IMPORTANCE:Platinum-based chemotherapy is widely used in the management of solid tumors, and up to 60% of patients will experience ototoxic side effects, including hearing loss and tinnitus. The advent of novel otoprotective drugs generates a need for more flexible and innovative real-time approaches to drug-induced ototoxicity monitoring in cancer patients. OBJECTIVE:To assess the feasibility of recruitment and compliance to monitoring of ototoxicity with mobile out-of-booth audiometry, Distortion Product Otoacoustic Emission (DPOAE) testing, and serum biomarker measurements. DESIGN:Prospective observational study. SETTING:Cancer ambulatory care. PARTICIPANTS:Patients undergoing platinum-based chemotherapy for bone or testicular cancer. RESULTS:Twenty-three patients were enrolled between February 2021 and June 2023. The mean age was 17.5 years (range: 13 to 36 y). Average compliance to monitoring assessments was 86.5% for audiometry, 84.8% for DPOAEs, and 84.3% for serum biomarker measurements. End-of-treatment audiometric changes indicative of ototoxicity were found in 18 (78.2%) of the participants. CONCLUSIONS AND RELEVANCE:High compliance with serial mobile audiometry and otoacoustic emissions testing, and serum biomarker measurement suggests that ototoxicity monitoring can be successfully performed in cancer ambulatory care rather than in an audiology department setting. Further investigation is required to understand the role of serum prestin and otolin-1 in monitoring ototoxicity.

Background:Patients with advanced or recurrent endometrial cancer have poor survival outcomes because effective treatment options remain limited. We investigated the efficacy and safety of cediranib plus weekly paclitaxel or olaparib versus weekly paclitaxel alone after prior platinum-based chemotherapy. Methods:COPELIA was an open-label, randomised, phase 2 trial undertaken at 15 centres in the United Kingdom. Eligible participants were aged 16 years or older with histologically confirmed endometrial cancer, Eastern Cooperative Oncology Group performance status 0-1, a life expectancy greater than 16 weeks, and at least one previous line of platinum-based chemotherapy. Patients were randomly assigned to paclitaxel 80 mg/m2 on days 1, 8, and 15 of a 28-day cycle (arm 1, control); cediranib 20 mg daily plus the same paclitaxel schedule (arm 2, experimental); or cediranib 20 mg daily plus olaparib 300 mg twice daily (arm 3, experimental). Up to six cycles of paclitaxel were permitted. Patients in arm 2 with a Response Evaluation Criteria in Solid Tumours (RECIST) version 1.1 complete or partial response, or stable disease, after paclitaxel plus cediranib continued cediranib monotherapy. The primary endpoint was the proportion of patients free from RECIST-defined progression or death at 3 months (PFS at 3 months). Secondary endpoints included RECIST response, PFS at 6 months, median PFS, median overall survival (OS), safety, and quality of life. Translational analyses assessed vascular response, defined as a reduction of 5% or more in plasma Tie2 within 9 weeks of treatment initiation. The trial was registered with EudraCT (2016-004617-28), ISRCTN (16320634), and ClinicalTrials.gov (NCT03570437). Findings:Between May 1, 2018, and January 11, 2022, 124 patients were enrolled and randomised to arm 1 (n = 41), arm 2 (n = 41), or arm 3 (n = 42). Median follow-up in arms 1, 2, and 3 was 34.4 months (interquartile range [IQR] 11.6-37.6), 26.3 months (11.8-not evaluable), and 23.7 months (15.0-not evaluable), respectively. PFS at 3 months was significantly higher in arm 2 than in arm 1 (73.2% versus 48.8%; adjusted odds ratio [aOR] 3.2, lower limit of one-sided 80% confidence interval [CI] 2.1; p = 0.01). RECIST response was also higher in arm 2 versus arm 1 (56.4% versus 28.2%; aOR 5.7, 95% CI 1.8-17.6; p < 0.001). No differences were observed between arm 3 and arm 1 for PFS at 3 months (aOR 1.0, lower limit of one-sided 80% CI 0.71; p = 0.46) or RECIST response (aOR 0.8, 95% CI 0.3-2.6; p = 0.73). Median OS was 12.8 months (95% CI 7.3-16.8) in arm 1, 18.1 months (9.5-26.4; log-rank p = 0.42 versus arm 1) in arm 2, and 13.9 months (11.2-18.3; log-rank p = 0.86 versus arm 1) in arm 3. There was no difference in PFS at 6 months and median PFS between arm 1 and arm 2 or arm 3. Grade 3 adverse events occurring in ≥10% of patients in arm 2 included hypertension (15%), neutropenia (12%), and diarrhoea (10%). Quality-of-life differences favoured arm 1 over arm 2 for diarrhoea and gastrointestinal symptoms (p < 0.001). In multivariable analyses, cediranib-treated patients who achieved a Tie2-defined vascular response had significantly improved PFS (hazard ratio 0.54, 95% CI 0.33-0.88; p = 0.014). Interpretation:Paclitaxel plus cediranib improved 3-month PFS and RECIST response compared with paclitaxel alone in advanced or recurrent endometrial cancer. However, this effect diminished over time; with no difference in PFS observed at 6 months, nor median PFS or median OS. Plasma Tie2 identified cediranib-treated patients with improved PFS. Further evaluation of paclitaxel plus cediranib in patients previously treated with platinum-based chemotherapy and immunotherapy is warranted, ideally incorporating plasma Tie2 as a response biomarker to guide treatment continuation. Funding:AstraZeneca.

Introduction Head and neck cancer is the eighth most common cancer in the UK. Current standard of care treatment for patients with recurrent/metastatic squamous cell head and neck carcinoma (HNSCC) is platinum-based chemotherapy combined with the anti-epidermal growth factor receptor (anti-EGFR) monoclonal antibody, cetuximab. However, most patients will have poor median overall survival (OS) of 6–9 months despite treatment. HNSCC tumours exhibit an immune landscape poised to respond to immunotherapeutic approaches, with most tumours expressing the immunosuppressive receptor programmed death-ligand 1 (PD-L1). We undertook the current study to determine the safety and efficacy of avelumab, a monoclonal antibody targeting the interaction between PD-L1 and its receptor on cytotoxic T-cells, in combination with cetuximab.Methods and analysis This is a multi-centre, single-arm dose de-escalation phase II safety and efficacy study of avelumab combined with cetuximab; the study was to progress to a randomised phase II trial, however, the study will now complete after the safety run-in component. Up to 16 participants with histologically/cytologically recurrent/metastatic squamous cell carcinoma (including HNSCC) who have not received cetuximab previously will be recruited. All patients will receive 10 mg/kg avelumab and cetuximab (500, 400 or 300 mg/m2 depending on the cohort open at time of registration) on days 1 and 15 of 4-week cycles for up to 1 year, (avelumab not given cycle 1 day 1). A modified continual reassessment method will be used to determine dose de-escalation. The primary objective is to establish the safety of the combination and to determine the optimum dose of cetuximab. Secondary objectives include assessing evidence of antitumour activity by evaluating response rates and disease control rates at 6 and 12 months as well as progression-free and OS.Ethics and dissemination Approval granted by City and East REC (18/LO/0021). Findings will be published in peer-reviewed journals and disseminated at conferences.Trial registration number NCT03494322.

Background: Obe-cel is an autologous CD19 chimeric antigen receptor (CAR) T cell product designed to reduce toxicity and improve persistence through a fast off-rate CD19 binding domain. The clinical activity of obe-cel has been explored in adults with R/R B-ALL in a Phase I study (ALLCAR19, NCT02935257; Roddie C et al. J Clin Oncol 2021) and a Phase Ib/II study (FELIX, NCT04404660; Roddie C et al. J Clin Oncol 2023;41[16 Suppl]:7000). Additionally, obe-cel has been tested in pts with R/R B-cell chronic lymphocytic leukemia (B-CLL) and R/R B-cell non-Hodgkin lymphoma (B-NHL) (ALLCAR19 extension; Roddie C et al. Blood 2022;140[1 Suppl]:7452-3). Pts from the ALLCAR19 and FELIX Phase Ib studies are in long-term follow up (≥22 mos), and the ALLCAR19 extension has been recruiting for 3 years. We report an analysis of long-term efficacy and safety data from the ALLCAR19 and FELIX Phase Ib studies, as well as data from the ALLCAR19 extension. Methods: ALLCAR19 is a multicenter, non-randomized, open-label Phase I study in pts aged ≥16 years with B-cell malignancies. ALLCAR19 initially recruited pts with R/R B-ALL but was then amended (extension study) to also include pts with R/R B-CLL and R/R B-NHL. FELIX is a global, single-arm Phase Ib/II study enrolling pts aged ≥18 years with R/R B-ALL.Study designs have been presented previously. Obe-cel was administered as a split dose in pts with B-ALL (target dose 410 × 10 6 CAR T cells) and pts with CLL (target dose 230 × 10 6 CAR T cells), and as a single infusion in pts with B-NHL (target dose 200 × 10 6 CAR T cells); the pt populations in the two studies were similar. Pts with B-ALL from the ALLCAR19 and FELIX Phase Ib studies are being followed long term for disease progression and survival. For this analysis, data in pts with B-ALL from the ALLCAR19 and FELIX Phase Ib studies were pooled. Data in pts with CLL or B-NHL are presented from the ALLCAR19 extension study. Results: Outcomes in pts with R/R B-ALL: Data in pts with B-ALL were pooled (20 pts from ALLCAR19 [data cut-off Jun 26, 2023] and 16 from FELIX Phase Ib [data cut-off Mar 16, 2023]). The median age of the pooled cohort was 41.5 (range 18 to 74) years and pts had received a median of 3 (range 2 to 6) prior lines of treatment. Twenty-nine of the 36 pts (81%) achieved complete remission (CR)/CR with incomplete hematologic recovery post obe-cel infusions, per investigator assessment. The event-free survival rate was 64% at 6 mos and 49% at 12 mos. With a median follow up of 43 (range 19 to 62) mos, 13/36 pts (36%) remain in remission (8 from ALLCAR19; 5 from FELIX Phase Ib). Among these 13 ongoing responders, 2 (15%) had consolidation with allogeneic hematopoietic stem cell transplantation (allo-HSCT). Ten of the 11 ongoing responders (91%) who did not receive allo-HSCT still had detectable CAR T cells at the last follow up. All ongoing remissions were measurable residual disease negative at last available assessment. The estimated 2-, 3- and 4-year overall survival rates were 44%, 39% and 39%, respectively. Outcomes in pts with R/R B-CLL/B-NHL: The extension phase of the ALLCAR19 study enrolled 35 pts with B-CLL or B-NHL, of which 26 (B-CLL n=5; B-NHL n=21) received obe-cel (data cut-off Jun 26, 2023). The median age of this combined cohort was 61 (range 39 to 79) years and pts had received a median of 3 (range 2 to 8) prior lines of treatment. At a median follow up of 24 mos, the overall response rate for this cohort was 92% (n=24), and 58% of responders (n=14) were alive without disease progression at last follow up. Late toxicity: Of the 11 long-term R/R B-ALL responders who had not received consolidation allo-HSCT, 10 have ongoing B-cell aplasia. Of the 14 ongoing responders in the R/R B-CLL/B-NHL cohort, 12 have ongoing B-cell aplasia (<20 B cells/µl). Of note, ongoing B-cell aplasia did not correlate with an increased risk of late serious infection. No other long-term toxicity ascribed to obe-cel was reported. Conclusions: The combined analysis of data from the ALLCAR19 and FELIX Phase Ib studies shows long-term efficacy and safety of obe-cel in pts with R/R B-ALL, with approximately one-third of pts still in remission without consolidative allo-HSCT after a median follow up of >3 years. Durable responses of >2 years were also seen in pts with R/R B-CLL and R/R B-NHL.B-cell aplasia was commonly found in long-term follow up of obe-cel recipients, but without a corresponding rise in serious late infections. Obe-cel can effect durable long-term remissions in B-cell malignancies.

<p>Table S1: Treatments undertaken with timing of imaging; S2: Changes in CgA at first post-treatment time-point (3-5 weeks) compared to baseline and effect on OS.</p>

Background: Relapsed/refractory (r/r) primary central nervous system lymphoma (PCNSL) has a median overall survival of 2-8 months and few therapeutic options. CD19 CAR-T therapy is highly effective for systemic B-cell lymphoma, but associated neurotoxicity has led to exclusion of PCNSL patients from pivotal clinical trials. We have previously described AUTO1, a CD19 CAR with a fast off-rate CD19 binding domain, designed to reduce immune toxicity and improve engraftment. Its clinical activity has been tested in r/r paediatric and adult B-ALL, NHL and CLL, with high remission rates, low incidence of CRS/ICANS despite CAR trafficking into the cerebrospinal fluid (CSF) and long-term CAR-T engraftment. Aims: Based on these favourable characteristics, we initiated the CAROUSEL study of AUTO1 in PCNSL (NCT04443829), testing both intravenous (IV) and intraventricular (I-VEN) routes of CAR-T delivery. We incorporated anti-PD1 into the conditioning strategy with the aim of enhancing CAR expansion and preventing PD1-mediated CAR silencing in the PCNSL microenvironment. Methods: Manufacturing: CAR-T products were generated using a semi-automated, closed process from non-mobilised patient leukapheresate. Study Design: Subjects>/=16y underwent lymphodepletion with fludarabine (Flu;30mg/m2x3) and cyclophosphamide (Cy;60mg/kgx1) and Pembrolizumab (200mgx1) prior to IV AUTO1 infusion (250x10^6 CAR-T). In the event of failure to achieve PR or a frank relapse event, a 2nd dose of AUTO1 was administered into the CSF via ommaya reservoir (25x10^6 CAR-T) following Flu/Cy at an interval of >/=28d post-IV dosing. Study endpoints include manufacture feasibility, grade 3-5 toxicity and remission rates at 1 and 3 months. Results: As of 14 February 2022, we enrolled 6 patients, apheresed and manufactured 5 products and infused 5 patients with IV AUTO1 and 1 patient with I-VEN AUTO1. Of enrolled patients, median age was 43y (range 23-50), median ECOG was 1 (range 1-2) and median prior lines of therapy was 2 (range 1-3). All patients had parenchymal disease and all received bridging therapy. Following IV CAR-T, grade 1 and 2 CRS affected 1 and 3 patients respectively and any grade ICANS was observed in 2 patients, with 2 grade 3 events reported. AUTO1 engraftment and response was evaluable in 4 patients at month 1 following IV infusion. Excellent AUTO1 engraftment was observed in all patients in blood and CSF, and all 4 patients had ongoing CAR-T persistence at last follow-up. CSF and blood cytokines were also evaluated. 2/4 evaluable patients had no measurable disease at 2 and 6 months of follow-up respectively. 1 patient had PD at month 1 and was unable to receive further trial treatment. 1 patient had SD at month 1 but progressed at month 2. This patient had an ommaya reservoir implanted to receive I-VEN AUTO1, following which the patient developed grade 2 CRS for 5 days, without overt ICANS. The patient achieved PR at month 1 but progressed at month 2. Summary/Conclusion: In r/r PCNSL, AUTO1 shows encouraging response rates and excellent CAR-T engraftment/expansion in the blood and CSF. I-VEN administration was well-tolerated and AUTO1 demonstrated activity via direct CSF infusion in a patient who failed IV therapy. Additional patients, updated biological data and longer follow-up will be presented.

Background Epidermal growth factor receptor (EGFR) is a therapeutic target to which HER2/HER3 activation may contribute resistance. This Phase I/II study examined the toxicity and efficacy of high-dose pulsed AZD8931, an EGFR/HER2/HER3 inhibitor, combined with chemotherapy, in metastatic colorectal cancer (CRC). Methods Treatment-naive patients received 4-day pulses of AZD8931 with irinotecan/5-FU (FOLFIRI) in a Phase I/II single-arm trial. Primary endpoint for Phase I was dose limiting toxicity (DLT); for Phase II best overall response. Samples were analysed for pharmacokinetics, EGFR dimers in circulating exosomes and Comet assay quantitating DNA damage. Results Eighteen patients received FOLFIRI and AZD8931. At 160 mg bd, 1 patient experienced G3 DLT; 160 mg bd was used for cohort expansion. No grade 5 adverse events (AE) reported. Seven (39%) and 1 (6%) patients experienced grade 3 and grade 4 AEs, respectively. Of 12 patients receiving 160 mg bd, best overall response rate was 25%, median PFS and OS were 8.7 and 21.2 months, respectively. A reduction in circulating HER2/3 dimer in the two responding patients after 12 weeks treatment was observed. Conclusions The combination of pulsed high-dose AZD8931 with FOLFIRI has acceptable toxicity. Further studies of TKI sequencing may establish a role for pulsed use of such agents rather than continuous exposure. Trial registration number ClinicalTrials.gov number: NCT01862003.

PURPOSE:To assess the safety and tolerability of a vandetanib-eluting radiopaque embolic (BTG-002814) for transarterial chemoembolization (TACE) in patients with resectable liver malignancies.MATERIALS AND METHODS:The VEROnA clinical trial was a first-in-human, phase 0, single-arm, window-of-opportunity study. Eligible patients were aged ≥18 years and had resectable hepatocellular carcinoma (HCC) (Child-Pugh A) or metastatic colorectal cancer (mCRC). Patients received 1 mL of BTG-002814 transarterially (containing 100 mg of vandetanib) 7-21 days prior to surgery. The primary objectives were to establish the safety and tolerability of BTG-002814 and determine the concentrations of vandetanib and the N-desmethyl vandetanib metabolite in the plasma and resected liver after treatment. Biomarker studies included circulating proangiogenic factors, perfusion computed tomography, and dynamic contrast-enhanced magnetic resonance imaging.RESULTS:Eight patients were enrolled: 2 with HCC and 6 with mCRC. There was 1 grade 3 adverse event (AE) before surgery and 18 after surgery; 6 AEs were deemed to be related to BTG-002814. Surgical resection was not delayed. Vandetanib was present in the plasma of all patients 12 days after treatment, with a mean maximum concentration of 24.3 ng/mL (standard deviation ± 13.94 ng/mL), and in resected liver tissue up to 32 days after treatment (441-404,000 ng/g). The median percentage of tumor necrosis was 92.5% (range, 5%-100%). There were no significant changes in perfusion imaging parameters after TACE.CONCLUSIONS:BTG-002814 has an acceptable safety profile in patients before surgery. The presence of vandetanib in the tumor specimens up to 32 days after treatment suggests sustained anticancer activity, while the low vandetanib levels in the plasma suggest minimal release into the systemic circulation. Further evaluation of this TACE combination is warranted in dose-finding and efficacy studies.

Background: We have previously described AUTO1, a CD19 CAR with a fast off-rate CD19 binding domain, designed to reduce CAR T-cell immune toxicity and improve engraftment. Its clinical activity has been tested in r/r paediatric and adult B-ALL (Ghorashian S et al., Nat Med 2019; Roddie C et al., JCO 2021). This data confirms the intended function of the receptor, with low levels of CRS/ICANS and long-term engraftment of CAR T-cells observed in both patient groups. Aims: We have initiated testing of AUTO1 in the setting of B-NHL and CLL/SLL (NCT02935257). Methods: Manufacturing: CAR T-cell products were generated using a semi-automated closed process from non-mobilised patient leukapheresate. Study design: Subjects ≥ 16y underwent lymphodepletion with fludarabine (30mg/m x3) and cyclophosphamide (60mg/kg x1) prior to AUTO1 infusion, with the exception of the DLBCL cohort who additionally received a single dose of pembrolizumab (200mg) on day -1 to potentiate CAR-T expansion. AUTO1 dose varies based on the indication. Split dosing of 230 x106 CD19 CAR T-cells at day 0 and day 9 is employed in the CLL cohort. A single dose of 200 x106 CD19 CAR T-cells is delivered to patients with B-NHL. Study endpoints include feasibility of manufacture, grade 3-5 toxicity and remission rates at 1 and 3 months. Results: As of 8th February 2022, we enrolled 23 patients: 11 low grade NHL (LG-NHL:7 with FL and 3 with MCL), 7 DLBCL and 5 CLL. Apheresis was successful in all 23 patients and product manufacture was successful in 22 (pending in the last). 19 patients were infused: 10 with LG-NHL, 6 with DLBCL and 3 with CLL. 1 CLL patient was pending infusion at time of data cut-off and 2 patients died pre-infusion: 1 MCL patient, from COVID-19 and 1 CLL patient, from intracerebral haemorrhage. Patients treated with AUTO1 had a median age of 60 years (range 39-79), had received a median of 3 prior lines of treatment (range 2-8). Grade 1 CRS was reported in 6/19 and Grade 2 CRS in 3/19. No ICANS was observed in the B-NHL and CLL cohorts. CAR engraftment was observed in 13/13 patients evaluated by qPCR with ongoing persistence in 12/13 patients at last follow-up. In the LG-NHL and DLBCL cohorts 10/10 and 4/5 evaluable patients respectively were in CMR by 18FDG PET-CT post-treatment. Responses were ongoing in 9/10 LG-NHL at 12 months and in 4/4 DLBCL at months 1, 3, 3 and 6. In the CLL cohort, 2/3 evaluable patients achieved MRD negative remission in the bone marrow with residual small volume lymph nodes by CT at 6 and 3 months of follow-up respectively. 1 CLL patient did not engraft and had SD at month 1. Summary/Conclusion: AUTO1 has a tolerable safety profile in patients with r/r B-NHL and CLL despite high disease burden. Early data shows excellent complete remission rates and excellent CAR engraftment/expansion. Additional patients, updated data and longer follow up will be presented.

INTRODUCTION: AUTO1 is a fast off-rate CD19 binding domain CAR, designed to reduce immune toxicity and improve engraftment. Its clinical activity has been tested in r/r paediatric and adult B-ALL (Ghorashian S et al., Nat Med 2019; Roddie C et al., JCO 2021) and now in adult B-NHL and CLL/SLL (NCT02935257). Here we present data from adult B-NHL and CLL cohorts treated with AUTO1, and report on long-term follow-up of adult B-ALL patients post-AUTO1. METHODS:Manufacturing: AUTO1 products were generated using a closed process from non-mobilised patient leukapheresis. Study design: Subjects ≥ 16y received fludarabine (30mg/m2 x3) and cyclophosphamide (60mg/kg x1) conditioning. DLBCL patients additionally received pembrolizumab (200mg) on day -1. In B-ALL, patients received split dose AUTO1 (day 0: ≥20% Bone Marrow (BM) blasts, AUTO1 dose=10 x106; <20% BM blasts, AUTO1 dose=100 x106. At day+9: if no grade 3-5 CRS/ICANS, dose 2 is administered to a total AUTO1 dose of 410 x106. Split dosing is employed in the CLL cohort (day 0 AUTO1 dose= 30 x106; day 9 AUTO1 dose= 200 x106). B-NHL patients receive a single AUTO1 dose of 200 x106. Study endpoints include manufacture feasibility, grade 3-5 toxicity and remission rates at 1 and 3 months. RESULTS:NHL/CLL cohort: as of 27th July 2022, 28 patients were enrolled: 9 with Follicular Lymphoma (FL), 4 with Mantle Cell Lymphoma (MCL), 8 with DLBCL and 7 with CLL. Apheresis/ manufacture was successful in 26 (1x pending; 1x not harvested due to intracerebral haemorrhage). Median age was 61 years (range 39-79), and patients received a median of 3 prior treatment lines (range 2-8). To date, 23 patients have been infused: 7 FL, 3 MCL, 8 DLBCL and 5 CLL. 2 patients died pre-infusion (1x MCL, COVID-19; 1x CLL, intracerebral haemorrhage). Grade 1 CRS was reported in 7/23 and Grade 2 CRS in 7/23 patients. No ICANS was observed. AUTO1 engraftment was demonstrated in 18/18 patients evaluated by qPCR with ongoing persistence in 17/18 patients at last follow-up. The ORR at month 1 in the FL, MCL and DLBCL cohorts was 7/7 (100%), 3/3 (100%) and 7/8 (88%) respectively, and ongoing responses at last follow-up (FU) were observed in 5/7 FL (FU range, 5-21 months), 2/3 MCL (range, 12-18 months), and 7/8 DLBCL (range, 1-12 months). 2 patients died in remission from COVID-19. In the CLL cohort, 4/5 patients achieved flow negative remission in the bone marrow with ongoing response in all 4 at last FU. B-ALL cohort: of the 20 B-ALL patients treated previously, 8/20 (40%) are in ongoing CR at a median FU of 35 months (IQR 24-36) post-AUTO1, with ongoing B-cell aplasia in 7/8 (89%). CONCLUSIONS: AUTO1 has a tolerable safety profile in patients with r/r B-cell cancers despite high disease burden. In the B-ALL cohort, long-term follow-up indicates that 40% of patients continue in remission post-AUTO1. In both indolent and aggressive NHL and in CLL, AUTO1 shows excellent ORR and CAR engraftment/persistence. Additional patients, updated data and longer follow up will be presented. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

PURPOSE Prognosis for adult B-cell acute lymphoblastic leukemia (B-ALL) is poor, and there are currently no licensed CD19 chimeric antigen receptor (CAR) therapeutics. We developed a novel second-generation CD19-CAR (CAT19-41BB-Z) with a fast off rate, designed for more physiologic T-cell activation to reduce toxicity and improve engraftment. We describe the multicenter phase I ALLCAR19 ( NCT02935257 ) study of autologous CAT19-41BB-Z CAR T cells (AUTO1) in relapsed or refractory (r/r) adult B-ALL. METHODS Patients age ≥ 16 years with r/r B-ALL were eligible. Primary outcomes were toxicity and manufacturing feasibility. Secondary outcomes were depth of response at 1 and 3 months, persistence of CAR-T, incidence and duration of hypogammaglobulinemia and B-cell aplasia, and event-free survival and overall survival at 1 and 2 years. RESULTS Twenty-five patients were leukapheresed, 24 products were manufactured, and 20 patients were infused with AUTO1. The median age was 41.5 years; 25% had prior blinatumomab, 50% prior inotuzumab ozogamicin, and 65% prior allogeneic stem-cell transplantation. At the time of preconditioning, 45% had ≥ 50% bone marrow blasts. No patients experienced ≥ grade 3 cytokine release syndrome; 3 of 20 (15%) experienced grade 3 neurotoxicity that resolved to ≤ grade 1 within 72 hours with steroids. Seventeen of 20 (85%) achieved minimal residual disease–negative complete response at month 1, and 3 of 17 underwent allogeneic stem-cell transplantation while in remission. The event-free survival at 6 and 12 months was 68.3% (42.4%-84.4%) and 48.3% (23.1%-69.7%), respectively. High-level expansion (Cmax 127,152 copies/µg genomic DNA) and durable CAR-T persistence were observed with B-cell aplasia ongoing in 15 of 20 patients at last follow-up. CONCLUSION AUTO1 demonstrates a tolerable safety profile, high remission rates, and excellent persistence in r/r adult B-ALL. Preliminary data support further development of AUTO1 as a stand-alone treatment for r/r adult B-ALL.

Background: We have previously described AUTO1 (CAT19), a CD19 CAR with a fast off-rate CD19 binding domain, designed to reduce CAR T-cell immune toxicity and improve engraftment. Its clinical activity has been tested in r/r paediatric and adult B-ALL. Cumulatively, this data confirms the intended fucntion of the receptor, with low levels of CRS/ ICANS and long-term engraftment of CAR T-cells observed in both patient groups. Recently, CAR therapy has been explored in indolent lymphomas such as follicluar (FL) and mantle cell lymphoma (MCL), but a high incidence of toxicity inluding Grade 3-4 ICANS has been reported. Aims: We have initiated testing of AUTO1 (CAT19) in the setting of indolent B-cell lymphoma (NCT02935257). Methods: Manufacturing: CAR T-cell products were generated using a semi-automated closed process from non-mobilised leucapheresate. Study design: subjects>/=16y underwent lymhpodepletion with fludarabine (30mg/m2 x3) and cyclophosphamide (60mg/kg x1) followed by a single CAR T-cell infusion of 200 x10

Background: Circulating tumor cells (CTCs) are detectable in patients with neuroendocrine tumors (NETs) and are accurate prognostic markers although the optimum threshold has not been defined.Objective: This work aims to define optimal prognostic CTC thresholds in PanNET and midgut NETs.Patients and Methods: CellSearch was used to enumerate CTCs in 199 patients with metastatic pancreatic (PanNET) (90) or midgut NETs (109). Patients were followed for progression-free survival (PFS) and overall survival (OS) for a minimum of 3 years or until death.Results: The area under the receiver operating characteristic curve (AUROC) for progression at 12 months in PanNETs and midgut NETs identified the optimal CTC threshold as 1 or greater and 2 or greater, respectively. In multivariate logistic regression analysis, these thresholds were predictive for 12-month progression with an odds ratio (OR) of 6.69 (P <.01) for PanNETs and 5.88 (P <.003) for midgut NETs. The same thresholds were found to be optimal for predicting death at 36 months, with an OR of 2.87 (P <.03) and 5.09 (P <.005) for PanNETs and midgut NETs, respectively. In multivariate Cox hazard regression analysis for PFS in PanNETs, 1 or greater CTC had a hazard ratio (HR) of 2.6 (P <.01), whereas 2 or greater CTCs had an HR of 2.25 (P <.01) in midgut NETs. In multivariate analysis OS in PanNETs, 1 or greater CTCs had an HR of 3.16 (P <.01) and in midgut NETs, 2 or greater CTCs had an HR of 1.73 (P <.06).Conclusions: The optimal CTC threshold to predict PFS and OS in metastatic PanNETs and midgut NETs is 1 and 2, respectively. These thresholds can be used to stratify patients in clinical practice and clinical trials.

INTRODUCTION We have previously described AUTO1, a CD19 CAR with a fast off-rate CD19 binding domain, designed to reduce CAR-T immune toxicity and improve engraftment. Its clinical activity has been tested in r/r paediatric and adult B-ALL. Cumulatively, this data confirms the intended function of the receptor, with low levels of CRS/ICANS and long-term engraftment of CAR T-cells observed in both patient groups. Recently, CAR-T therapy has been explored in indolent lymphomas such as follicular (FL) and mantle cell lymphoma (MCL), but a high incidence of toxicity including Grade 3-4 ICANS has been reported. We have initiated testing of AUTO1 in the setting of indolent and high-grade B-NHL and CLL (NCT02935257). METHODS Manufacturing: CAR T-cell products were generated using a semi-automated closed process from non-mobilised leukapheresate. Study design: Subjects ≥ 16y underwent lymphodepletion with fludarabine (30mg/m 2 x3) and cyclophosphamide (60mg/kg x1) prior to AUTO1 infusion, with the exception of the DLBCL cohort who additionally received a single dose of pembrolizumab (200mg) on day -1 to potentiate CAR-T expansion. AUTO1 dose varies based on the indication. Split dosing of 230 x10^6 CD19 CAR T-cells at day 0 and day 9 is employed in the CLL cohort. A single dose of 200 x10^6 CD19 CAR T-cells is delivered to patients with B-NHL. Study endpoints include feasibility of manufacture, grade 3-5 toxicity and remission rates at 1 and 3 months. RESULTS As of 17th May 2021, we recruited 13 patients: 7 with FL, 4 with MCL, 1 DLBCL and 1 CLL. Apheresis and product manufacture was successful in all 13 patients and 9 patients were infused: 7 with FL and 2 with MCL. Three patients (1 DLBCL, 1 CLL and 1 MCL) were pending infusion at time of data cut-off and 1 patient (MCL) died due to Covid-19 prior to infusion. Patients treated with AUTO1 had a median age of 56 years (range 39-68y), had received a median of 3 prior lines of treatment (range 2-5) and all patients had stage IV disease at screening. Grade 1 CRS was reported in 4/9 and Grade 2 CRS in 1/9. 1/9 developed MAS which resolved with anakinra/dexamethasone. No ICANS was observed on study. Excellent CAR engraftment was observed and 9/9 patients were in CMR by 18FDG PET-CT post-treatment. At a median of 6.1 months (range 4.0-8.1m), 8/9 patients were disease free at last follow-up. One patient died in CMR at month 5.6 of COVID-19. CONCLUSION AUTO1 has a tolerable safety profile in adult patients with r/r B-NHL despite high disease burden. Early data shows 100% complete remission rates and excellent CAR engraftment/expansion. Additional MCL, CLL and DLBCL patients, updated data and longer follow up will be presented. Roddie: Celgene: Consultancy, Speakers Bureau; Novartis: Consultancy; Gilead: Consultancy, Speakers Bureau. Hartley: Astra Zeneca: Ended employment in the past 24 months; ADC Therapeutics: Consultancy, Current equity holder in publicly-traded company, Current holder of stock options in a privately-held company. Farzaneh: Autolus: Consultancy, Current equity holder in publicly-traded company. Lowdell: Autolus: Consultancy, Current equity holder in publicly-traded company. Linch: Autolus: Consultancy, Current equity holder in publicly-traded company. Pule: Autolus: Current Employment, Current equity holder in publicly-traded company. Peggs: Autolus: Consultancy, Current equity holder in publicly-traded company.

Single-cell profiling of circulating tumor cells (CTCs) as part of a minimally invasive liquid biopsy presents an opportunity to characterize and monitor tumor heterogeneity and evolution in individual patients. In this study, we aimed to compare single-cell copy number variation (CNV) data with tissue and define the degree of intra- and inter-patient genomic heterogeneity. We performed next-generation sequencing (NGS) whole-genome CNV analysis of 125 single CTCs derived from seven patients with neuroendocrine neoplasms (NEN) alongside matched white blood cells (WBC), formalin-fixed paraffin-embedded (FFPE), and fresh frozen (FF) samples. CTC CNV profiling demonstrated recurrent chromosomal alterations in previously reported NEN copy number hotspots, including the prognostically relevant loss of chromosome 18. Unsupervised hierarchical clustering revealed CTCs with distinct clonal lineages as well as significant intra- and inter-patient genomic heterogeneity, including subclonal alterations not detectable by bulk analysis and previously unreported in NEN. Notably, we also demonstrated the presence of genomically distinct CTCs according to the enrichment strategy utilized (EpCAM-dependent vs size-based). This work has significant implications for the identification of therapeutic targets, tracking of evolutionary change, and the implementation of CTC-biomarkers in cancer.

Introduction: Prognosis for adult B-cell Acute Lymphoblastic Leukaemia (B-ALL) is poor and there is currently no licensed CD19 Chimeric Antigen Receptor (CAR) therapeutic. We developed a novel CD19 CAR (CAT-41BBz CAR) with a fast off-rate, designed to result in more physiological T-cell activation, reduce toxicity and improve engraftment. We describe updated data from the Phase I ALLCAR19 (NCT02935257) study of AUTO1 in relapsed/refractory adult B-ALL. Methods: Manufacturing: AUTO1 utilises non-mobilised autologous leukapheresate. The first 6 products were generated using a standard dynabead/WAVE bioreactor process and subsequent products using a semi-automated closed process. Study design: Patients aged >16y underwent lymphodepletion with fludarabine (30mg/m2 x3) and cyclophosphamide (60mg/kg x1) followed by split dose CAR T-cell infusion (Day 0: if ≥20% Bone Marrow (BM) blasts, infuse 10 x 106 CAR T-cells; if <20% BM blasts, 100 x 106 CAR T-cells. At Day+9: if no grade 3-5 Cytokine Release Syndrome (CRS)/immune effector cell associated neurotoxicity syndrome (ICANS), infuse Dose 2, to a total dose of 410 x106 CAR T-cells). Study endpoints include feasibility of manufacture, grade 3-5 toxicity and remission rates at 1 and 3 months. Results: As of 13 May 2020, 24 patients have been leukapheresed, 23 products manufactured and 19 patients received at least 1 dose of AUTO1. The median age was 43y (range 18-62), 26% had prior blinatumomab, 47% had prior inotuzumab ozogamicin and 63% had prior hematopoietic stem cell transplantation (HSCT). At the time of pre-conditioning, 42% had ≥50% BM blasts. No patients experienced ≥Grade 3 CRS (Lee criteria), 3/19 (16%) experienced Grade 3 ICANS that swiftly resolved with steroids. Of 19 infused patients, 16/19 (84%) achieved Minimal Residual Disease (MRD) negative complete response (CR). Currently 6 patients have died, none related to AUTO1. 11/19 (58%) patients remain on study and continue in MRD negative remission at a median follow up of 12.2 months (range 0.6-24.4m). To date, only 2 patients underwent HSCT whilst in remission. For all treated patients, the event-free survival (EFS) at 6 months was 62% and 76% for those whose products were manufactured using the closed process. Patients exhibited robust CAR expansion (mean peak CAR T levels 716,769 copies/µg DNA). Conclusions: AUTO1 has a tolerable safety profile in adult patients with r/r B-ALL despite high disease burden. Early data shows high remission rates with 84% achieving MRD negative CR. This preliminary data supports the further development of AUTO1 as a standalone treatment in patients with r/r B-ALL. Data from additional patients and longer follow up will be presented. Furthermore, data from extension cohorts of patients with low- and high- grade B-cell Non-Hodgkin's lymphoma and chronic lymphocytic leukaemia will be presented. Figure Disclosures Roddie: Celgene: Honoraria; Gilead: Honoraria; Novartis: Honoraria. O'Reilly:Gilead: Honoraria; Novartis: Honoraria, Other: Travel support. Hartley:ADC Therapeutics: Consultancy, Current equity holder in publicly-traded company, Research Funding. Linch:Autolus: Consultancy. Pule:UCLB: Patents & Royalties; Mana Therapeutics: Other: entitled to share of revenue from patents filed by UCL; Autolus: Current Employment, Other: owns stock in and receives royalties, Patents & Royalties. Peggs:Autolus: Consultancy.

Patients with R/M SCC have low response rates to second line therapies, including PD-1 inhibitors nivolumab and pembrolizumab, representing an area of unmet clinical need. Cetuximab has modest activity as a single agent but potentiates the activity of radiotherapy in locally advanced head & neck SCC (HNSCC) and chemotherapy in R/M HNSCC. Cetuximab initiates Natural Killer cell antibody-dependent cell-mediated cytotoxicity, resulting in an anti-tumour immune response and the potential to augment the activity of PD-1/PD-L1 inhibition. Trial entry required histologically confirmed R/M SCC of any site, unselected by PD-L1 expression, considered incurable by local therapies and no previous treatment with cetuximab for recurrent/metastatic disease. Prior therapy with anti-PD-1, anti-PD-L1 or anti-PD-L2 was excluded. Patients had avelumab 10 mg/kg + cetuximab 500 mg/m2 intravenously every 2 weeks, for up to 1 year. Primary endpoint was occurrence of dose-limiting toxicity within 42 days of treatment starting, graded using CTCAE v5. Secondary endpoints were objective response (ORR) and disease control rate (DCR) at 6 and 12 months using iRECIST. 16 patients, median age 58 years (range 34 – 88), were enrolled from 2 UK hospitals between July 2018 and October 2019. The trial stopped after completing the safety run-in. 5 patients remain on treatment, 9 stopped treatment early (7 disease progression, 1 patient choice, 1 due to risk of COVID-19). 2 patients died whilst on treatment (both unrelated to trial treatment). Grade 3 AEs were seen in 4 patients and grade 5 in 1 patient. None were related to trial treatment. No patients experienced dose-limiting toxicity. Of 10 patients evaluable for response by iRECIST 2 (20%) had complete response, 3 (30%) had partial response and 4 (40%) had stable disease as their best response, representing an ORR of 50%. One patient had confirmed disease progression. In 6 patients who remained on trial for >6 months, all 6 had disease control at 6 months (2 CR, 1 PR, 3 SD). Avelumab + cetuximab is safe and tolerable, and demonstrates promising efficacy in R/M SCC patients.

Patients with recurrent/metastatic head and neck squamous cell carcinoma (R/M HNSCC) often present with loco-regional disease; approximately 14% of patients present with metastatic disease. Despite aggressive primary therapy over 50% of newly diagnosed patients treated radically with either surgery and/or radiotherapy with/without chemotherapy will develop recurrent disease, with the majority of recurrences occurring locally. Standard therapy for these patients is a platinum-containing regime (PCCC) or PD-1 inhibitor.