Veterinary oncology (London, England)最新文献

Background: Doxorubicin (DOX) is an anthracycline chemotherapeutic used for many canine malignancies, and its adverse event (AE) profile has been well-described in dogs. A limited sampling (LS) pharmacokinetic model was recently developed in canine patients to predict hematologic exposure to DOX. The primary goal of this study was to evaluate within-patient and between-patient variability in DOX exposure over three consecutive doses using the LS model. A secondary goal was to determine if there is a correlation between DOX exposure and gastrointestinal (GI) AEs utilizing a standardized owner questionnaire.

Methods: We performed a prospective evaluation of DOX exposure in seven tumor-bearing dogs across three cycles of treatment and compared the coefficient of variation (%CV) of the between- and within-patient exposures.

Results: This data set corroborated the ability of the DOX LS model to predict absolute neutrophil count for patients whose absolute neutrophil counts are lower at seven days compared to baseline. Dose normalized within-patient variability (4.7%) was significantly lower (p < 0.001) than between-patient variability (25.4%) in DOX exposure. Decreased appetite (p = 0.005) and increased nausea (p = 0.02) were significantly correlated with DOX exposure.

Conclusions: Together, these data suggest wide interpatient variability in dose-normalized DOX exposure but much more consistent exposure within an individual patient across treatment cycles. Future studies can attempt to target specific DOX exposures and absolute neutrophil counts to assess if DOX dosing can be personalized to increase efficacy and decrease GI AEs.

Supplementary information: The online version contains supplementary material available at 10.1186/s44356-025-00038-z.

Background: Canine osteosarcoma (OSA) and melanoma are highly aggressive cancers with a high propensity for lung metastasis. A recent phase 1 trial demonstrated that inhaled recombinant human interleukin-15 (rhIL-15) immunotherapy is safe and may be effective against lung metastases in dogs with metastatic OSA or melanoma. Notably, baseline lymphopenia correlated with clinical benefit in that trial. Building on these findings, this study evaluates whether preconditioning with doxorubicin to induce transient lymphodepletion before inhaled rhIL-15 immunotherapy improves the overall response rate in dogs with metastatic melanoma or tumors of bone.

Methods: Dogs with established pulmonary metastases from melanoma or bone tumors were treated with doxorubicin (30 mg/m² or 1 mg/kg) 7 days before starting inhaled rhIL-15 (50 µg twice daily x 14 days). Response rate was the primary objective of this study. Secondary objectives included toxicity assessment, immune correlative analyses via hematological monitoring, and NanoString transcriptomics analysis.

Results: Ten dogs were enrolled, with eight reaching the Day 35 evaluable response assessment. One dog achieved a complete response lasting over 1 year, and two had stable disease, yielding an overall clinical benefit rate (CBR) of 30% and an overall response rate (ORR) of 10%. Doxorubicin preconditioning did not alter the toxicity of inhaled rhIL-15 therapy. While doxorubicin treatment decreased absolute lymphocyte counts (ALC), as expected, this reduction did not improve the CBR or ORR compared to the prior study evaluating inhaled rhIL-15 monotherapy. Transcriptomic analysis of peripheral blood mononuclear cells did not reveal a significant increase in natural killer or cytotoxic T-cell population frequencies following doxorubicin and inhaled rhIL-15 therapy.

Conclusions: Preconditioning with standard doses of doxorubicin does not alter the clinical response or safety profile of inhaled rhIL-15 therapy in dogs with advanced pulmonary metastatic disease. Further research is needed to evaluate combinatorial treatment strategies that enhance the efficacy of inhaled rhIL-15 in dogs with advanced metastatic disease.

Supplementary information: The online version contains supplementary material available at 10.1186/s44356-025-00040-5.

Background: Canine cutaneous mast cell tumors (MCTs) are a common, yet clinically challenging tumor type given their variable biological behavior. Although patients with low grade MCTs can often be effectively managed with surgery alone, most dogs with high grade MCTs succumb to their disease despite multimodal therapy. An improved understanding of the immune tumor microenvironment (TME) may help identify novel prognostic and therapeutic targets.

Methods: In this study, we interrogated the immune transcriptional profiles of the TME in low and high grade MCTs, and quantified intratumoral T cells. Twelve client-owned dogs with MCTs (6 Kiupel low grade with clinically benign behavior and 6 Kiupel high grade with clinically aggressive behavior) that underwent curative-intent surgery were selected. Tumor grade was confirmed by a single veterinary pathologist. RNA was extracted from all tumors followed by immune transcriptional profiling utilizing the NanoString Canine IO panel and analysis using the ROSALIND platform. T cell density was determined by immunohistochemical staining for CD3 and quantified using ImageScope software (Leica Biosystems) following digital slide capture. Lymphocytic infiltrate was further characterized in the TME of one high grade MCT using co-immunofluorescence.

Results: Immune transcriptional profiling identified 9 differentially expressed genes between low and high grade MCTs (p-adj < 0.05). Programmed cell death protein 1 (PDCD1) and inducible T-cell costimulator ligand (ICOSLG) gene expression were significantly higher in a subset of high grade MCTs. ICOSLG expression positively correlated with T cell score (r_s = 0.6434, p = 0.0278). Although the T cell density was not significantly different between low (mean of 76.42 CD3 + /mm², SD 12 CD3 + /mm²) and high grade MCTs (mean of 129.1 CD3 + /mm², SD 96.06 CD3 + /mm²), greater variation of T cell densities was observed across high grade MCTs compared to low grade (p = 0.0059). Immunofluorescence of one high grade MCT with marked T cell infiltration revealed organized aggregates of T and B cells consistent with tertiary lymphoid structures (TLS).

Conclusions: Our data revealed significant differences in the immune TME of low and high grade MCTs and provides rationale to further investigate potential prognostic and therapeutic roles of immune checkpoints in canine MCTs.

Supplementary information: The online version contains supplementary material available at 10.1186/s44356-025-00020-9.

Canine malignant insulinoma is a rare, highly metastatic and life-threatening neuroendocrine tumour of pancreatic beta cells. To map the single-cell transcriptomic landscape of canine insulinoma for the first time, transcriptomic profiles of 5,532 cells were captured from two spontaneous insulinomas (Patient 1 and 2) and one associated metastasis (Patient 2) in two Boxer dogs. Distinct cancer, endocrine, and immune cell populations were identified. Notably, all three tumour samples contained two transcriptionally distinct insulin-expressing tumour cell populations (INS⁺ and INS⁺FOS ^low ), characterised here for the first time. These two cancer cell populations significantly differed by ~ 8,000 differentially expressed genes (DEGs), particularly tumour suppressor genes (e.g. TP53, EGR1) and cancer-related pathways (e.g., MAPK, p53). In contrast, COX7A2L was one of a few genes ubiquitously expressed and significantly upregulated (> 20-fold) in both insulin-expressing tumour populations compared to other captured populations. Both populations were also characterised by expression of chromogranin/secretogranin neuroendocrine tumour marker genes (e.g. CHGA, SCGN). There were far fewer gene expression differences observed between insulin-expressing tumour cells from the two patients (~ 600 DEGs) than between the two cancer cell populations within each patient. These DEGs included CLTRN, TMSB4X, CSRP2, LGALS2, and C15orf48. Unexpectedly for a tumour of endocrine origin, the metastasis in Patient 2 exhibited > 20-70 fold upregulation of exocrine pancreatic genes including CLPS, PRSS2, PRSS and CTRC. Immune cell analyses identified distinct infiltrating immune populations, including memory T cells and macrophages and revealed likely tumour-immune interactions, including the CD40-CD40L interaction. This study provides the first single-cell RNA sequencing (scRNA-seq) analysis of naturally occurring insulinoma in any species, revealing tumour cell heterogeneity, novel immune microenvironment features, and potential therapeutic targets. Despite its small scale, the findings highlight the utility of scRNA-seq in veterinary oncology and its translational potential for pancreatic neuroendocrine tumours across species.

Supplementary information: The online version contains supplementary material available at 10.1186/s44356-025-00026-3.

Immunotherapeutic approaches to cancer treatment have gained significant traction in recent years, due in large part to the success of immune checkpoint inhibitors and T cell-based therapies. Comparative oncology is the study of naturally-occurring cancer in companion (pet) animals, mainly dogs, and is a powerful tool in cancer research and drug development. Given their intact, educated immune systems and natural co-evolution of tumor, microenvironment and stromal components, tumor-bearing pet dogs are an attractive species in which to explore these cellular interactions and test novel therapeutic approaches. Moreover, similarities between the canine and human immune systems support assessment of a wide variety of approaches, including antagonistic or agonistic antibodies directed at specific cellular targets, tumor vaccines, cell-based therapies, and combinations of these with conventional cancer treatments such as chemotherapy and radiotherapy. This manuscript provides specific examples of how canine immunotherapeutic studies informed an approach destined for human use, with an emphasis on study design, correlative immune assay development and application, and definition of biologic effect.

Radiation therapy (RT) is a common treatment modality for dogs with locally advanced head and neck tumors. Most dogs experience a clinical benefit secondary to RT, however, long term remissions are rare. This study evaluates the feasibility and safety profile of intratumoral CBC101 (a proprietary hydrogel-based injectable resiquimod formulation), a toll-like receptor (TLR) 7/8 agonist with immunomodulatory properties, when used in combination with radiation therapy. Three dogs with histologically confirmed head/neck cancers were prospectively enrolled. A baseline CT scan was performed. Dogs received palliative radiation therapy (8 Gy × 4) in conjunction with intratumoral CB101. A follow-up CT scan was performed at week 12 to assess tumor response and to evaluate for metastatic disease. Intratumoral CB101 was well-tolerated, feasible, and produced minimal adverse effects. Only one grade 1 adverse event was attributable to CB101; all other adverse events were expected radiation therapy side effects. The data obtained from this preliminary study will be used for further investigation into appropriate dosing and timing of intratumoral resiquimod or other TLRs, with eventual escalation into phase II and III clinical trials.

Background: Insulinoma is the most common pancreatic neuroendocrine tumour in dogs and humans. The understanding of driving factors and critical survival genes in insulinomas is limited and overall survival is poor for canine and human malignant insulinoma. This study aimed to use single-cell RNA-sequencing to conduct a multispecies analysis of insulinoma cell lines to understand their single-cell transcriptomic landscape. Secondly, the impact of freeze-thawing on the pancreatic beta single-cell transcriptome was investigated, to determine whether cryoarchiving of primary insulinoma samples may be feasible in future studies.

Methods: Single-cell transcriptomic analysis was performed using fresh and cryopreserved multispecies insulinoma cell lines (canINS, CM, INS-1 and MIN6). R and Seurat were used to perform cell clustering and specific cluster marker genes were identified by the FindMarkers function. Metascape was used to identify statistically enriched pathways for specific cell clusters. Differentially expressed genes between fresh and cryopreserved single-cell transcriptome profiles, were defined as genes with a log2 fold change > 0.25 and a Bonferroni-adjusted P < 0.05, based on the Wilcoxon rank sum test.

Results: Based on the specific cell line single-cell transcriptome profiles, five or six cell clusters were constructed per cell line. All cell lines expressed neuroendocrine markers and additionally INS-1 and MIN6 displayed a gene signature indicative of mature/functional pancreatic islet/beta-cells. DEPTOR, BICC1, GHR, CCNB2, CENPA, LMO4, VANGL1, and L1CAM were identified as cross-species conserved insulinoma cluster marker genes. Little effect was found of cryopreservation and thawing on overall gene expression at the single-cell level in insulinoma cell lines: only 6 and 29 genes had a log2 fold change > 1 in cryopreserved versus fresh canINS and CM, respectively.

Conclusions: canINS, CM, INS-1 and MIN6 are all principally relevant as insulinoma models and the demonstrated differences in their single-cell transcriptomic profiles could aid researchers in selecting the appropriate cell lines for specific study objectives. Cross-species conserved insulinoma cluster marker genes were identified that harbour oncogenes and their involvement in insulinoma tumourigenesis should be investigated in future studies. The good comparability between cryopreserved and fresh insulinoma cells allows for inclusion of cryopreserved insulinoma patient samples in future studies, which allows for reduced assay-based variability.

Supplementary information: The online version contains supplementary material available at 10.1186/s44356-025-00025-4.

Muscle-invasive urothelial carcinoma (MIUC) is the most common type of bladder malignancy in dogs, but the treatments used in the clinic are relatively ineffective for most of them. Dogs represent a naturally- occurring model for human MIUC and the advances in veterinary oncology could benefit human oncology as well. The field of epigenetics presents unique opportunities for new cancer therapeutics or biomarkers, as epigenetic modification of key genes can regulate tumor initiation and progression. This review summarizes the existing literature on epigenetic changes in canine MIUC as compared to human MIUC and provides suggestions for future studies that could benefit both human and canine patients.