Leveraging single-cell transcriptomic data to uncover immune suppressive cancer cell subsets in triple-negative canine breast cancers.

Introduction: Single-cell RNA sequencing (scRNA-seq) has become an essential tool for uncovering the complexities of various physiological and immunopathological conditions in veterinary medicine. However, there is currently limited information on immune-suppressive cancer subsets in canine breast cancers. In this study, we aimed to identify and characterize immune-suppressive subsets of triple-negative canine breast cancer (TNBC) by utilizing integrated scRNA-seq data from published datasets. Methods: Published scRNA-seq datasets, including data from six groups of 30 dogs, were subjected to integrated bioinformatic analysis. Results: Immune modulatory TNBC subsets were identified through functional enrichment analysis using immune-suppressive gene sets, including those associated with anti-inflammatory and M2-like macrophages. Key immune-suppressive signaling, such as viral infection, angiogenesis, and leukocyte chemotaxis, was found to play a role in enabling TNBC to evade immune surveillance. In addition, interactome analysis revealed significant interactions between distinct subsets of cancer cells and effector T cells, suggesting potential T-cell suppression. Discussion: The present study demonstrates a versatile and scalable approach to integrating and analyzing scRNA-seq data, which successfully identified immune-modulatory subsets of canine TNBC. It also revealed potential mechanisms through which TNBC promotes immune evasion in dogs. These findings are crucial for advancing the understanding of the immune pathogenesis of canine TNBC and may aid in the development of new immune-based therapeutic strategies.

Clinical Outcome of Multicentric Lymphoma Treated with Cyclophosphamide, Doxorubicin, Vincristine, and Prednisolone (CHOP) in Small Breed Dogs.

Lymphoma is one of the most common malignant tumors in dogs. Combination chemotherapy with vincristine, cyclophosphamide, doxorubicin, and prednisolone (CHOP) is the most effective treatment for multicentric lymphoma. Previous studies have evaluated the response of large dogs to CHOP treatment and identified prognostic factors; however, studies on small dogs are lacking. In this study, we investigated the outcomes and prognostic factors for small dogs with multicentric lymphoma treated with CHOP. The responses of patients to CHOP treatment were assessed, and 54.3% were evaluated as being in complete remission (CR), 31.4% in partial remission (PR), and 14.3% in no remission (NR). The overall response rate was 85.7%. The median survival times for CR, PR, and NR patients were 683 days (85-1496 days), 241 days (15-777 days), and 119 days (61-308 days), respectively. Among the CR patients, survival was longer under the following conditions: age under 10 years (p = 0.011), no cardiovascular heart disease (p = 0.046), and no history of hospitalization due to side effects from chemotherapy (p = 0.002). These results might help clinicians build treatment plans for multicentric lymphoma in small breed dogs.

Intravenous injection of allogenic canine mesenchymal stem cells in 40 client-owned dogs: a safety assessment in veterinary clinical trials.

BACKGROUND: The aim of this study was to evaluate the adverse effects of allogeneic mesenchymal stem cells (MSCs) transplanted via intravenous infusion in dogs and examine their safety. We performed a retrospective analysis of various clinical assessments, including physical examination, blood tests, and radiographs, and monitored the formation of neoplasms during a 6-month follow-up period in 40 client-owned dogs that received intravenous infusion of adipose tissue-derived MSCs (AT-MSCs) for the treatment of various underlying diseases between 2012 and 2018. RESULTS: No significant adverse effects of MSC therapy were detected by clinical assessment, blood tests, or radiographic examination in the 6-month follow-up period after the first MSC treatment. Additionally no new neoplasms were observed during this period. CONCLUSIONS: To our knowledge, this study is the first to evaluate the safety aspects (≥ 6 months) associated with intravenous allogeneic AT-MSC infusion. These results suggest that allogenic AT-MSC infusion could be a useful and relatively safe therapeutic approach in canines.

Pyogenic granuloma of the hard palate leading to alveolar cleft: a case report.

This case report describes a rare occurrence of pyogenic granuloma (PG) in the hard palate deviating from its typical gingival location that led to the formation of an alveolar cleft. The aggressive growth pattern of the lesion, with atypical progression from a pedunculated nodule to an alveolar cleft, raised concern. The diagnosis was based on magnetic resonance imaging and computed tomography findings, which revealed a tadpole-shaped lesion originating from the midline hard palate. The differential diagnosis included a minor salivary gland tumor. Surgical excision was performed under general anesthesia and resulted in a mucosal defect without nasolabial fistula formation or bone exposure. The palatal defect was packed with oxidized regenerated cellulose and closed with Vicryl Rapide sutures, both of which contributed to the patient's successful outcomes. Our comprehensive approach, extending across the stages of surgical planning, execution, and postoperative care, demonstrated the advantages of a multidisciplinary strategy for the accurate diagnosis and effective treatment of palatal PGs. This report makes a meaningful contribution to the existing literature on common oral lesions by emphasizing the importance of a broad differential diagnosis and a systematic approach to oral pathologies. It also raises clinical awareness of PGs with atypical presentations and the diagnostic challenge that they pose.

Using 3D Bioprinted Autologous Minimally Manipulated Homologous Adipose Tissue for Limb Salvage in Treating Diabetic Foot Ulcer.

Reconstructive surgeons face challenges when considering limb salvage methods for the treatment of diabetic foot ulcers (DFUs). In this article, we present our experience with autologous fat grafting as a viable alternative in cases where flap reconstruction is difficult. We encountered a 78-year-old female patient with a nonhealing DFU who had multiple comorbidities, including renal failure and severe peripheral arterial disease. During the initial multidisciplinary meeting, due to extensive necrosis and osteomyelitis, amputation was recommended. However, the patient expressed a strong preference for a salvage procedure and refused amputation. After careful consideration, we opted to reconstruct the patient's foot using three-dimensional bioprinted autologous minimally manipulated homologous adipose tissue. The AMHAT was engrafted well without complications such as autolysis, graft failure, or infection. After the operation, the large defect with partial bone exposure was covered with healthy granulation tissue. The size of the wound decreased to less than half its original size after 6 weeks of surgery, and it decreased to less than 25% after 12 weeks of surgery. The AMHAT may be an appealing treatment option for diabetic foot patients who are unsuitable for flap reconstruction due to comorbidities.

Mitigating Gas Evolution in Electron Beam-Induced Gel Polymer Electrolytes Through Bi-Functional Cross-Linkable Additives.

The current high-capacity lithium-ion batteries (LIBs), reliant on flammable liquid electrolytes (LEs) and nickel-rich cathodes, are plagued by safety hazards, especially the risk of hazardous gas release stemming from internal side reactions. To address these safety concerns, an electron beam (E-beam)-induced gel polymer electrolyte (E-Gel) is introduced, employing dipentaerythritol hexaacrylate (DPH) as a bi-functional cross-linkable additive (CIA). The dual roles of DPH are exploited through a strategically designed E-beam irradiation process. Applying E-beam irradiation on the pre-cycled cells allows DPH to function as an additive during the initial cycle, establishing a protective layer on the surface of the anode and cathode and as a cross-linker during the E-beam irradiation step, forming a polymer framework. The prepared E-Gel with CIA has superior interfacial compatibility, facilitating lithium-ion diffusion at the electrode/E-Gel interface. The electrochemical assessment of 1.2 Ah pouch cells demonstrates that E-Gel substantially reduces gas release by 2.5 times compared to commercial LEs during the initial formation stage and ensures superior reversible capacity retention even after prolonged cycling at 55 °C. The research underscores the synergy of bifunctional CIA with E-beam technology, paving the way for large-scale production of safe, high-capacity, and commercially viable LIBs.

Predictive factors of lymph node metastasis in papillary thyroid cancer.

This study aimed to evaluate factors that predict lymph node metastasis (LNM) in papillary thyroid cancer (PTC). This retrospective cross-sectional study compared the demographic, clinical, and ultrasonographic findings of patients with PTC with and without LNM. Subgroup analysis was conducted for micro-PTCs (<1 cm). Among total (n = 512; mean age, 47.3 ± 12.7 years) and micro-PTC patients (n = 312), 35.7% and 19.6% had LNM, respectively. Younger age, male sex, tumor size, bilaterality, and suspicious ultrasound features of the tumor were associated with LNM. In multiple logistic regression analysis, among all patients, age, tumor size, and extrathyroidal extension were independent risk factors for LNM (all p<0.05). In the micro-PTC subgroup, age, extrathyroidal extension, bilaterality of tumor, and presence of autoimmune thyroid disease were independent risk and protective factors for LNM (all p<0.05). In the receiver operating characteristic analysis, the accuracy of the multivariable logistic regression model for predicting LNM among all patients and micro-PTC was acceptable (area under the curve = 0.729 and 0.733, respectively). Age, sex, tumor size, and extrathyroidal extension can assist in predicting LNM in PTC patients. Additionally, the bilaterality of tumors and presence of autoimmune thyroid disease can assist in predicting LNM in micro-PTCs.

Off-Label Use of Crisdesalazine (GedaCure) in Meningoencephalitis in Two Dogs.

An 8-year-old, castrated male Shih-tzu dog (Case 1) showing ataxia and gait disorder was referred for neurological examination and magnetic resonance imaging. Through comprehensive examinations, the patient was tentatively diagnosed with meningoencephalitis of unknown origin (MUO) and treatment with prednisolone and cytosine arabinoside was started. The symptoms were improving with immunosuppressive treatment. However, severe bacterial cystitis occurred and we could not avoid tapering off prednisolone. Then, neurological signs recurred. Therefore, we added crisdesalazine, which allowed us to reduce the daily dosage of immunosuppressants easily. In another case, a 4-year-old, spayed female Yorkshire terrier dog (Case 2) was referred to our hospital showing a head tilt, circling, and loss of the menace reflex. The patient was tentatively diagnosed with MUO and treatment with some immunosuppressants was attempted. The clinical symptoms improved, but the alleviation was inadequate. Thus, we added crisdesalazine. The neurological signs then markedly improved. Moreover, the drugs could be tapered off more easily than before. Crisdesalazine is a novel drug that has antioxidant and anti-inflammatory action in brain disease and is used particularly for dementia. In this paper, we tried an off-label use of this drug in canine MUO patients, and found that it had, in these two patients, additional therapeutic effects on the MUO.

A propensity score-matched analysis of advanced energy devices and conventional monopolar device for colorectal cancer surgery: comparison of clinical and oncologic outcomes.

Purpose: The safety, efficiency, and versatility of novel surgical energy devices have been proved by recent studies. This study aims to investigate the impact of surgical energy devices on operative and oncologic outcomes of minimally invasive colorectal cancer surgery. Methods: The study group included 80 patients who underwent minimally invasive colorectal cancer surgery with a conventional monopolar device and 217 patients with advanced surgical energy devices between August 2015 and December 2017. The propensity score matching for tumor lesion, preoperative level of CEA, and operation technique produced 63 matched pairs. Results: In patient characteristics, there was no significant difference between the groups after the propensity score matching. The amount of blood loss (72 mL vs. 54 mL, P = 0.123) and conversion cases to another surgery (11.1% vs. 4.8%, P = 0.187) tended to be higher in monopolar group, while operation time and intraoperative complications were not significantly different. The short-term clinical outcomes including time to soft diet, the length of hospital stays, and the morbidity within 30 days after surgery or pathologic outcomes were comparable between the groups. During the median follow-up of 52.9 and 51.1 months in each study group, the 5-year overall survival rates of the monopolar and advanced energy groups were 84.6% and 91.6% (P = 0.276), and the 5-year disease-free survival rates were 78.0% and 84.6% (P = 0.328), respectively. Conclusion: The use of surgical energy devices based on surgeons' preference did not show significant impact on operative and long-term outcomes compared with conventional monopolar devices in minimally invasive colorectal cancer surgery.

Comparison of Patients Satisfaction with Direct to Implant versus Latissimus Dorsi Flap with Implant Breast Reconstruction Using Breast-Q.

Background The latissimus musculocutaneous flap (LD flap) is a useful option for breast reconstruction following mastectomy. It has the advantage of obtaining sufficient tissue padding and natural shape by using autologous tissue. However, with the emergence of the skin-sparing mastectomy technique and artificial dermis matrix, direct-to-implant (DTI) breast reconstruction has become the first choice of surgery. The purpose of this study was to compare the satisfaction levels of patients who underwent DTI and LD flap with implant using patient-reported Breast-Q results. Methods A retrospective study was performed reviewing the records of 49 women who underwent immediate breast reconstruction with DTI or LD flap with implant and responded to the BREAST-Q questionnaire after the operation. The patient-reported breast-Q results were analyzed and correlated to the demographic information and intraoperative information. Results A total of 26 patients who underwent reconstruction with LD flap with implant and 23 patients with DTI were identified and responded to the questionnaire after an average of 32.3 and 10.4 months postoperation, respectively. According to the patient response to the breast-q values, satisfaction with breast was 60.0 and 57.0 points, psychosocial well-being 61.0 and 60.0 points, and sexual well-being 41.0 and 43.0 points in the two groups. Overall, there was no significant difference in the breast-Q score between the two groups. Conclusion Patients who underwent DTI breast reconstruction seemed equally satisfied with the appearance and outcome of their breast reconstruction compared with LD flap with implant. Therefore, it appears that DTI is adequately replacing LD with implant.

A Comparative Analysis of Patient Satisfaction and Cosmetic Outcomes after Breast Reconstruction through BREAST-Q and the Judgment of Medical Panels: Does it Reflect Well in Terms of Aesthetics in Korean Patients?

Background Currently, the BREAST-Q can effectively measure patient's satisfaction on the quality of life from the patient's perspective in relation to different type of breast reconstruction. However, evaluation of patient satisfaction and cosmetic outcomes in breast reconstruction may have potential to led bias. Methods To maximize the benefits of using BREAST-Q to evaluate clinical outcome, we performed comparative study focused on the correlation between postoperative BREAST-Q and cosmetic outcomes assessed by medical professionals. For the current analysis, we used three postoperative BREAST-Q scales (satisfaction with breast, psychosocial well-being, and sexual well-being). The Ten-Point Scale by Visser et al was applied to provide reproducible grading of the postoperative cosmetic outcomes of the breast. The system includes six subscales that measured overall aesthetic outcome, volume, shape, symmetry, scarring, and nipple-areolar complex. The photographic assessments were made by five medical professionals who were shown photographs on a computer screen in a random order. Obtained data were stored in Excel and evaluated by Spearman's correlations using SPSS Statistics. Results We enrolled 92 women in this study, 10 did not respond to all scales of postoperative BREAST-Q, the remaining 82 women had undergone breast reconstruction. The correlation between BREAST-Q score and aesthetic score measured by Ten-Point Scale for the three BREAST-Q scales all show positive values in Spearman's correlation coefficient. Conclusion A significant correlation without any bias observed was found between the patient's satisfaction measured by BREAST-Q after breast reconstruction and the medical expert's aesthetic evaluation.

Adenosine-Prefabricated Adipose Tissue Improves Fat Graft Survival by Promoting VEGF-Dependent Angiogenesis.

BACKGROUND: Angiogenesis plays an important role in determining the fat graft survival. However, clinical preconditioning techniques that target angiogenesis during fat grafting have not been established so far. Adenosine has emerged as a regulator of angiogenesis under hypoxic conditions; therefore, the aim of this study was to investigate the effects and underlying mechanisms of adenosine prefabrication on fat graft survival. METHODS: In the first animal study, a total of 32 mice were transplanted with fat prefabricated with vehicle (Control, N = 16) or adenosine (Adenosine, N = 16). In the second animal study, 24 mice were divided into three groups based on the type of fat graft: Control (N = 8), Adenosine (N = 8), and Axitinib (cotreatment of adenosine with axitinib, N = 8). At 1- and 4-weeks post-transplantation, grafts were evaluated by histopathological and biochemical assessment. Adenosine-induced vascular endothelial growth factor (VEGF) production and angiogenesis were determined using cell cultures. RESULTS: The retention volumes of fat grafts in the adenosine group were significantly increased until 4 weeks. Fat grafts from the adenosine group exhibited greater structural integrity, reduced fibrosis, and increased blood vessels. The expression levels of angiogenesis-related genes, Vegfa, Vegfr1, Vegfr2, and Vwf, were elevated in the adenosine group. Furthermore, adenosine upregulated VEGF production in preadipocytes, thereby enhancing the migration of endothelial cells. Treatment with the axitinib, VEGF receptor inhibitor, abrogated the adenosine-induced angiogenesis in the fat grafts. CONCLUSION: Adenosine prefabrication in fat improved the graft survival by enhancing angiogenesis through the VEGF/VEGFR axis in the preadipocytes and endothelial cells. Therefore, this method may be used as a novel strategy to increase the retention rate in fat grafts.

Prevalence of Reticulocytosis in the Absence of Anemia in Dogs with Cardiogenic Pulmonary Edema Due to Myxomatous Mitral Valve Disease: A Retrospective Study.

Myxomatous mitral valve disease (MMVD) is the most common heart disease in small breed dogs. Dogs with MMVD commonly show clinical signs of dyspnea due to cardiogenic pulmonary edema (CPE). Reticulocytosis in the absence of anemia (RAA) is a hematological finding in hypoxic conditions. We aimed to assess the prevalence of RAA in dogs with CPE due to MMVD, and evaluate whether RAA is reversible with amelioration of dyspnea. Twenty-nine client-owned dogs with CPE due to MMVD were included. Dogs who died within 6 weeks of the onset of CPE were included in the non-survival group, while the others comprised the survival group. Of the 21 dogs, RAA was observed in 17 dogs (80.9%). In the RAA group, the absolute reticulocyte count significantly decreased as CPE resolved (p < 0.001). The mean absolute reticulocyte count in the RAA group was 163.90 ± 50.77 on the first measurement and 78.84 ± 25.64 after resolution of CPE. In the RAA group, no significant differences in mean absolute reticulocyte count were observed between the survival and non-survival groups at either the first or second measurement. Our results indicate that RAA occurs in dogs with MMVD-related CPE and can resolve after resolution of CPE.

A rare case of cecal malignant peripheral nerve sheath tumor in a dog.

A 12-year-old mixed-breed dog presented with a 2-month history of abdominal distension. Radiographic examination, abdominal ultrasonography, and computed tomography revealed a mass in the cecum (15.0 × 11.9 × 4.5 cm). The cecal mass was surgically removed and examined histopathologically. Immunohistochemically, the neoplastic cells expressed S-100 and neuron specific enolase but not α-smooth muscle actin and CD117 (c-kit). These histologic and immunohistochemical features indicated that the mass was consistent with a malignant peripheral nerve sheath tumor (MPNST). In dogs, most MPNSTs arise from the brachial plexus, spinal nerve root, and skin of the extremities. However, gastrointestinal MPNSTs in dogs have not been described previously. To the best of our knowledge, this is the first report to describe cecal MPNST in a dog.

Expression of the hedgehog signalling pathway and the effect of inhibition at the level of smoothened in canine osteosarcoma cell lines.

Osteosarcoma (OSA) is the most common malignant bone cancer in dogs. Canine and human OSA share several features, including tumour environments, response to traditional treatment, and several molecular pathways. Hedgehog (Hh) signalling is known to contribute to tumorigenesis and progression of various cancers, including human OSA. This study aimed to identify the role of the Hh signalling pathway in canine OSA cell lines, including Abrams, D17, and Moresco, focusing on the signal transducer Smoothened (SMO). mRNA and protein levels of Hh pathway components, including SHH, IHH, SMO, and PTCH1, were aberrant in all examined OSA cell lines compared with canine osteoblast cells. The SMO inhibitor cyclopamine significantly decreased cell viability and colony-forming ability in the canine OSA cell lines in a dose-dependent manner. Moresco cells, which expressed the highest level of SMO protein, were the most sensitive to the anticancer effect of cyclopamine among the three canine OSA cell lines tested. Hh downstream target gene and protein expression in canine OSA cell lines were downregulated after cyclopamine treatment. In addition, cyclopamine significantly increased apoptotic cell death in Abrams and Moresco cells. The findings that Hh/SMO is activated in canine OSA cell lines and cyclopamine suppresses OSA cell survival via inhibition of SMO suggest that the Hh/SMO signalling pathway might be a novel therapeutic target for canine OSA.

Big data analysis of the risk factors and rates of perioperative transfusion in immediate autologous breast reconstruction.

Patients undergoing autologous breast reconstruction (ABR) are more likely to require perioperative transfusions due to the increased intraoperative bleeding. In addition to the mastectomy site, further incisions and muscle dissection are performed at the donor sites, including the back or abdomen, increasing the possibility of transfusion. The purpose of this study was to evaluate perioperative transfusion rates and risk factors according to the type of ABR through analysis of big data. Patients who underwent total mastectomy for breast cancer between 2014 and 2019 were identified. The patients were divided into mastectomy only and immediate ABR groups. The transfusion rate was 14-fold higher in the immediate ABR group (16.1%) compared to the mastectomy only group (1.2%). The transfusion rate was highest with the pedicled transverse rectus abdominis myocutaneous flap (24.2%). Performance of the operation in medical institutions located in the provinces and coronary artery disease (CAD) were significant risk factors for the need for transfusion. The perioperative transfusion risk among patients undergoing immediate ABR was related to the flap type, location of medical institution, and CAD. Based on the higher transfusion rate in this study (16.1%) compared to previous studies, the risk factors for the need for transfusion should be determined and evidence-based guidelines should be developed to reduce the transfusion rates.

Transient thrombocytopenia in a cat following G-CSF treatment.

A 4-year-old, castrated male, Russian blue cat with idiopathic epilepsy was diagnosed with neutropenia. The neutropenia was classified as idiopathic after blood tests and abdominal imaging did not reveal an infectious, inflammatory or neoplastic aetiology. As a treatment trial for idiopathic neutropenia, the cat was administered granulocyte colony-stimulating factor by subcutaneous injection once daily for 3 days. Two weeks after completion of granulocyte colony-stimulating factor therapy, the cat developed severe thrombocytopenia, with the granulocyte colony-stimulating factor therapy considered to be the most likely cause. No treatment was initiated, and the thrombocytopenia had resolved spontaneously by 2 weeks after diagnosis. This is the first reported case of transient severe thrombocytopenia in a cat following granulocyte colony-stimulating factor treatment.

Anti-tumor effects of rivoceranib against canine melanoma and mammary gland tumour in vitro and in vivo mouse xenograft models.

BACKGROUND: Rivoceranib, a novel tyrosine kinase inhibitor, exhibits anti-tumour effects by selectively blocking vascular endothelial growth factor receptor-2 (VEGFR2) in cancer cells. Recently, the therapeutic effects of rivoceranib on solid tumours have been elucidated in human patients. However, the anti-tumour effects of rivoceranib against canine cancer remain unclear. Here, we investigated the anti-tumour effects of rivoceranib using in vitro and in vivo mouse xenograft models. METHODS: We performed cell proliferation, cell cycle, and migration assays to determine the effects of rivoceranib on canine solid tumour cell lines in vitro. Furthermore, apoptosis and angiogenesis in tumour tissues were examined using a TUNEL assay and immunohistochemistry methods with an anti-cluster of differentiation-31 antibody, respectively. Additionally, the expression levels of cyclin-D1 and VEGFR2 activity were determined using western blot analysis. RESULTS: Rivoceranib treatment showed anti-proliferative effects and mediated cell cycle arrest in the canine melanoma cell line (LMeC) and the mammary gland tumour (MGT) cell line (CHMp). In animal experiments, rivoceranib decreased the average volume of LMeC cells compared to that following control treatment, and similar results were observed in CHMp cells. Histologically, rivoceranib induced apoptosis and exerted an anti-angiogenic effect in tumour tissues. It also downregulated the expression of cyclin-D1 and inhibited VEGFR2 activity. CONCLUSION: Our results show that rivoceranib inhibits proliferation and migration of tumour cells. These findings support the potential application of rivoceranib as a novel chemotherapeutic strategy for canine melanoma and MGTs.

Extracellular vesicles derived from DFO-preconditioned canine AT-MSCs reprogram macrophages into M2 phase.

BACKGROUND: Mesenchymal stem/stromal cells (MSCs) are effective therapeutic agents that ameliorate inflammation through paracrine effect; in this regard, extracellular vesicles (EVs) have been frequently studied. To improve the secretion of anti-inflammatory factors from MSCs, preconditioning with hypoxia or hypoxia-mimetic agents has been attempted and the molecular changes in preconditioned MSC-derived EVs explored. In this study, we aimed to investigate the increase of hypoxia-inducible factor 1-alpha (HIF-1α)/cyclooxygenase-2 (COX-2) in deferoxamine (DFO)-preconditioned canine MSC (MSCDFO) and whether these molecular changes were reflected on EVs. Furthermore, we focused on MSCDFO derived EVs (EVDFO) could affect macrophage polarization via the transfer function of EVs. RESULTS: In MSCDFO, accumulation of HIF-1α were increased and production of COX-2 were activated. Also, Inside of EVDFO were enriched with COX-2 protein. To evaluate the transferring effect of EVs to macrophage, the canine macrophage cell line, DH82, was treated with EVs after lipopolysaccharide (LPS) stimulation. Polarization changes of DH82 were evaluated with quantitative real-time PCR and immunofluorescence analyses. When LPS-induced DH82 was treated with EVDFO, phosphorylation of signal transducer and transcription3 (p-STAT3), which is one of key factor of inducing M2 phase, expression was increased in DH82. Furthermore, treated with EVDFO in LPS-induced DH82, the expression of M1 markers were reduced, otherwise, M2 surface markers were enhanced. Comparing with EVDFO and EVnon. CONCLUSION: DFO preconditioning in MSCs activated the HIF-1α/COX-2 signaling pathway; Transferring COX-2 through EVDFO could effectively reprogram macrophage into M2 phase by promoting the phosphorylation of STAT3.

3D-culture models as drug-testing platforms in canine lymphoma and their cross talk with lymph node-derived stromal cells.

BACKGROUND: Malignant lymphoma is the most common hematopoietic malignancy in dogs, and relapse is frequently seen despite aggressive initial treatment. In order for the treatment of these recurrent lymphomas in dogs to be effective, it is important to choose a personalized and sensitive anticancer agent. To provide a reliable tool for drug development and for personalized cancer therapy, it is critical to maintain key characteristics of the original tumor. OBJECTIVES: In this study, we established a model of hybrid tumor/stromal spheroids and investigated the association between canine lymphoma cell line (GL-1) and canine lymph node (LN)-derived stromal cells (SCs). METHODS: A hybrid spheroid model consisting of GL-1 cells and LN-derived SC was created using ultra low attachment plate. The relationship between SCs and tumor cells (TCs) was investigated using a coculture system. RESULTS: TCs cocultured with SCs were found to have significantly upregulated multidrug resistance genes, such as P-qp, MRP1, and BCRP, compared with TC monocultures. Additionally, it was revealed that coculture with SCs reduced doxorubicin-induced apoptosis and G2/M cell cycle arrest of GL-1 cells. CONCLUSIONS: SCs upregulated multidrug resistance genes in TCs and influenced apoptosis and the cell cycle of TCs in the presence of anticancer drugs. This study revealed that understanding the interaction between the tumor microenvironment and TCs is essential in designing experimental approaches to personalized medicine and to predict the effect of drugs.