Stephen A. MorseIntroduction
The Biosafety and Biosecurity Section of Frontiers in Bioengineering and Biotechnology began in 2014 to serve as the forum to address many of the biosafety and biosecurity challenges posed by bioengineering and biotechnology. Although there are many areas to highlight in a Grand Challenges article, captured herein are a few topics that are of current and future concern to the field.
Frontier 1: expansion of high-containment laboratories
Anticipating the biosafety and biosecurity challenges to the global expansion of high containment laboratories was subject of an international workshop held more than a decade ago (National Academy of Sciences and National Research Council, 2012). The meeting covered: 1) Technological options to meet diagnostic and research needs; 2) Laboratory construction and commissioning; 3) Operational maintenance to provide sustainable capabilities, safety, and security; and 4) Measures for encouraging a culture of responsible conduct. Since that meeting, new, emerging and re-emerging infectious diseases such as SARS-CoV, MERS, Ebola, Zika, H5N1 avian influenza, SARS-CoV-2, and monkeypox have resulted in a further increase of high containment laboratories (Lentzos and Koblentz, 2023). The proliferation of the number of BSL-4 and BSL-3+ facilities have elevated risks as well as demand for improved training, standards, and oversight (Morrison and Simoneau, 2023).
Many of the high containment labs are in or planned for urban centers where an accidental or intentional release could be problematic (Klotz and Sylvester, 2014). In fact, several outbreaks have been linked or suggested to be the result of pathogens escaping from a laboratory: influenza H1N1 in 1977 (Palese, 2004), SARS-CoV in 2004 (Manheim and Lewis, 2022), and possibly SARS-CoV-2 (Gostin and Gronvall, 2023). It is also particularly concerning that several countries that have built or plan to build BSL-4 and BSL-3+ laboratories scored low on the metrics used to assess their biosafety and biosecurity capabilities (Lentzos and Koblentz, 2023).
From a global perspective, biorisk management has been proposed to address both biosafety and biosecurity in laboratories (Salerno and Gaudioso, 2015; Rodgers et al., 2021). Because biorisk management is oriented around performance, a laboratory in a low-resource country can implement it as effectively as one in a developed country. To help inexperienced laboratories as well as those with experience, the International Organization for Standardization (ISO) released ISO 35001:2019, a standard on biorisk management for laboratories that work with dangerous pathogens (https://www.iso.org/standard/71293.html). To further address these concerns, the World Health Organization (WHO) developed the “Global Guidance Framework for the Responsible Use of the Life Sciences: Mitigating Biorisks and Governing Dual-Use Research” (WHO, 2022). This biorisk management framework encompasses three core pillars: laboratory biosafety, laboratory biosecurity and the oversight of dual-use research (WHO, 2022).
Frontier 2: dual-use-research of concern
Dual-Use Research of Concern (DURC) is defined as “life science research that, based on current understanding, can be reasonably anticipated to provide knowledge, information, products, or technologies that could be misapplied to pose a significant threat with broad potential consequences to public health and safety, agricultural crops and other plants, animals, the environment, materiel, or national security” (Casadevall et al., 2015). DURC presents unique biosecurity hazards when compared to less risky biological research. Policies developed in the U.S., mandate risk-benefit assessments, oversight mechanisms, transparency, and collaborative efforts between government entities and research institutions to ensure the responsible conduct of government-funded high-risk research. Unfortunately, government funding now accounts for less than half of life science funding (https://www.economicstrategygroup.org/publication/seven-recent-developments/). Therefore, a new paradigm is needed to capture non-government-funded DURC such as the construction of infectious horsepox virus, a virus in the Orthopox genus, which is related to smallpox virus, funded by a U. S. pharmaceutical company (Noyce et al., 2018).
In the aftermath of the 2001 anthrax attack (Jernigan et al., 2002), significant concern about dual-use research was raised in scientific and government circles following the publication of two papers. In the first, Jackson et al. (2001) introduced the gene for interleukin 4 (IL-4) into mousepox virus in an effort to produce a contraceptive vaccine to control the wild mouse population. Unexpectedly, the mice inoculated with the recombinant virus died even though they were genetically resistant to mousepox virus infection or had been immunized against it. Thus, instead of creating a contraceptive, they created a virus with enhanced pathogenicity. It was feared that the findings could be a blueprint to construct a more virulent smallpox virus that would evade the immunity established by vaccination. The second paper (Cello et al., 2002) described the synthesis of full-length poliovirus cDNA by assembling oligonucleotides of plus and minus strand polarity. The synthetic poliovirus cDNA was transcribed by RNA polymerase into viral RNA, which translated and replicated in a cell-free extract resulting in the de novo synthesis of infectious poliovirus. The media framed this work as a recipe for the synthesis of viruses for use as a biological weapon. Others argued that poliovirus was relatively simple and that the synthesis of more complex viruses like influenza or smallpox would be far more challenging. However, technology advances rapidly, and within 3 years, the influenza virus responsible for the 1918 pandemic, which resulted in an estimated 20 million deaths, was reconstructed using reverse genetics (Trumpy et al., 2005).
In 2005, a National Science Advisory Board for Biosecurity (NSABB) was established to provide advice, guidance, and leadership to the U. S. government regarding oversight of dual use life sciences research. Among its accomplishments, the NSABB developed a definition for DURC and identified seven specific categories of experiments that were likely to be worrisome (Imperiale and Casadevall, 2015). These are experiments that would: “1) Enhance the harmful consequences of a biological agent or toxin; 2) Disrupt immunity or the effectiveness of an immunization without clinical and/or agricultural justification; 3) Confer to a biological agent or toxin resistance to clinically and/or agriculturally useful prophylactic or therapeutic interventions against that agent or toxin or facilitates its ability to evade detection methods; 4) Increases the stability of, transmissibility of, or ability to disseminate a biological agent or toxin; 5) Alters the host range or tropism of a biological agent or toxin; 6) Enhances the susceptibility of a host population; and 7) Generates a novel pathogenic agent or toxin or reconstitutes an eradicated or extinct biological agent” (Casadevall et al., 2014a). These categories of experiments could help scientists, editors, and reviewers in deciding whether their proposed, ongoing work, or submitted manuscript is DURC.
In 2012, research groups in the Netherlands (Herfst et al., 2012), U.S. and Japan (Imai et al., 2012) produced mutated variants of the highly pathogenic H5N1 avian influenza virus that could infect ferrets via the airborne route. Though their stated aim was to determine what mutations could alter the host range of the virus, in order to be better prepared if such changes occurred in nature, the work provoked a heated debate in public and scientific circles, as to whether it should have been carried out at all, both for biosafety (the virus could escape the laboratory) and biosecurity (a blueprint for those with nefarious intent) reasons (Wain-Hobson, 2014; Nixdorf, 2024). This type of DURC has been termed gain-of-function (GOF) research because the modified H5N1 virus gained the ability to infect ferrets, which it did not have before (Casadevall et al., 2014b).
To address the biosecurity issues raised by these articles, some governments, academic institutions, professional societies, and journals subsequently developed policies concerning the publication of DURC. Science, Nature Publishing Group and the American Society for Microbiology journal group developed dual-use review policies (Resnick et al., 2011; Casadevall et al., 2015). However, results of a 2011 survey of 155 journals indicated that relatively few (N = 12) had a written dual-use policy and only nine said they had experience reviewing dual-use research in the past 5 years (Resnick et al., 2011). It is not known if the situation has improved over the ensuing years.
A more recent and problematic development for biosecurity is Open Science, which is a set of practices that aim to improve the reliability and efficiency of scientific research and are generally characterized by increased transparency (Smith and Sandbrink, 2022). Although, Open Science may encourage the exercise of best practices, there are also instances where it may also contribute to biosafety and biosecurity risk. Of particular importance to DURC, is the use of preprints, which are author-formatted articles publicly deposited in a repository. These preprints are a recent and novel way to disseminate microbiology research prior to formal peer review (Schloss, 2017). Preprints now account for 4% of all research articles (Yoshizawa et al., 2024). In one study, Malicki et al. (2020) found that only 68% of preprint servers provided some form of screening or moderation before the article was made public though not all of the screening involved dual-use, safety, or biosecurity-related criteria to mitigate the risk from publishing DURC.
Frontier 3: synthetic biology
In 2017, the U. S. Department of Defense requested that the National Academies of Sciences, Engineering, and Medicine (NASEM) address the changing nature of the biodefense threat in the age of synthetic biology. The final report was published in 2018 (NASEM, 2018). Synthetic biology [now called engineering biology (Di Euliis et al., 2024)] is a branch of science within biotechnology that comprises a broad range of methodologies from various disciplines that enable the modification of biological organisms. These engineering biology approaches have the potential to be used in ways that could change the presentation of a bioterrorist attack. The final report (NASEM, 2018) stated that “engineering biology expands what is possible by creating new bioweapons and also expands the range of actors who could undertake such efforts and decreases the time required.” Three potential capabilities were considered to be most concerning: (1) re-creating known pathogenic viruses, (2) making existing bacteria more dangerous, and (3) making harmful biochemicals via in situ synthesis. With regard to biochemicals, engineering biology blurs the line between biological and chemical weapons. It can allow the delivery of biochemicals by a biological agent (Galanie et al., 2015). It may also be possible to modulate human physiology and behavior in novel ways by potentially allowing the engineering of microorganisms, the microbiome, or immune system (Borzenkov et al., 1994). More recently, the integration of AI with engineering biology has introduced biosecurity challenges that will need to be addressed (De Haro, 2024).
DNA synthesis technologies are catalyzing rapid advances in engineering biology. While the promise of this technology is immense, so is its potential for intentional or accidental misuse. Nucleic acids underpin much of research and development in the life sciences and serves as a critical control point. In 2010, in the interest of biosecurity, the U. S. Department of Health and Human Services (HHS) issued screening guidance for commercial providers of synthetic double-stranded DNA (dsDNA), which called on providers to voluntarily screen all orders (as cited by Diggans and Leproust, 2019). Subsequently, members of the International Gene Synthesis Consortium (IGSC), which represents about 80% of global gene synthesis capacity, implemented the Harmonized Screening Protocol (HSP). IGSC members screen every gene order against the DNA sequences in a common curated Regulated Pathogen Data Base, and against all entries found in internationally coordinated sequence databases (e.g., NCBI/GenBank) (IGSC, 2017).
In October 2023, Executive Order 14110 was issued and included a section, in response to reducing the risks of misuse of synthetic nucleic acids by improving associated biosecurity measures (Presidential Documents, 2023). In response, the new guidance expanded the definition of Sequences of Concern (SOCs) to include all sequences that contribute to pathogenicity or toxicity, whether from regulated (e.g., Select Agents) or unregulated agents. To ensure that synthetic nucleic acids are distributed and used responsibly, the guidance recommends that providers and third-party vendors verify the legitimacy of each recipient of nucleic acids containing SOCs, and that all parties maintain records of SOC transfers. The guidance also recommends that the manufacturers of benchtop nucleic acid synthesis equipment verify the legitimacy of their customers. The framework incorporates and supplements the 2023 HHS guidance (HHS, 2023).
Conclusion
Advances in biotechnology and developments in artificial intelligence (AI) have resulted in a rapidly evolving threat landscape, which has presented new challenges for preventing the malicious, reckless, or accidental misuse of the life sciences.
Author contributions
SM: Writing–original draft, Writing–review and editing.
Funding
The author(s) declare that no financial support was received for the research, authorship, and/or publication of this article.
Conflict of interest
Author SM was employed by IHRC, Inc.
Generative AI statement
The author(s) declare that no Generative AI was used in the creation of this manuscript.
Publisher’s note
All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.
References
Borzenkov, V. M., Pomerantsev, A. P., Pomerantseva, O. M., and Ashmarin, I. P. (1994). A study of nonpathogenic Francisella, Brucella, and Yersinia strains as producers of recombinant β-endorphin. Bull. Exp. Biol. Med. 117 (6), 615–618. doi:10.1007/bf02444334
Casadevall, A., Dermody, T. S., Imperiale, M. J., Sandri-Goldin, R. M., and Shenk, T. (2014a). On the need for a national board to assess dual use research of concern. J. Virol. 88 (12), 6535–6537. doi:10.1128/JVI.00875-14
Casadevall, A., Dermody, T. S., Imperiale, M. J., Sandri-Goldin, R. M., and Shenk, T. (2015). Dual-use research of concern (DURC) review at American Society for Microbiology journals. mBio 6 (4), 012366–e1315. doi:10.1128/mBio.01236-15
Casadevall, A., Howard, D., and Imperiale, M. J. (2014b). An epistemological perspective on the value of gain-of-function experiments involving pathogens with pandemic potential. MBio 5, e01875–e01814. doi:10.1128/mBio.01875-14
Cello, J., Paul, A. V., and Wimmer, V. (2002). Chemical synthesis of poliovirus cDNA: generation of infectious virus in the absence of natural template. Science 297, 1016–1018. doi:10.1126/science.1072266
De Haro, L. P. (2024). Biosecurity risk assessment for the use of artificial intelligence in synthetic biology. Appl. Biosaf. 29 (2), 96–107. doi:10.1089/apb.2023.0031
DiEuliis, D., Imperiale, M. J., and Berger, K. M. (2024). Biosecurity assessments for emerging transdisciplinary biotechnologies: revisiting biodefense in an age of synthetic biology. Appl. Biosaf. 29 (3), 123–132. doi:10.1089/apb.2024.0005
Diggans, J., and Leproust, E. (2019). Next steps for access to safe, secure DNA synthesis. Front. Bioeng. Biotechnol. 7, 86. doi:10.3389/fbioe.2019.00086
Galanie, S., Thodey, K., Trenchard, I. J., Interrante, M. F., and Smolke, C. D. (2015). Complete biosynthesis of opioids in yeast. Science 349 (6252), 1095–1100. doi:10.1126/science.aac9373
Gostin, L. O., and Gronvall, G. K. (2023). The origins of covid-19 – why it matters (and why it doesn’t). New Eng. J. Med. 388, 2305–2308. doi:10.1056/NEJMp2305081
Herfst, S., Schrauwen, E. J., Linster, M., Chutinimikul, S., de Wit, E., Munster, V. J., et al. (2012). Airborne transmission of influenza A/H5N1 virus between ferrets. Science 336, 1534–1541. doi:10.1126/science.1213362
Imai, M., Watanabe, T., Hatta, M., Das, S. C., Ozawa, M., Shinya, K., et al. (2012). Experimental adaptation of an influenza H5 HA confers respiratory droplet transmission to a reassortant H5 HA/H1N1 virus in ferrets. Nature 486, 420–428. doi:10.1038/nature10831
Imperiale, M. J., and Casadevall, A. (2015). A new synthesis for dual use research of concern. PLoS Med. 12 (4), e1001813. doi:10.1371/journal.pmed.1001813
International Gene Synthesis Consortium (2017). Harmonized screening Protocol v2.0. Available at: https://genesynthesisconsortium.org/wp-content/uploads/IGSCHarmonizedProtocol11-21-17.pdf (Accessed November 12, 2024).
Jackson, R. J., Ramsay, A. J., Christensen, C. D., Beaton, S., Hall, D. F., and Ramshaw, I. A. (2001). Expression of mouse interleukin-4 by a recombinant ectromelia virus suppresses cytolytic lymphocyte responses and overcomes genetic resistance to mousepox. J. Virol. 75, 1205–1210. doi:10.1128/jvi.75.3.1205-1210.2001
Jernigan, D. B., Raghunathan, P. L., Bell, B. P., Brechner, R., Bresnitz, E., Butler, J. C., et al. (2002). Investigation of bioterrorism-related anthrax, United States, 2001: epidemiologic findings. Emerg. Infect. Dis. 8 (10), 1019–1028. doi:10.3201/eid0810.020353
Klotz, L. C., and Sylvester, E. J. (2014). The consequences of a lab escape of a potential pandemic pathogen. Front. Public Health 2, 116. doi:10.3389/fpubh.2014.00116
Lentzos, F., and Koblentz, G. D. (2023). High consequence bio labs: growing risks and lagging governance. Release Glob. BioLabs 2023 Rep. Available at: https://static1.squarespace.com/static/62fa334a3a6fe8320f5dcf7e/t/6414862df5bf8f14cec76598/1679066680422/Global-Biolabs-2023-Report.pdf (Accessed March 16, 2023).
Malicki, M., Jeroncic, A., ter Riet, G., Bouter, L. M., Ioannidis, J. P. A., Goodman, S. N., et al. (2020). Preprint servers’ policies, submission requirements, and transparency in reporting and research integrity recommendations. JAMA 324, 1901–1903. doi:10.1001/jama.2020.17195
Manheim, D., and Lewis, G. (2022). High-risk human-caused pathogen exposure events from 1975-2016 [version 2; peer review: 2 approved]. F1000 Res. 10, 752. doi:10.12688/f1000research.55114.2
Morrison, J. S., and Simoneau, M. (2023). Eight commonsense actions on biosafety and biosecurity. CSIS Briefs. Available at: www.CSIS.org.
National Academies of Sciences, Engineering, and Medicine (2018). Biodefense in the age of synthetic biology. Washington, D.C.: The National Academies Press.
National Academy of Sciences and National Research Council (2012). Biosecurity challenges of the global expansion of high-containment biological laboratories: summary of a workshop. Washington, D. C.: The National Academies Press.
Nixdorff, K. (2024). “The problem of dual use in the twenty-first century, p. 69-81,” in Essentials of biological security. A global perspective. Editors L. Shang, W. Zhang, and M. Dando 1st Edn (Hoboken, New Jersey: John Wiley and Sons, Inc.).
Noyce, R. S., Lederman, S., and Evans, D. H. (2018). Construction of an infectious horsepox virus vaccine from chemically synthesized DNA fragments. PLoS ONE 13, e0188453. doi:10.1371/journal.pone.0188453
Presidential Documents (2023). Executive order of october 30, 2023: safe, secure, and trustworthy development and use of artificial intelligence. Fed. Reg. 88 (210), 75191–75226.
Resnick, D., Barner, D. D., and Dinse, G. E. (2011). Dual-use review policies of biomedical research journals. Biosecur. Bioterror. 9 (1), 49–54. doi:10.1089/bsp.2010.0067
Rodgers, J., Lentzos, F., Koblentz, G. D., and Ly, M. (2021). How to make sure the labs researching the most dangerous pathogens are safe and secure. Bull. At. Sci. Available at: https: //thebulletin.org/2021/07/how-to-make-sure-the-labs-researching-the-most-dangerous-pathogens-are safe-and-secure/.
R. M. Salerno, and J. Gaudioso (eds) (2015). Laboratory biorisk management: biosafety and biosecurity. CRC Press, Florida: Boca Raton.
Smith, J. A., and Sandbrink, J. B. (2022). Biosecurity in an age of open science. PLoS Biol. 20 (4), E3001600. doi:10.1371/journal.pbio.3001600
Trumpy, T. M., Basler, C. F., Aguilar, P. V., Zeng, H., Solorzano, A., Swayne, D. E., et al. (2005). Characterization of the reconstructed 1918 Spanish influenza pandemic virus. Science 310, 77–80. doi:10.1126/science.1119392
U. S. Department of Health and Human Services (2023). Screening framework guidance for providers and users of synthetic nucleic acids. Available at: https://aspr.hhs.gov/Preparedness/legal/guidance/syndna/Pages/default.aspx.
Wain-Hobson, S. (2014). The irrationality of GOF avian influenza virus research. Front. Public Health 2, 77. doi:10.3389/fpubh.2014.00077
World Health Organization (2022). Global guidance framework for the responsible use of the life sciences: mitigating biorisks and governing dual-use research. Geneva: World Health Organization.
Keywords: high-containment laboratories, dual-use-research, biorisk management, synthetic biology, nucleic acid synthesis
Citation: Morse SA (2025) Grand challenge in biosafety and biosecurity. Front. Bioeng. Biotechnol. 12:1538723. doi: 10.3389/fbioe.2024.1538723
Received: 03 December 2024; Accepted: 13 December 2024;
Published: 07 January 2025.
Edited and reviewed by:
Ranieri Cancedda, Independent Researcher, Genova, ItalyCopyright © 2025 Morse. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
*Correspondence: Stephen A. Morse, c21vcnNlODhAYW9sLmNvbQ==