PROJECT SUMMARY / ABSTRACT The broad goal of Project 3 is to devise and implement novel strategies of effective, low-toxicity EBV-specific T cell (EBVST) therapy for EBV-positive lymphomas; approximately 30% of all human lymphomas. The lymphoma environment lacks the positive signaling required to maintain the anti-tumor activities of EBVSTs and instead expresses inhibitory signals producing EBVST anergy. Our constitutive interleukin 7 receptor (C7R) not only provides intrinsic cytokine signaling but activates an anti-apoptotic program that provides resistance to tumor induced T-cell dysfunction. C7R produced expansion of GD2.CAR T-cells that was similar to lymphodepleting chemotherapy (LD) in patients with neuroblastoma, and unlike LD, did not induce cytokine release syndrome (CRS). C7R expression in EBVSTs produced more rapid clearance of autologous EBV-transformed B-cell tumors in an NSG xenograft model and in Aim 1, we will test the hypothesis that C7R will enhance the expansion and persistence of EBVSTs in patients with lymphoma, will provide resistance to the immunosuppressive tumor microenvironment and provide a non-toxic alternative to LD. EBV-reactivation produces exponential expansion of infused autologous EBVSTs and tumor clearance in patients with EBV+ lymphoma. In Aim 2 we will clinically evaluate an EBV latency reversal agent, panobinostat (PBST) to enhance the expansion and function of C7R-EBVSTs. PBST also has direct anti-lymphoma activity by downregulating STAT5 and increasing the expression of pro-apoptotic caspases. These inhibitory activities will be counteracted in EBVSTs by the C7R that upregulates STAT5. Thus, our second hypothesis (Aim 2) is that PBST will simultaneously increase antigen stimulation of C7R-EBVST while selectively increasing susceptibility of the tumor cells to C7R-EBVST killing. The ideal therapy for EBV+ lymphoma would be off-the-shelf, partially-HLA matched C7R-EBVSTs that are potent, immediately available, and relatively inexpensive. However, graft versus host disease (GVHD) and graft rejection caused by alloreactive T-cells curtail the clinical efficacy of allogeneic cell therapies. Aim 3 will evaluate CD30.CAR-modified C7R-EBVSTs to resolve both problems, EBVSTs have not produced GVHD in hundreds of allogeneic recipients in the stem cell transplant setting, and since alloactivated T-cells upregulate CD30, they should be eliminated by the CD30.CAR. Indeed banked CD30.CAR-EBVSTs have safely produced clinical responses (3 complete), in 6 of 8 patients with CD30+lymphoma. However, CD30.CAR-EBVSTs did not persist. Hence Aim 3 will test the hypothesis that C7R will enhance CD30.CAR-EBVST persistence and anti-tumor activity in a murine allo-rejection model. These 3 hypotheses will be tested in the three specific Aims: Aim 1. Can C7R enhance expansion, and long-term activity of EBVSTs in patients with EBV+ lymphoma? Aim 2. Can panobinostat boost EBVST expansion and anti-tumor activity in patients? Aim 3. Can C...