Comparative biology is big in veterinary science
It helps us understand, it helps us develop new solutions
So that, when we see studies on Extracellular Vescicles and CDB, we have to smile real big.
Because it means change is coming, and even the newst things in human research are coming to veterinary science.
This week’s big shoutout goes to Thierry Olivry and his team at Nextmune for their new Multiplex Molecular Macroarray and to Tania Gutiérrez Riquelme and colleagues for their great proteomics work on EV!
Enjoy this week’s edition!
In Vitro
Investigation on the In Vitro Effects of Resveratrol on Peripheral Blood Mononuclear Cells Harvested from Healthy and Atopic Dogs
In this study Domenico Santoro from the University of Florida, USA, evaluates resveratrol’s potential as a safer anti-inflammatory treatment for canine atopic dermatitis (AD) compared to conventional therapies, which often have side effects and high costs. The in vitro effects of resveratrol were examined on peripheral blood mononuclear cells (PBMCs) from nine healthy and 11 atopic dogs. PBMCs were exposed to resveratrol at various concentrations (1.5–9 μg/mL), with and without phytohemagglutinin stimulation. Cytotoxicity, host defense peptides (HDPs), oxidative stress markers (catalase and superoxide dismutase), and pro-inflammatory cytokines were assessed after 24 hours. Results indicated no cytotoxicity in either group across all conditions. Low concentrations of resveratrol increased catalase levels only in healthy PBMCs, but HDP levels remained unaffected. Significant reductions in monocyte chemotactic protein-1 and interleukin-6 were observed at the highest concentration (9 μg/mL) in both unstimulated healthy and stimulated atopic PBMCs. Overall, resveratrol demonstrated minimal effects on HDP secretion and pro-inflammatory cytokine levels at the tested concentrations, while proving non-toxic to PBMCs from both healthy and atopic dogs.
Santoro D, Nadeau P., Archer L..Investigation on the in vitro effects of resveratrol on peripheral blood mononuclear cells harvested from healthy and atopic dogs. Research in Veterinary Science. 2024, 105441, ISSN 0034-5288, https://doi.org/10.1016/j.rvsc.2024.105441.
Proteomic Analysis of Extracellular Vesicles Derived from Canine Mammary Tumour Cell Lines Identifies Protein Signatures Specific for Disease State
This study from Tania Gutiérrez Riquelme, from the Freie Universität Berlin, Germany, explores the proteomic profiles of extracellular vesicles (EVs) derived from canine mammary tumour (CMT) cell lines to identify non-invasive diagnostic markers for CMT. EVs, containing unique protein and RNA cargo, were isolated from five distinct canine mammary cell lines representing non-neoplastic, adenoma, and carcinoma states. Whole-cell lysates (WCLs) and EV lysates underwent LC-MS/MS analysis, with protein abundance compared between groups. Distinct protein signatures were observed in WCLs and EVs, particularly for proteins related to cell adhesion, migration, and RNA processing. Weighted gene correlation network analysis (WGCNA) identified co-abundant EV protein clusters specific to CMT stages, revealing distinct adenoma and carcinoma EV signatures, absent in WCLs. Key proteins like biglycan emerged as potential biomarkers for canine mammary carcinomas, offering a foundation for developing a liquid biopsy approach for CMT diagnosis, minimizing the need for invasive tissue biopsies.
Gutierrez-Riquelme, T., Karkossa, I., Schubert, K. et al. Proteomic analysis of extracellular vesicles derived from canine mammary tumour cell lines identifies protein signatures specific for disease state. BMC Vet Res 20, 488 (2024). https://doi.org/10.1186/s12917-024-04331-1
In Vitro Antitumor Effect of Oils Rich in CBD and THC Cannabis Extract in Canine Prostate Carcinoma Cell Lines
Calheiros et al, Sao Paulo State University- The Universidade Estadual Paulista (UNESP), Brazil, investigates the antitumor potential of cannabidiol (CBD) and Δ9-tetrahydrocannabinol (THC) in canine prostate carcinoma cell lines, PC1 and PC2. Prostate cancer in dogs presents similarities to androgen-independent prostate cancer in humans, making it a valuable model for comparative research. Results demonstrated that both CBD- and THC-rich cannabis extracts significantly inhibited cell proliferation, with half-maximal inhibitory concentrations (IC50) at 3.43 and 3.57 μM for CBD and 4.90 and 4.48 μM for THC. Furthermore, cell death was confirmed via Annexin V and Propidium iodide staining. These findings suggest a promising direction for further research on the therapeutic applications of cannabis extracts in treating canine prostate carcinoma, with potential implications for human medicine.
Calheiros, L.G.R.d.M.; Pedro, G.; Oliveira da Silva, T.; Amorim, R.M.; Alves, C.E.F.; Laufer-Amorim, R. In Vitro Antitumor Effect of Oils Rich in CBD and THC Cannabis Extract in Canine Prostate Carcinoma Cell Lines. Vet. Sci. 2024, 11, 501. https://doi.org/10.3390/vetsci11100501
Diagnostics & Biomarkers
Validation of a Multiplex Molecular Macroarray for the Determination of Allergen-Specific IgE Sensitizations in Dogs
Thierry Olivry and his team at Nextmune, Sweden, validated the Pet Allergy Xplorer (PAX), a novel multiplex molecular macroarray designed to detect IgE sensitizations in dogs. Developed by Nextmune, this platform incorporates both allergen extracts and molecular components to provide detailed insights into allergen cross-reactivity. The validation process included testing the specificity and accuracy of IgE detection, as well as evaluating confounding factors such as hemolysis and lipemia. Notably, the PAX includes a cross-reactive carbohydrate determinants (CCD) detection and blocking feature, increasing result reliability. Data from over 23,000 European dogs revealed sensitization rates across 145 environmental and insect allergens, showcasing PAX as an effective, reproducible tool for diagnosing canine allergies.
Olivry, T.; Fontao, A.M.; Aumayr, M.; Ivanovova, N.P.; Mitterer, G.; Harwanegg, C. Validation of a Multiplex Molecular Macroarray for the Determination of Allergen-Specific IgE Sensitizations in Dogs. Vet. Sci. 2024, 11, 482. https://doi.org/10.3390/vetsci11100482
Immunohistochemical Expression of Tenascin-C in Canine Meningiomas
This study from Sant’Ana et al, Universidade de Brasília, Brazil, investigated the expression of tenascin-C (TN-C), a marker associated with more aggressive central nervous system tumors in humans, within different grades of canine meningiomas. The study analyzed 21 cases of canine meningiomas classified as grade I (12 cases), grade II (6 cases), and grade III (3 cases). Immunohistochemistry for TN-C, along with epithelial membrane antigen (EMA), Ki-67, pan-cytokeratin (Pan CK), and vimentin, was performed to characterize TN-C distribution. Results showed increased TN-C immunoreactivity corresponding with tumor grade, from mild in grade I to moderate in grade II, and moderate to severe in grade III. TN-C positivity was primarily noted in the tumor stroma and perivascular spaces, particularly in higher-grade tumors. The findings suggest that TN-C may serve as a valuable stromal marker in higher-grade canine meningiomas, with potential diagnostic applications in identifying more aggressive forms of the disease.
Sant’Ana, F.J.F.d.; Blasco, E.; Pumarola, M. Immunohistochemical Expression of Tenascin-C in Canine Meningiomas. Vet. Sci. 2024, 11, 462. https://doi.org/10.3390/vetsci11100462
Nesfatin-1 and Phoenixin-14 in Female Dogs with Uterine Disease: Potential Biomarkers for CEH and Pyometra”
This study from Marta Rybska from the @Poznan University of Life Sciences, Poland, explored the roles of nesfatin-1, a neuropeptide influencing reproductive function, and its interaction with phoenixin-14 in female dogs with cystic endometrial hyperplasia (CEH) and pyometra. Results showed elevated nesfatin-1 levels in both the uterine tissue and blood plasma of overweight and CEH/pyometra-affected dogs, while phoenixin-14 levels were lower, regardless of body condition. The findings suggest a contrasting interaction between nesfatin-1 and phoenixin-14, potentially contributing to the development of these conditions and aiding in biomarker discovery for early detection.
Rybska, M., Skrzypski, M., Billert, M. et al. Nesfatin-1 expression and blood plasma concentration in female dogs suffering from cystic endometrial hyperplasia and pyometra and its possible interaction with phoenixin-14. BMC Vet Res 20, 486 (2024). https://doi.org/10.1186/s12917-024-04336-w
Lymphoid Aggregates in Canine Cutaneous and Subcutaneous Sarcomas: Immunohistochemical and Gene Expression Evidence for Tertiary Lymphoid Structures
This study from Rugh et al from the Colorado State University , USA, investigates lymphoid aggregates (LAs) in canine cutaneous and subcutaneous soft-tissue sarcomas (STS), a type of mesenchymal tumor with potential for recurrence and metastasis. In human sarcomas, LAs that resemble tertiary lymphoid structures (TLS) are associated with better prognoses, prompting examination of LAs in canine sarcomas. Using a criterion of LAs with >700 cells, researchers measured RNA expression in large LAs, aggregate-adjacent sarcoma tissue (AAS), aggregate-free sarcoma, and lymph nodes. Immunohistochemistry (IHC) was used to detect markers CD3, CD20, MUM-1, and PNAd, revealing significant upregulation of CD3 and CD20 mRNA in LAs. Additionally, large LAs displayed increased expression of TLS-characteristic chemokines, including CXCR5, CCL19, CCL21, and CXCL13. Plasma cells and high endothelial venules were also identified in tumors with large LAs, indicating TLS presence. Large LAs were found in 12% of tumors, while LAs of any size were present in 30%. These findings suggest that large LAs in canine STS resemble TLS, potentially offering prognostic insights.
Rugh, K., Ashton, L., Schaffer, P. and Olver, C. (2024), Lymphoid Aggregates in Canine Cutaneous and Subcutaneous Sarcomas: Immunohistochemical and Gene Expression Evidence for Tertiary Lymphoid Structures. Vet Comp Oncol. https://doi.org/10.1111/vco.13020
Clinical
Treatment with Oclacitinib, a Janus Kinase Inhibitor, Down-Regulates and Up-Regulates CD25 and Foxp3 Expression, Respectively, in Peripheral Blood T Cells of Dogs with Atopic Dermatitis
Agnieszka Jasiecka-Mikołajczyk et al from the Uniwersytet Warmińsko-Mazurski w Olsztynie, Poland, examined the effects of oclacitinib (OCL), a Janus kinase inhibitor used to treat canine atopic dermatitis (AD), on T-cell subsets in dogs over 28 days. Eight dogs were treated, and blood samples were analyzed to track immune modulation and safety. Results showed that while overall CD4+ and CD8+ T-cell counts remained stable, OCL treatment decreased CD25+ activated T cells and increased Foxp3+ regulatory T cells, suggesting a shift towards immune regulation. Additionally, OCL reduced eosinophil levels, supporting its effectiveness and safety as an immune-modulating treatment for canine AD.
Jasiecka-Mikołajczyk, A., Socha, P. Treatment with oclacitinib, a Janus kinase inhibitor, down-regulates and up-regulates CD25 and Foxp3 expression, respectively, in peripheral blood T cells of dogs with atopic dermatitis. BMC Vet Res 20, 489 (2024). https://doi.org/10.1186/s12917-024-04340-0
Age at Tumor Diagnosis in 14,636 Canine Cases from the Pathology-Based UNIPI Animal Cancer Registry, Italy: One Size Doesn’t Fit All
This large-scale study from Niccolò Fonti et al, from the Università di Pisa, Italy, evaluates the age at cancer diagnosis across 14,636 canine patients in Italy, highlighting the need for personalized cancer screening approaches in dogs. Data was gathered from the UNIPI Animal Cancer Registry, coding cases by tumor type, breed, sex, neutering status, body size, and other factors. The median age at diagnosis was 9 years for benign tumors and 10 for malignant tumors, with earlier diagnoses in intact and purebred dogs. Tumor types like lymphomas and mast cell tumors had the earliest median diagnostic age. Larger body size, specific head shapes (brachy- and dolichocephalic), and intact female status were associated with earlier diagnoses. These findings suggest that a breed-specific approach to cancer screening in dogs may improve early detection, challenging the current “one-size-fits-all” screening standard.
Fonti, N.; Parisi, F.; Lachi, A.; Dhein, E.S.; Guscetti, F.; Poli, A.; Millanta, F. Age at Tumor Diagnosis in 14,636 Canine Cases from the Pathology-Based UNIPI Animal Cancer Registry, Italy: One Size Doesn’t Fit All. Vet. Sci. 2024, 11, 485. https://doi.org/10.3390/vetsci11100485
