Phase I-II Study of Denileukin Diftitox (ONTAK®) in Patients with Advanced Refractory Breast Cancer.
Authors: Salazar L. , Higgins D. , Childs J. , Bates N. , Dang Y. , Slota M. , Coveler A. , Waisman J. , Disis M. 
Published: December 2010

Abstract
Background: CD4+CD25+Foxp3+ regulatory T cells (Tregs) are potent suppressors of CD4+ and CD8+ T cells, produce the immunosuppressive cytokine TGF-β; and as such, may down-regulate immune responses to tumor antigens. Additionally, Tregs are increased in the peripheral blood (PB) and tumors of breast cancer patients; and are associated with poor prognosis. Depletion of PB and tumor-associated Tregs may induce anti-tumor immunity by augmenting anti-tumor effector T cells and enhancing endogenous tumor specific immunity. ONTAK®, a diphtheria/IL-2R fusion protein depletes PB Tregs when given intravenously (IV) and selectively targets tumor cells that overexpress IL-2R. Breast tumors have been shown to overexpress IL-2R which is associated with their malignant potential. We hypothesized that ONTAK® could (1) have direct anti-tumor activity in breast cancers that overexpress IL-2R, and (2) deplete Tregs resulting in generation of functional immune effector cells and enhanced anti-tumor immunity. A phase I-II study was conducted to evaluate the safety of IV ONTAK® and assess its effect on Tregs and endogenous immunity in patients with advanced refractory breast cancer.Materials and Methods: 15 patients with progressive stage IV breast cancer following standard therapy were sequentially enrolled and received IV ONTAK® 18 mcg/kg/day on Days1-5 every 21 days for a total of 6 cycles and/or maximal tumor response. Toxicity was evaluated on Days 1 8, and 14 of each cycle per CTEP CTCAE v3.0. Tumor response was evaluated per RECIST at baseline, and after cycles 3 and 6. PB was collected at baseline and after cycles 2, 4, and 6 for evaluation of Tregs, sIL-2R, and endogenous tumor-antigen specific T cell immunity to HER-2/neu (HER2), CEA, and MAGE-3 via RT-PCR, LUMINEX and IFN-γ ELISPOT assay, respectively. Expression of IL-2R in patient paraffin embedded tumor samples was analyzed by IHC analysis.Results: 15 subjects have been enrolled and 14/15 have completed treatment; median age is 58 years (range, 32-69) and median salvage regimens is 3 (range, 2-8). 7/14 subjects had triple negative tumors. 7 subjects completed 1-2 and 7 completed 3-6 ONTAK® cycles, respectively. 4 subjects who completed 6 cycles of ONTAK® had SD or PR per RECIST. ONTAK®-related toxicities have been primarily grade I and II fatigue, nausea, and headache; and transient grade 3 hypoalbuminemia and lymphopenia. Preliminary data in 2 subjects shows enhanced tumor-antigen specific T cell immunity defined as mean tumor antigen-specific T cell precursors:PBMC to CEA (pre- ONTAK® 1:250,000; post- ONTAK® 1:15,000) and HER2 (pre- ONTAK® 1:63,000; post- ONTAK® 1:6,312). Immunologic analyses are ongoing and will be presented along with clinical data on all patients.Conclusions: ONTAK® is well-tolerated when used as a salvage regimen in heavily pretreated breast cancer patients. Additionally, ONTAK® treatment can enhance endogenous immunity to known breast cancer antigens and potentially lead to more effective eradication of tumor.
Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 4130.