Cross-species oncogenomics offers insight into human muscle-invasive bladder cancer.

BACKGROUND: In humans, muscle-invasive bladder cancer (MIBC) is highly aggressive and associated with a poor prognosis. With a high mutation load and large number of altered genes, strategies to delineate key driver events are necessary. Dogs and cats develop urothelial carcinoma (UC) with histological and clinical similarities to human MIBC. Cattle that graze on bracken fern also develop UC, associated with exposure to the carcinogen ptaquiloside. These species may represent relevant animal models of spontaneous and carcinogen-induced UC that can provide insight into human MIBC. RESULTS: Whole-exome sequencing of domestic canine (n = 87) and feline (n = 23) UC, and comparative analysis with human MIBC reveals a lower mutation rate in animal cases and the absence of APOBEC mutational signatures. A convergence of driver genes (ARID1A, KDM6A, TP53, FAT1, and NRAS) is discovered, along with common focally amplified and deleted genes involved in regulation of the cell cycle and chromatin remodelling. We identify mismatch repair deficiency in a subset of canine and feline UCs with biallelic inactivation of MSH2. Bovine UC (n = 8) is distinctly different; we identify novel mutational signatures which are recapitulated in vitro in human urinary bladder UC cells treated with bracken fern extracts or purified ptaquiloside. CONCLUSION: Canine and feline urinary bladder UC represent relevant models of MIBC in humans, and cross-species analysis can identify evolutionarily conserved driver genes. We characterize mutational signatures in bovine UC associated with bracken fern and ptaquiloside exposure, a human-linked cancer exposure. Our work demonstrates the relevance of cross-species comparative analysis in understanding both human and animal UC.

'Know thyself' - host factors influencing cancer response to immune checkpoint inhibitors.

Immune checkpoint inhibitors (ICIs) have revolutionised oncology and are now standard-of-care for the treatment of a wide variety of solid neoplasms. However, tumour responses remain unpredictable, experienced by only a minority of ICI recipients across malignancy types. Therefore, there is an urgent need for better predictive biomarkers to identify a priori the patients most likely to benefit from these therapies. Despite considerable efforts, only three such biomarkers are FDA-approved for clinical use, and all rely on the availability of tumour tissue for immunohistochemical staining or genomic assays. There is emerging evidence that host factors - for example, genetic, metabolic, and immune factors, as well as the composition of one's gut microbiota - influence the response of a patient's cancer to ICIs. Tantalisingly, some of these factors are modifiable, paving the way for co-therapies that may enhance the therapeutic index of these treatments. Herein, we review key host factors that are of potential biomarker value for response to ICI therapy, with a particular focus on the proposed mechanisms for these influences. © 2022 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

Boosting antitumor response with PSMA-targeted immunomodulatory VLPs, harboring costimulatory TNFSF ligands and GM-CSF cytokine.

Therapeutic strategies based on immunomodulation have improved cancer therapy. Most approaches target co-stimulatory pathways or the inhibition of immunosuppressive mechanisms, to enhance immune response and overcome the immune tolerance of tumors. Here, we propose a novel platform to deliver targeted immunomodulatory signaling, enhancing antitumor response. The platform is based on virus-like particles derived from lentiviral capsids. These particles may be engineered to harbor multifunctional ligands on the surface that drive tropism to the tumor site and deliver immunomodulatory signaling, boosting the antitumor response. We generated virus-like particles harboring a PSMA-ligand, TNFSF co-stimulatory ligands 4-1BBL or OX40L, and a membrane-anchored GM-CSF cytokine. The virus-like particles are driven to PSMA-expressing tumors and deliver immunomodulatory signaling from the TNFSF surface ligands and the anchored GM-CSF, inducing T cell proliferation, inhibition of regulatory T cells, and potentiating elimination of tumor cells. The PSMA-targeted particles harboring immunomodulators enhanced antitumor activity in immunocompetent challenged mice and may be explored as a potential tool for cancer immunotherapy.

Aptamer-mediated transcriptional gene silencing of Fox p 3 inhibits regulatory T cells and potentiates antitumor response.

The inhibition of immunosuppressive mechanisms may switch the balance between tolerance and surveillance, leading to an increase in antitumor activity. Regulatory T cells play an important role in the control of immunosuppression, exhibiting the unique property of inhibiting T cell proliferation. These cells migrate to tumor sites or may be generated at the tumor site itself from the conversion of lymphocytes exposed to tumor microenvironment signaling. Because of the high similarity between regulatory T cells and other lymphocytes, the available approaches to inhibit this population are nonspecific and may antagonize antitumor response. In this work we explore a new strategy for inhibition of regulatory T cells based on the use of a chimeric aptamer targeting a marker of immune activation harboring a small antisense RNA molecule for transcriptional gene silencing of Fox p 3, which is essential for the control of the immunosuppressive phenotype. The silencing of Fox p 3 inhibits the immunosuppressive phenotype of regulatory T cells and potentiates the effect of the GVAX antitumor vaccine in immunocompetent animals challenged with syngeneic tumors. This novel approach highlights an alternative method to antagonize regulatory T cell function to augment antitumor immune responses.

Extracellular vesicles produced by immunomodulatory cells harboring OX40 ligand and 4-1BB ligand enhance antitumor immunity.

Genetically modified tumor cells harboring immunomodulators may be used as therapeutic vaccines to stimulate antitumor immunity. The therapeutic benefit of these tumor vaccines is extensively investigated and mechanisms by which they boost antitumor response may be further explored. Tumor cells are large secretors of extracellular vesicles (EVs). These EVs are able to vehiculate RNA and proteins to target cells, and engineered EVs also vehiculate recombinant proteins. In this study, we explore immunomodulatory properties of EVs derived from antitumor vaccines expressing the TNFSF ligands 4-1BBL and OX40L, modulating immune response mediated by immune cells and eliminating tumors. Our results suggest that the EVs secreted by genetically modified tumor cells harboring TNFSF ligands can induce T cell proliferation, inhibit the transcription factor FoxP3, associated with the maintenance of Treg phenotype, and enhance antitumor activity mediated by immune cells. The immunomodulatory extracellular vesicles have potential to be further engineered for developing new approaches for cancer therapy.

Development of a flow cytometry assay which allows to evaluate the efficiency of immunomodulatory vaccines to enhance T cell-mediated antitumor response.

Immunotherapy has revolutionized the treatment of cancer. Since tumor cells exhibit low immunogenicity and can induce several mechanisms of tolerance, the use of monoclonal antibodies or other immunomodulators, targeting costimulation of T cells may mediate the inhibition of immunosuppressive mechanisms, favouring immune surveillance and enhancing the detection and elimination of tumor cells. We developed a new in vitro assay, based on flow cytometry, which allows exploring the therapeutic potential of tumor-derived immunomodulatory lineages, enhancing anti-tumor response. We generated tumor-derived cells that simultaneously co-express eGFP and one immunomodulatory molecule (OX40L, 4-1BBL or GM-CSF). These genetically modified tumor-derived cells are irradiated and then incubated with primary T cells to evaluate the killing activity, which can be estimated by a decrease in the eGFP positive cells. The results have shown correlation with in vivo experiments. This model may contribute to the development of high-throughput assays for the screening of immunomodulators and a reduction in the use of experimental animals.

Exploring Synergy in Combinations of Tumor-Derived Vaccines That Harbor 4-1BBL, OX40L, and GM-CSF.

Recent studies have demonstrated that combination of modulatory immune strategies may potentiate tumor cell elimination. Most strategies rely on the use of monoclonal antibodies that can block cell surface receptors to overcome tumor-induced immunosuppression or acting as costimulatory ligands to boost activation of T cells. In this study, we evaluate the use of combinations of genetically modified tumor-derived cell lines that harbor the costimulatory T cell ligands 4-1BB ligand, OX40L, and the cytokine GM-CSF. The aim of these treatments is to boost the activation of T cells and the elimination of cancer cells. These tumor-derived cells are able to activate or reinforce T cell activation, thereby generating a potent and specific antitumor response. We developed a high-content in vitro imaging assay that allowed us to investigate synergies between different tumor-derived cells expressing modulatory immune molecules, as well as the influence on effector T cells to achieve tumor cell death. These results were then compared to the results of in vivo experiments in which we challenged immunocompetent animals using the B16F10 syngeneic model of melanoma in C57BL6 mice. Our results suggest that there is a substantial therapeutic benefit to using combinations of syngeneic tumor vaccines that express immune modulators. In addition, we observed that combinations of tumor-derived cells that expressed costimulatory ligands and GM-CSF induced a long-term protective effect by preventing cancer development in both cured and rechallenged animals.

Marcadores genéticos en pacientes con síndrome de ovario poliquístico y dos de sus familiares en primer grado de consanguinidad: estudio de casos y controles / Genetic markers in patients with polycystic ovary syndrome and in two relatives in the first degree of consanguinity: case control study

Objetivo: estimar la asociación entre los marcadores genéticos D19S884 y UCSNP-19 y el síndrome de ovario poliquístico (SOP).Materiales y métodos: estudio de 50 casos con SOP según criterios de Rotterdam y 100 controles: dos familiares femeninos en primer grado de consanguinidad sin la enfermedad. Se evaluaron características sociodemográficas y clínicas. Los marcadores genéticos D19S884 y UCSNP-19 se identificaron por reacción en cadena de la polimerasa. Las variables cuantitativas se presentan con media ± desviación estándar; se utilizaron las pruebas de t de Student y de McNemar. Se calcularon los OR y el IC 95%. Resultados: la edad promedio en los casos fue 23 ± 6 años y en los controles de 39 ± 18 años. Los casos se asociaron de manera significativa a hirsutismo OR = 3,6 (IC 95%: 1,3-12,8) y acné OR = 4,3 (IC 95%: 1,4-17,4). Los polimorfismos del ins e ins/ins de UCSNP-19 tuvieron las mayores proporciones en los dos grupos de estudio, siendo el primero más frecuente en casos y el segundo en controles; sin embargo, esta diferencia no fue estadísticamente significativa. Se identificaron 14 alelos de D19S884 que van desde 215 a 242 pb. Conclusiones: no se encontró asociación entre el polimorfismo UCSNP-19 del gen CAPN10 y del marcador D19S994 con el SOP en la población estudiada

Objective: To estimate the association between genetic markers D19S884 and UCSNP-19 and Polycystic Ovary Syndrome (PCOS).Materials and methods: Study of 50 cases of PCOS consistent with the Rotterdam criteria and of 100 controls and two first-degree female relatives without the disease. Social, demographic and clinical characteristics were assessed, and genetic markers D19S884 and UCSNP-19 were identified using polymerase chain reaction. Quantitative variables are expressed as mean ± standard deviation; the Student t test and the McNemar test were used. Odds ratios and 95% confidence intervals were estimated. Results: Mean ages were 23 ± 6 years and 39 ± 18 years for the cases and controls, respectively. Cases showed a significant association with hirsutism OR = 3.6 (CI 95%: 1.3-12.8) and acne OR = 4.3 (CI 95%: 91.4-17.4). Del/ins and ins/ins polymorphisms of UCSNP-19 were found in the highest proportions in the two study groups, the former being more frequent among the cases and the latter among the controls. However, this difference was not statistically significant. Fourteen alleles of D19S884 were identified, ranging from 215 to 242 bp. Conclusions: No association was found between the CAPN10 gene UCSNP-19 polymorphism and the D19S994 marker with PCOS in the population studied