CD19/CD22 targeting with cotransduced CAR T cells to prevent antigen-negative relapse after CAR T-cell therapy for B-cell ALL.

ABSTRACT: CD19-negative relapse is a leading cause of treatment failure after chimeric antigen receptor (CAR) T-cell therapy for acute lymphoblastic leukemia. We investigated a CAR T-cell product targeting CD19 and CD22 generated by lentiviral cotransduction with vectors encoding our previously described fast-off rate CD19 CAR (AUTO1) combined with a novel CD22 CAR capable of effective signaling at low antigen density. Twelve patients with advanced B-cell acute lymphoblastic leukemia were treated (CARPALL [Immunotherapy with CD19/22 CAR Redirected T Cells for High Risk/Relapsed Paediatric CD19+ and/or CD22+ Acute Lymphoblastic Leukaemia] study, NCT02443831), a third of whom had failed prior licensed CAR therapy. Toxicity was similar to that of AUTO1 alone, with no cases of severe cytokine release syndrome. Of 12 patients, 10 (83%) achieved a measurable residual disease (MRD)-negative complete remission at 2 months after infusion. Of 10 responding patients, 5 had emergence of MRD (n = 2) or relapse (n = 3) with CD19- and CD22-expressing disease associated with loss of CAR T-cell persistence. With a median follow-up of 8.7 months, there were no cases of relapse due to antigen-negative escape. Overall survival was 75% (95% confidence interval [CI], 41%-91%) at 6 and 12 months. The 6- and 12-month event-free survival rates were 75% (95% CI, 41%-91%) and 60% (95% CI, 23%-84%), respectively. These data suggest dual targeting with cotransduction may prevent antigen-negative relapse after CAR T-cell therapy.

The outcomes of constrained total hip arthroplasty for recurrent dislocation of hip hemiarthroplasty.

INTRODUCTION: The incidence of hemiarthroplasty dislocation for fracture neck of femurs ranges between 1 and 15% and the one-year mortality is 49- 70%. Revision of hemiarthroplasty to total hip replacement using a constrained liner has shown to improve the morbidity and mortality rates. The aim of the study was to assess whether conversion of dislocated hemiarthroplasty to total hip replacement improve functional and one year mortality. METHODS: A retrospective analysis of the number of patients who had recurrent dislocations of hemiarthroplasty for fracture neck of femurs were carried out. The data were obtained from NHFD (National Hip Fracture Database) and internal hospital computer systems (Medway, Theatre notes and PACS) between Dec 2008 and Dec 2020. Patient demographics including age, sex, Abbreviated Mental Test Score (AMTS), functional assessment, mortality at one and two years were documented. The risk factors which led to dislocations such as Parkinsons disease, Cerebrovascular accidents, Musculo-neuropathies and Alzheimer`s disease was also noted. RESULTS: A total of 3994 patients were admitted during the study period of which 1735 (43.4%) patients had hemiarthroplasty. Fifty-six (3.23%) patients had dislocation of hemiarthroplasty. The mean age was 81.4 years (range - 61 to 95). There were 40 (71.4%) females and 16 males (28.6%). The average AMTS score was 5.3. All 56 patients had closed manipulative reduction under anaesthesia within in 12 h of admission. Thirty-one patients (55.4%) went on to have recurrent dislocations of which 18 patients (58.4%) had total hip replacement using captive cup, 6 patients (19.4%) had open reduction,3 patients (9.7%) had excision arthroplasty procedure and four patients (12.5%) had no intervention, Eighteen patients who had total hip replacement with constrained captive for followed up to a minimum of two years (range2- 12 years). There were no intraoperative complications, dislocation or periprosthetic fractures in the follow up period. There was no mortality at the end of two years of follow up in this group, two-year mortality for the patients with alternative management for dislocated hemiarthroplasty was 76.67. CONCLUSION: Treatment of recurrent hemiarthroplasty dislocation by revising to a total hip replacement with a constrained liner gives good functional and mortality outcomes.

Clinically Applicable Assessment of Tisagenlecleucel CAR T Cell Treatment by Digital Droplet PCR for Copy Number Variant Assessment.

Chimeric antigen receptor (CAR) T cell therapy is an innovative immunotherapy for treating cancers in both children and adults with proven utility in numerous clinical trials. Significantly, some CAR T cell therapies have now been approved by relevant national regulatory bodies across numerous countries for clinical therapeutic use outside of clinical trials. One such recently licensed product is tisagenlecleucel, a CAR T therapy approved for the treatment of B-cell acute lymphoblastic leukemia (B-ALL) using autologous T cells from the patient. The genetically engineered T cells target a protein called CD19, common to B cells, through a CAR incorporating a 4-1BB costimulatory domain to improve response. Since tisagenlecleucel is now a standard of care treatment for B-ALL, it is clinically essential to be able to accurately monitor these CAR T cells in patients. Assessment of the copy number variant (CNV) of the CAR T cell products allows this within a clinically acceptable timeframe for optimal patient benefit. However, no standardized method with high reproducibility and efficiency has been described within a routine clinical laboratory setting. Here, we demonstrated a novel digital droplet PCR (ddPCR)-based methodology for the study of CNV (ddPCR-CNV) in 4-1BB CD19-specific CAR T cells with universal applicability across clinical diagnostic laboratories.

Screening of Neonatal UK Dried Blood Spots Using a Duplex SMN1 Screening Assay.

Spinal muscular atrophy (SMA) is an autosomal inherited neuromuscular genetic disease caused, in 95% of cases, by homozygous deletions involving the SMN1 gene exon 7. It remains the leading cause of death in children under 2 years of age. New treatments have been developed and adopted for use in many countries, including the UK. Success of these treatments depends on early diagnosis and intervention in newborn babies, and many countries have implemented a newborn screening (NBS) or pilot NBS program to detect SMN1 exon 7 deletions on dried blood spots. In the UK, there is no current NBS program for SMA, and no pilot studies have commenced. For consideration of adoption of NBS for a new condition, numerous criteria must be satisfied, including critical assessment of a working methodology. This study uses a commercially available real-time PCR assay to simultaneously detect two different DNA segments (SMN1 exon 7 and control gene RPP30) using DNA extracted from a dried blood spot. This study was carried out in a routine clinical laboratory to determine the specificity, sensitivity, and feasibility of SMA screening in a UK NBS lab setting. Just under 5000 normal DBSs were used alongside 43 known SMA positive DBSs. Study results demonstrate that NBS for SMA using real-time PCR is feasible within the current UK NBS Laboratory infrastructure using the proposed algorithm.