Poor Persistence Research Articles

Development of universal CAR-T cells

The efficacy of adoptive immunotherapy using CD19-targeting chimeric antigen receptor (CAR)-engineered T cells against B-cell malignancies has already been established in the clinic. However, high economic costs and heterogeneous quality of CAR-T cells derived from individual patients hinder further expansion of their applicability to various cancer types, including solid tumors. Mass CAR-T cell production from healthy donors is a promising approach to overcome these problems, given that allogeneic immunity elicited against donor CAR-T cells by the recipient's immune system is controlled. CAR-T cells genetically ablated with T-cell receptor and human leukocyte antigen molecules, referred to as universal CAR-T cells, may enable the use of allogeneic T cells for off-the-shelf adoptive cancer immunotherapy. However, several concerns, such as poor persistence of infused CAR-T cells and chromosomal abnormalities due to genome editing, remain to be addressed. Thus, recent clinical trials on universal CAR-T cells are summarized and future perspectives to overcome current challenges are discussed in this review.

Tangential Immersion: Increasing Persistence in Boring Consumer Behaviors

AbstractConsumers’ lives are filled with myriad behaviors that can be successfully executed with minimal attention. Many such low-attention behaviors benefit from persistence but are often not performed long enough (e.g., hygiene, exercise). The current work examines consumer persistence-failures through an attentional lens. Specifically, drawing on boredom and resource-matching frameworks, we suggest one key driver of poor consumer persistence is that many behaviors demand less attention than consumers have available, leaving excess attention that leads to boredom and premature abandonment. The current research thus proposes an attention-matching framework for persistence and suggests that concurrently performing a task that engages excess attention will improve the match between attentional demands and available resources, thereby increasing persistence. Five experiments across a range of low-attention behaviors (e.g., toothbrushing, coordination exercise) demonstrate that concurrently performing a task that occupies excess attention (e.g., reading, listening), delays boredom and increases persistence. Moreover, two important boundary conditions arise. First, the focal behavior must require minimal attention, leaving excess attention available to attend to the tangential task. Second, the tangential task must engage excess attention without exceeding attentional capacity. This research provides important theoretical and practical contributions, offering the potential to improve consumer well-being by increasing persistence in low-attention behaviors.

Women’s Inheritance Rights and Child Health Outcomes in India

Does a legal change in women’s inheritance rights have long-term effects on child health outcomes? This paper examines the effect of an improvement in women’s inheritance rights on child nutritional health outcomes in India using a difference-in-differences estimation approach. We use the staggered implementation of the Hindu Succession (Amendment) Act, 2005 to investigate the impact of the reform on anthropometric indictors of child health: being underweight, stunted, and wasted. The findings of this study reveal that an improvement in women’s inheritance rights has a positive impact on children’s health and reduces the probability of nutritional deficiency in the child. We identify mechanisms such as increased educational levels, better marital outcomes, and improved intrahousehold bargaining power of women as potential pathways through which inheritance rights affect child nutritional health outcomes. The results of the paper lend credence to growing evidence that legal recognition of women’s inheritance rights can have sustained and second-generation effects, in spite of poor enforcement mechanisms and persistence of deep-rooted societal bias against women holding property.

Switch receptor T3/28 improves long-term persistence and antitumor efficacy of CAR-T cells

BackgroundChimeric antigen receptor (CAR) T cells have been successfully used in tumor immunotherapy due to their strong antitumor responses, especially in hematological malignancies such as B cell acute lymphoid leukemia....

Real‐world persistence of erenumab for preventive treatment of chronic and episodic migraine: Retrospective real‐world study

ObjectiveTo describe the real‐world treatment persistence (defined as the continuation of medication for the prescribed treatment duration), demographics and clinical characteristics, and treatment patterns for patients prescribed erenumab for migraine prevention in Canada.BackgroundThe effectiveness of prophylactic migraine treatments is often undermined by poor treatment persistence. In clinical trials, erenumab has demonstrated efficacy and tolerability as a preventive treatment, but less is known about the longer term treatment persistence with erenumab.MethodsThis is a real‐world retrospective cohort study where a descriptive analysis of secondary patient data was conducted. Enrollment and prescription data were extracted from a patient support program for a cohort of patients prescribed erenumab in Canada between September 2018 and December 2019 and analyzed for persistence, baseline demographics, clinical characteristics, and treatment patterns. Descriptive analyses and unadjusted Kaplan–Meier (KM) curves were used to summarize the persistence and dose escalation/de‐escalation at different timepoints.ResultsData were analyzed for 14,282 patients. Median patient age was 47 years, 11,852 (83.0%) of patients were female, and 9443 (66.1%) had chronic migraine at treatment initiation. Based on KM methods, 71.0% of patients overall were persistent to erenumab 360 days after treatment initiation. Within 360 days of treatment initiation, it is estimated that 59.3% (KM‐derived) of patients who initiated erenumab at 70 mg escalated to 140 mg, and 4.4% (KM‐derived) of patients who initiated at 140 mg de‐escalated to 70 mg.ConclusionsThe majority of patients prescribed erenumab remained persistent for at least a year after treatment initiation, and most patients initiated or escalated to a 140 mg dose. These results suggest that erenumab is well tolerated, and its uptake as a new class of prophylactic treatment for migraine in real‐world clinical practice is not likely to be undermined by poor persistence when coverage for erenumab is easily available.

A Data-Driven Intervention Framework for Improving Adherence to Growth Hormone Therapy Based on Clustering Analysis and Traffic Light Alerting Systems.

Recombinant human growth hormone (r-hGH) is an established therapy for growth hormone deficiency (GHD); yet, some patients fail to achieve their full height potential, with poor adherence and persistence with the prescribed regimen often a contributing factor. A data-driven clinical decision support system based on "traffic light" visualizations for adherence risk management of patients receiving r-hGH treatment was developed. This research was feasible thanks to data-sharing agreements that allowed the creation of these models using real-world data of r-hGH adherence from easypod™ connect; data was retrieved for 11,015 children receiving r-hGH therapy for ≥180 days. Patients' adherence to therapy was represented using four values (mean and standard deviation [SD] of daily adherence and hours to next injection). Cluster analysis was used to categorize adherence patterns using a Gaussian mixture model. Following a traffic lights-inspired visualization approach, the algorithm was set to generate three clusters: green, yellow, or red status, corresponding to high, medium, and low adherence, respectively. The area under the receiver operating characteristic curve (AUC-ROC) was used to find optimum thresholds for independent traffic lights according to each metric. The most appropriate traffic light used the SD of the hours to the next injection, with an AUC-ROC value of 0.85 when compared to the complex clustering algorithm. For the daily adherence-based traffic lights, optimum thresholds were >0.82 (SD, <0.37), 0.53-0.82 (SD, 0.37-0.61), and <0.53 (SD, >0.61) for high, medium, and low adherence, respectively. For hours to next injection, the corresponding optimum thresholds were <27.18 (SD, <10.06), 27.18-34.01 (SD, 10.06-29.63), and >34.01 (SD, >29.63). Our research indicates that implementation of a practical data-driven alert system based on recognised traffic-light coding would enable healthcare practitioners to monitor sub-optimally-adherent patients to r-hGH treatment for early intervention to improve treatment outcomes.

AMV564 Depletes Myeloid-Derived Suppressor Cells and Expands T Cells: Potential to Address Key Limitations of Cellular Therapies

BackgroundMyeloid-derived suppressor cells (MDSC) contribute to an immunosuppressive tumor environment and are a barrier to immune therapeutic approaches, including cell-based therapies such as chimeric antigen receptor T cells (CAR T). Despite good overall response rates with certain subsets of B cell leukemias and lymphomas, a significant percentage of patients treated with CAR T therapy do not respond or subsequently relapse. Poor CAR T expansion, poor persistence of infused cells, and clinical treatment failure are associated with tumor and systemic immune dysregulation including high blood levels of peripheral blood monocytic MDSC (M-MDSCs) and interleukin-6, both of which are associated with lack of durable responses 1. In addition, CAR T therapy has been limited by the occurrence of severe cytokine release syndrome (CRS), which is associated with high IL-6 production 2 by myeloid cells such as MDSC.AMV564 is a potent T cell engager that selectively depletes MDSC while promoting T cell activation and proliferation without significant IL-6 induction 3. In phase 1 studies in acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), and solid tumors, AMV564 has been demonstrated to be clinically safe and active with some patients achieving complete remissions.MethodsCell lines, primary human cells, and patient samples were analyzed using flow cytometry with appropriate marker panels. T cell activation and cytotoxicity assays were conducted using primary human T cells from healthy donors and target cells (3:1 ratio) for 72 hours. T cell activation using ImmunoCult Human CD3/CD28 served as an assay reference.ResultsAnalysis of patients treated with AMV564 demonstrated statistically significant selective depletion of M-MDSC by cycle 2 (Fig. 1A). While on AMV564 therapy, median IL-6 levels remained below 100 pg/mL despite robust T cell activation and expansion. Granzyme B production by CD8 T cells increased significantly between Cycle 1 and Cycle 2 in patients on therapy, and effector CD8 T cells expand over the course of treatment (Fig. 1B-C). These data collectively support the finding that AMV564 both removes a key source of immune suppression and is a potent agonist of T cell function and differentiation in patients.AMV564 potently activates and expands primary T cells ex vivo. Across donors, peak proliferation was significantly higher with AMV564 than with the CD3/CD28 reference (Fig. 2A). Importantly, T cell viability remained significantly higher with AMV564 when compared to reference control (CD3/CD28), and there was no evidence of activation-induced cell death (AICD) in AMV564-treated samples (Fig. 2B).ConclusionsAMV564 depletes MDSC and stimulates expansion and longevity of T cells without significant IL-6 induction, suggesting a possible strategy for improvement in efficacy of cell-based therapy such as CAR T approaches. As circulating M-MDSC both at baseline and after CAR T infusion correlate with poor clinical efficacy 4, AMV564 may have beneficial effects during the conditioning phase of cell therapy, after re-infusion of CAR T products into patients, or both. Ex vivo studies using donor T cells and ongoing in vitro studies using CAR T molecules suggest that AMV564 may provide dual benefit with respect to both depletion of MDSC and T cell agonism.

Overexpression of Rorγt in CAR T Cells Improves Persistence and Reduces Exhaustion

Although T cells modified with a chimeric antigen receptor (CAR) are highly effective, durability of remissions may be limited by the poor persistence of adoptively transferred CAR T cells in patients. Growing evidence supports the idea that the pre-infusion phenotype of CAR T cells affects the efficacy of CAR T cell therapies, in part due to the increased ability of certain subsets of CAR T cells to persist long-term. Studies using conventional T cells have suggested that adoptive transfer of T cells programmed to have Th17/Tc17 effector functions can increase the anti-tumor efficacy and persistence of transferred cells when compared to adoptive transfer of non-Th17/Tc17 cells. However, standard CAR formats currently in clinical use that contain either a CD28 or 4-1BB costimulatory domain fail to generate substantial numbers of Th17/Tc17-like cells. Thus, few studies have investigated whether Th17/Tc17 CAR T cells could enhance the efficacy of CAR T cell therapies.We generated a multi-cistronic lentiviral construct encoding ROR-related Orphan Receptor gamma (RORγt), a key lineage-specifying transcription factor involved in Th17/Tc17 differentiation, followed by a murine CAR targeting CD19 that contains a CD28 costimulatory domain (1928z). RORγt expression did not hinder degranulation in response to CAR antigen upon co-culture with E2aPbx, a murine acute lymphoblastic leukemia cell line. Significantly higher numbers of RORγt-1928z CAR T cells produce IL-17 compared to 1928z CAR T cells (p<0.0001) suggesting that RORγt expression is sufficient to program type-17 effector functions in CAR T cells.To determine the impact of RORγt overexpression on CAR T cell phenotype and persistence in vivo, we adoptively transferred 1x10 6 1928z or RORγt-1928z CAR T cells into leukemia-bearing C57Bl6/J recipients and analyzed CAR T cells from the bone marrow at days 4, 11, and 31. As early as day 4 post-transfer, RORγt-1928z CAR T cells were present at significantly higher numbers than 1928z CAR T cells, and remained higher through Day 31 (0.16% of the bone marrow for 1928z CAR T cells vs 0.84% of the bone marrow for RORγt-1928z CARs; p= 0.0012, n=5 mice per group. The RORγt-1928z CAR T cells were enriched for an effector/effector memory phenotype (CD62L-) compared to 1928z CAR T cells, which were mostly of a central memory phenotype (CD62L+). Interestingly, the RORγt-1928z CAR T cells were enriched for IL-7Rα+ cells and expressed more IL7Rα than their 1928z CAR T cell counterparts (p=0.0012), suggesting an increased capacity for self-renewal in RORγt-1928z CAR T cells.Previous work from our lab has demonstrated that CAR T cells that persist after leukemia clearance have an exhausted phenotype, with increased expression of PD1 and a reduced functionality 1. After leukemia clearance, RORγt-1928z CAR T cells expressed lower levels of PD1 than their 1928z counterparts (p<0.0001). Finally, 1928z CAR T cells form a population of Eomes hiPD1 hi cells, associated with terminal exhaustion, that is absent in the RORγt-1928z CAR T cells.Taken together, our preliminary data demonstrates that overexpression of RORγt is sufficient to program type-17 effector functions in CAR T cells and suggests that RORγt overexpression can enhance the ability of CAR T cells to persist and self-renew, and prevent terminal exhaustion. RORγt overexpression may enhance persistence of CAR T cells and durability of remissions in hematologic malignancies. Furthermore, RORγt overexpression may also be a strategy to enhance CAR T cell efficacy against solid tumors, where an immunosuppressive microenvironment contributes to T cell dysfunction.1.Yinmeng Yang, Mark Eric Kohler, Terry J. Fry; Effect of Chronic Endogenous Antigen Stimulation on CAR T Cell Persistence and Memory Formation. Blood2017; 130 (Supplement 1): 166. doi: https://doi.org/10.1182/blood.V130.Suppl_1.166.16 DisclosuresFry: Sana Biotechnology: Current Employment, Current equity holder in publicly-traded company.

P.026 Erenumab associated with high persistence among Canadian patients for preventive treatment of chronic and episodic migraine in real-world practice

Background: Real world use of oral prophylactic migraine therapies is often limited by poor patient tolerance. The objective of this study was to describe the demographics and clinical characteristics of patients prescribed erenumab following its launch in Canada (September 2018) and to evaluate the real-world treatment persistence and dose management. Methods: This was a retrospective, descriptive analysis of de-identified secondary patient data that includes baseline demographics, clinical characteristics, plus erenumab treatment management, collected through Novartis’ Go Program® (Patient Support Program). Only data collected from patients with a documented informed consent were included in the analysis. Results: 14,282 patients met eligibility criteria. The mean age of patients was 46.3 years, 83.0% were female, and 66.1% reported having ≥15 monthly migraine days. 52.5% were initiated on the 140 mg dose of erenumab and 59.3% of those who initiated the 70 mg dose escalated to 140 mg within 360 days. After 360 and 450 days, the KM-derived persistence was 71.0% and 63.4%, respectively. Conclusions: The high persistence reported here suggests that erenumab has a meaningful degree of tolerability in the real-world setting and increases confidence that the real-world use and benefits of erenumab will not be undermined by the poor persistence observed with traditional migraine prophylactic agents.

Real-world use and outcomes of hypomethylating agent therapy in higher-risk myelodysplastic syndromes: why are we not achieving the promise of clinical trials?

Myelodysplastic syndromes are hematological malignancies characterized by ineffective hematopoiesis and a high risk of progression to acute myeloid leukemia. Hypomethylating agents (HMAs), azacitidine and decitabine, are standard of care therapy for higher-risk myelodysplastic syndromes. However, outcomes reported for real-world studies fall short of those achieved in clinical trials. We conducted a targeted literature review exploring real-world utilization, persistence and outcomes with intravenous and subcutaneous HMA therapies to better understand barriers to achieving optimal outcomes in clinical practice. The potential benefits of oral HMA therapy were also explored. Underutilization and poor persistence with HMA therapy are associated with suboptimal outcomes, highlighting the need for approaches to improve utilization and persistence, so that patients achieve the optimum benefit from HMA therapy.

Immunization effect and persistence of hepatitis B vaccine in HIV-infected patients with different CD4+T cell levels

Objective: To explore the immunogenicity and persistence of hepatitis B vaccine in HIV-infected patients with different CD4+T cell (CD4) levels, and analyze the influence effect of CD4 levels on immunization response. Methods: A total of 182 HIV-infected patients who participated in a randomized controlled trial of 20 µg and 60 µg hepatitis B vaccination at month 0, 1, and 6 in 2014 by Guangxi Zhuang Atonomous Region CDC and Ningming county CDC were surveyed. Six months later after the first dose and 1 month, 6 months, 1 year, and 3 years later after the full course of the vaccination, 5 ml of the venous blood of the patients was collected, and the anti-HBs was detected by Chemiluminescent Microparticle Immunoassay (CMIA). On the basis of previous studies, this study focused on analyzing the immunogenicity and persistence of hepatitis B vaccine under different CD4 levels. Results: One month later after the whole course of hepatitis B vaccination, the anti-HBs geometric mean concentration (GMC), anti-HBs positive rate (≥10 mIU/ml) and strong positive rate (≥100 mIU/ml) in HIV patients with CD4 <350 cells/µl were 442.50 mIU/ml, 71.05% (27/38) and 44.74% (17/38), respectively, which were significantly lower than those HIV-infected patients with CD4 ≥350 cells/µl [583.90 mIU/ml, 92.13% (117/127) and 77.95% (99/127)] (P<0.05). After controlling the confounding factors, the probability of being anti-HBs positive induced by hepatitis B vaccine in patients with CD4 <350 cells/µl was 0.14 times higher than in those with CD4≥350 cells/µl (95%CI: 0.03-0.62), and patients with CD4 <350 cells/µl had higher risk of no response. From 6 months to 3 years after the whole course of the vaccination, the anti-HBs GMC (195.00-27.55 mIU/ml vs. 300.10-45.81 mIU/ml), the positive rate (56.67%-36.67% vs. 78.57%- 51.58%) and the strong positive rate (33.33%-6.67% vs.44.64%-15.79%) in patients with CD4 <350 cells/µl gradually declined, lower than the levels in those with CD4 ≥350 cells/µl. Conclusions: HIV-infected patients with CD4 <350 cells/µl have high risk of no response to hepatitis B vaccination and poor immune persistence. It is necessary to strengthen the anti-HBs monitoring in HIV-infected patients, with special attention to those with CD4 <350 cells/µl. When anti-HBs is negative, hepatitis B vaccine should be injected as early as possible.

A novel and efficient tandem CD19- and CD22-directed CAR for B cell ALL

CD19-directed chimeric antigen receptor (CAR) Tcells have yielded impressive response rates in refractory/relapse B cell acute lymphoblastic leukemia (B-ALL); however, most patients ultimately relapse due to poor CAR Tcell persistence or resistance of either CD19+ or CD19- B-ALL clones. CD22 is a pan-B marker whose expression is maintained in both CD19+ and CD19- relapses. CD22-CAR Tcells have been clinically used in B-ALL patients, although relapse also occurs. Tcells engineered with a tandem CAR (Tan-CAR) containing in a single construct both CD19 and CD22 scFvs may be advantageous in achieving higher remission rates and/or preventing antigen loss. We have generated and functionally validated using cutting-edge assays a 4-1BB-based CD22/CD19 Tan-CAR using in-house-developed novel CD19 and CD22 scFvs. Tan-CAR-expressing Tcells showed similar invitro expansion to CD19-CAR Tcells with no increase in tonic signaling. CRISPR-Cas9-edited B-ALL cells confirmed the bispecificity of the Tan-CAR. Tan-CAR was as efficient as CD19-CAR invitro and invivo using B-ALL cell lines, patient samples, and patient-derived xenografts (PDXs). Strikingly, the robust antileukemic activity of the Tan-CAR was slightly more effective in controlling the disease in long-term follow-up PDX models. This Tan-CAR construct warrants a clinical appraisal to test whether simultaneous targeting of CD19 and CD22 enhances leukemia eradication and reduces/delays relapse rates and antigen loss.

Clinical outcomes associated with anti-obesity medications in real-world practice: A systematic literature review.

SummaryAnti‐obesity medications (AOMs) are efficacious and well tolerated in randomized controlled trials, but findings may not be generalizable to routine clinical practice. This systematic literature review aimed to identify real‐world (RW) evidence for AOMs to treat adults ( ≥ 18 years) with obesity or overweight (BMI ≥ 27 kg/m2). Searches conducted in MEDLINE, Embase, Health Technology Assessment (HTA) Database, National Health Service (NHS) Economic Evaluation Database, and Cochrane Central Register of Controlled Trials for studies of relevant FDA‐approved AOMs yielded 41 publications. Weight loss (WL) was consistently observed, with 14% to 58.6% of patients achieving ≥ 5% WL on orlistat, phentermine/topiramate, naltrexone/bupropion, phentermine, or liraglutide in studies of 3–6 months' duration where this was measured. When cardiometabolic risk factors were assessed, AOMs reduced or had no impact on blood pressure, lipids, or glycemia. RW data on the impact of AOMs on existing obesity‐related comorbidities and mortality were generally lacking. AOMs were associated with various adverse events, but these were of mild to moderate severity and no unexpected safety signals were reported. A pattern of poor adherence and persistence with AOMs was observed across studies. Overall, the review confirmed the effectiveness of AOMs in RW settings but demonstrated large gaps in the evidence base.

Mitochondria as Playmakers of CAR T-cell Fate and Longevity.

The development of chimeric antigen receptor (CAR) T-cell therapy has led to a paradigm shift in cancer treatment. However, patients often do not benefit from CAR T-cell therapy due to poor persistence of the adoptively transferred cells. Development of strategies based on the generation and maintenance of long-lasting memory T cells may expand the therapeutic effects of CAR T cells. Mitochondrial metabolic pathways play crucial roles in regulating the fate, function, and longevity of T cells. Here, we discuss how reprogramming of mitochondrial metabolic pathways influences function, persistence, and determination of CAR T-cell fate toward a memory phenotype. Moreover, we explore how mitochondrial activity determines persistence and the clinical outcome of CAR T-cell therapy. In addition, we review some strategies for manipulating CAR T-cell mitochondria to improve the survival of CAR T cells.

Gene-Edited Interleukin CAR-T Cells Therapy in the Treatment of Malignancies: Present and Future.

In recent years, chimeric antigen receptor T cells (CAR-T cells) have been faced with the problems of weak proliferation and poor persistence in the treatment of some malignancies. Researchers have been trying to perfect the function of CAR-T by genetically modifying its structure. In addition to the participation of T cell receptor (TCR) and costimulatory signals, immune cytokines also exert a decisive role in the activation and proliferation of T cells. Therefore, genetic engineering strategies were used to generate cytokines to enhance tumor killing function of CAR-T cells. When CAR-T cells are in contact with target tumor tissue, the proliferation ability and persistence of T cells can be improved by structurally or inductively releasing immunoregulatory molecules to the tumor region. There are a large number of CAR-T cells studies on gene-edited cytokines, and the most common cytokines involved are interleukins (IL-7, IL-12, IL-15, IL-18, IL-21, IL-23). Methods for the construction of gene-edited interleukin CAR-T cells include co-expression of single interleukin, two interleukin, interleukin combined with other cytokines, interleukin receptors, interleukin subunits, and fusion inverted cytokine receptors (ICR). Preclinical and clinical trials have yielded positive results, and many more are under way. By reading a large number of literatures, we summarized the functional characteristics of some members of the interleukin family related to tumor immunotherapy, and described the research status of gene-edited interleukin CAR-T cells in the treatment of malignant tumors. The objective is to explore the optimized strategy of gene edited interleukin-CAR-T cell function.

Single and ternary competitive adsorption-desorption and degradation of amphenicol antibiotics in three agricultural soils

The widespread usage of veterinary antibiotics results in antibiotic contamination and increases environmental risks. This study was evaluated the single and ternary competitive adsorption-desorption and degradation of three amphenicol antibiotics (AMs): chloramphenicol (CAP), thiamphenicol (TAP), and florfenicol (FF) in three agricultural soils. The adsorption capacity of amphenicol antibiotics in the soil was weak, and the Kf value was in the range of 0.15–3.59 μg1-1/nL1/n kg−1. In the single adsorption-desorption experiment, the ranked order of adsorption capacity was TAP > FF > CAP. However, in the ternary competitive adsorption experiment, the order was changed to be CAP > FF > TAP. The degradation of AMs in soils was performed at various conditions. All AMs were vulnerable to microbial degradation in soils. A higher initial concentration would reduce the degradation rate and enhance the persistence of AMs in soil. The degradation of AMs was positively influenced by changes in soil moisture content and culture temperatures up to 30 °C and decreased at higher temperatures. An equation was used to predict the leachability of AMs in soils and assess their risk to the water environment. The weak adsorption capacity and poor persistence of FF indicated that it may have a strong effect on groundwater based on the equation. It is imperative to further assess the biological impacts of FF at environmentally relevant concentrations given its mobility and extensive use in the livestock industry.

Squamous and Respiratory Metaplasia After Olfactory Mucosal Resection.

Resection of the olfactory mucosa (OM) is sometimes unavoidable during surgery; however, it is not known whether the OM can completely recover thereafter. The aim of this study was to uncover whether the OM fully recovers after mucosal resection and describe the process of OM regeneration. 8-week-old male Sprague–Dawley rats (n = 18) were subjected to OM resection at the nasal septum; six rats were euthanized for histological examination 0, 30, and 90 days after surgery. Immunohistochemistry was performed to identify olfactory receptor neuron (ORN) lineage cells [mature and immature ORNs and ORN progenitors, and olfactory ensheathing cells (OECs)], as well as dividing and apoptotic cells. Squamous and respiratory metaplasia and inflammatory cell infiltration were also assessed. On day 30 after resection, the mucosa had regenerated, and mainly contained thin nerve bundles, basal cells, and immature ORNs, with a few mature ORNs and OECs. On day 90, the repaired nasal mucosa had degenerated into stratified squamous or ciliated pseudostratified columnar epithelia, with reducing ORNs. The lamina propria contained numerous macrophages. Partial regeneration was observed within 1 month after OM resection, whereas subsequent degeneration into squamous and respiratory epithelia occurred within 3 months. Given the poor persistence of ORNs and OECs, OM resection is likely to result in olfactory impairment. Overall, surgeons should be cautious not to injure the OM during surgery.

Abstract CT007: CD19/CD20 bispecific chimeric antigen receptor (CAR) in naive/memory T-cells for the treatment of relapsed or refractory B-cell lymphomas

Abstract 50% of patients with relapsed/refractory (R/R) B-cell lymphomas (BCL) who initially respond to CD19 CAR T therapy will relapse either due to poor CAR-T persistence or CD19 antigen escape. Preclinical data demonstrate that engineering of bispecific anti-CD19/CD20 CAR in naïve/memory T-cells (TN/MEM) via lentiviral transduction effectively targets tumor cells, overcomes antigen escape and enhances CART persistence (Zah E et al., Cancer Immunol Res, 2016). We conducted a first-in-human phase 1 clinical trial with bispecific CD19/CD20 CAR T-cells for patients with R/R BCL (NCT04007029). Patients with measurable disease after 2 or more lines of therapy for DLBCL and PMBCL, and 3 or more lines of therapy for MCL, FL and CLL were included. Autologous leukocytes obtained by leukapheresis were sorted for CD14-/CD25-/CD62L+ TN/MEM cells, followed by lentiviral transduction of the bispecific CD19/CD20 CAR. Following Cy-Flu lymphodepletion, patients are infused with CD19/CD20 CAR TN/MEM cells, at doses ranging from 5 x 107 and 2 x 108 CAR positive cells. 5 patients are included in the analysis, 4 in the first cohort and 1 in the second cohort. All patients had CD19/CD20+ B cell malignancy on tissue biopsy prior to CAR-T. Patients had previously received a median of 4 prior lines of therapy, and 4 received bridging therapy. Infusions were well tolerated, and no dose limiting toxicities were identified. All patients had grade 1 CRS, with median onset 8 days (range 7-11) after infusion. No patient had immune effector cell-associated neurotoxicity syndrome (ICANS). Peak expansion was noted on day 14. Median follow up is 11.1 months (95% CI 2.1-14.8), with 4 of patients in ongoing complete remission. The 1 patient who did not respond had early disease progression, with CD19 and CD20 negative mediastinal lymphoma at day 14 after CAR infusion. Median progression-free survival (PFS) and overall survival (OS) were not reached and all responders demonstrate ongoing CAR T-cell persistence and B-cell aplasia by data cutoff. Bispecific CD19/CD20 CAR in naïve/memory T-cells are safe and effective in patients with R/R BCL. To our knowledge, this is the first study that potentially obviates the challenges of the commonest causes of relapse after CAR T-cell therapy, poor CAR-T persistence and antigen escape mechanisms, by means of modifying naïve/memory T cells and dual antigen targeting, respectively. Patient and Treatment CharacteristicsID12345DiagnosisMCLFLPMBCLFLFLSexMMFMFAge6058296235Prior lines of therapy44343Stage44442EECOG01100Bridging therapy?YesYesYesYesNoDosage, x 106 CART50505050200Max CRS/ICANS gradeGrade 1 CRS. No ICANS.Grade 1 CRS. No ICANS.Grade 1 CRS. No ICANS.Grade 1 CRS. No ICANS.Grade 1 CRS. No ICANS.Required treatment for CRS/ICANS?NoNoNoNoNoBest Response to CARTCRCRNone; refractoryCRCRPFS (months)14.811.10.54.82.1OS (months)14.811.16.24.82.1Subsequent Follow-upAlive, ongoing remissionAlive, ongoing remissionNo response to CART; died from infectious complications after subsequent therapy with nivolumab and brentuximab.Alive, ongoing remissionAlive, ongoing remission Citation Format: Sanaz Noelle Ghafouri, Christopher Walthers, Mobina Roshandell, Brenda Ji, Jacqueline Trent, Jia Ming Chen, Jacob Naparstek, Caitlin Harris, Thomas Schweppe, Karla K. Nawaly, Monica Mead, Sven de Vos, Patricia Young, Caspian Oliai, Gary Schiller, John M. Timmerman, Yvonne Y. Chen, Sarah Larson. CD19/CD20 bispecific chimeric antigen receptor (CAR) in naive/memory T-cells for the treatment of relapsed or refractory B-cell lymphomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr CT007.

VTE Recurrence and Safety of Anticoagulants Among Patients with Cancer Treated for Venous Thromboembolism

Introduction:Anticoagulation (AC) is effective for the treatment of venous thromboembolism (VTE) in cancer patients, but is associated with an increased risk of bleeding. Currently, LMWH monotherapy is the recommended treatment for cancer patients with VTE; however, emerging data from Medicare patients suggests that rivaroxaban had a significantly lower risk of VTE recurrence and similar risk of major bleeding compared to LMWH or warfarin. In this study, we compared the effectiveness and risk of major bleeding associated with these anticoagulants in a younger cohort of patients from a commercial claims database.Methods: Truven Health Analytics MarketScan databases were used to find patients aged ≥18 newly diagnosed with cancer who then had their first VTE (deep vein thrombosis (DVT) or pulmonary embolism (PE); excluding upper VTE) between 1/1/2013 - 9/30/2016 and AC start ≤ 7 days. Patients were categorized into LMWH, warfarin, or rivaroxaban cohorts based on index AC. The warfarin cohort included patients who started LMWH and warfarin ≤ 7 days of their first VTE. Inverse probability of treatment weights based on propensity score were used to adjust for treatment differences in the comparison of LMWH vs. rivaroxaban, warfarin vs. rivaroxaban, and LMWH vs. warfarin. Patients were observed until the earliest of end of eligibility or data end. VTE recurrences were defined conservatively as hospitalizations with a primary diagnosis of VTE ≥7 days after the first VTE. Major bleeding events were identified using validated criteria (Cunningham et al., 2011) and evaluated while patients were on treatment. Kaplan-Meier rates at 6 and 12 months were used to compare VTE recurrence between cohorts. Time on treatment was approximately 30-50% of total eligibility time, so rates at 3 and 6 months were used to compare major bleeding between cohorts. Cox proportional hazard models were used to compare VTE recurrence and major bleeding risk between cohorts.Results: A total of 12,457 patients (LMWH: N=4,313; warfarin; N=4,774; rivaroxaban: N=3,370) were included. After weighting the cohorts, baseline demographic and clinical characteristics were well balanced. Mean patient age ranged from 61.3 to 63.6 years. Median observation time with LMWH was shorter than with rivaroxaban (6.8 vs. 8.3 months, p<0.001) and with warfarin (7.4 vs. 9.8 months, p<0.001). Time on treatment was significantly shorter as well (Figures 1A-1C). VTE recurrence rates were significantly lower with rivaroxaban than with LMWH users at 6 months (8.7% vs. 11.7%; p=0.003) and 12 months (11.9% vs.14.7%; p=0.006), respectively, with a hazard ratio (HR [95%CI]) of 0.83 (0.73-0.96; p=0.010; Figure 1A). Major bleeding rates for rivaroxaban and LMWH users were similar at 3 months (3.2% vs. 3.5%; p=0.592) and 6 months (4.4% vs. 4.9%; p=0.438), respectively [HR 0.91 (0.71-1.17; p=0.455]. VTE recurrence rates for rivaroxaban and warfarin users were similar at 6 months (8.2% vs 8.8%; p=0.530) and 12 months (11.3% vs 11.6%; p=0.675), respectively [HR 0.95 (0.83-1.09; p=0.456; Figure 1B)]. Major bleeding rates for rivaroxaban and warfarin users were also similar at 3 months (3.2% vs. 2.8%; p=0.199) and 6 months (4.2% vs. 3.8%; p=0.362), respectively [HR 1.08 (0.86-1.37; p=0.500)]. Recurrence rates were significantly lower for warfarin users than for LMWH users at 6 months (10.2% vs. 12.4%; p=0.006) and 12 months (13.3% vs. 15.3%; p=0.011), respectively, but the rate of recurrence over the entire follow up was not significantly different between the two cohorts [HR of 0.91 (0.81-1.02; p=0.103; Figure 1C)]. Major bleeding rates for warfarin and LMWH users were similar at 3 months (3.1% vs. 3.7%; p=0.114) and 6 months (4.3% vs. 5.1%; p=0.109), respectively [HR of 0.86 (0.68-1.08; p=0.187)].Conclusion: In this large cohort of cancer patients treated for VTE, time on treatment was shorter for patients treated with LMWH than oral agents. Rivaroxaban was associated with a significantly lower rate of VTE recurrence than LMWH and a similar rate of VTE recurrence to warfarin; major bleed rates were similar for all. While LMWH has been associated with greater reductions in recurrent VTE compared to warfarin in trials, poor persistence on LMWH in real world settings may explain its lack of comparative effectiveness in this analysis. [Display omitted] DisclosuresKhorana:Bayer: Consultancy, Honoraria; Amgen: Consultancy, Honoraria, Research Funding; Sanofi: Consultancy, Honoraria; Janssen Scientific Affairs, LLC: Consultancy, Honoraria, Research Funding; Leo: Consultancy, Honoraria, Research Funding; Halozyme: Consultancy, Honoraria; Pfizer: Consultancy, Honoraria; Roche: Consultancy, Honoraria. Milentijevic:Janssen Scientific Affairs, LLC: Employment, Equity Ownership. McCormick:Janssen Scientific Affairs, LLC: Research Funding. Laliberté:Janssen Scientific Affairs, LLC: Research Funding. Crivera:Janssen Scientific Affairs, LLC: Employment, Equity Ownership. Lefebvre:Janssen Scientific Affairs, LLC: Research Funding. Lejeune:Janssen Scientific Affairs, LLC: Research Funding. Rozjabek:Janssen Scientific Affairs, LLC: Other: Fellowship supported by Janssen Scientific Affairs, LLC. Schein:Janssen Scientific Affairs, LLC: Employment, Equity Ownership, Other: Own in excess of $10,000 of J&J stock. Streiff:Roche: Research Funding; CSL Behring: Consultancy, Research Funding; Janssen Scientific Affairs, LLC: Consultancy, Research Funding; Portola: Research Funding.

CAR-T Cells Targeting BAFF-Receptor for B-Cell Malignancies: A Potential Alternative to CD19

Background: Chimeric antigen receptor (CAR) T-cell adoptive therapies have the potential to revolutionize the treatment of cancers. Particularly in hematological malignancies, there are reports of promising clinical outcomes in advanced non-Hodgkin lymphomas (NHLs) with CD19-CAR T-cell therapy. However, disease relapse is problematic, and is thought to be caused by poor long-term persistence of the CAR T-cells, and loss of the CD19 target on tumors. Thus, there is an urgent need for improved novel CAR T-cell therapies directed at alternative targets.The CAR T-cell platform relies on antibody-derived single chain fragments (scFv), which are genetically engineered into chimeric T-cell receptors, and that recognize target cell surface proteins on tumors. One potential target is B-cell activating factor receptor (BAFF-R), a tumor necrosis factor receptor superfamily protein (TNFRSF13C) specifically involved in B lymphocyte development and mature B-cell survival, that is primarily expressed on B cells and various subtypes of B-cell NHLs and ALL. We have recently developed a humanized therapeutic BAFF-R antibody with strong anti B-cell tumor activity.Method and Results: We adapted a scFv based on our humanized anti-BAFF-R antibody onto a second generation CAR platform containing CD3ζ and 4-1BB intracellular signaling domains. In response to BAFF-R-expressing malignant human B cells (NHLs, acute lymphoblastic leukemias, and chronic lymphocytic leukemias), our CAR T cells readily proliferated and secreted cytotoxic cytokines. We demonstrated both significant levels of BAFF-R CAR T-cell activation and malignant B-cell killing in vitro .Established human NHL tumors in xenogeneic models were eliminated following BAFF-R CAR T-cell treatments in vivo . Remarkable tumor-free survival was repeatedly observed in human lymphoma xenograft models including JeKo-1 (mantle cell lymphoma) and Raji (Burkitt lymphoma) in NSG mice. Moreover, upon tumor re-challenge and without any further treatment tumors failed to develop, due to persisting CAR T cells conferring continued anti-lymphoma activity. We pursued optimization of CAR T-cell persistence by comparing three subsets of early stage T cells (central memory, TCM; memory stem, TSCM; and naïve, TN) as potential starting material for CAR T cell generation. Our in vivo studies show CAR T cells from the TN population retained highest potency eradicating established tumors at a minimal therapeutic dose compared to other subtypes; 80% of TN CAR treated mice achieved long-term survival compared to 20% of TCM and 40% of TSCM CAR treated mice. We also observed the long term anti-tumor effects conferred by CD8+ CAR T cells required the addition of CD4+ CAR T cells. Finally, we performed a head-to-head comparison of BAFF-R CAR T cells with CD19 CAR T cells in the Raji model (Figure). Comparable expression levels of BAFF-R and CD19 were observed on the tumor cells (A), and both treatments elicited significant antitumor effects compared to T-cell and saline controls (B-C). However, the BAFF-R CAR T-cell treatment demonstrated long-term tumor free survival in all treated mice compared to the CD19 CAR T-cell treated cohort, which only showed delayed tumor growth (P<0.01, B-C).Conclusion: BAFF-R CAR T cells demonstrate remarkable efficacy against B-cell malignancies. Unexpectedly, we found that naïve T cells were superior to TCM or TSCM subpopulations as starting material in vivo . Naïve T-cell starting products, after transduction and expansion, generated sufficient numbers of TCM and TSCM CAR T cells for activity. Targeting BAFF-R combined with our strategy to improve CAR T-cell persistence potentially addresses unmet clinical needs in B-cell NHLs and ALL, particularly in the setting of CD19 CAR-T-resistance or CD19-negative relapse. [Display omitted] DisclosuresNeelapu:Novartis: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Bristol-Myers Squibb: Research Funding; Karus: Research Funding; Merck: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Celgene: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Kite Pharma: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Cellectis Inc.: Research Funding; Poseida Therapeutics, Inc: Research Funding. Kwak:InnoLifes: Consultancy, Equity Ownership; Pepromene Bio: Consultancy, Equity Ownership; Celltrion, Inc: Consultancy.