Animal Models of Human Pathology: Revision, Relevance and Refinements (4th Edition)

A special issue of Biomedicines (ISSN 2227-9059). This special issue belongs to the section "Molecular and Translational Medicine".

Deadline for manuscript submissions: 15 July 2026 | Viewed by 7387

Editor


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Guest Editor
Institute of Pathology, Faculty of Medicine, University of Ljubljana, Ljubljana, Slovenia
Interests: challenges in animal model research; translation; unbiased reporting of animal model characteristics and results; ethical justification
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Special Issue Information

Dear Colleagues,

The use of animal models of human pathology is accepted on the assumption that it benefits humans. However, recent publications have shown that research on animals faces serious challenges. The increasing number of potential targets, molecular pathways, and treatment strategies that have been recognized as promising in animal models have failed when translated into human trials. We have reached the point where the clinical relevance of animal models needs urgent clarification.

Multiple methodological problems in animal research have already been exposed, such as poor experimental design, inadequate use of fundamental statistical principles (i.e., randomization, blinding, inadequate power, inadequate sample size, pseudo-replication, etc.), and nontransparent reporting, which results in low scientific validity and irreproducibility of results. However, research on animal models also requires comprehensive knowledge about the model, as well as an understanding of the complex pathogenesis of diseases, which involves both local and systemic effects in the body. Every animal model has its own characteristics, advantages and limitations. There are factors specific to the disease or animal model that can influence not only the severity of the disease but also underlying mechanisms, and, when these factors are not taken into account, research may result in the discovery of new targets and disease pathways that are of no scientific or clinical value. There are also animal-model-specific factors that can seriously affect the results and lead to false conclusions and failed translation. Although the overall animal health and welfare issues—such as animal clinical state, morbidity, mortality, humane endpoints and humane interventions, whole body necropsy findings, sampling principles and pathohistological diagnosis—are of vital importance in the interpretation of the molecular mechanisms or treatment strategies in an organism, this information is usually lacking or rarely properly addressed in animal model studies.

The purpose of this Special Issue is thus to promote high-quality papers of basic research using animal models to understand diseases and underlying mechanisms or to investigate new treatment strategies in various human diseases such as cancer, bowel diseases, kidney injury, and Parkinson’s disease, among others. New approaches towards the use of animal models or refinements of particular animal models of human pathology, as well as methodological and welfare principles (such as experimental design and welfare or supportive measures in animal models), are also welcome.

Dr. Martina Perše
Guest Editor

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Keywords

  • animal models
  • biomarkers
  • human disease
  • pathology
  • translation
  • mechanisms
  • nephrology
  • gastroenterology
  • urology
  • neuroscience
  • cancer
  • experimental design
  • reproducibility
  • refinements

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Published Papers (7 papers)

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Research

Jump to: Review

23 pages, 22213 KB  
Article
Comparative Evaluation of Analgesics in a Murine Bile Duct Ligation Model
by Emily Leitner, Tim Schreiber, Hanna Krug, Praveen Vasudevan, Simone Kumstel, Lisa Ernst, René Hany Tolba, Brigitte Vollmar and Dietmar Zechner
Biomedicines 2025, 13(12), 3034; https://doi.org/10.3390/biomedicines13123034 - 10 Dec 2025
Viewed by 914
Abstract
Background: Reliable analgesia is essential to ensure animal welfare and experimental validity in preclinical disease models. However, evidence on the efficacy and side effects of analgesics remains limited. This study investigated the effects of three commonly used analgesics on animal well-being in a [...] Read more.
Background: Reliable analgesia is essential to ensure animal welfare and experimental validity in preclinical disease models. However, evidence on the efficacy and side effects of analgesics remains limited. This study investigated the effects of three commonly used analgesics on animal well-being in a murine model of cholestasis. Methods: Thirty male C57BL/6J mice underwent transmitter implantation followed by bile duct ligation (BDL) and received continuous metamizole (3 g/L), tramadol (1 g/L), or carprofen (0.15 g/L) via drinking water before and after surgery. Welfare was evaluated using multiple parameters, including body weight, a distress score, drinking volume, burrowing and nesting behavior, mouse grimace scale (MGS), and telemetric data (heart rate, heart rate variability: SDNN and RMSSD, core body temperature, and locomotion). Additionally, liver and gastrointestinal tissues were analyzed histologically for necrosis and immune cell infiltration. Results: Even prior to surgery, analgesic-specific reductions in body weight, drinking behavior, and burrowing and nesting activity were observed. After transmitter implantation, metamizole treatment led to significantly reduced body weight, drinking volume, and locomotion compared to the other two analgesics. Following BDL, all treatment groups exhibited pronounced distress, weight loss, and reduced activity. Tramadol treatment resulted in slightly improved MGS and SDNN values, indicating minor benefits without sustained welfare restoration. In contrast, carprofen treatment was associated with reduced survival and inflammatory alterations in the forestomach. Conclusions: None of the tested analgesic regimens fully restored animal welfare after BDL. However, tramadol provided modest advantages, suggesting it may represent the most suitable option among the tested analgesics for the BDL model. Full article
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28 pages, 8479 KB  
Article
Multiparametric Detection of Effects of TILs and Oncolytic Virotherapy on Xenograft Mouse Model of Glioblastoma
by Gaukhar M. Yusubalieva, Daria A. Chudakova, Polina G. Shirokikh, Diana V. Yuzhakova, Elena B. Kiseleva, Daria A. Sachkova, Varvara V. Dudenkova, Daria P. Kirsova, Maria S. Myzina, Elvira P. Yanysheva, Alexander V. Panov, Natalia F. Zakirova, Anastasia V. Poteryakhina, Alexander S. Semikhin, Alexander A. Kalinkin and Vladimir P. Baklaushev
Biomedicines 2025, 13(12), 2977; https://doi.org/10.3390/biomedicines13122977 - 4 Dec 2025
Viewed by 1243
Abstract
Background/Objectives: Glioblastoma (GBM) is an aggressive primary brain tumor with dismal prognosis and limited treatment options. Immunotherapy, including personalized approaches using tumor-infiltrating lymphocytes (TILs) and allogeneic natural (NK) or engineered killer cells (chimeric antigen receptor NK, NK-CAR), and oncolytic viruses (OV), has shown [...] Read more.
Background/Objectives: Glioblastoma (GBM) is an aggressive primary brain tumor with dismal prognosis and limited treatment options. Immunotherapy, including personalized approaches using tumor-infiltrating lymphocytes (TILs) and allogeneic natural (NK) or engineered killer cells (chimeric antigen receptor NK, NK-CAR), and oncolytic viruses (OV), has shown some potential in GBM. Combining different therapeutic strategies may enhance treatment efficacy. Here, we present a xenograft GBM mouse model with multiparametric detection for various immunotherapy research applications. Methods: In a xenograft GBM NOD-Prkdcs scid Il2rgem1/Smoc (NSG) mouse model based on orthotopic transplantation of patient-derived GBM cultures retaining tumor heterogeneity, intravenous and intratumor immunotherapeutic interventions by TIL and OV therapy were performed. Xenograft engraftment was evaluated using intravital MRI; delivery of OV and TILs to the tumor and changes in the tumor and peritumoral space were assessed using intravital confocal microscopy; and metabolic and structural changes in the tumor and peritumoral environment were assessed via fluorescence lifetime imaging microscopy (FLIM) and optical coherence tomography (OCT). The intravital imaging data were compared with the results of preliminary and final histological and immunocytochemical data. Results: Both OV and TILs demonstrated tumor-specific targeting and delivery across the blood–brain barrier. Further, we showed that in this model the xenograft response to both therapeutic treatments can be assessed using FLIM and OCT. Conclusions: Overall, this work presents an optimized mouse model suitable for assessing the effect of combined TIL immunotherapy and OV on GBM in translational studies. Full article
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26 pages, 6236 KB  
Article
Genomic Organization, Evolutionary Conservation and Expression of Ataxin-2 and Ataxin-2-like Genes Underscore the Suitability of Zebrafish as a Model Organism for SCA2 and Related Diseases
by Franz Vauti, Lukas Eilers, Anneke Kroll and Reinhard W. Köster
Biomedicines 2025, 13(12), 2974; https://doi.org/10.3390/biomedicines13122974 - 3 Dec 2025
Cited by 1 | Viewed by 1288
Abstract
Background/Objectives: The Ataxin-2 protein (ATXN2) plays an essential role in RNA metabolism and many cellular processes. Dysregulation or mutation of the Ataxin-2 gene (ATXN2) can lead to neurodegenerative diseases such as spinocerebellar ataxia type 2 (SCA2) and amyotrophic lateral sclerosis (ALS). [...] Read more.
Background/Objectives: The Ataxin-2 protein (ATXN2) plays an essential role in RNA metabolism and many cellular processes. Dysregulation or mutation of the Ataxin-2 gene (ATXN2) can lead to neurodegenerative diseases such as spinocerebellar ataxia type 2 (SCA2) and amyotrophic lateral sclerosis (ALS). Despite numerous efforts in this field in other animal models, little is known about Atxn2 in zebrafish. In this study, we aim to investigate the potential suitability of zebrafish as a model for Atxn2-related diseases by performing basic analyses on Atxn2. Methods: We performed a bioinformatic protein analysis of Atxn2 from zebrafish and its paralog Atxn2l in relation to human and other vertebrate homologues. Based on a structural analysis of the atxn2 and atxn2l genes, the expression of the predicted transcripts was detected by RT-PCR and the spatiotemporal expression pattern was determined by whole-mount in situ hybridization. Results: We found similarities between the protein sequences of Atxn2 and Atxn2l in zebrafish and their functional domains with those of orthologs in humans and other vertebrates. The predicted transcripts of atxn2 and atxn2l were experimentally verified and their spatiotemporal expression patterns were determined during zebrafish development. Splicing variants were detected for both genes, suggesting a different role for the isoforms in different tissues. Conclusions: These findings provide new insights into the atxn2 and atxn2l genes, suggesting the zebrafish as a suitable animal model for functional studies and research on disease modeling of SCA2 and ALS. Full article
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15 pages, 3315 KB  
Article
Shortcoming of the Mouse Model of Postoperative Ileus: Small Intestinal Lengths Have Similar Variations in In- and Outbred Mice and Cannot Be Predicted by Allometric Parameters
by Maximiliane von Stumberg, Ejder Akinci, Berkan Ertim and Christina Oetzmann von Sochaczewski
Biomedicines 2025, 13(12), 2948; https://doi.org/10.3390/biomedicines13122948 - 30 Nov 2025
Cited by 1 | Viewed by 958
Abstract
Background/Objectives: The mouse model of postoperative ileus separates the gastrointestinal tract into 15 sections, 10 of which are in the small intestine, to measure intestinal transit time. Usually, mice are standardised according to age or body weight. This inherently assumes that intestinal [...] Read more.
Background/Objectives: The mouse model of postoperative ileus separates the gastrointestinal tract into 15 sections, 10 of which are in the small intestine, to measure intestinal transit time. Usually, mice are standardised according to age or body weight. This inherently assumes that intestinal lengths are similar among the included mice irrespective of the method of standardisation. We aimed to test this assumption by comparing intestinal lengths, measuring their variability in commonly used out- and inbred strains. Methods: Mice were humanely killed, and their intestines were removed and measured in a standardised fashion. We compared the coefficients of variability via the modified signed-rank likelihood test. Results: We included 125 mice of the Crl:CD1(ICR) background and 10 mice of the C57Bl/6J and C57Bl/6NCrl substrains. The mean small intestinal length of Crl:CD1(ICR) mice was 437 mm (standard deviation 54), while it was 473 mm (standard deviation 29) in C57Bl/6J mice and 419 mm (standard deviation 57) in C57Bl/6NCrl mice. The respective coefficients of variation were 12.4%, 6.1%, and 13.6% and did not differ between the out- and inbred strains (modified signed likelihood ratio 5.878, p = 0.053). This was not the case for caecal and large intestinal lengths. Conclusions: Due to substantial variation in small intestinal length, the separation of the small intestine into ten equally sized segments to measure intestinal transit time might not be warranted. This could be addressed by measuring small intestinal transit time in absolute values and relative to the intestinal length. Full article
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Review

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48 pages, 1144 KB  
Review
Strategies for Optimizing Genetic Mouse Models to Enhance the Understanding of Parkinson’s Disease
by Zhiqiang Shen, Linlin Ma and George D. Mellick
Biomedicines 2026, 14(5), 1162; https://doi.org/10.3390/biomedicines14051162 - 20 May 2026
Viewed by 475
Abstract
Background: Parkinson’s disease (PD) has become the fastest-growing neurodegenerative disorder worldwide. A valuable approach for unraveling the disease’s mechanisms and new therapeutic targets involves investigating the PD-causing genes identified in families exhibiting the Mendelian inheritance of parkinsonism. Methods: In this article, [...] Read more.
Background: Parkinson’s disease (PD) has become the fastest-growing neurodegenerative disorder worldwide. A valuable approach for unraveling the disease’s mechanisms and new therapeutic targets involves investigating the PD-causing genes identified in families exhibiting the Mendelian inheritance of parkinsonism. Methods: In this article, we review how genetically modified mouse models can be employed to decipher the genetic architecture of PD. Results: We first discuss how well the human motor and non-motor symptoms of PD are currently evaluated in these PD mouse models, highlighting limitations. The pathogenic roles of five inherited PARK genes in PD are then extensively examined through their respective genetic mouse models in terms of phenotypic and cellular impacts. Furthermore, we discuss the strengths and weaknesses of existing transgenic mouse models and highlight significant accomplishments and advancements in this field from 2018 to the present. Conclusions: Building upon the current understanding of PD, we propose potential directions for enhancing genetic mouse models to further unveil the underlying mechanisms of PD and advance therapeutic research. Full article
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39 pages, 3908 KB  
Review
DSS Colitis Model: Traps, Tricks, and Reporting Recommendations
by Martina Perše
Biomedicines 2026, 14(4), 928; https://doi.org/10.3390/biomedicines14040928 - 18 Apr 2026
Cited by 1 | Viewed by 925
Abstract
The dextran sodium sulfate (DSS) colitis model is the most widely used experimental model of inflammatory bowel disease (IBD) due to its simplicity and versatility, with over 7000 PubMed entries in the last decade and an exponential rise in recent years. Since its [...] Read more.
The dextran sodium sulfate (DSS) colitis model is the most widely used experimental model of inflammatory bowel disease (IBD) due to its simplicity and versatility, with over 7000 PubMed entries in the last decade and an exponential rise in recent years. Since its initial description in 1985, DSS colitis has been extensively evaluated across species, most notably in mice and rats, and has yielded substantial insights into IBD pathogenesis. However, the model’s multifactorial nature poses a dual challenge: it offers an opportunity but complicates study design, interpretation, and translational relevance. This complexity is worsened by inconsistent reporting, which hampers reproducibility and comparability across studies. The broad use of the DSS-induced colitis model yields numerous insights about the model, which help better understand its complexity, characteristics and limitations. Although DSS colitis is induced locally, inflammation in the colon and gut barrier destruction may also affect other organs (such as the liver and brain) and their metabolism and molecular responses, which, in turn, may interfere with colitis-underlying mechanisms and drug response, and may influence the interpretation of results. These intrinsic (intra-experimental) characteristics of the DSS model are summarised in the paper (colitis, gut–brain axis, gut–liver axis). In addition, the DSS model is heavily influenced by numerous extrinsic (inter-experimental) factors (environmental, microbiological, genetic), which may further complicate the colitis model, the study outcomes, and data interpretation, and these are also discussed in the paper. As science advances and new data accumulate, understanding the intricate interplay among internal mechanisms, external factors, and technical variables becomes increasingly essential for the accurate interpretation of DSS outcomes. This review synthesises the complexity and interdependence of factors shaping the DSS model, emphasising the need for meticulous reporting and consideration of methodological nuances to enhance reproducibility, interpretation, and translational value in DSS colitis research. In addition, the review provides practical guidance through a “traps and tricks” subsection and checklist table designed to provide a framework and practical recommendations to better understand, apply, and interpret DSS model results in the context of broader systemic and methodological considerations. Full article
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22 pages, 1963 KB  
Review
Pre-Clinical Models of Traumatic Brain Injury—A Narrative Review Towards “Animal Neuro-ICUs”
by Franziska Münz, Andrea Hoffmann, Michael Gröger, Ohad Sharon, Magnus Scheer, Sandra Kress, Maximilian Feth, Peter Radermacher and Thomas Kapapa
Biomedicines 2026, 14(3), 688; https://doi.org/10.3390/biomedicines14030688 - 17 Mar 2026
Viewed by 822
Abstract
The presence of traumatic brain injury (TBI) is a critical determinant of post-traumatic mortality and morbidity. Not only is TBI one of the leading causes of death among severely injured patients, but it also substantially impacts long-term outcomes following severe trauma. Neurocritical care [...] Read more.
The presence of traumatic brain injury (TBI) is a critical determinant of post-traumatic mortality and morbidity. Not only is TBI one of the leading causes of death among severely injured patients, but it also substantially impacts long-term outcomes following severe trauma. Neurocritical care has a profound effect on outcomes following brain injury; nevertheless, its application in preclinical studies remains infrequent. This review therefore discusses strategies to improve the translational relevance of experimental TBI research, including the integration of neurocritical care principles in animal models. The review further addresses the impact of observation periods after injury and the selection of appropriate animal models (large vs. small animal models). In addition, commonly used injury induction methods—including controlled cortical impact (CCI), fluid percussion injury (FPI), weight-drop models, and blast injury paradigms—are discussed in terms of their reproducibility and clinical relevance. Finally, the review explores whether age, comorbidities, and sex influence TBI outcomes—and, if so, how these variables should be incorporated into experimental designs to improve translational fidelity. Full article
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