This proposal includes a career development plan and research plan that will provide Dr. Mullinax with the expertise and data necessary to reach his long-term career goal of becoming an independent surgeon- scientist. The short-term career goals identified in this plan include developing expertise in the conduct of early phase clinical trials, the bioinformatics analysis of genomic datasets, and the immunobiology of T-cell subsets present in soft tissue sarcoma. The career development plan and scientific aims are in alignment to ensure efficient progress during this award, leveraging both the institutional strength of Moffitt Cancer Center and scientific expertise of his mentors in the delivery of cellular immunotherapy. The clinical trial included on this proposal is based on the preclinical work that Dr. Mullinax has completed over the past three years since joining the faculty from fellowship in surgical oncology. His work has focused on methods to expand tumor infiltrating lymphocytes (TIL) from soft tissue sarcoma (STS). After demonstrating the feasibility of expanding tumor-reactive TIL, a fundamental requirement for adoptive cell therapy (ACT), he completed an FDA investigational new drug (IND) application, developed a clinical trial protocol, and obtained funding for activation. With these efforts successful, he is now the principal investigator of a clinical trial which will assess the safety and feasibility of ACT in patients with metastatic STS. The scientific work he has proposed includes a deeper study of the interaction between the tumor and T-cells within STS. The molecular pathogenesis of STS is different than most epithelial tumors where copy number variation or reciprocal chromosomal translocation results in oncogenesis of mesenchymal tissue, as opposed to the point mutations commonly seen in epithelial malignancies. Dr. Mullinax has outlined a scope of work that will utilize bioinformatics analysis to understand the relationship between genomic instability and the expansion of TIL from STS. He will also complete experiments to understand the tumor-specific activity of T- cell subsets, specifically tissue resident memory (TRM) T-cells, as a method of selecting optimal clonal expansion in the cellular infusion product delivered to patients in ACT clinical trials. Completion of this work will be overseen closely by his primary mentor Shari Pilon-Thomas, PhD and co- mentor James J Mulé, PhD. He will meet weekly with his primary mentor, bi-weekly with his clinical trial advisors (Damon Reed, MD and Amod Sarnaik MD), and quarterly with his complete advisory committee which adds Jose Conejo-Garcia, MD, PhD and Jamie Teer, PhD to the above faculty. He has described a rigorous schedule of didactic course work to address training the ethics of trial design, grant writing, and biostatistics. Completion of this work will position Dr. Mullinax well for R01 submission during the penultimate year of this award and ensure the expertise needed to conduct early phase clinical trials which translate the findings of his laboratory to therapeutic options for the STS patients that he treats in his clinical practice.

PROJECT NARRATIVE Adoptive cell therapy (ACT) has demonstrated durable objective response in patients with solid tumors but has not been previously tested in patients with metastatic soft tissue sarcoma (mSTS). I will conduct a phase 1 trial of ACT in patients with mSTS where the safety of this regimen will be assessed as the primary endpoint and use tissue from this trial for laboratory investigations that evaluate the T-cell subsets responsible for tumor- specific activity and the genomic events of these tumors relative to the T-cell infiltration. I have also described career development goals that align with the scope of scientific work to ensure my successful transition from mentored junior faculty to independent surgeon-scientist with the ability to safely deliver clinical trials designed from my laboratory research.