Steven Suter

This randomized, prospective, controlled clinical trial is designed to obtain data to evaluate the effectiveness and safety of Autologous Prescription Product, Code 95A7.50 (unlicensed), also referred to as the ELIAS Cancer Immunotherapy (ECI���������) as a treatment for dogs with appendicular osteosarcoma. All dogs will undergo amputation of the affected limb and the tumor will be submitted to ELIAS. Dogs will be randomized into one of two cohorts: one cohort will receive the experimental immunotherapy, while the other cohort will receive standard of care chemotherapy, consisting of four (4) doses of carboplatin. The outcomes of the two cohorts will be compared for effectiveness and any adverse events occurring during the study will be noted and assessed.

The supplement funds supports projects for Dr. Steven Suter: (1) Phase 2 of the MDH Pipeline-Bortezomib as First Line Therapy for Osteosarcoma and (2) Testing a new Prodrug Nanoparticle Formulation of Niclosamide as Novel Therapy for Osteosarcoma. The 3rd project these funds support is for Dr. Mariani (3) Establishing Laser Ablation as a Treatment for Canine Giloma and Modeling the Human Condition

The purpose of these studies are to: 1)Investigate the antimetastatic effects of rapamycin when added to an SOC backbone for the treatment of canine osteosarcoma, as measured by statistically significant improvement in disease-free interval (DFI) over SOC alone in a prospective, randomized clinical trial setting, and 2)Identify key factors related to tolerability and clinical efficacy of rapamycin when studied in the Minimal Residual Disease (MRD) setting, such as PK parameters, evidence of target modulation within surrogate tissues, and patient/tumor related factors

The purpose of these studies are to: 1)Investigate the antimetastatic effects of rapamycin when added to an SOC backbone for the treatment of canine osteosarcoma, as measured by statistically significant improvement in disease-free interval (DFI) over SOC alone in a prospective, randomized clinical trial setting, and 2)Identify key factors related to tolerability and clinical efficacy of rapamycin when studied in the Minimal Residual Disease (MRD) setting, such as PK parameters, evidence of target modulation within surrogate tissues, and patient/tumor related factors

Canine diffuse large B-cell lymphoma (DLBCL) is a common, aggressive cancer in dogs. The average survival time after initial diagnosis is one year. Unfortunately, the diagnosis of DLBCL is often made late in disease and after the cancer is advanced, which negatively impacts the survival of the dog. Therefore, there is a need to 1) develop non-invasive screening methods to diagnose this aggressive cancer in its early stages and 2) identify novel therapies to treat this cancer. In human oncology, the discovery of a new type of gene called long non-coding RNA (lncRNA) has led to the development of non-invasive screening methods for certain cancers. These lncRNAs are also being explored for their use as new cancer targets for drug development, which are expected to have fewer side effects than currently treatments. Since these lncRNA can be detected in blood of cancer patients, they can be used in non-invasive, early detection assays for some cancers. Recently, in human patients with DLBCL, lncRNA HOX transcript antisense RNA (HOTAIR) is highly expressed in human DLBCLs, as well as other cancers, and its expression is predictive of a poor prognosis; it is also being explored as a new target for cancer therapy. We will investigate canine lncRNA HOTAIR to determine if it is expressed in canine DLBCL and can be detected in the blood of cancer patients. If successful, this proposal will open new lines of research improving the detection and treatment of a common and devastating cancer of dogs.

While genetic modification of mice has resulted in the generation of transgenic lines that have helped elucidate the role of a myriad of molecules in biological and pathological processes, the same is not true in swine. Technically, the mouse can be genetically manipulated not only inexpensively but also with exquisite precision. Xenotransplantation of pig organs into humans has the potential to significantly alleviate the problems associated with organ donor availability. Unfortunately, before swine can be used as an organ donor for humans it will be necessary to diminish or abolish all or most rejection phenomena including hyperacute rejection, acute vascular rejection, cellular rejection, and chronic rejection. This will require not only a thorough understanding of the rejection phenomena in xenotransplantation, but equally important a series of complex genetic modifications to modify swine molecules involved in each of the rejection steps. We propose here the continuation of the development of technology that will allow such precise genetic manipulation. To reach this overall goal we propose to: (a) Develop mechanism for enhancing homologous recombination in somatic cells. (b) Extend our abilities to do complex genetic modifications in somatic cells by extension of cellular life span. (c) Generate cloned swine using non-transgenic and genetically modified fetal fibroblasts. (d). Develop transgenic pigs that can be used in xenotrasnplantation and stem cell biology. Completion of the proposed aims will result not only in the generation of swine that can be used to facilitate the development of xenotransplant organs, but equally important it will result in the development of technology to allow the application of precise genetic manipulations to swine.

T-cell lymphomas (TCL), which account for approximate 10-15% of human lymphomas, have a much poorer prognosis than B-cell lymphomas. Therapy development for TCL has been slow because of the relative rarity of the disease, subtype heterogeneity within the broader category of TCL, and, importantly, the lack of faithful animal models. Recent data suggests that the vast majority of boxer T-cell lymphomas are comprised of TCRαβ+ CD4+ T-cells. This is in stark contrast to T-cell lymphomas in other breeds of dogs where TCRαβ and CD4/CD8 expression is highly variable. Similar to human T-cell lymphoma patients, canine T-cell lymphoma patients, including boxers, have a much shorter overall survival than dogs with BCL. We hypothesize that the development of this very specific type of T-cell lymphoma (CD4+ helper T-cell) in a highly inbred breed of dogs, such as boxers, is a heritable disease. Since human TCL is difficult to study because of its rarity and heterogeneity, boxers may provide a robust animal model to elucidate the drivers of T-cell lymphomagenesis. The gene(s) we identify could provide essential information about the biology of TCL that are relevant to both canine and human TCL, reinforcing the One Health concept.

Thrombocytopenia is a common blood abnormality defined by a decreased platelet number. Platelets are key contributors to hemostasis, or the ability to form a stable blood clot to prevent bleeding. Patients with critically low platelet counts are at increased risk for developing spontaneous, life-threatening hemorrhage. Guidelines in human medicine advocate immediate and aggressive therapeutic intervention to avoid life-threatening bleeding by using platelet transfusions. Our institution recently evaluated the use of a bed-side analyzer (thromboelastography or TEG), platelet count, and standard coagulation profile to predict bleeding in a thrombocytopenic population of canines and found TEG, specifically the maximum amplitude (MA), to be the best indicator of bleeding (manuscript in progress). This tool is already used in human medicine to assess hemostasis and guide transfusions in thrombocytopenic patients undergoing surgical procedures. This study attempts to identify the most efficacious platelet transfusion product to prevent and potentially treat bleeding episodes in the same aforementioned population of thrombocytopenic canines. The investigators anticipate that this data will serve as a foundation for future studies to improve management and reduce mortality due to life-threatening hemorrhage in thrombocytopenic dogs. Also, financial resources can be directed towards the most efficacious product available, as pet owners? financial restrictions often dictate whether or not animals continue with treatment.

Receptor tyrosine kinase (RTK) inhibitors are an emerging treatment for dogs with mast cell tumors. Approximately 30% of dogs with mast cell tumors have a mutation in a RTK protein called c-kit. Therefore, these dogs may respond better to a drug called Palladia, which directly inhibits this protein when it is mutated. We have also recently found that a canine cell line called GL-1 has a mutation in another RTK called FLT3 and that ~25% of dogs with acute lymphocytic leukemia harbor this mutation. Preliminary data suggests that Palladia, and perhaps another commercially available RTK inhibitor called masitinib, also bind to and inhibit FLT3. The goal of this study is to utilize canine cell lines that either do or don't express FLT3 to determine if Palladia or masitinib have growth inhibitory effects. If so, these findings may lay the groundwork for future studies whereby these drugs can be tested in dogs who have acute lymphocytic leukemia.

Thrombocytopenia, or decreased platelet number, is a common hemostatic disorder and one of the most common causes of spontaneous bleeding in canines encountered by veterinarians. A recent retrospective study of thrombocytopenia in a referral hospital canine population found an incidence of 6.7 percent. Patients with critically low platelet counts are at increased risk for developing spontaneous, life-threatening hemorrhage. This can be devastating for critically ill patients and results in prolonged intensive care unit (ICU) stays and increased mortality rates. Guidelines in human medicine advocate immediate and aggressive therapeutic intervention to avoid life-threatening bleeding by using platelet transfusions. However, no clinical study has been published to date evaluating the efficacy of the available fresh and stored platelet products in a population of thrombocytopenic dogs. Our institution recently evaluated the use of a bed-side analyzer (thromboelastography or TEG), platelet count, and standard coagulation profile to predict bleeding in a thrombocytopenic population of canines and found TEG, specifically the maximum amplitude (MA), to be the best indicator of bleeding (manuscript in progress). This tool is already used in human medicine to assess hemostasis and guide transfusions in thrombocytopenic patients undergoing surgical procedures. As platelet transfusions become more common in clinical veterinary medicine, it will be important to provide evidence-based information regarding the efficacy of platelet transfusion products. This study attempts to identify the most efficacious platelet transfusion product to prevent and potentially treat bleeding episodes in a population of dogs with total body irradiation (TBI) induced thrombocytopenia using TEG. The results of this study may have immediate clinical impact on the selection of platelet transfusion products in critically ill thrombocytopenic patients, and provide a foundation for larger-scale research projects in the areas of transfusion medicine and management of thrombocytopenic patients.