Adoptive T Cell Therapy for Malignancies in Dogs—A Model of Hope Edmund Sullivan, DVM Bellingham, WA Thank you for attending our presentation today on the topic of allogeneic stem cell transplantation for the treatment of canine lymphoma. Dr. Sullivan and I have been working for the last 3 ½ years to make available to client-owned dogs the transplantation techniques that were developed over many years by researchers at the Fred Hutchinson Cancer Research Center in Seattle. Through their generous participation and sharing of knowledge, we have been able to apply these techniques to our patients with lymphoma.

Conflict of Interest Disclosures Edmund Sullivan, DVM Affiliations: Fred Hutchinson Cancer Research Center Hematologics Corporation Engene Oncology The Perseus Foundation The NorthWest Veterinary Blood Bank Advanced Veterinary Service, Inc. Cellphire, Inc. We would like to acknowledge and thank our affiliate institutions and members of the biotechnology industry that have contributed to our efforts.

Dogs are the model for transplantation techniques Demonstrated and expanded upon at Fred Hutch by Thomas, Storb, Applebaum and others Wide variation in phenotype is the result of a small difference in genotype Share environment with humans—diet, home, yard, car, and most recently identified, biome Life-time outcome of treatment known in brief time In humans, allogeneic hematopoietic stem cell transplants represent an important therapeutic tool for the treatment of otherwise incurable malignant and nonmalignant diseases. Because of the similarities in the immune systems of dogs and humans, dogs are an excellent large animal model for transplantation procedures, and over the last 40 years a huge volume of data on transplantation techniques in dogs has accumulated. Much of this information is applicable to dogs with naturally occurring diseases, and our efforts have been to assimilate this data into protocols that allow treatment of dogs. There are several recent advances that have contributed to the feasibility of completing these transplants.

Are Dogs the model for Adoptive Immunotherapy? Selection, expansion, stimulation, and infusion of CD8+ T Cells Anti tumor whole-cell vaccines prior to cell harvest Mobilization and apheresis—enhances numbers and quality(?) of T cells Pilot study with dogs using T cells after CHOP chemotherapy—no consolidation In humans, allogeneic hematopoietic stem cell transplants represent an important therapeutic tool for the treatment of otherwise incurable malignant and nonmalignant diseases. Because of the similarities in the immune systems of dogs and humans, dogs are an excellent large animal model for transplantation procedures, and over the last 40 years a huge volume of data on transplantation techniques in dogs has accumulated. Much of this information is applicable to dogs with naturally occurring diseases, and our efforts have been to assimilate this data into protocols that allow treatment of dogs. There are several recent advances that have contributed to the feasibility of completing these transplants.

”Comet” 6 Yr Golden Retreiver T cell lymphoma—treated with an allogeneic stem cell transplant Secondary Malignancy—Fibrosarcoma of distal tibia Treated with Adoptive T Cells from allogeneic donor after the donor was primed with a whole cell tumor vaccine made from the fibrosarcoma In humans, allogeneic hematopoietic stem cell transplants represent an important therapeutic tool for the treatment of otherwise incurable malignant and nonmalignant diseases. Because of the similarities in the immune systems of dogs and humans, dogs are an excellent large animal model for transplantation procedures, and over the last 40 years a huge volume of data on transplantation techniques in dogs has accumulated. Much of this information is applicable to dogs with naturally occurring diseases, and our efforts have been to assimilate this data into protocols that allow treatment of dogs. There are several recent advances that have contributed to the feasibility of completing these transplants.

”Tucker” 5 yr Visla with B cell lymphoma that was non- responsive to induction chemotherapy Allogeneic stem cell transplant from brother induced a brief remission Adoptive T cell transfer (selected, expanded, stimulated) from the donor resulted in 4 year remission (and counting) In humans, allogeneic hematopoietic stem cell transplants represent an important therapeutic tool for the treatment of otherwise incurable malignant and nonmalignant diseases. Because of the similarities in the immune systems of dogs and humans, dogs are an excellent large animal model for transplantation procedures, and over the last 40 years a huge volume of data on transplantation techniques in dogs has accumulated. Much of this information is applicable to dogs with naturally occurring diseases, and our efforts have been to assimilate this data into protocols that allow treatment of dogs. There are several recent advances that have contributed to the feasibility of completing these transplants.

”Jasper” 9 yr Golden Retreiver with B cell lymphoma Autologous stem cell transplant, with subsequent relapse and disease progression Adoptive T cell infusions Molecular remission for 3 years—no evidence of lymphoma at the time of death In humans, allogeneic hematopoietic stem cell transplants represent an important therapeutic tool for the treatment of otherwise incurable malignant and nonmalignant diseases. Because of the similarities in the immune systems of dogs and humans, dogs are an excellent large animal model for transplantation procedures, and over the last 40 years a huge volume of data on transplantation techniques in dogs has accumulated. Much of this information is applicable to dogs with naturally occurring diseases, and our efforts have been to assimilate this data into protocols that allow treatment of dogs. There are several recent advances that have contributed to the feasibility of completing these transplants.

”Bleu” 9 yr Black Lab with Acute Myeloid Leukemia Induction chemotherapy, allogeneic stem cell transplant from niece Disease progression post transplant Allogeneic T cells—selected, expanded, stimulated—infused Molecular remission at time of death In humans, allogeneic hematopoietic stem cell transplants represent an important therapeutic tool for the treatment of otherwise incurable malignant and nonmalignant diseases. Because of the similarities in the immune systems of dogs and humans, dogs are an excellent large animal model for transplantation procedures, and over the last 40 years a huge volume of data on transplantation techniques in dogs has accumulated. Much of this information is applicable to dogs with naturally occurring diseases, and our efforts have been to assimilate this data into protocols that allow treatment of dogs. There are several recent advances that have contributed to the feasibility of completing these transplants.

”Miles Davis”, “Chad”, and “Rosco” B Cell lymphoma Induction Chemotherapy Consolidation with high dose Cytoxan Molecular Remission—3 years and counting In humans, allogeneic hematopoietic stem cell transplants represent an important therapeutic tool for the treatment of otherwise incurable malignant and nonmalignant diseases. Because of the similarities in the immune systems of dogs and humans, dogs are an excellent large animal model for transplantation procedures, and over the last 40 years a huge volume of data on transplantation techniques in dogs has accumulated. Much of this information is applicable to dogs with naturally occurring diseases, and our efforts have been to assimilate this data into protocols that allow treatment of dogs. There are several recent advances that have contributed to the feasibility of completing these transplants.

Are Dogs a model for Adoptive Immunotherapy? In humans, allogeneic hematopoietic stem cell transplants represent an important therapeutic tool for the treatment of otherwise incurable malignant and nonmalignant diseases. Because of the similarities in the immune systems of dogs and humans, dogs are an excellent large animal model for transplantation procedures, and over the last 40 years a huge volume of data on transplantation techniques in dogs has accumulated. Much of this information is applicable to dogs with naturally occurring diseases, and our efforts have been to assimilate this data into protocols that allow treatment of dogs. There are several recent advances that have contributed to the feasibility of completing these transplants.

YES, they are. Huge area of interest in human medicine Few regulatory hurdles, rapid disease progression, lifetime consequences known in relatively short time Discoveries made in dogs will illuminate the path for people In humans, allogeneic hematopoietic stem cell transplants represent an important therapeutic tool for the treatment of otherwise incurable malignant and nonmalignant diseases. Because of the similarities in the immune systems of dogs and humans, dogs are an excellent large animal model for transplantation procedures, and over the last 40 years a huge volume of data on transplantation techniques in dogs has accumulated. Much of this information is applicable to dogs with naturally occurring diseases, and our efforts have been to assimilate this data into protocols that allow treatment of dogs. There are several recent advances that have contributed to the feasibility of completing these transplants.

The Path Forward No shortage of patients—B cell lymphoma and other malignancies are common in dogs Better techniques for growing the APC cells. Currently using flask-based systems Faster, better techniques for collecting and expanding T cells. Currently using flask-based systems which are slow (average 32 days) and susceptible to contamination In humans, allogeneic hematopoietic stem cell transplants represent an important therapeutic tool for the treatment of otherwise incurable malignant and nonmalignant diseases. Because of the similarities in the immune systems of dogs and humans, dogs are an excellent large animal model for transplantation procedures, and over the last 40 years a huge volume of data on transplantation techniques in dogs has accumulated. Much of this information is applicable to dogs with naturally occurring diseases, and our efforts have been to assimilate this data into protocols that allow treatment of dogs. There are several recent advances that have contributed to the feasibility of completing these transplants.

The Perseus Foundation Our non-profit partner that is providing a research and development grant to establish adoptive immunotherapies in dogs with the hope and expectation of translation to people. In humans, allogeneic hematopoietic stem cell transplants represent an important therapeutic tool for the treatment of otherwise incurable malignant and nonmalignant diseases. Because of the similarities in the immune systems of dogs and humans, dogs are an excellent large animal model for transplantation procedures, and over the last 40 years a huge volume of data on transplantation techniques in dogs has accumulated. Much of this information is applicable to dogs with naturally occurring diseases, and our efforts have been to assimilate this data into protocols that allow treatment of dogs. There are several recent advances that have contributed to the feasibility of completing these transplants.