 2001/18/EC related to the deliberate release of genetically modified organisms.

The Biotechnology Advisory Board would not like to see the concept of “genetically modified organism” watered down and supports, therefore, the position that the use of DNA vaccines and gene therapy on animals should not, as a rule, be regarded as genetic modification. However, the Board would like to retain the possibility of exercising discretion in case-by-case evaluations and would, therefore, advise against stipulating an absolute requirement of heredity for the animal to be termed genetically modified. If it can be shown probable either 1) that the added genetic material will be inherited by the offspring, 2) that the genetic material will pose a risk to health or the environment if it is inherited, 3) that the genetic material, through recombination, can result in organisms with new, unwanted properties, or 4) that the genetic material will give the organism properties that will lead to a public outcry, the Board would recommend that one allows for the possibility of defining the organism as genetically modified, with the subsequent manda-

1) Section 5 of the Animal Welfare Act. Breeding It is prohibited to alter the heritable material of animals by means of gene technology methods or by means of traditional breeding methods if: 1. it makes the animal unfit for carrying out normal behaviour or if it adversely affects its physiological functions; 2. the animal is caused unnecessary suffering; 3. the alteration arouses general ethical concerns. It is prohibited to breed animals covered by the first paragraph.

2 tory application of labelling requirements. Subject to these criteria, most DNA vaccine plasmids would not render animals genetically modified, whereas certain forms of gene therapy would be covered by the criteria. Furthermore, DNA treatment, when used in the production of a genetically modified organism, would not be exempted from the rules currently in force.

The deliberate release of genetically modified organisms must be assessed for possible risk and, if necessary, the precautionary principle must be applied. This holds true under Norwegian law, the EU Directive 2001/18/EC and the Cartagena Protocol on biosafety. If Norway were to determine that the same rules should apply to DNA-treated animals, but without simultaneously classifying them as genetically modified organisms, this might be perceived as an exceptional regulation. Hence, the Board would recommend that the regulatory system is drafted in such a way that the requirement of risk assessment may be linked to the need to evaluate whether treatment would render the animal genetically modified or not. In this way, the risk assessment is covered by international rules currently in force.

The regulatory system could be worded in such way that it would also apply to the deliberate release of DNA-treated animals that have been imported alive after having undergone treatment. There should also be the possibility of assessing whether animals having received DNA vaccines and gene therapy in the form of living, genetically modified viruses or bacteria meet the criteria for a genetically modified organism, in order to avoid the use of genetically modified viruses instead of plasmids in instances where plasmids would involve a lesser environmental risk.

Regulating DNA treatment in relation to medicinal product regulation The Biotechnology Advisory Board would recommend regulation of the treatment itself, i.e. the use of DNA vaccines and gene therapy on animals. Since Norway is linked to the EU’s system for the evaluation and approval of medicinal products (EMEA), Norway cannot impose restrictions on marketing permission for DNA vaccines and gene therapy products that have been approved as veterinary medicinal products. However, their use may be restricted, if required.

DNA vaccines and gene therapy may also be administered in the form of genetically modified viruses or bacteria. Medicinal products that are also genetically modified organisms are exempted from the provisions of EU Directive 2001/18/EC on the placing on the market of GMOs, but an equivalent environmental risk assessment is required. This type of assessment also covers the animals and environment in which GMO medicinal products are to be used. Furthermore, it is stipulated in the related food and feed regulation that animals treated with genetically modified medicinal products are not to be considered as genetically modified.

DNA vaccines and gene therapy products that are not GMOs are not covered by the same statutory requirement for an environmental impact assessment. Hence, the animals to be treated and other organisms in their environment would not be subject to an environmental risk assessment equivalent to that of the GMO medicinal products. The majority of the Biotechnology Advisory Board holds that it is important to establish in law the requirement for an environmental risk assessment that relates to medicinal products based on nucleic acids, and that such assessments should be conducted by the same bodies that today evaluate the deliberate release of GMOs. The Board’s majority calls on the environmental authorities to strive to ensure that such a regime is implemented in the EU as well. This would ensure regard for the environment as well as confidence in producers.

Board members Grethe Evensen and Christina Abildgaard maintain that Norway, out of consideration for Norwegian trade and industry and its competitiveness, should await and follow the European Union’s coming practice for the regulation of animals treated with EMEA-authorised DNA vaccines and gene therapy products. A harmonized regulatory practice in Norway would be crucial to preserving the industrial competence that has been built up in Norway, as well as to

3 ensuring further efforts to develop products with the aim of obtaining permission for their use and marketing in Norway. Norway has a large fish-farming industry and approximately one half of the world market for fish vaccines. Hence, Norway should proceed with caution in this matter.

Application requirements Applications for the deliberate release of DNA-treated animals or for the treatment of animals that are in the environment will, in principle, require an environmental impact assessment and will be subject to a case-by-case evaluation, in accordance with the same principles as for the deliberate release of genetically modified organisms. As experience with DNA-treated animals gradually accumulates, the requirements might possibly be adjusted by means of administrative regulations. So as to enable a practical, case-by-case evaluation of the use of marketed products, the Board would suggest that the producer, or the importer or the potential users should be allowed to apply for authorisation to use the product. To maintain proper control of the number of animals receiving treatment, a reporting system could be set up. For experimental activities and clinical trials using non-approved products, the project manager would be the applicant.

For DNA-treatments of animals, it is necessary to have information about the duration and spread of the DNA in the animal, the risk of integration, the number of animals receiving treatment, etc. Environmental impact assessment requirements in applications for DNA treatment could be based on the guidelines already developed by the EMEA, WHO and FDA for DNA vaccines and gene therapy, and on the environmental impact assessment applying to the deliberate release of genetically modified organisms.

Labelling The Biotechnology Advisory Board wishes to avoid a “watering down” of the concept of GMO and favours, therefore, the position that animals having received DNA vaccines and gene therapy for veterinary medical purposes would not be covered by the GMO concept unless they fulfilled the criteria for classifying an animal as a genetically modified organism, as outlined above.

The introduction of a separate labelling category for DNA-treated animals might be a relevant option. In that connection, the Biotechnology Advisory Board believes that emphasis should be placed on the wish of consumers for relevant information – not only about the end product, but also about the production method. The implications of the EEA agreement should also be taken into account. It should further be considered whether a separate labelling category or a negative labelling ought to be internationally recognized before being implemented in practice. Information could also be made available to consumers in ways other than through direct labelling of the products. Furthermore, to ensure consumer confidence, requirements should be specified concerning the level of residual quantities of the added genetic material in the end product, similar to the rules on maximum residual limits of veterinary medicinal products in products of animal origin.

Yours sincerely

Werner Christie Sissel Rogne Chairman Director

Enclosure: Official in charge: Grethe S. Foss Discussion paper (manuscript 26.02.03) Senior advisor

4

Address: P.O.Box 522 Sentrum, NO-0105 Oslo, Norway • Tel: +47 24 15 60 20• Fax: +47 24 15 60 29 • e-mail: [email protected] • www.bion.no

The Norwegian Biotechnology Advisory Board