B-NHL Patients Research Articles

Auto Hematopoietic Stem Cell Transplantation Combined with Another Target Humanized CAR-T Cells for Refractory/Relapsed B-Cell Non-Hodgkin Lymphoma after Failure of Murinized CD19-CAR-T Therapy

Background: The prognosis of refractory/relapsed aggressive B-cell non-Hodgkin lymphoma (r/r B-NHL) is extremely poor especially for the patients who failed to CD19-Specific chimeric antigen receptor-T (CAR-T) cells therapy.Even sequentially autologous hematopoietic stem cell transplantation(ASCT) could not maintain a durable remission in most patients.Aims: To prolong relapse-free survival, we combined ASCT and another target humanized CAR-T cells to treat r/r B-NHL patients failed to murinized CD19-CAR-T cells therapy with either CD22 or CD20 antigen expression on tumor cell.The safety and efficacy will be evaluated.Methods: From December 2019 to March 2021, 12 patients were enrolled. The median age was 38 (16-68) years old. The diagnosis included DLBCL (n=8) ,BL(n=3) and PMBCL (n=1). The median IPI score was 3 (range,2-4).There were 9 patients(9/12,75%) with extranodal lesions. Six cases(6/12,50%) were with TP53 mutations. The disease status was progressive disease in all patients who failed to multi-line therapies and murinized CD19-CAR-T cells therapy.In order to further reduce the tumor burden, all patients were treated with combined chemotherapy before transplantation. Before the trial, the expression of CD20 and CD22 antigen in tumor tissue was positive confirmed by immunohistochemistry,and the target was selected according to the antigen expression. Conditioning with BEAM-based regimen was applied. The kinetics and function of CAR-T cells was monitored by quantitative PCR and flow cytometry. The efficacy was evaluated by PET-CT every 3 month after transplantation. Results: The autologous peripheral blood stem cells were infused with the median CD34 + cells 3.91(range,0.46-9.46)×10 6/kg.Humanized CAR-T cells with the median 1.85 (range,0.13-3.26)×10 6/kg were infused 2 day after stem cells,including target antigen CD20(7/12,58.3%) and CD22(5/12,41.7%). Cytokine release syndrome (CRS) occurred in 11 cases with 5 cases in grade I, 3 case in grade II and 3 cases in grade III.One case developed immune effector cell-associated neurotoxicity syndrome (ICANS) in grade IV. The peak of cytokine IFN-γ and IL-6 post baseline in patients with grade III CRS were significantly higher than those in patients with grade I-II CRS,especially in ICANS patient.Six cases with grade II and III were relieved with glucocorticoid. The neutrophil and platelets engraftment was achieved in all cases on median days 14 (range,9-22) and 14(range,8-35) respectively post-transplant .Seven cases of bacterial enteritis were seen. Pneumonia occurred in 7 cases.For CAR-T cells expansion,the peak time in vivo was on median 11(range,7-28) days after CAR-T cells infusion.The median peak lever of CAR-T cells was 20.3 (range,0.13-60.4)×10 6/L, which was positively correlated with the number of CART infused. The tumor burden before transplantation was not significantly associated with CAR-T cells expansion.The median duration of CAR-T cells in vivo was 30 days, and the longest lasting time was 139 days post-transplant so far. B-cell aplasia was documented in all cases(7/7,100%) of CD20-CART group and two cases(2/5,40%) of CD22-CART group during the follow-up. With the median follow-up 266 (range,118-565) days, 9/12(75%) patients survived,seven cases(7/12,58.3%) achieved complete remission(CR),2 cases(2/12,16.7%) achieved PD and survival with tumor.Kaplan-Meier survival analysis showed that OS and PFS rates were 71.3% and 66.6% respectively at 9 months after transplantation.Two cases(2/12,16.7%) with BL and one with DLBCL (1/12,8.3%)died of PD.Significantly lower levels of Cumulative CAR T cell levels (AUC) during the first 1 month post transplantation were observed in patients with disease progression compared with those who had durable responses (P<0.0001).Conclusion: CRS is manageable and has no influence on hematopoiesis reconstitution.With current protocol, complication was mild and encouraging disease control was found. ASCT combined with another target humanized CAR-T therapy is a safe and effective salvage strategy for r/r B-NHL after failure of murinized CD19-CAR-T. Long-term follow-up is needed.[Key words] refractory/relapsed B-cell non-Hodgkin lymphoma; failure of CD19-CAR-T; another target CAR-T cell; autologous hematopoietic stem cell transplantation DisclosuresNo relevant conflicts of interest to declare.

Anti-CD19 CART (C-CAR011) Therapy for Relapsed or Refractory B-Cell Non-Hodgkin Lymphoma

To evaluate the safety and efficacy of C-CAR011 in the treatment of relapsed or refractory B-cell non-Hodgkin lymphoma (R/R B-NHL) patients. B-NHL patients treated with C-CAR011 infusion following lympho-depletion were enrolled. All the patients were followed up for 1 year after C-CAR011 treatment(5.0×106 /kg). The primary endpoint of this research was safety; and the secondary endpoints included objective response rate (ORR), progression-free survival (PFS), duration of overall response (DOR), overall survival (OS) at Week 4, 12 and Month 6 and 12. The ratio of the male and female of 6 patients was 1∶1, and the patients were treated with C-CAR011 at a dose of 5.0×106 /kg. The median age was 46.5 years old (37-62 years); 5 patients were diagnosed as diffuse large b-cell lymphoma (DLBCL) and 1 patient was diagnosed as follicular lymphoma (FL). The most common adverse events (AEs) were hematological toxicities (100%, Grade 3-4), cytokine release syndrome (CRS) (66.7%, Grade 1-3), neurotoxicity (16.7%, Grade 3). All these AEs were reversible and could be conversed. The ORR at Week 4, 12 and Month 6, 12 were 50%(3/6), 50%(3/6), 16.7%(1/6) and 16.7%(1/6), respectively. Three patients (50%) achieved complete response (CR) and one of them stayed evaluated as CR until Month 12. The median PFS was 178 days (91->365 days). For 3 patients with response, the DOR were 150 days, 154 days and >12 months, respectively. The OS rates of the patients at Week 12, Month 6, 12 were 100%, 100%, 83.3%, respectively, while the median OS was not reached. C-CAR011 is a safe treatment option for R/R B-NHL; some patients could achieve long-term sustained responses after C-CAR011 infusion（ClinicalTiral.gov number, NCT03483688）.

Abstract 1574: Preclinical evaluation of epcoritamab combined with standard of care therapies for the treatment of B-cell lymphomas

Abstract B-cell non-Hodgkin lymphoma (B-NHL) is a heterogeneous disease with an unmet medical need for new efficacious, well tolerated, off-the-shelf therapies that can combine with standard of care (SOC) regimens. Epcoritamab (GEN3013; DuoBody®-CD3×CD20) is a novel subcutaneously administered bispecific antibody with a manageable safety profile and promising preliminary anti-tumor activity in both aggressive and indolent B-NHL. Epcoritamab simultaneously binds to CD3 on T cells and to CD20-expressing tumor cells to induce potent T-cell-mediated killing. SOC treatments for B-NHL include rituximab in combination with chemotherapy regimens such as CHOP (cyclophosphamide, doxorubicin, vincristine and prednisone), bendamustine, or immunomodulatory agents such as lenalidomide. These therapies have mechanisms of action distinct from that of epcoritamab, and mostly non-overlapping adverse event profiles. Previous preclinical studies demonstrated that epcoritamab can induce effective anti-tumor activity in the presence of a rituximab analog, supporting the combination of the two in clinical trials. Here, we present in vitro studies conducted to evaluate whether epcoritamab can be used in combination with current SOC therapies for B-NHL. Healthy donor T cells as effector cells and/or CD20-expressing B-NHL tumor cell lines were pre-treated with each individual SOC component (lenalidomide, cyclophosphamide, doxorubicin, vincristine, prednisone) to evaluate the impact on T cells and target cell lines. Next, co-cultures of pre-treated T and B-NHL cells were incubated with the same SOC component in the presence of epcoritamab, and T-cell mediated cytotoxicity and associated T-cell activation were assessed by flow cytometry. Bendamustine was added together with epcoritamab during the T-cell activation and cytotoxicity assay to assess potential antagonizing effect. Lenalidomide enhanced T-cell activation induced by CD3 crosslinking with immobilized anti-CD3 or epcoritamab, resulting in higher potency of these T cells to exert epcoritamab-induced cytotoxicity of CD20-expressing tumor cells. T cells pre-treated with individual CHOP components were capable of mediating epcoritamab-induced cytotoxicity. Finally, bendamustine did not antagonize T-cell activation and had an additive effect on T-cell-mediated cytotoxicity by epcoritamab. These preclinical data indicate that epcoritamab-can be combined with these SOC agents. In conclusion, these data warrant clinical evaluation of epcoritamab combinations with multiple SOC therapies. Given the promising single-agent activity of epcoritamab, these combinations may lead to deep and durable responses that can translate into improved long-term outcomes for B-NHL patients. Epcoritamab combination therapies are planned to be evaluated in a clinical trial sponsored by Genmab and Abbvie. Citation Format: Christopher W. Chiu, Ida H. Hiemstra, Wessel ten Hagen, Rajaa Snijdewint-Nkairi, Bart de Jong, Roberto S. Oliveri, Brian Elliot, Edith Szafer-Glusman, Danita Schuurhuis, Julie Blaedel, Tahamatan Ahmadi, Esther Breij, A. Kate Sasser, Maria Jure-Kunkel. Preclinical evaluation of epcoritamab combined with standard of care therapies for the treatment of 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 1574.

Predicting Clinical Response and Safety of JCAR017 in B-NHL Patients: Potential Importance of Tumor Microenvironment Biomarkers and CAR T-Cell Tumor Infiltration

▪Introduction:JCAR017 is a CD19-directed 4-1BB CAR T-cell product administered in a defined composition at a precise dose of CD8 and CD4 CAR T cells. TRANSCEND NHL 001 is the first multicenter phase 1 trial of JCAR017 in R/R B-cell NHL (NCT02631044). Interim results (Abramson, 14-ICML 2017) demonstrated high complete response rates and low incidence of cytokine release syndrome (CRS) and neurotoxicity (NT). Tumor cell density, T cell density and detection of known immunosuppressive pathways were assessed retrospectively in tumor biopsies from TRANSCEND patients to examine relationships with clinical efficacy, CAR T PK, and safety endpoints.Method:Patients with R/R DLBCL or MCL were eligible. Patients received lymphodepletion followed by JCAR017 at one of two dose levels (DL1, 5 × 107 cells; DL2, 1 × 108 cells). Tumor biopsies were collected prior to treatment and 7 to 20 days after JCAR017 administration. Pathology review of H&E staining confirmed tissue quality and identified tumor areas. In this July 7th, 2017 data cut, 43 biopsies (26 pretreatment; 17 post-treatment; 15 matched pairs) from 25 DLBCL and 3 MCL patients were examined. The best overall response (BOR), CRS and NT rates of this cohort were 71% (20/28), 36% (10/28; grade 1, 2) and 18% (5/28; grade 2-4). CAR T cell infiltration was quantified using in situ hybridization (ISH) probes specific to JCAR017 CAR mRNA. CAR T cells, non-CAR T cells and B cells were enumerated using a 5-plex immunofluorescence (IF) assay (EGFR, CD4, CD8, CD19, CD20). Immunosuppressive pathways were assessed using a second 5-plex IF assay (CD73, FOXP3, CD163, IDO, PD-L1). Images were analyzed using HALO software. Univariate t-tests were used for statistical analysis, and all reported p-values are 2-sided without multiplicity adjustment.Results:Pretreatment tumors had varying cellular compositions: tumor cells (median: 77%; range 5 - 96%), CD4+ cells (0.90%; 0.02-15%), and CD8+ cells (1.5%; 0-23%). Preliminary data showed patients with a CR or PR at month 3 had a higher percentage of endogenous CD4+ cells in pretreatment tumors than those with a PD (CR, PR median: 7.9%; PD median: 0.38%; p < 0.0001). Percentages of CD8+ cells in pretreatment tumors did not differ between month 3 response groups (CR, PR median: 1.9%; PD median: 0.47%; p = 0.6496).Following JCAR017 therapy, CAR T cells infiltrated the tumor and comprised up to 22% of cells. The level of tumor infiltration trended higher in patients achieving a CR (median: 3.9%) or PR (median: 1.1%) compared to those with a BOR of SD, PD (median: 0.51%). Although both CD4+ and CD8+ CAR T cells were able to infiltrate the tumor area, patients with a CR had a higher ratio of CAR T cells that were CD8+ (as compared to CD4+) than those with a BOR of SD or PD (CR median: 0.83; SD,PD median: 0.14; p = 0.0097).Comparing matched pre- and post-treatment biopsies, patients achieving a BOR of CR or PR trended toward having a larger increase in CD8+ cells (CAR T and non-CAR T) in tumors as compared to patients achieving a BOR of SD or PD (CR, PR median change: +5.3%; SD, PD median change: +0.06%; p = 0.1225). Although expression of immunosuppressive factors varied widely among patients at pre-treatment (CD73 (median: 1.5%; range 0-42%), FOXP3 (0.10%; 0-1.5%), IDO (0.06%; 0-11%), CD163 (1.2%; 0-24%) and PD-L1 (0.16%; 0-56%)) and post-treatment (CD73 (1.6%; 0-53%), FOXP3 (0.09%; 0-4.3%), IDO (0.28%; 0-15%), CD163 (3.6%; 0-22%) and PD-L1 (3.3%; 0-65%)), increases in CD8+ cells in matched biopsies were associated with increases in IDO (R2 = 0.64) and PD-L1 (R2 = 0.61) expression.Conclusions:The state of the tumor microenvironment prior to treatment may impact the efficacy of CAR T cell therapy. This initial analysis suggests durable response at month 3 is associated with higher levels of CD4+ cells in pretreatment tumors. Following JCAR017 therapy, CAR T cells, both CD4+ and CD8+, infiltrated the tumor and adjacent tissue. BOR was associated with an increase in CAR T cells. Increasing levels of CD8+ cells in the tumor were associated with increases in IDO and PD-L1, suggesting that therapies targeting these pathways may enhance JCAR017 activity. Detailed analyses are ongoing to examine additional associations of tumor burden, T cell infiltration, and immunosuppressive factor expression with clinical efficacy, PK and safety, and will be presented. DisclosuresSwanson:Juno Therapeutics: Employment, Equity Ownership. Do:Juno Therapeutics: Employment, Equity Ownership. Merrigan:Juno Therapeutics: Employment, Equity Ownership. Lonning:Juno Therapeutics: Employment, Equity Ownership. Prentiss:Juno Therapeutics: Employment. Sutherland:Juno Therapeutics: Employment, Equity Ownership. Xie:Juno Therapeutics: Employment. Pham:Juno Therapeutics: Employment, Equity Ownership. Li:Juno Therapeutics: Employment, Equity Ownership. Albertson:Juno Therapeutics: Employment, Equity Ownership, Patents & Royalties. Stern:Juno Therapeutics: Employment, Equity Ownership. Moulis:Juno Therapeutics: Employment, Equity Ownership.

Preliminary Safety Profile of the CD19-Directed Defined Composition CAR T Cell Product JCAR017 in Relapsed/Refractory Aggressive B-NHL Patients: Potential for Outpatient Administration

Preliminary Safety Profile of the CD19-Directed Defined Composition CAR T Cell Product JCAR017 in Relapsed/Refractory Aggressive B-NHL Patients: Potential for Outpatient Administration

Pharmacokinetic, Pharmacodynamic and Blood Analytes Associated with Clinical Response and Safety in Relapsed/Refractory Aggressive B-NHL Patients Treated with JCAR017

Introduction:JCAR017 is a CD19-directed 4-1BB CAR T cell product administered in a defined composition at a precise dose of CD8 and CD4 CAR T cells. TRANSCEND NHL 001 is the first multicenter Phase 1 trial of JCAR017 in relapsed/refractory (R/R) B-cell NHL (NCT02631044). In previously reported interim results (Abramson et al, 14-ICML 2017), JCAR017 demonstrated high CR rates and low rates of cytokine release syndrome (CRS) and neurotoxicity (NT). The accompanying cellular pharmacokinetics (PK), pharmacodynamics (PD), and blood analyte evaluations are reported herein.Methods:Patients (pts) on the TRANSCEND NHL 001 trial received lymphodepletion with fludarabine and cyclophosphamide, followed by a single flat dose of JCAR017 at one of two dose levels (DL1, 5x107 CAR T cells; DL2, 1x108 CAR T cells). A small cohort received a 2-dose schedule at DL1. Blood samples were collected and evaluated for PK, PD, and cytokine levels. CAR T cell expansion was evaluated in bone marrow at day 11 (±3 days) after infusion. PK was measured using validated flow cytometry and qPCR-based assays to detect EGFRt (a marker encoded by the CAR transgene and expressed separately) or host cell integrated CAR transgene, respectively. B cell aplasia was assessed by CD19 flow cytometry. Plasma cytokines were evaluated. Cytokines and additional plasma analytes are being evaluated on additional samples and will be presented. Correlations were observed with clinical outcomes, including best overall response, durable response, and all grade NT and CRS. Univariate nonparametric tests were used for statistical analysis, and all reported p-values are 2-sided without multiplicity adjustment.Results:Using a data cut-off of May 7, 2017, 55 pts in the DLBCL cohort were evaluable for safety; 54 pts were evaluable for efficacy. Four MCL pts were also evaluated for safety. Blood samples from 59 pts were included in this analysis. Pts receiving DL2 relative to DL1 had higher median Cmax and median AUC0-28 for CD3+/CAR+, CD4+/CAR+, and CD8+ CAR+ T cell subsets in peripheral blood (AUC: DL2 vs DL1 was 1836 vs 461, 350 vs 182, and 1628 vs 114, for CD3, CD4, and CD8, respectively; p < 0.05 for CD8; Cmax: DL2 vs DL1 was 99.8 vs 27.9, 15.1 vs 5.2, and 73.1 vs 5.5 cells/μl, respectively). Median time to maximum CD3+ CAR T cell expansion was 15 days (range 8-29) and did not differ between dose levels. JCAR017 cells homed to the bone marrow at relatively similar levels of CD4+ and CD8+ CAR+ T cells to blood.Median Cmax and median AUC0-28 of CD8+ CAR+ T cells were higher in responding patients and with durable response at month 3 (CD8 Cmax median = 20.8 vs 5.5; CD8 AUC median = 235 vs 55 in CR/PR at Month 3 vs PD at Month 3). Of pts evaluable for CAR T cell persistence, 90% and 93% of 29 pts had detectable CD8+ and CD4+ CAR+ T cells, respectively, at month 3; 63% and 58% of 19 pts had detectable CD8+ and CD4+CAR+ T cells, respectively, at month 6. At months 3 and 6, no statistically significant differences in the persistence of CAR T cells were seen in those with durable response or relapse. Most striking, CAR T cells were detectable at time of relapse in 89% of 11 pts with PK, even though B cell aplasia (<1 cell/µl) was demonstrated in nearly all pts 97% (34/35) at month 3, 100% (24/24) at month 6.Higher Cmax and AUC0-28 at DL2 did not increase CRS or NT. For any NT or for > Gr2 CRS, both CD4+/CAR+ and CD8+/CAR+ median AUCs were 5-10 fold and 3-5 fold higher, respectively, than median DL2 AUC. Higher disease burden and baseline levels of inflammatory cytokines appeared to be associated with higher CAR T cell peak levels, higher cytokine peak levels, and higher incidences of CRS and NT. Preliminary findings suggest that patient factors may result in higher expansion of CAR T cells and the associated CRS and NT.Conclusions:Based on current data, JCAR017 has demonstrated increased CAR T cell expansion and persistence and higher durability of response at 3 months at higher dose levels, without increased toxicity. Exploratory analyses suggest that high peak levels of CAR T cells and cytokines in the blood may be associated with NT and CRS and may be influenced by baseline patient factors. CAR T cells are present at the time of relapse, suggesting opportunities for combination approaches. DisclosuresHeipel:Juno Therapeutics: Employment, Equity Ownership. Smith:Juno Therapeutics: Employment, Equity Ownership. Brown:Juno Therapeutics: Employment. Karimi:Juno Therapeutics: Employment, Equity Ownership. Xie:Juno Therapeutics: Employment. Li:Juno Therapeutics: Employment, Equity Ownership. Albertson:Juno Therapeutics: Employment, Equity Ownership, Patents & Royalties. Garcia:Juno Therapeutics, Inc.: Employment, Equity Ownership. Devries:Juno Therapeutics: Employment. Sutherland:Juno Therapeutics: Employment, Equity Ownership.

Association of PET/CT response assessment prior to CAR T-cell infusion with outcomes after CAR T-cell therapy in aggressive B-cell lymphomas.

e19568 Background: Anti-CD19 chimeric antigen receptor modified T-cell therapies (CAR-T) have expanded the treatment possibilities for many patients with relapsed/refractory (R/R) aggressive large B-cell lymphomas (B-NHL) but fails to achieve remissions in a considerable proportion of patients. In this study, we examined whether response to systemic therapy prior to CAR T-cell infusion is associated with outcome. Methods: We analyzed a retrospective dataset of R/R B-NHL patients (pts) treated with commercially available CAR-T at the University of Pennsylvania treated from April 2018 to August 2020. 18F-FDG PET/CT scans obtained directly prior to and after CAR-T therapy were re-evaluated for response by an independent radiologist using the Lugano 2014 criteria. Association between pre-therapy and post-therapy responses were examined using Fisher’s exact test. Results: 50 pts with aggressive R/R B-NHL were enrolled. 34 pts received tisagenlecleucel and 16 received axi-cel. Median age was 61 (range 29-82), and 27 pts (54%) were men. Most pts (n = 48, 96%) had ECOG performance status of 0-1, with a median number of 3 prior therapies (range 1-11). 14 (28%) of pts had LDH elevation at time of CAR T cell infusion. Median time from pre-therapy PET/CT to infusion was 47 days (range: 6-155). Median PFS was 3.4 months. Prior to CAR T cell infusion, responses were as follows: 12 complete response (CR), 9 partial response (PR), 8 stable disease (SD) and 21 progressive disease (PD). Post-therapy responses were: 20 CR, 5 PR, 2 SD, and 23 PD. 67% of pts with a CR pre-therapy had continued CR after CAR T-cell infusion, whereas 32% pf pts without a CR had a CR after infusion (p &lt; 0.05). There was a trend toward CR/PR immediately prior to CAR T cell infusion being significantly associated with CR after CAR-T. We found that high metabolic tumor volume ( &gt; 350cc vs ≤350cc) was associated with poorer PFS (log rank, p = 0.05). We did not find elevated LDH or use of bridging therapy to be associated with response to CAR-T. Conclusions: This is the largest report of lymphoma patients treated with CAR-T while in remission to date and suggests the importance of optimizing clinical response prior to CAR-T. Although patients in CR were excluded from the registrational trials, it is notable that patients in CR prior to CAR-T had a high rate of CR post CAR T cells despite the absence of active lymphoma on imaging. Longer follow-up is needed to confirm these early observations.

Real-life experience with the combination of polatuzumab vedotin, rituximab, and bendamustine in aggressive B-cell lymphomas.

Transplant-ineligible relapsed/refractory (rr) diffuse large B-cell lymphoma (DLBCL) patients represent an unmet medical need. Polatuzumab vedotin (Pola), an anti-CD79b antibody-drug-conjugate (ADG), with bendamustine- rituximab(BR) has recently gained approval for these patients, both in the USA and Europe, based on the GO29365 phase IIb trial. Real-life data with Pola are extremely limited. We report the outcomes of 61 Greek patients, who received Pola-(B)R mainly within a compassionate use program. Treatment was given for up to six 21-day cycles. Bendamustine was omitted in three cases due to previous short-lived responses. Fourty-nine rrDLBCL(efficacy cohort-EC) and 58 rr aggressive B-NHL (safety cohort-SC) patients received at least 1 Pola-BR cycle. Twenty-one (43%) patients of the EC responded with 12/49 (25%) CR and 9/49 (18%) PR as best response. Median progression-free survival, overall survival and duration of response were 4.0, 8.5, and 8.5 months respectively, while 55% of patients experienced a grade ≥3 adverse event, mainly hematologic. Treatment discontinuations and death during treatment were mainly due to disease progression. Twenty-two (41%) patients received further treatment; 11/22 are still alive, including one after CAR-T cells, and two after stem cell transplantation. Our data confirm that Pola-BR is a promising treatment for rrDLBCL patients, inducing an adequate response rate with acceptable toxicity. Pola-BR could be used as bridging therapy before further consolidative treatments.

Epcoritamab induces potent anti-tumor activity against malignant B-cells from patients with DLBCL, FL and MCL, irrespective of prior CD20 monoclonal antibody treatment

Epcoritamab (DuoBody-CD3xCD20, GEN3013) is a novel bispecific IgG1 antibody redirecting T-cells toward CD20+ tumor cells. Here, we assessed the preclinical efficacy of epcoritamab against primary tumor cells present in the lymph node biopsies from newly diagnosed (ND) and relapsed/refractory (RR) B-NHL patients. In the presence of T-cells from a healthy donor, epcoritamab demonstrated potent activity against primary tumor cells, irrespective of prior treatments, including CD20 mAbs. Median lysis of 65, 74, and 84% were achieved in diffuse large B-cell lymphoma (n = 16), follicular lymphoma (n = 15), and mantle cell lymphoma (n = 8), respectively. Furthermore, in this allogeneic setting, we discovered that the capacity of B-cell tumors to activate T-cells was heterogeneous and showed an inverse association with their surface expression levels of the immune checkpoint molecule Herpesvirus Entry Mediator (HVEM). In the autologous setting, when lymph node (LN)-residing T-cells were the only source of effector cells, the epcoritamab-dependent cytotoxicity strongly correlated with local effector cell-to-target cell ratios. Further analyses revealed that LN-residing-derived or peripheral blood-derived T-cells of B-NHL patients, as well as heathy donor T-cells equally mediated epcoritamab-dependent cytotoxicity. These results show the promise of epcoritamab for treatment of newly-diagnosed or relapsed/refractory B-NHL patients, including those who became refractory to previous CD20-directed therapies.

Genetic Variants of the NKG2C/HLA-E Receptor-Ligand Axis Are Determinants of Progression-Free Survival and Therapy Outcome in Aggressive B-Cell Lymphoma.

Simple SummaryNKG2C and its ligand HLA-E represent key molecules for NK cell-mediated immune responsiveness. However, the impact of genetic variants in NKG2C and HLA-E on clinical outcomes of aggressive B-cell non-Hodgkin lymphoma patients (B-NHL) has not been clarified. In this study, we analyzed the distribution of NKG2C deletion status and HLA-E variants in 441 patients and 192 healthy individuals. Homozygous deletion of NKG2C (NKG2C−/−) was more often found in high-risk patients compared to patients with a lower risk and consequently was associated with reduced 2-year progression-free survival. The HLA-E*01:01 allele frequency was increased in B-NHL patients and was strongly related with complete remission. Our results show that absence of NKG2C and HLA-E allelic variations is predictive for B-NHL outcome; while carriers of HLA-E*01:01 are characterized by high, complete remission rates, NKG2C−/− was rare, but associated with poorer outcome. Prospective validation of our results identifies patients that may benefit from risk-adapted therapy.Aggressive B-cell lymphomas account for the majority of non-Hodgkin lymphomas (B-NHL). NK cells govern the responses to anti-CD20 monoclonal antibodies and have emerged as attractive targets for immunotherapy in subtypes of B-NHL. NKG2C and its cognate ligand HLA-E represent key molecules for fine-tuning of NK cell-mediated immune responses. Here, we investigated the impact of genetic variants of NKG2C and HLA-E on clinical outcomes of 441 B-NHL patients. Homozygous deletion of NKG2C (NKG2C−/−) was three-fold increased in patients compared to 192 healthy controls. Among studied patients, NKG2C−/− was more abundant in International Prognostic Index (IPI) high-risk patients compared to patients with a lower IPI (p = 0.013). Strikingly, NKG2C−/− was associated with a significantly reduced 2-year PFS (progression-free survival) (p = 0.0062) and represented an independent risk factor for 2-year PFS in multivariate analysis (p = 0.005). For HLA-E, the cognate ligand of NKG2C, the HLA-E*01:01 allele frequency was increased in B-NHL patients compared to controls (p = 0.033) and was associated with complete remission in univariate (p = 0.034) and multivariate (p = 0.018) analysis. Our data suggest that NKG2C and HLA-E genotyping is a promising tool for both defining risk groups of aggressive B-NHL and predicting response to immune therapeutic approaches.

PROGNOSIS OF PEDIATRIC NON HODGKIN LYMPHOMAS TREATED WITH FAB/LMB96 IN TERTIARY CENTER, SAUDI ARABIA

Purpose: This study aimed at describing prognosis of pediatric B-NHL patients treated with FAB LMB-96 based protocol without dose modification, and to identify possible prognostic factors influencing the outcome and the treatment related toxicities.Methodology: Through a retrospective study design, the researchers traced 48 pediatric cases who had been diagnosed with Mature B-NHL and treated at Prince Nora Oncology Center (PNOC) in the period from January 2007 to December 2016. Data were retrieved from the medical records, radiology images and pathology specimen for cases who met the inclusion criteria. For operational definition, cases were defined according to WHO classification, while staging was performed according to Murphy’s classification. Kaplan Meier survival function analysis was used to determine overall survival and Event Free Survival; and Chi Square test was used to verify significance in the differences according to characteristics of the cases. Findings: Out of all cases (n=48), Saudis formed (85.4%) and male to female ratio was 2.4:1, with a median age at diagnosis of 5.6 years. Most of the cases were diagnosed as Burkitt’s lymphoma (87.5%), mainly in stage III (37.5%) and stage IV (39.6%), chiefly as primary abdominal retro-peritoneal tumors (47.9%). Almost all cases (97.9%) developed chemotherapy related hematologic toxicity and fever neutropenia, and one third (34.8%) had septic shock. At the end of follow-up (median=112 months), there were 40 patients (83.3%) achieved remission, out of them, 6 (12.5%) relapsed. Death was attributed to disease recurrence (3 cases) and treatment related toxicities (4 patients). No statistically significant difference detected in the overall and Event Free survival rates according to their age, gender, histology, biochemical markers, disease locations, staging and FAB group classifications (P &gt;0.05).Conclusion and recommendations: Childhood non-Hodgkin lymphomas are almost all high grade and frequently extranodal. They fall mainly into the categories Burkitt lymphoma, lymphoblastic lymphoma and anaplastic large cell lymphoma. Our findings are comparable with those reported in other international trials

Evaluation of CD160 and CD200 Expression as Differentiating Markers between Chronic Lymphocytic Leukemia and Other Mature B-Cell Neoplasms

Background: The present work aimed to investigate the expression of CD160/ CD200 in CLL and other mature B-cell neoplasms (MBN) and their use as an additional diagnostic tool for differentiating CLL from other MBN. Materials and Methods: Using flow cytometry, we detected the expression of CD160 &CD200 on B-cells from 30 CLL patients, 30 other MBN patients in addition to 20 controls. CDs160/200 measurements were determined as a percentage expression (≥20% was considered positive) and as a ratio of the mean fluorescence intensities (MFIR) of leukemic cells/controls and were considered positive when the ratios were ≥2 and 20, respectively. Results: 90% and 100% of the CLL group expressed CDs160/200 in comparison to 60% and 63.3% of other MBN (p=0.007, p<0.001), respectively. By MFIR, 96.7% and 50% of our CLL group expressed CDs160/200 in comparison to 76.7% and 30% of other MBN, respectively. CDs160/ 200 were not expressed on the controls. Positive co-expression of CD160 and CD200 was found in 90% of the CLL cases, 60% of HCL patients and only in 40% of B-NHL. However, double negative expression of both markers was found only in 24% of the B-NHL patients. Conclusion : CD160 with CD200 can be used as additional diagnostic markers to the available routine panel to differentiate between B-CLL and other non-specified B-NHL patients.

Cellular Kinetics and Anti-Therapeutic Antibody in Relapsed/Refractory B-NHL Patients Treated with JWCAR029

Cellular Kinetics and Anti-Therapeutic Antibody in Relapsed/Refractory B-NHL Patients Treated with JWCAR029

DOSE‐ADJUSTED EPOCH FOR LYMPHOMA‐ASSOCIATED HEMOPHAGOCYTIC LYMPHOHISTIOCYTOSIS: A STONE TWO BIRDS

Purpose: Lymphoma-associated hemophagocytic lymphohistiocytosis (LA-HLH) has a high fatality rate and the worst outcome. The major cause of LA-HLH is aggressive non-Hodgkin's lymphoma (NHL), especially T/NKT cell lymphomas. Until now, there is no recommended therapeutic schedule for this fatal disease. Dose-adjusted (DA) EPOCH (etoposide, prednisone, vincristine, cyclophosphamide, and doxorubicin) shows effective activity in patients with aggressive NHL, which also contains the critical drugs for HLH of HLH-94/04. The investigators therefore developed DA-EPOCH regimen to treat non-Hodgkin's lymphoma with hemophagocytic lymphohistiocytosis and assess its clinical outcome including safety and efficacy. Methods: We did a single-arm, open-label, phase 2 clinical trial in previously untreated patients diagnosed as NHL with HLH (NCT01818908). We planned for B-NHL patients with HLH to receive six cycles of the DA-EPOCH-R regimen while for T/NK-NHL patients with HLH to receive six cycles of the DA-EPOCH regimen. Also, after achieving more than partial response (PR) after six cycles of DA-EPOCH±R regimen, autologous stem cell transplantation (ASCT) for B-NHL or allogeneic stem cell transplantation (allo-SCT) for T/NK-NHL was followed as consolidation therapies. The DA-EPOCH±R regimen included etoposide (50 mg/m2), vincristine (0.4 mg/m2), and doxorubicin (10 mg/m2) at daily doses by continuous intravenous infusion over 96 hours (days 1−4); cyclophosphamide (750 mg/m2, intravenous bolus, day 5), prednisone (60 mg/m2 bid, days 1−5) with or without rituximab (375 mg/m2, day 0). The cycle was repeated every 3 weeks. The primary endpoint was the overall response rate (ORR). Conclusions: DA-EPOCH-R regimen as front-line treatment followed by ASCT as consolidation treatment demonstrates a highly effective and safe strategy for B-NHL patients with HLH while DA-EPOCH cannot improve outcomes for T/NK-NHL patients with HLH. Keywords: DA-R-EPOCH; histiocytes; non-Hodgkin lymphoma (NHL).

Concentration and Glycoform of Rituximab in Plasma of Patients with B Cell Non-Hodgkin's Lymphoma.

Therapeutic antibodies have heterogeneities in their structures, although its structural alteration in the body is unclear. Here, we analyzed the change of amino acid modifications and carbohydrate chains of rituximab after administration to patients. Twenty B cell non-Hodgkin's lymphoma patients who were treated with rituximab for the first time or after more than one year's abstinence were recruited. Structural analysis of rituximab was carried out at 1h after administration and at the trough by using liquid chromatography/time-of-flight-mass spectrometry. Plasma rituximab concentration and pharmacodynamic markers were also determined. Of recruited twenty, 3 patients exhibited rapid rituximab clearance. Nine types of carbohydrate chains were detected in rituximab isolated from the blood. The composition ratios in some glycoforms were significantly different between at 1h after administration and at the trough, although consisted amino acids remained unchanged. The patients with high clearance showed extensive alterations of glycoform composition ratios. However, pharmacodynamics makers were not different. Inter-individual variations in plasma concentrations of rituximab were found in some B-NHL patients. We could analyze a change in glycoforms of rituximab in the patients, and this finding may affect the pharmacokinetics of rituximab.

Dose-Adjusted EPOCH Regimen as First-Line Treatment for Non-Hodgkin's Lymphoma with Hemophagocytic Lymphohistiocytosis: A Single-Arm, Open-Label, Phase 2 Trial

Background: The treatment of malignancy-associated HLH especially NHL-associated HLH has not been prospectively studied. The objective of this study was to evaluate the efficacy, safety and feasibility of DA-EPOCH±R regimen which contains etopooside, cyclophosphamide, doxorubicin, vincristine, prednisone with or without rituximab as a first-line treatment for NHL with HLH. Methods: In this prospective, single-arm, multicenter phase II clinical trial, we enrolled patients with previously untreated, histologically confirmed NHL aged 17-80 years and diagnosed as HLH according to HLH 2004 standard. A total of 6 cycles were planned if the patient achieved at least partial remission after the interim assessment. The cycle was repeated every 3 weeks. The primary endpoint was the overall response rate (ORR). Also, after achieving more than partial response (PR) after six cycles of DA-EPOCH±R regimen, autologous stem cell transplantation (ASCT) for B-NHL or allogeneic stem cell transplantation (allo-SCT) for T/NK-NHL was followed as consolidation therapies. This study was registered with Clinical Trials.gov, number NCT01818908. Findings: Fifty-nine patients were enrolled and evaluated in the study. Twenty-six patients (44.1%) were B-NHL with HLH and the other 33 patients (55.9%) were T/NK-NHL with HLH. The median cycle was six (range 1-6) for all B-NHL patients and three (range 1-6) for T/NK-NHL patients. The ORR at the end of the treatment was 80.7% for B-NHL with HLH while only 12.1% was for T/NK-NHL with HLH. With a median follow-up of 54 months (30-72 months) for B-NHL with HLH, 5-year progression-free survival (PFS) was 56.7 ± 9.9% and 5-year overall survival (OS) was 73.1 ± 8.7%. For the T/NK-NHL with HLH, 3-month PFS and OS rate was 30.3 ± 8.0% and 51.5 ± 8.7%. For the 21 B-NHL patients who achieved more than PR, 12 patients (57.1%) received ASCT as the consolidation therapy. B-NHL patients who received ASCT have significantly superior PFS (P<0.001) and OS (P=0.033) than patients who did not. Grade 3/4 neutropenia toxicities occurred on 43% of all cycles. Grade 3/4 thrombocytopenia occurred on 14% of all cycles. Interpretations: DA-EPOCH-R regimen as front-line treatment followed by ASCT as consolidation treatment demonstrates a highly effective and safe strategy for B-NHL patients with HLH while DA-EPOCH cannot improve outcomes for T/NK-NHL patients with HLH. Funding: National Natural Science Foundation of China Declaration of Interest: We declare no potential conflicts of Interest. Ethical Approval: All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Presence of autoimmune disease affects not only risk but also survival in patients with B-cell non-Hodgkin lymphoma.

Although autoimmune diseases (AIDs) are known to predispose to non-Hodgkin lymphoma (NHL), their association with NHL prognosis has rarely been investigated. We examined associations between autoimmunity and B-cell NHL onset by comparing AID history (determined by self-report and medication review and supplemented by chart review where possible) among 435 adult B-NHL patients in Hadassah-Hebrew University Medical Center, diagnosed 2009-2014, and 414 age-and-sex frequency-matched controls. We examined AIDs as a whole, B- and T-cell-mediated AIDs, and autoimmune thyroid diseases. Among cases, we used Kaplan-Meier and Cox regression models to assess the association of AID with overall survival and relapse-free survival, adjusting for prognostically important patient and disease characteristics such as Ki67% staining, International Prognostic Index, rituximab treatment, and histological subgroup. Autoimmune diseases were associated with B-NHL (odds ratio [OR]=1.95; 95% confidence interval (CI), 1.31-2.92), especially AIDs mediated by B-cell activation (OR=5.20; CI, 1.90-14.3), which were particularly associated with marginal zone lymphoma (OR=19.3; CI, 4.59-80.9). We found that time to relapse for all B-NHL patients with AIDs was significantly shorter (mean of 49.21mo [±3.22]) than among patients without AID (mean of 59.74mo [±1.62]), adjusted hazard ratio [HRadj ]=1.69 (CI, 1.03-2.79). Specifically, in patients with diffuse large B-cell lymphoma, of whom 91.8% had received rituximab, a history of B-cell-mediated AIDs was associated with shorter relapse-free survival and overall survival, HRadj =8.34 (CI, 3.01-23.1) and HRadj =3.83 (CI, 1.20-12.3), respectively. Beyond confirming the well-known association between AIDs and B-NHL, we found that AID is an adverse prognostic factor in B-cell lymphoma, associated with a shortened time to relapse, suggesting that there are specific therapeutic challenges in the subgroup of patients suffering from both these diseases. Further work is required to address mechanisms of resistance to standard treatment in the setting of AID-associated B-NHL. In the era of immunotherapy, these findings have particular relevance.

High durable CR rates and preliminary safety profile for JCAR017 in R/R aggressive b-NHL (TRANSCEND NHL 001 Study): A defined composition CD19-directed CAR T-cell product with potential for outpatient administration.

120 Background: JCAR017 is a defined composition, CD19-directed 4-1BB CAR T cell product administered at a precise dose of CD8 and CD4 CAR T cells in a seamless design Phase 1 pivotal trial of relapsed/refractory (R/R) B-cell NHL (TRANSCEND NHL 001; NCT02631044). Methods: Patients (pts) with R/R DLBCL NOS, PMBCL, FL grade 3B, or MCL and adequate organ function are eligible. The FULL dataset includes all pts in the DLBCL cohort (i.e. excludes MCL). The CORE dataset includes pts meeting inclusion for a planned pivotal DLBCL cohort (DLBCL NOS [ de novo or transformed from follicular lymphoma], ECOG 0-1, no prior allo-SCT). Results: As of July 7, 2017, 69 pts were treated in the DLBCL cohort and evaluable for safety. In the FULL dataset, 21 pts (30%) had cytokine release syndrome (CRS), with 1 serious CRS event (1%; Gr 4). Neurotoxicity (NT) at any grade developed in 14 (20%), including 10 (14%) with Gr 3-4 events. There were no Gr 5 CRS or NT events. Median time to onset of first CRS and NT was 5 days and 10 days, respectively, 13 (19%) received anti-cytokine therapy and only one required vasopressor support. In the CORE dataset (n = 49), similar rates of CRS and NT were shown. The majority of pts, 64% (44/69), had no CRS or NT, and median onset of CRS and NT were 5 and 11 days, respectively, suggesting outpatient delivery of JCAR017 may be feasible. In the DLBCL cohort, 68 pts were evaluable for efficacy; best overall response, 3-month, and 6-month response rates in the FULL dataset were 75% (51/68), 49% (27/55), and 40% (14/35), respectively; and, in the CORE dataset, were 84% (41/49), 65% (26/40), and 57% (13/23). The best overall, 3-month, and 6-month CR rates were 61% (30/49), 53% (21/40), and 52% (12/23), respectively, in the CORE dataset. Among 16 double/triple hit pts, best ORR was 81%, and 3-month CR rate was 60%. A trend toward improved response rate at 3 months was observed in pts treated at DL2 compared to DL1. Conclusions: At the symposium, we will report updated efficacy and the preliminary safety profile for JCAR017, which supports potential outpatient administration in R/R Aggressive B-NHL patients. Clinical trial information: NCT02631044.

Dynamic Analysis of Human Natural Killer Cell Response at Single-Cell Resolution in B-Cell Non-Hodgkin Lymphoma.

Natural killer (NK) cells are phenotypically and functionally diverse lymphocytes that recognize and kill cancer cells. The susceptibility of target cancer cells to NK cell-mediated cytotoxicity depends on the strength and balance of regulatory (activating/inhibitory) ligands expressed on target cell surface. We performed gene expression arrays to determine patterns of NK cell ligands associated with B-cell non-Hodgkin lymphoma (b-NHL). Microarray analyses revealed significant upregulation of a multitude of NK-activating and costimulatory ligands across varied b-NHL cell lines and primary lymphoma cells, including ULBP1, CD72, CD48, and SLAMF6. To correlate genetic signatures with functional anti-lymphoma activity, we developed a dynamic and quantitative cytotoxicity assay in an integrated microfluidic droplet generation and docking array. Individual NK cells and target lymphoma cells were co-encapsulated in picoliter-volume droplets to facilitate monitoring of transient cellular interactions and NK cell effector outcomes at single-cell level. We identified significant variability in NK-lymphoma cell contact duration, frequency, and subsequent cytolysis. Death of lymphoma cells undergoing single contact with NK cells occurred faster than cells that made multiple short contacts. NK cells also killed target cells in droplets via contact-independent mechanisms that partially relied on calcium-dependent processes and perforin secretion, but not on cytokines (interferon-γ or tumor necrosis factor-α). We extended this technique to characterize functional heterogeneity in cytolysis of primary cells from b-NHL patients. Tumor cells from two diffuse large B-cell lymphoma patients showed similar contact durations with NK cells; primary Burkitt lymphoma cells made longer contacts and were lysed at later times. We also tested the cytotoxic efficacy of NK-92, a continuously growing NK cell line being investigated as an antitumor therapy, using our droplet-based bioassay. NK-92 cells were found to be more efficient in killing b-NHL cells compared with primary NK cells, requiring shorter contacts for faster killing activity. Taken together, our combined genetic and microfluidic analysis demonstrate b-NHL cell sensitivity to NK cell-based cytotoxicity, which was associated with significant heterogeneity in the dynamic interaction at single-cell level.

High prevalence of hepatitis B virus infection in patients with aggressive B cell non-Hodgkin's lymphoma in China.

We retrospectively analyzed a large study to investigate the association of hepatitis B virus (HBV) with aggressive B cell non-Hodgkin's lymphoma (aggressive B-NHL) in China, where HBV is endemic. HBV was present in 39 aggressive B-NHL patients (10.46%), 13 indolent B-NHL patients (5.09%), 12 multiple myeloma (MM) patients (3.67%), and 5 solitary plasmacytoma (SP) patients (6.67%). HBV infection was significantly associated with increased risks for aggressive B-NHL (P < 0.01). HBV seems to have a very important role in the pathogenesis of aggressive B-NHL in China.