Quality in Preclinical Research
We study factors which affect the translational predictiveness of preclinical research in general, and stroke research in particular
Ulrich Dirnagl ORCID
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A substantial fraction of published preclinical research results cannot be reproduced, while translation of spectacularly successful novel treatments in experimental models of disease too often fail when tested in clinical trials. After decades of own research in experimental stroke research, and a number of futile efforts to translate our findings into benefit for our patients I have turned my attention towards the complex causes of the failures, and to developing potential remedies. To this end we use meta-research – that is we quantitatively investigate the prevalence and effects of low internal, external, construct, and statistical validity (e.g. via meta analytic approaches) of preclinical research, design and implement tools to improve research quality in preclinical medicine (e.g. novel statistical approaches, quality management systems, error management, electronic laboratory notebook, etc.), evaluate the effectiveness of such implementations in the field (using quantitative and qualitative methods), and perform animal experiments to investigate how external validity of stroke research can be improved (e.g. by using ‘Wildlings’) and made thus more predictive for informing clinical translation.
SPF husbandry as a challenge to external and construct validity in preclinical neuroscience research
To date, preclinical stroke research did not succeed in translating any drug-based or non-pharmacological interventions for the treatment of brain ischemia into clinical practice. Consequently, there is an urgent need to develop effective therapies. Stroke is a vascular disease, and vascular remodelling plays an important role in both its acute and chronic phase. Importantly, immunological processes play a significant role in the pathophysiology of ischemic cascades (1), from the acute events following arterial occlusion to regenerative processes underlying post-ischemic tissue repair (2). Thus, mouse models of vascular diseases need to recapitulate human immune responses (3). However, the current standard laboratory mice used in basic research and preclinical trials are specific-pathogen-free animals (SPF) with immature immune systems when compared to free-living mice or humans due to lack of natural interactions with microbiota and pathogens that humans experience on a daily basis. Yet feral mice or laboratory animals with mature immune systems have not been investigated in the context of stroke. We speculate that raising mice under (too) clean SPF conditions is an important reason for translational failure, in particular with respect to vascular mechanisms as they are critically modulated by the immune system. Work by Stephan Rosshart and others has shown that laboratory mice are too far removed from natural, “wild” environmental conditions to faithfully mirror the physiology of free-living mammals like humans. Mammals and their immune systems evolved to survive and thrive in a microbial world and consequently behave differently in an unnatural, ‘clean’ environment. This distorts how the immune system of laboratory mice develops and functions, leading to false assumptions of how our own “wild” immune system works (4-8).
Thus, any treatment strategy targeting immunological mechanisms developed in rodents may fail if we continue to use conventional SPF mice that do not resemble fundamental human traits such as a mature immune system. We speculate that vascular and neuronal remodelling after stroke in conventional SPF mice is heavily biased by the absence of natural host-microbiota and pathogen interactions, thereby causing the high degree of discrepancy in outcomes between humans and SPF rodents.
We posit that wildling mice are better suited to study disease pathophysiology than conventional SPF mice, at the very least in the context of immunological research questions. Given the broad impact of microbiota on overall host physiology (9-11) as well as the clear link between immunity and vascular diseases such as stroke, we hypothesize that the ‘stroke-phenotype’ (infarct volume, mortality, sensory-motor deficit and recovery from it, post-stroke angiogenesis and response to microbiota) of wildlings will better resemble that of humans and in contrast significantly deviate from that of standard laboratory mice. Our proposal might not only lead to relevant results in the field of vascular diseases, but also open up a promising window of opportunity of better translational prediction for other CNS diseases, e.g. caused by neuroinflammation or/and neurodegeneration.
(in collaboration with AG Molecular Stroke Research, funded by Excellence Cluster Neurocure)
RE-PLACE Replacing paper note books with electronic ones
RE-PLACE studies the process of shifting from analogue documentation of research processes in paper lab books to so-called electronic laboratory notebooks (ELN). ELNs do not only digitalize the documentation of researchers’ experiments, but also link their notes directly to research data generated in experiments. On the initiative of BIH QUEST Center, the Berlin Institute of Health ( BIH ) is currently implementing an institutional programme to replace analogue research documentations with ELN Labfolder. To evaluate this implementation process, Charité - Universitätsmedizin Berlin together with Otto-von-Guericke-University Magdeburg started the BMBF -funded research project RE-PLACE in August 2020
Vollert J, Macleod M, Dirnagl U, Kas MJ, Michel MC, Potschka H, Riedel G, Wever KE, Würbel H, Steckler T, Rice ASC. The EQIPD framework for rigor in the design, conduct, analysis and documentation of animal experiments. Nat Methods. 2022 Sep 5. doi: 10.1038/s41592-022-01615-y.
Hülsemann M, Wiebach J, Drude NI, Kniffert S, Behm L, Hönzke K, Baumgardt M, Hippenstiel S, Hocke AC, Dirnagl U, Tölch U. Introducing quality measures in an academic research consortium: Lessons and recommendation from implementing an ad hoc quality management system for organ model research: Lessons and recommendation from implementing an ad hoc quality management system for organ model research. EMBO Rep. 2022 Aug 3;23(8):e55095. doi: 10.15252/embr.202255095. Epub 2022 Jul 25.
Dirnagl U, Duda GN, Grainger DW, Reinke P, Roubenoff R. Reproducibility, relevance and reliability as barriers to efficient and credible biomedical technology translation. Adv Drug Deliv Rev. 2022 Mar;182:114118. doi: 10.1016/j.addr.2022.114118
Dirnagl U, Bannach-Brown A, McCann S. External validity in translational biomedicine: understanding the conditions enabling the cause to have an effect. EMBO Mol Med. 2022 Feb 7;14(2):e14334. doi: 10.15252/emmm.202114334. Epub 2021 Dec 20.
Bespalov A, Bernard R, Gilis A, Gerlach B, Guillén J, Castagné V, Lefevre IA, Ducrey F, Monk L, Bongiovanni S, Altevogt B, Arroyo-Araujo M, Bikovski L, de Bruin N, Castaños-Vélez E, Dityatev A, Emmerich CH, Fares R, Ferland-Beckham C, Froger-Colléaux C, Gailus-Durner V, Hölter SM, Hofmann MC, Kabitzke P, Kas MJ, Kurreck C, Moser P, Pietraszek M, Popik P, Potschka H, Prado Montes de Oca E, Restivo L, Riedel G, Ritskes-Hoitinga M, Samardzic J, Schunn M, Stöger C, Voikar V, Vollert J, Wever KE, Wuyts K, MacLeod MR, Dirnagl U, Steckler T. Introduction to the EQIPD quality system. Elife. 2021 May 24;10:e63294. doi: 10.7554/eLife.63294
Drude NI, Martinez Gamboa L, Danziger M, Dirnagl U, Toelch U. Improving preclinical studies through replications. Elife. 2021 Jan 12;10:e62101. doi: 10.7554/eLife.62101.
Emmerich CH, Gamboa LM, Hofmann MCJ, Bonin-Andresen M, Arbach O, Schendel P, Gerlach B, Hempel K, Bespalov A, Dirnagl U, Parnham MJ. Improving target assessment in biomedical research: the GOT-IT recommendations. Nat Rev Drug Discov. 2020 Nov 16:1-18. doi: 10.1038/s41573-020-0087-3
Percie du Sert N, Hurst V, Ahluwalia A, Alam S, Avey MT, Baker M, et al. The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research. PLOS Biol. 2020 Jul 14 [cited 2020 Jul 15];18(7):e3000410. https://dx.plos.org/10.1371/journal.pbio.3000410
Dirnagl, U. (2020). Resolving the Tension Between Exploration and Confirmation in Preclinical Biomedical Research. In Handbook of Experimental Pharmacology (Vol. 257, pp. 71–79). Springer. https://doi.org/10.1007/164_2019_278
Piper SK, Grittner U, Rex A, Riedel N, Fischer F, Nadon R, Siegerink B, Dirnagl U.Exact replication: Foundation of science or game of chance? PLoS Biol. 2019 Apr 9;17(4):e3000188 doi: 10.1371/journal.pbio.3000188
Dirnagl U. Rethinking research reproducibility. EMBO J. 2019 Jan 15;38(2). pii: e101117.
Dirnagl U, Kurreck C, Castaños-Vélez E, Bernard R. (2018) Quality management for academic laboratories: burden or boon? Professional quality management could be very beneficial for academic research but needs to overcome specific caveats. EMBO Rep.19. pii: e47143.
Yarborough M, Bredenoord A, D'Abramo F, Joyce NC, Kimmelman J, Ogbogu U, Sena E, Strech D, Dirnagl U. The bench is closer to the bedside than we think: Uncovering the ethical ties between preclinical researchers in translational neuroscience and patients in clinical trials. PLoS Biol. 2018 Jun 6;16(6):e2006343.
Neumann K, Grittner U, Piper S, Rex A, Florez-Vargas O, Karystianis G, Schneider A, Wellwood I, Siegerink B, Ioannidis JPA, Kimmelman J, Dirnagl U (2017) Increasing efficiency of preclinical research by group sequential designs. Plos Biol 15(3):e2001307 DOI: 10.1371/journal.pbio.2001307
Dirnagl U, Przesdzing I, Kurreck C, Major S. (2016) A laboratory critical incident and error reporting system for experimental biomedicine. Plos Biol 14: e2000705
Dirnagl U (2016) Thomas Willis Lecture: Is translational stroke research broken, and if so, how can we fix it? Stroke 47:2148-53
Holman C, Piper SK, Grittner U, Diamantaras AA, Kmmelman J, Siegerink B, Dirnagl U (2016) Where Have All the Rodents Gone? The Effects of Attrition in Experimental Research on Cancer and Stroke. Plos Biol 4(1):e1002331 DOI: 10.1371/journal.pbio.1002331
Begley CG, Buchan AM, Dirnagl U (2015). Robust research: Institutions must do their part for reproducibility. Nature 525:25-7. Kimmelman J, Mogil JS, Dirnagl U (2014) Distinguishing between exploratory and confirmatory preclinical research will improve translation. Plos Biol PLoS Biol 12(5): e1001863.
Dirnagl U, Endres M (2014) Found in translation - Preclinical stroke research predicts human pathophysiology, clinical phenotypes, and therapeutic outcomes. Stroke 2014; 45: 1510-1518