PD1 Expression in EGFRvIII-Directed CAR T Cell Infusion Product for Glioblastoma Is Associated with Clinical Response.

The epidermal growth factor receptor variant III (EGFRvIII) has been investigated as a therapeutic target for chimeric antigen receptor (CAR) T cell therapy in glioblastoma. Earlier research demonstrated that phenotypic and genotypic characteristics in T cells and CAR T product predicted therapeutic success in hematologic malignancies, to date no determinants for clinical response in solid tumors have been identified. We analyzed apheresis and infusion products from the first-in-human trial of EGFRvIII-directed CAR T for recurrent glioblastoma (NCT02209376) by flow cytometry. Clinical response was quantified via engraftment in peripheral circulation and progression-free survival (PFS), as determined by the time from CAR T infusion to first radiographic evidence of progression. The CD4+CAR T cell population in patient infusion products demonstrated PD1 expression which positively correlated with AUC engraftment and PFS. On immune checkpoint inhibitor analysis, CTLA-4, TIM3, and LAG3 did not exhibit significant associations with engraftment or PFS. The frequencies of PD1+GZMB+ and PD1+HLA-DR+ CAR T cells in the CD4+ infusion products were directly proportional to AUC and PFS. No significant associations were observed within the apheresis products. In summary, PD1 in CAR T infusion products predicted peripheral engraftment and PFS in recurrent glioblastoma.

BET bromodomain protein inhibition reverses chimeric antigen receptor extinction and reinvigorates exhausted T cells in chronic lymphocytic leukemia.

Chimeric antigen receptor (CAR) T cells have induced remarkable antitumor responses in B cell malignancies. Some patients do not respond because of T cell deficiencies that hamper the expansion, persistence, and effector function of these cells. We used longitudinal immune profiling to identify phenotypic and pharmacodynamic changes in CD19-directed CAR T cells in patients with chronic lymphocytic leukemia (CLL). CAR expression maintenance was also investigated because this can affect response durability. CAR T cell failure was accompanied by preexisting T cell-intrinsic defects or dysfunction acquired after infusion. In a small subset of patients, CAR silencing was observed coincident with leukemia relapse. Using a small molecule inhibitor, we demonstrated that the bromodomain and extra-terminal (BET) family of chromatin adapters plays a role in downregulating CAR expression. BET protein blockade also ameliorated CAR T cell exhaustion as manifested by inhibitory receptor reduction, enhanced metabolic fitness, increased proliferative capacity, and enriched transcriptomic signatures of T cell reinvigoration. BET inhibition decreased levels of the TET2 methylcytosine dioxygenase, and forced expression of the TET2 catalytic domain eliminated the potency-enhancing effects of BET protein targeting in CAR T cells, providing a mechanism linking BET proteins and T cell dysfunction. Thus, modulating BET epigenetic readers may improve the efficacy of cell-based immunotherapies.

Usefulness of EZFluence software for radiotherapy planning of breast cancer treatment.

This study compared the EZFluence planning technique for irradiation of the breast with commonly used Field-in-Field (FiF) technique by analyzing the dose uniformity, the dose to the lung, heart, and other organs at risk, the total Monitor Unit (MU), and the time spent for planning. Two different 3-dimensional conformal dose plans were created for 20 breast cancer patients. Six patients were treated to a dose of 5000 cGy in 25 fractions and 14 were treated to a dose of 4256 cGy in 16 fractions. Average breast volume was 800 cc (range 128 to 1892 cc). For the FiF technique, the planner manually created between 2 to 4 subfields per gantry angle and sequentially blocked the 115% and 110% isodose line until a homogenous dose distribution was achieved. For the EZFluence technique, the planner implemented the EZFluence script that created an optimal fluence pattern, which was then imported into Eclipse where dose was calculated. Both techniques were optimized to make sure 95% of the breast planning target volume (PTV) received at least 95% of the prescribed dose. Compared to FiF technique, the plans produced by using EZFluence technique, showed the MU increased by 36.9% (p = 0.0002), whereas the planning time decreased significantly by 84.6% (p = 0.00001). The mean heart dose and the relative volume of the heart receiving ≥ 30 Gy (V30) were similar for both techniques. The mean lung dose and the relative volume of lung receiving ≥ 20 Gy (V20) were also comparable between 2 techniques. The contralateral breast mean dose and its relative volume receiving ≥ 3 Gy (V3) and ≥10 Gy (V10) were equally spared and avoided. EZFluence planning technique yielded a 4.6% (p = 0.04) reduction in PTV receiving 105% of the prescribed dose (V105) for the large breast with separation > 22 cm and PTV volume > 650 cc. The EZFluence planning technique yielded the overall comparable or improved dosimetry while significantly reducing planning time.