Double-negative T cells utilize a TNFα-JAK1-ICAM-1 cytotoxic axis against acute myeloid leukemia.

ABSTRACT: Allogeneic double-negative T cells (DNTs) are a rare T-cell subset that effectively target acute myeloid leukemia (AML) without inducing graft-versus-host disease in an allogeneic setting. A phase 1 clinical trial demonstrated the feasibility, safety, and potential efficacy of allogeneic DNT therapy among patients with relapsed AML. However, the molecular mechanisms of DNT-mediated cytotoxicity against AML remain elusive. Thus, we used a flow cytometry-based high throughput screening to compare the surface molecule expression profile on DNTs during their interaction with DNT-susceptible or -resistant AML cells and identified a tumor necrosis factor α (TNFα)-dependent cytotoxic pathway in DNT-AML interaction. TNFα secreted by DNTs, upon encountering susceptible AML targets, sensitized AML cells to DNT-mediated killing, including those otherwise resistant to DNTs. Mechanistically, TNFα upregulated ICAM-1 on AML cells through a noncanonical JAK1-dependent pathway. DNTs then engaged with AML cells more effectively through an ICAM-1 receptor, lymphocyte function-associated antigen 1, leading to enhanced killing. These results reveal a TNFα-JAK1-ICAM-1 axis in DNT-mediated cytotoxicity against AML to improve therapeutic efficacy.

CRISPR screen identifies genes that sensitize AML cells to double-negative T-cell therapy.

Acute myeloid leukemia (AML) remains a devastating disease in need of new therapies to improve patient survival. Targeted adoptive T-cell therapies have achieved impressive clinical outcomes in some B-cell leukemias and lymphomas but not in AML. Double-negative T cells (DNTs) effectively kill blast cells from the majority of AML patients and are now being tested in clinical trials. However, AML blasts obtained from ∼30% of patients show resistance to DNT-mediated cytotoxicity; the markers or mechanisms underlying this resistance have not been elucidated. Here, we used a targeted clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) screen to identify genes that cause susceptibility of AML cells to DNT therapy. Inactivation of the Spt-Ada-Gcn5-acetyltransferase (SAGA) deubiquitinating complex components sensitized AML cells to DNT-mediated cytotoxicity. In contrast, CD64 inactivation resulted in resistance to DNT-mediated cytotoxicity. Importantly, the level of CD64 expression correlated strongly with the sensitivity of AML cells to DNT treatment. Furthermore, the ectopic expression of CD64 overcame AML resistance to DNTs in vitro and in vivo. Altogether, our data demonstrate the utility of CRISPR/Cas9 screens to uncover mechanisms underlying the sensitivity to DNT therapy and suggest CD64 as a predictive marker for response in AML patients.

Evaluating patient preparedness for cataract surgery and satisfaction with preoperative care.

OBJECTIVE: To evaluate preoperative preparedness and patient satisfaction after implementation of the Cataract Screening Preprocedural Questionnaire (CSPQ) at Hamilton Regional Eye Institute. DESIGN: Single-centred, prospective, cross-sectional study. PARTICIPANTS/METHODS: One-hundred fifty-one adult patients undergoing elective cataract surgery completed the CSPQ questionnaire. Based on the responses, patients were triaged to the preoperative anaesthesia clinic or booked directly for surgery. Outcome measures included anxiety level, delays or cancellations in surgery, preoperative blood pressure, glucose levels, and satisfaction with the quality of preoperative instructions provided. Results were compared between patients who attended and those who bypassed the preoperative anaesthesia clinic. RESULTS: Of the study population, 87 patients were female (57.6%) and the mean age was 72.0 ± 10.5 years. Only 11 patients (7.43%) were referred for preoperative consultation. Patients reported receiving preoperative instructions via handouts (94%) and verbally in combination with handouts (59.33%). Patients felt that adequate information was provided regarding preoperative medications (96.69%), eye drops (99.34%), fasting guidelines (98.68%), arrival time (99.34%), and instructions for accompaniment/drivers postoperatively (100%). All patients were compliant with the fasting guidelines. Patient satisfaction with the information received regarding cataract surgery and anaesthesia were 4.39 ± 0.88 and 3.80 ± 0.95, respectively. There was no difference in the anxiety level between patients who attended and those who bypassed the preoperative clinic (4.09 ± 2.92 and 5.18 ± 2.57, p = 0.14). There were no cancellations, delays, or immediate postoperative systemic complications. CONCLUSIONS: The CSPQ model can be effectively used to streamline the preoperative preparation of patients for cataract surgery while maintaining a high degree of patient satisfaction with the perioperative experience.