NHL Patients Research Articles

Phase I Trial of ‘Re-Priming’ Radiation Therapy for Relapsed/Refractory Non-Hodgkin Lymphoma Patients in Incomplete Response after Chimeric Antigen Receptor T-Cell (CAR-T) Therapy

Inpatients with relapsed/refractory non-Hodgkin lymphoma (R/R NHL) treated with CD19-directed CAR-T, only ∼40% achieve complete response (CR) by day 30 PET/CT evaluation. Of those who do not, the large majority (∼70%) ultimately fail, providing an ideal target for early therapeutic intervention to 're-prime' CAR-T. Preclinical and early clinical studies suggest potential synergy and immune augmentation when combining RT with CAR-T. Here we report the phase I results of a prospective phase I/II clinical trial hypothesizing that early salvage focal RT to poor responding sites of disease after CAR-T in R/R NHL patients is safe (phase I) and will improve conversion to CR by day 90 post-CAR-T PET/CT from 29% (historical control) to 58% (phase II). Weopened a single-arm open-label phase I/II prospective clinical trial at our institution for R/R NHL patients treated with CD19-directed CAR-T with incomplete response on day 30 post-CAR-T PET/CT scan (defined as Lugano > = 4). The phase I component used a 'Rolling 6' design with 6 patients enrolled concurrently at the "definitive" dose level (40-50 Gy EQD2 [i.e., 30 Gy in 5 fractions], with de-escalation to "palliative" dose level (20-32.5 Gy EQD2 [i.e., 20 Gy in 5 fractions]) if >2 dose-limiting toxicities (DLT) observed. Hypofractionated regimens (i.e., 5 fractions) directed only to residual FDG-avid disease were recommended to minimize lymphopenia and potentially result in a more favorable immune microenvironment. DLT rate was defined within 60 days of RT by CTCAE v5.0 grade 4+ hematologic, grade 3+ dermatitis/burn, pneumonitis, enteritis, or other toxicity attributable to RT, as well as new grade 3+ cytokine release syndrome (CRS) per ASTCT consensus guidelines or grade 3+ neurotoxicity per ASTCT ICANS consensus guidelines for adults. BetweenApril 2021 and July 2022, 6 patients were enrolled. All 6 patients had diffuse large B-cell lymphoma (DLBCL), with 3/6 (50%) transformed from low-grade follicular lymphoma. 2/6 had primary refractory DLBL, while the other 4/6 had median 2.5 lines of treatment prior to CAR-T. No patient had prior RT to a site of residual FDG-avid disease on day 30 post-CAR-T PET/CT. 5/6 patients were treated to 30 Gy in 5 fractions, with the remainder patient treated to 36 Gy in 10 fractions. No grade 3+ DLTs related to RT were observed in the 60-day post-RT period. RT related toxicities included grad 1 alopecia, grade 1 radiation pneumonitis, grade 1 nausea & vomiting, and grade 2 skin infection. Early salvage focal "definitive" dose RT to sites of incomplete response on day 30 post-CAR-T PET/CT for R/R/ NHL patients was safe with no de-escalation of dose needed. This dose will used in the subsequent phase II component of the trial.

HLA-A*02:06 allele may be susceptible to myelodysplastic syndrome in Zhejiang Han population, China.

The association between HLA loci and haematological malignancy has been reported in certain populations. However, there are limited data for HLA loci at a high-resolution level with haematological malignancy in China. In this study, a total of 1115 patients with haematological malignancies (including 490 AML, 410 acute lymphoblastic leukaemia (ALL), 122 myelodysplastic syndrome [MDS] and 93 non-Hodgkin's lymphoma [NHL]) and 1836 healthy individuals as a control group in the Han population of Zhejiang Province, China, were genotyped for HLA-A, HLA-C, HLA-B, HLA-DRB1 and HLA-DQB1 loci at high resolution. The possible association between HLA alleles and haplotypes and haematologic malignancy was analysed. The allele frequencies (AFs) of HLA-A*02:05, HLA-A*02:06, HLA-A*32:01, HLA-B*35:03, HLA-B*54:01, HLA-B*55:07, HLA-DRB1*04:05, HLA-DRB1*15:01, HLA-DQB1*04:01 and HLA-DQB1*06:02 in the MDS patients were much higher than those in the control group (P<0.05), while the AFs of HLA-C*07:02, HLA-DRB1*03:01, HLA-DRB1*14:54, HLA-DQB1*02:01 and HLA-DQB1*05:03 were obviously lower than those in the control group (p < .05). Interestingly, the differences in these HLA alleles in patients with MDS were not significant after applying Bonferroni correction (Pc>.05), except for HLA-A*02:06 (Pc<.01). There were 13, 6 and 10 HLA alleles with uncorrected significant differences (p < .05) among patients with AML, ALL and NHL, respectively, compared with those in the control group, but the differences in these HLA alleles were not significant after correction (Pc>.05). Compared to those of the control group, there were some haplotypes over 1.00% frequency in patients with AML, MDS and NHL patients with uncorrected significant differences (p < .05). However, none of them showed a significant difference after correction as well (Pc>.05). The study reveals that HLA-A*02:06 may lead to susceptibility to MDS, but none of the HLA alleles were associated with AML, ALL or NHL after correction. These data will help to further understand the role of HLA loci in the pathogenesis of haematological malignancy in China.

Inhibitory NKG2A+ and absent activating NKG2C+ NK cell responses are associated with the development of EBV+ lymphomas.

Epstein-Barr virus (EBV) is a ubiquitous herpesvirus, which infects over 90% of the adult human population worldwide. After primary infections, EBV is recurrently reactivating in most adult individuals. It is, however, unclear, why these EBV reactivations progress to EBV+ Hodgkin (EBV+HL) or non-Hodgkin lymphomas (EBV+nHL) only in a minority of EBV-infected individuals. The EBV LMP-1 protein encodes for a highly polymorphic peptide, which upregulates the immunomodulatory HLA-E in EBV-infected cells, thereby stimulating the inhibitory NKG2A-, but also the activating NKG2C-receptor on natural killer (NK) cells. Using a genetic-association approach and functional NK cell analyses, we now investigated, whether these HLA-E-restricted immune responses impact the development of EBV+HL and EBV+nHL. Therefore, we recruited a study cohort of 63 EBV+HL and EBV+nHL patients and 192 controls with confirmed EBV reactivations, but without lymphomas. Here, we demonstrate that in EBV+ lymphoma patients exclusively the high-affine LMP-1 GGDPHLPTL peptide variant-encoding EBV-strains reactivate. In EBV+HL and EBV+nHL patients, the high-expressing HLA-E*0103/0103 genetic variant was significantly overrepresented. Combined, the LMP-1 GGDPHLPTL and HLA-E*0103/0103 variants efficiently inhibited NKG2A+ NK cells, thereby facilitating the in vitro spread of EBV-infected tumor cells. In addition, EBV+HL and EBV+nHL patients, showed impaired pro-inflammatory NKG2C+ NK cell responses, which accelerated the in vitro EBV-infected tumor cells spread. In contrast, the blocking of NKG2A by monoclonal antibodies (Monalizumab) resulted in efficient control of EBV-infected tumor cell growth, especially by NKG2A+NKG2C+ NK cells. Thus, the HLA-E/LMP-1/NKG2A pathway and individual NKG2C+ NK cell responses are associated with the progression toward EBV+ lymphomas.

Peripherally inserted central catheter insertion and management in Hodgkin and non-Hodgkin lymphomas: a 13-year monocentric experience

BackgroundNon-Hodgkin’s lymphoma (NHL) and Hodgkin’s lymphoma (HL) are two of the most common hematologic diseases that require an infusion of immunochemotherapies in conjunction with radiotherapy, often in an outpatient setting. For relapsed/refractory disease, autologous peripheral hematopoietic stem cell transplantation and sometimes allogeneic transplantation (HSCT) are considered standard treatment options. Recently, chimeric antigen receptor (CAR) T cells and bispecific antibodies have emerged as an important and effective option for the treatment of relapsed/refractory patients. These medical approaches deserve effective, safe, and durable vascular access, especially for the ambulatory population undergoing discontinuous treatment associated with high rates of complications and life-threatening toxicities. Peripherally inserted central catheters (PICCs) are vascular devices with an intermediate-to-long-term lifespan that are inserted ultrasonically into a peripheral brachial vein. Their ease of insertion by trained nurses and low rate of catheter-related infectious and thrombotic complications make them ideal devices for treating oncology and hematology patients.PurposeIn this study, we aim to demonstrate that PICCs are an essential tool for the treatment of HL and NHL patients in terms of efficiency and safetyMethods and resultsFrom March 2007 to June 2020, 316 PICC implantations were performed by our PICC team in 276 HL patients and 363 PICC in 322 NHL patients. The total lifespan of the PICCs was 50,660 days in HL and 43,919 days in NHL patients. Most PICCs were removed at the end of therapy, and the rate of mechanical complications was low. Only one and four episodes of confirmed PICC-related catheter-related bloodstream infections (CRBSIs) (0.3%; 0.02/1,000 days/PICC and 1.2%; 0.07/1,000 days/PICC) were recorded in HL and NHL patients, respectively. There were only 11 (3.6%; 0.25/1,000 days/PICC) and nine (2.6%; 0.17/1,000 days/PICC) episodes of symptomatic PICC-related thrombotic complications in HL and NHL patients, respectively, without removal.ConclusionOur data indicate that the PICC can be considered the device of choice for treating HL and NHL patients because it is easy to insert, safe to use, long-lasting, and has a low complication rate, especially in the outpatient setting.

17th International Conference on Malignant Lymphoma, Palazzo dei Congressi, Lugano, Switzerland, 13 - 17 June, 2023.

Background: Axi-cel was approved by NMPA to treat R/R LBCL adult patients who had received 2 or more prior treatments in 2021 June. For better understanding the efficacy and safety of commercial Axi-cel in Chinese R/R NHL patients in real world setting, we conducted this multi-center, non-interventional study (ChiCTR2100047990). The accrual goal is 200 patients, and the primary endpoint is median OS. Here we made an interim analysis and reported clinical outcomes of Chinese patients treated by commercial CAR-T products in real world for the first time. Methods: R/R NHL patients treated with commercial Axi-cel in 17 authorized treatment centers from 11/2021 to 02/2023 were included. All patients signed written informed consents. We reported best objective response rate (bORR), best complete response (bCR) rate and adverse events. Results: A total of 101 R/R NHL patients were efficacy-evaluable. The median age was 56.8 years old, and 25 (24.8%) patients were ≥65 years. Fifty-nine patients were male. Baseline characteristics, including 80 (79.2%) with DLBCL, 4 (4.0%) with PMBCL, and 6 (5.9%) with high grade B-cell lymphoma. Forty-two (44.2%) patients had IPI ≥3 and 18 (24.3%) patients ECOG PS were ≥2. The median prior lines of therapy was 2, including 36 (35.6%) patients got ≥3 previous treatments and 10 patients had history of ASCT. Eighty-four patients were resistant to previous therapy, and the primary refractory subgroup reached 47 (46.5%). Bridging therapy was given in 56 (55.4%) patients, while combination therapy in 29 (28.7%) patients. The median follow-up was 9.2 months. The bORR and bCR was 83.2% (95% CI, 74.4 to 89.9) and 58.4% (95% CI, 48.2 to 68.1) respectively (Figure 1A). Response rates were consistent across key covariates, including disease type, disease stage, IPI score, cell-of-origin subtype, etc. Patients ≥65 years had favorable ORR [92.0% (95% CI, 74.0–99.0)] and CR rate [80.0% (95% CI, 59.3–93.2)]. Median PFS was 12.0 months (95% CI, 7.3 to NA). The median DOR and OS were not reached. No new safety signal was observed in Chinese patients. The most common ≥ grade 3AE were white-cell count decreased (in 87.6% of the patients), neutropenia (in 82.9%), pyrexia (in 73.3%). Eighty-one patients experienced cytokine release syndrome (CRS) of any grade, and 16 (15.2%) patients occurred grade 3 or higher. Any grade of neurologic events (NE) occurred in 17 (16.2%) patients, and only 1 (1.0%) patients were grade 4. No grade 5 CRS or NE appeared (Figure 1B-C). Fifty-five percent received tocilizumab and 52% received glucocorticoids to manage CRS and/or NE. The median cumulative cortisone-equivalent corticosteroid dose was 2000 mg, which was similar to ZUMA-1 cohort6 and much smaller than ZUMA-1 cohort1+2. The research was funded by: The research was funded by Fosun Kite Biotechnology Co., Ltd., Shanghai, China. Keywords: Aggressive B-cell non-Hodgkin lymphoma, Cellular therapies No conflicts of interests pertinent to the abstract.

CLINICAL EFFICACY OF CD34 POSITIVE SELECTION EX‐VIVO PURGING DURING AUTOLOGOUS STEM CELL TRANSPLANTATION IN PERIPHERAL T CELL LYMPHOMA

Background: Autologous stem cell transplantation (ASCT) is the standard treatment for peripheral T-cell lymphomas (PTCLs). Strategies that reduce the relapse rate and treatment-related mortality are essential for successful ASCT. Because in vivo/in vitro purging methods using rituximab are not appropriate for T-cell lymphomas, an ex vivo CD34+ selective purging system using CliniMACS is the most reliable method to reduce autograft tumor cell contamination in these tumors. In the present study, we retrospectively analyzed the clinical outcomes of HDT/ASCT after ex vivo purging via the CD34+ selective method in T-cell NHL patients. Methods: We retrospectively investigated the influence of ex vivo purging with CD34+ selection to maximize the effects of ASCT. Of 67 consecutive PTCL patients, 32 and 35 underwent purged and unpurged ASCT. Results: The purged group had improved overall survival (OS), disease-free survival (DFS), and cumulative incidence of relapse (hazard ratio [HR] = 2.68, p = 0.016; HR = 3.97, p = 0.002; and HR = 3.65, p = 0.004, respectively), compared to the unpurged group. Prognostic factor analysis showed that unpurged ASCT, chromosomal abnormalities at initial diagnosis, high risk, and pre-ASCT disease status were associated with poor survival outcomes. Subgroup analysis demonstrated that purging was most appropriate for International Prognostic Index high-risk patients who underwent upfront ASCT (HR = 3.35 [OS] and = 6.59 [DFS]). In purged ASCT group, NK cell activity and the lymphocyte-to-monocyte ratio known as an independent prognostic factor were increased after ASCT with statistical significance; the lymphocyte-to-monocyte ratio (LMR) was significantly increased after ASCT in the purged group (mean difference = 1.484, 95% CI = 0.407–2.560; p = 0.009). Also, Engraftment was achieved in all patients with no need for unpurged autologous stem cell backup in purged group. There were no engraftment failures or differences in adverse events including of transplant-related viral infections between the two groups. The research was funded by: The research was funded by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (NRF-2020R1G1A1099654). Keywords: Aggressive T-cell non-Hodgkin lymphoma, Stem Cell Transplant No conflicts of interests pertinent to the abstract.

Comparative Pre-Clinical Analysis of CD20-Specific CAR T Cells Encompassing 1F5-, Leu16-, and 2F2-Based Antigen-Recognition Moieties.

Over the past decade, CAR T cell therapy for patients with B cell malignancies has evolved from an experimental technique to a clinically feasible option. To date, four CAR T cell products specific for a B cell surface marker, CD19, have been approved by the FDA. Despite the spectacular rates of complete remission in r/r ALL and NHL patients, a significant proportion of patients still relapse, frequently with the CD19 low/negative tumor phenotype. To address this issue, additional B cell surface molecules such as CD20 were proposed as targets for CAR T cells. Here, we performed a side-by-side comparison of the activity of CD20-specific CAR T cells based on the antigen-recognition modules derived from the murine antibodies, 1F5 and Leu16, and from the human antibody, 2F2. Whereas CD20-specific CAR T cells differed from CD19-specific CAR T cells in terms of subpopulation composition and cytokine secretion, they displayed similar in vitro and in vivo potency.

Evaluation of Hexokinase 2 and Serum LDH in B-cell Non-Hodgkin’s Lymphoma Patients

Background and Objective: Tumor cells produce energy by glycolysis and Lactate Dehydrogenase and Hexokinase are the key part of glycolysis, The present study aimed to evaluate the expression of Hexokinase 2 and serum LDH in B-cell Non-Hodgkin’s Lymphoma patients Method: A total of 84 B-cell Non-Hodgkin’s Lymphoma Patients were enrolled in the study. HK-2 Marker was studied by Immunohistochemistry and LDH analysis was done by Cobas 6000 analyser method and correlated with Clinico-Pathological Parameters and Diagnostic panel. RESULT:73% of B-cell NHL patients had Abnormal LDH. It was significantly correlated with Multiple Disease site (P=0.002). In correlation with Diagnostic Panel (MIb1, CD10, CD2, MUM1, BCL6, BCL2) a higher trend of Abnormal LDH was seen in BCL-6 positive and BCL-2 negative Patients. 64% of patients had HK-2 expression. In correlation with Clinical parameters, it was significantly correlated with &gt;50 age group, while in correlation with pathological parameters A significant correlation observed with single disease site as compared to multiple disease site (P=0.004), In correlation with Diagnostic panel A higher trend of HK-2 Expression was observed in CD10 Negative and BCL-2 Negative Patients. Further Univariate survival analysis of LDH and HK-2 showed similar Overall survival in Abnormal and normal LDH level patients and HK-2 Positive and negative Patients respectively. Further Intercorrelation of both markers, showed almost similar expression of HK-2 between patients with normal LDH level and abnormal LDH level. CONCLUSION: Abnormal LDH level was closely associated with patients having disease at multiple sites, suggesting its activity to be associated with the advancement of the disease, HK-2 positivity was related with older age group, strengthens the findings of increased altered metabolic activity with older age, They are also associated with BCL-2 negativity that indicates that anti-apoptotic activity of BCL-2 might be responsible for that Though there was no significant correlation of LDH,, HK-2 with overall survival, majority of the patients who were dead had abnormal LDH and half of them showed positive HK-2 expression Briefly our data indicates that LDH and HK-2 can be useful parameter in defining clinical and prognostic aspects in NHL patient but needs further evaluation with larger groups of patients Keywords: NHL, Altered metabolism, Hexokinase, LDH

MANIFASTATIONS OF COMPLICATION IN THE ANEMIC SYNDROME IN NON-HODGKIN LYMPHOMAS OF TRANSCARPATIA.

The aim: To investigate the main indicators characterising clinical and haematological manifestations in patients with NHL complicated by anaemia at the stages of the disease development for optimisation of diagnostics and prognosis of the disease. Materials and methods: Data from 40 patients were analysed, 40 (100%) of whom were diagnosed with NHL complicated by anaemia. The severity of anaemia was divided into: mild anaemia - haemoglobin 10 - 12 g/dl; moderate anaemia - 8 - 10 g/dl; severe anaemia - 6.5 - 8 g/dl; life-threatening anaemia - below 6.5 g/dl. Patients were divided into 2 groups: the first group included 27 patients with haemoglobin levels of 100-120 g/l; the second group consisted of 13 patients with haemoglobin levels of 80 to 99 g/l. All patients were examined using clinical, laboratory, instrumental and special research methods. Statistical processing of the results was performed using the methods of variation statistics using the Microsoft Excel XP. Results: The course of lymphoproliferative diseases in which the proliferation of a malignant clone is often complicated by anaemia. The prognostically unfavourable factors were identified. The article discusses possible pathophysiological mechanisms of the identified changes. Conclusions: The results of the study demonstrate that as the disease progresses and the tumour mass increases, NHL patients tend to develop more severe anaemia. The leading cause of anaemia in patients is the infiltration of the bone marrow by tumour cells and their negative impact on erythropoiesis.

A Single Center Analysis of Hematopoietic Recovery in Patients Undergoing Allogeneic BMT with Suboptimal CD34+ Doses

Background: Allogeneic stem cell transplant (allo-SCT) is the only curative treatment for many hematologic malignancies. Stem cells can be collected from bone marrow, peripheral blood, or cord blood. Peripheral blood is the most common utilized source due to its convenience and the ability to collect high numbers of stem cells, but studies suggest that bone marrow offers certain advantages over peripheral blood, especially lower rates of graft-vs-host disease (Holtick et. al. Cochrane 2014), leading to higher survival in patients with nonmalignant disorders . The ideal cell dose for SCT is 2 x 106 cells/kg CD34 cells. However, in cases of bone marrow graft, if a suboptimal cell dose is collected, it is often not feasible to repeat extraction. This is especially difficult for minority ethnic groups who already face systemic barriers to care. Our goal for this study was to assess the correlation of CD34 dose to hematopoietic recovery in a minority-rich population of fresh marrow allogeneic stem cell recipients. Prior studies have also suggested that the cell dosing for transplant should be calculated by ideal body weight (IBW) rather than the standard actual body weight (ABW) (Cilley et. al. Bone Marrow Transplantation 2004). We assessed the correlation of recovery for ABW vs IBW as well. Methods: From 2012-2022, eighteen patients were identified in the Montefiore health system who received a bone marrow graft for allo-SCT with suboptimal CD34 doses. Alongside CD34 dose, an additional predictor evaluated was total nucleated cell (TNC) dose. ABW-based and IBW-based doses were calculated for both CD34 and TNC doses. Several outcomes were tracked for the eighteen patients, including "Days to WBC 1000 cells/μL", "Days to ANC 500 cells/μL ", and "Days to Plt≥20,000 cells/μL." The relationship between cell doses (ABW and IBW-based) and outcomes were evaluated independently through calculation of Spearman's correlation. Analysis and modeling were performed in RStudio 4.1.2. Results: AML and NHL patients made up the largest proportion of the sample (7/18 and 5/18) (Table 1). The median ABW-based CD34 dose (0.530 x 106 CD34 cells/kg) was nearly four times smaller than the standard of 2 x 106 cells/kg CD34 cells. Fifteen of 18 patients achieved reconstitution goals. Of the three patients who did not, one had persistent disease at the time of transplant and developed multiorgan failure dying within 17 days prior to hematopoietic recovery. Another died at day 58 without any platelet recovery. A third maintained platelets above 20,000 cells/μL throughout transplant. Assessing ABW-based CD34 dosing and IBW-based CD34 dosing as independent predictors, none of the outcomes, were significantly correlated (P > 0.05) (Figure 2). However, the Spearman's correlation coefficients were consistently of larger magnitude for IBW-based CD34 dosing than for ABW-based CD34 dosing for all three outcomes. Assessing ABW-based TNC dosing and IBW-based TNC dosing as independent predictors yielded no significant correlation. Conclusion: Studies suggest that there are benefits to bone marrow grafts over that of peripheral stem cell grafts. However, bone marrow harvesting is a process that is not easily repeated in cases where doses are inadequate, especially if the recipient has already undergone conditioning. We reviewed 18 cases of allo-SCTs performed at a single center with suboptimal CD34+ doses. In most cases, hematopoietic reconstitution was achieved. In our cohort of patients with suboptimal CD34+ doses in allo-BM SCTs, we noted no significant correlation between the dosages and outcomes for hematopoietic reconstitution. These results suggest there may be utility in taking a more liberal approach to standards for bone marrow graft doses, but this requires further study, and a much larger sample size. Additionally, we looked at CD34 dosing based on ABW versus IBW and noted consistent increases in magnitude for the Spearman Correlation's. Given the small sample evaluated, it is worth looking at this further with larger samples to see if dosing for bone marrow harvest by IBW may be more correlated with hematopoietic reconstitution in allogeneic SCT than dosing by ABW. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

The Impact of Chimeric Antigen Receptor T Cell Quality to Clinical Safety and Efficacy in Non-Hodgkin Lymphomas

Background: Immunotherapy has become increasingly important in modern cancer treatments. It has become evident that chimeric antigen receptor-modified T (CAR-T) cells may be a promising treatment for hematological malignancies. Malignancies affecting B cells have been successfully treated using CAR-T cell therapies. Many studies have explored the influence of various factors on the efficacy and survival of CAR-T therapy from different aspects, including the patient's disease state, general condition, tumor burden, and even the dose of CAR-T cells infused. Few researchers have explored the effect of CAR-T cell quality on disease treatment impact on remission, survival, and prognosis. This study combines the essential condition of patients and quality data of CAR-T cells, such as transduction efficiency, killing efficiency, and the ratio of CD4+ and CD8+ subsets, to analyze CAR-T's safety and clinical efficacy in the treatment of non-Hodgkin's lymphoma in our center. Methods: This was a retrospective, single-center, investigator-initiated study. A total of 179 patients with NHL were included. Patients' baseline data were obtained from electronic medical records and telephone follow-ups. Laboratories provided data on the quality of patients' CAR-T cells. We conducted a multivariate analysis to determine the independent factors that influence treatment response and patient prognosis. Results: In our study, the median age of 179 NHL patients was 52 (range 17-78) years, and 51 (28.5%) were ≥60 years old. Before CAR-T treatment, 22 (12.3%) patients remained in complete remission (CR), and 83 (46.4%) patients were in progressive disease (PD). Among the available data, 105 (61.4%) patients had CAR-T cell killing efficiency (KE) <30%, while 66 (38.6%) patients had killing efficiency (KE) ≥30%. There were 101 (58%) patients with CAR-T cell transduction efficiency (TE) <40% and 73 (42%) with transduction efficiency (TE) ≥40%, respectively. CD4+/CD8+ ratio of CAR-T cells <1 in 32 (18.4%) patients and CD4+/CD8+ ratio of CAR-T cells ≥1 in 142 (81.6%) patients. Overall, the median event-free survival (PFS) was 10.6 months (95% CI: 4.064-17.136months), and the median overall survival has not yet been reached. The 6-month OS was 80.2%, and the 1-year OS was 70.6%. 2-year 0S is about 59.8%. In addition, we divided patients into different subgroups according to the TE and KE of CAR-T cells, and the results of multivariate logistic regression analysis were as follows. The subgroup with TE<40% had a higher complete remission rate than the subgroup with TE≥40%. And the subgroup with KE≥30% had a higher rate of complete remission than the subgroup with KE<30%, with p-values equal to 0.007 and 0.022, respectively. There was statistical significance, so TE and KE independently influenced efficacy factors. Multivariate cox regression analysis showed that the subgroup of CAR-T CD4+/CD8+<1 had better overall survival than CAR-T CD4+/CD8+≥1, p=0.039 (95% CI: 0.114-0.947). Therefore, the ratio of CAR-T CD4+/CD8+ was an independent factor for OS, while the subgroups of transduction efficiency and killing efficiency did not show statistical significance. Conclusion: In general, CAR-T therapy has shown significant efficacy in NHL, and the quality of CAR-T cells plays a vital role in treatment. Screening suitable CAR-T cells in future treatment will Make the treatment effect more satisfactory. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Pre-clinical efficacy of CD20-targeted chimeric antigen receptor T cells for non-Hodgkin's lymphoma

BackgroundA 4-1BB/CD3-ζ-costimulated CAR-T against CD20 (CAR-T20) was subjected to a systemic efficacy evaluation in a cell co-culture model, and NOD-SCID IL-2 receptor gamma null mice (short for NSG mice) were xenografted with human Burkitt's lymphoma Raji cells.MethodsCAR-T20 cells were incubated with target cells (K562, K562 CD20 or Raji cells) at ratios of 10:1 and 5:1 for 24 h, and the killing rate was estimated by an LDH cytotoxicity assay. To evaluate the effect of CAR-T20 on the survival time of tumor-bearing animals, 30 NSG mice were employed, and Raji-Luc cells (5 × 105 cells per mouse) were administered prior to CAR-T20 administration. The survival time, optical intensity of Raji-Luc cells, clinical symptoms, and body mass of the animals were observed. Another 144 male NSG mice were employed to investigate the proliferation and antitumor effects of CAR-T20. Human cytokine and murine cytokines were detected at 1, 7, 14, 21, 28, 42, 56 and 90 days post-CAR-T administration, while biochemistry index analysis, T-cell and CAR-T-cell detection in peripheral blood, and histopathological examination were performed at 14, 28, 56 and 90 days post-administration.ResultsCAR-T20 cells had a specific killing effect on CD20-expressing cells in vitro. At a dose of 1 × 106 per mouse or above, CAR-T20 prolonged the median survival time from 14 days to more than 3 months, inhibited the proliferation of Raji cells in mice, and alleviated the clinical manifestations and weight loss caused by the Raji-Luc cell load. CAR-T20 at a dose of 2 × 106 per mouse or above inhibited the proliferation of Raji cells in mice for up to 111 days post-administration without recurrence. The numbers of T cells and CAR-T cells in the animals administered CAR-T20 increased significantly when Raji cells were markedly proliferated and subsequently decreased when Raji cells were predominantly inhibited. CAR-T20 increased human IFN-γ, murine TNF and murine IL-6 levels and decreased human IL-10 levels in tumor-bearing mice. The incidences of xenografted tumors in organs/tissues were also reduced effectively by CAR-T20.ConclusionThe effective dose of CAR-T20 in mice starts from 1 × 106 per mouse, equivalent to a clinical dose of 5 × 106/kg. Together, our data support the clinical translation of CAR-T20 for R/R B-cell NHL patients.

Adapter CAR T Cell Therapy for the Treatment of B-Lineage Lymphomas.

CD19CAR T cells facilitate a transformational treatment in various relapsed and refractory aggressive B-lineage cancers. In general, encouraging response rates have been observed in B-lineage-derived non-Hodgkin’s lymphomas treated with CD19CAR T cells. The major cause of death in heavily pretreated NHL patients is lymphoma progression and lymphoma recurrence. Inefficient CAR T cell therapy is the result of the limited potency of the CAR T cell product or is due to loss of the targeted antigen. Target antigen loss has been identified as the key factor that can be addressed stringently by dual- or multitargeted CAR T cell approaches. We have developed a versatile adapter CAR T cell technology (AdCAR) that allows multitargeting. Screening of three different B-lineage lymphoma cell lines has revealed distinct immune target profiles. Cancer-specific adapter molecule combinations may be utilized to prevent antigen immune escape. In general, CD19CAR T cells become non-functional in CD19 negative lymphoma subsets; however, AdCAR T cells can be redirected to alternative target antigens beyond CD19, such as CD20, CD22, CD79B, and ROR-1. The capability to flexibly shift CAR specificity by exchanging the adapter molecule’s specificity broadens the application and significantly increases the anti-leukemic and anti-lymphoma activity. The clinical evaluation of AdCAR T cells in lymphoma as a new concept of CAR T cell immunotherapy may overcome treatment failure due to antigen immune escape in monotargeted conventional CAR T cell therapies.

CLL-393 Outcomes of COVID-19 Infections in Patients With Hematological Malignancies in Republic of North Macedonia

Context: Patients with hematological malignancies are at increased risk of complications and adverse outcomes associated with COVID-19, according to current available studies. These patients are particularly vulnerable due to impaired or compromised immune responses caused by treatment or the disease itself. Objective: Our aim was to examine the severity and outcomes of COVID-19 infections among patients with hematological malignancies in Republic of North Macedonia. Design: This is the first retrospective descriptive study on patients with COVID-19 and underlying hematological malignancies in Republic of North Macedonia. We focused on clinical characteristics and COVID-19 infection outcomes in different malignancy settings. Patients: We collected data on clinical characteristics, diagnosis, treatment, and outcomes in adult patients with hematologic malignancies and confirmed COVID-19 infections between June 2020 and April 2022. Parametric data were compared using Fisher's exact test. Results: Of 206 patients, 153 (74%) had lymphoid, and 53 (26%) had myeloid malignancies. Median age was 61 years (IQR 47–70 years), 118 (57%) patients were male, and 86 (74%) patients were older than 60 years and had ≥2 comorbidities, with only 16 (17%) patients younger than 60 years. Severe/critical clinical severity was identified in 77 (37%). Overall, 42 patients (20%) died, including 33 patients (78%) older than 60 years. Patients older than 60 years had significantly higher mortality (p=0.0015) than younger patients. The majority of patients (58%) became COVID-19–positive while on active treatment. Significantly worse outcomes were observed among patients who were on active treatment compared with patients who were in hematological remission (p=0.0028). Among lethal cases, the majority of patients were diagnosed with CLL and NHL. We found no significant differences in mortality between treated and untreated CLL and NHL patients. Among 37 patients treated with a cocktail of monoclonal antibodies (casirivimab/imdevimab), no significant difference in outcome was observed, although 86% of them survived. Conclusions: Patients with hematological malignancies and COVID-19 infection were associated with higher mortality associated with older age, more comorbidities, and active disease. Patients with B-cell malignancy were the most affected, but no significant difference in mortality rate was observed across different subgroups, which may suggest that B-cell impairment is a key factor leading to severe infection.

Characterization of anti-CD79b/CD3 bispecific antibody, a potential therapy for B cell malignancies.

High rates of relapse and poor prognosis confer an urgent need for novel therapeutic agents for B cell non-Hodgkin lymphomas (B-NHLs). Herein, we describe a human IgG-like anti-CD79b/CD3 bispecific antibody (IBI38D9-L) that selectively depletes antigen-positive malignant B cells as an alternative treatment option for relapsed or refractory NHL patients. The antitumor activity and mechanism of action of IBI38D9-L were investigated in vitro using B-NHL cell lines and human primary effector cells and in vivo using xenograft models reconstituted with human PBMCs (peripheral blood mononuclear cells). Pharmacokinetic (PK) properties and preclinical toxicology were evaluated in cynomolgus monkeys and HSC-NPG mice. IBI38D9-L exerted potent B cell killing as well as T cell activation and proliferation in a tumor cell-dependent manner in vitro and was active against B-NHL cell lines with various CD79b expression levels. Subcutaneous xenograft tumors in NOG mice engrafted with human PBMCs were eradicated by IBI38D9-L treatment. Moreover, IBI38D9-L-treated mice showed a strong infiltration of activated T cells. In HSC-NPG mice, IBI38D9-L resulted in potent B cell depletion in peripheral blood and induced only slight body weight loss and cytokine release syndrome without significant toxicological findings. In cynomolgus monkeys, IBI38D9-L was well tolerated with good pharmacokinetic profiles. Collectively, these preclinical efficacy and safety data provide strong scientific rationales for using anti-CD79b/CD3 bispecific antibody as a promising therapeutic agent for B cell malignancies.

Status of CMV IgM Seropositivity Before and After Starting Chemotherapy in Patients with Haematological Malignancies

Background: Even after significant improvement of management of haematological malignancy, post chemotherapy infection is still causing significant mortality and morbidity. Cytomegalovirus (CMV) is an important cause of morbidity and mortality in immunocompromised patients. This study was designed to assess prospectively the status of CMV IgM seropositivity in patients with haematological malignancy at diagnosis and after starting chemotherapy by serology test. In Bangladesh the prevalence IgG is high and IgM is low in general population but in patients with chemotherapy due to haematological malignancy status of CMV infection is not known. By finding out the infection rate we will be able to provide supportive care more effectively to patients of chemotherapy due to haematological malignancy. &#x0D; Methodology: This was a prospective type of observational study and patients were selected by purposive sampling. Assessment of CMV IgG and IgM antibody test had been done at the time of 1st diagnosis and IgM antibody for CMV reassessed 6 weeks after initial chemotherapy. Antibodies were detected by Chemiluminescence method from Virology department of BSMMU. Statistical analysis was done both manually and Windows based software device with Statistical Package for Social Science (SPSS). A p-value of &lt;0.05 was taken as significant during data calculation. Result: During 12 months study period a total 45 patients with haematological malignancy were included in the study. Out of 45 patients, 28 were male (62%) and 17 were female (38%), male female ratio was 1.65:1. The youngest of the participants was 15 and oldest was 70 years old. The mean age of the participants was 35.77 years with SD of 16.93. Among 45 patients of the study, ALL patients were 20 (44.4%), AML patients were 17 (37.8%), NHL patients were 5 (11.1%) and HD patients were 3 (6.7%). Before chemotherapy all participants were CMV IgG positive (100%) no one was CMV IgM Positive (0%). Six weeks after chemotherapy, 42 (93.3%) cases were CMV IgM antibody negative and only 3 (6.7%) cases were positive for CMV IgM antibody. McNemar test was done to measure the level of significance and p-value was 0.250, which was not significant statistically. Conclusion: This small study found that all participants were CMV IgG seropositive (100%). It indicates that higher percentages of haematological malignancy patients are previously infected with CMV so there is more risk of reactivation after chemotherapy or BMT. Post chemotherapy CMV IgM seropositivity (new infection or reactivation of CMV) was low 6.7% in this study. This percentage is low in comparison with other studies. Percentage could be high if the tests were carried out with more sensitive and specific tests for CMV like PCR, pp65 antigenaemia or CMV DNA.

P1166: REPEAT BIOPSY IN RELAPSED OR REFRACTORY DIFFUSE LARGE B CELL LYMPHOMA: A NATIONWIDE SURVEY AND RETROSPECTIVE STUDY

Background: Almost half of patients with diffuse large B-cell lymphoma (DLBCL) will have relapsed/refractory (R/R) disease after frontline immunochemotherapy. Although guidelines recommend histological confirmation of R/R disease, repeat biopsies are not always performed. Aims: To better appreciate the extent and reasoning for not performing repeat biopsies in R/R DLBCL. Methods: We conducted a two-part study: (1) using a nationwide electronic questionnaire consisting of 8 clinical vignettes, we evaluated the views of practicing hematologists in Israel regarding the need for repeat biopsies in suspected R/R DLBCL; (2) a single center retrospective study was utilized to describe real-life clinical practice patterns of performing procedures aimed at achieving tissue diagnosis for relapsed/refractory aggressive NHL (aNHL) patients: we included all consecutive adult (≥18 years of age) patients diagnosed with aNHL treated at a tertiary center in Israel, between 1/1/2013 and 1/1/2020, we identified patients who were diagnosed with R/R disease and analyzed data pertaining to utilization of histological confirmation via repeat biopsy. Each timepoint of management decision regarding biopsy performance was considered as a separate “episode of R/R lymphoma”. We extracted data regarding reasons for not performing the biopsies from the computerized system, according to the treating physician. Results: In the survey part, all 64 participating physicians opted not to perform a repeat biopsy in at least one clinical case scenario. Physicians’ age, gender, tenure or clinical experience with DLBCL patients did not correlate with the decision to perform a biopsy, whereas more physicians chose to perform biopsies in relapsed compared to refractory disease cases. In the retrospective part, 116 episodes of R/R aNHL among 61 patients were identified. In 72% of these episodes a repeat biopsy was not performed, mostly due to low likelihood of an alternative diagnosis or problematic location for biopsy. Patients with relapsed disease were more likely to undergo biopsy, as compared to those with refractory disease (47% vs 19%, p=0.002). Focusing on the group of patients with DLBCL (excluding primary CNS lymphoma, primary mediastinal B cell lymphoma and Burkitt lymphoma), there were 43 patients with refractory (n=26) or relapsed (n=17) DLBCL. In 61.4% out of the 83 episodes of R/R disease in these 43 patients, a biopsy was not performed: 74.5% of refractory DLBCL episodes versus 35.7% of relapsed DLBCL episodes (p=0.001). Stratifying biopsy utilization for each patient according to the timing of event, i.e first, second or third event of R/R disease, revealed that less biopsies were performed at more advanced relapse episodes. The most common reason for not performing a repeat biopsy was having stable radiological findings and other diagnoses deemed unlikely, as depicted in Figure 1. Image:Summary/Conclusion: Our study suggests that contrary to guideline recommendations, in clinical practice many patients do not undergo repeat biopsy in R/R DLBCL, especially in refractory cases. Since this is a common dilemma faced in clinical practice, future studies and recommendations should address the necessity of repeat biopsy, according to patient and disease related characteristics.

P1152: RESULTS FROM A PHASE I PHARMACOKINETIC (PK) AND SAFETY STUDY OF TRPH-222, A NOVEL CD22-TARGETING ANTIBODY-DRUG CONJUGATE, IN PATIENTS WITH RELAPSED/REFRACTORY B-CELL NON-HODGKIN LYMPHOMA (R/R NHL)

Background: TRPH-222 is a novel antibody-drug conjugate (ADC) comprised of a humanized anti-CD22 monoclonal antibody and maytansine, a potent microtubule inhibitor, joined by a novel third-generation linker-conjugation technology (SMARTag®). This approach enables site-specific conjugation of the maytansine payload while tightly controlling drug:antibody ratio (DAR), resulting in a highly stable anti-CD22 ADC with a non-cleavable linker designed to widen the therapeutic window. Aims: The primary objectives of this first in human study are to determine the safety, tolerability and pharmacokinetics (PK) of TRPH-222 monotherapy in patients with R/R NHL. Methods: TRPH-222-100 is an open-label, multicenter study comprised of dose-escalation and dose-expansion stages. TRPH-222 was administered IV once every 3 weeks. 22 patients were enrolled in dose-escalating cohorts of TRPH-222 (0.6 mg/kg to 10 mg/kg) from DLBCL, FL, TFL, MCL and MZL histologies, and 10 patients in a dose-expansion cohort (7.5 mg/kg) focusing on DLBCL and FL histologies. Results: As of January 7, 2022, 32 NHL patients have been enrolled: 15 indolent (14 FL and 1 MZL) and 17 aggressive histologies (15 DLBCL, 1 TFL and 1 MCL). Patients had a median age of 64.5 years, a median of 4 prior lines of therapy including 7 patients receiving prior CAR-T treatment. Three DLTs occurred in 2 patients during the study and comprised Grade 3 and 4 transaminase elevations; one each at 4.2 and 10 mg/kg and one Grade 3 thrombocytopenia at 4.2 mg/kg. Treatment-related serious adverse events (AEs) occurred in 2 patients (6.3%), caused by thrombocytopenia and pyrexia (both at 7.5 mg/kg TRPH-222). AEs were more prevalent at doses ≥7.5 mg/kg and less prevalent at lower doses. There was a trend to higher grade AEs in patients with aggressive histologies, compared to indolent ones. The most frequent (≥ 5%) treatment-emergent related AEs (Grade ≥3) included thrombocytopenia (34%), neutropenia (22%), ALT/AST elevation (6%), dry eye (6%) and blurred vision (6%). Cytopenias were non-febrile, infrequent, asymptomatic and resolved without significant intervention. Ocular findings were consistent with known epithelial keratopathy of ADCs and were generally low grade and resolved to ≤ Grade 1 with dose interruptions and/or reductions. Overall, TRPH-222 demonstrated a favourable safety profile with most AEs being predominantly low grade, tolerable, easily managed and reversible. Preliminary efficacy results suggest evidence of anti-tumor activity, most notably in patients with R/R FL. Of the 13 response-evaluable FL patients, 4 complete responses (CR) and 2 partial responses (PR) were observed, with an overall response rate (ORR) of 46% and a complete response rate (CRR) of 31%. Four patients with metabolic CRs maintained these CRs for long periods off treatment; 3 patients remain in CR with responses maintained for up to 25 months. Responses were generally early, durable and CRs were maintained off-therapy. Beyond FL, CRs were also observed in 1 DLBCL patient and in 1 MCL patient. Summary/Conclusion: TRPH-222 was found to be well tolerated at higher dose levels than evaluated for other ADCs. TRPH-222 monotherapy resulted in robust and durable CRs in FL across dose levels where patients were able to discontinue TRPH-222 while remaining in remissions. Collectively, these characteristics of TRPH-222 are favorable for further development in the indolent lymphoma setting either as monotherapy or in combination with other anti-tumor agents in B-cell lymphoma patients.

Upregulation of IL-1 and prostaglandin gene expression during and after induction of chemotherapy in the blood of NHL patients

Upregulation of IL-1 and prostaglandin gene expression during and after induction of chemotherapy in the blood of NHL patients

Abstract 2902: Development of PSB202, a bifunctional antibody pair that target CD20 and CD37, for the treatment of B-cell malignancies

Abstract CD20 and CD37 are co-expressed on most B-NHL and CLL tumors. Systemic depletion of normal and malignant B-cells by anti-CD20 antibodies (e.g. rituximab and obinutuzumab) and anti-CD37 antibodies (e.g. otlertuzumab and BI 836826) has been well tolerated in clinical studies. Depletion of B-cells is reversible, with the ability to repopulate B-cells from hematopoietic stem cells. Dual targeting CD20 and CD37 may improve the clinical efficacy and decrease drug resistance. PSB202 is a novel biological entity composed of two engineered monoclonal antibodies which are produced from a single stable CHO cell line - a humanized anti-CD20 antibody (PSB102) and a humanized anti-CD37 antibody (PSB107) at a 1:1 molecular ratio. In vitro binding studies demonstrated that PSB202 specifically binds to human CD20 and CD37. The antibodies’ binding to FcγRIIIa was enhanced by reducing the fucose level on each antibody component. PSB202 directly induced potent autonomous killing via target engagement. PSB202 mediated depletion of human and monkey blood B-cells and malignant human B-cell lines through ADCC activity at EC50 of double digit pM and CDC activity at EC50 of low single digit nM. Importantly, PSB202 effectively depleted B-cells in ex vivo cultures of blood cells from CLL and NHL patients who were refractive to rituximab treatment. The antitumor activity of PSB202 was demonstrated in human xenograft models in mice. PSB202 was able to reduce different human lymphoma tumors by up to ~97.9% in a dose-dependent manner, suggesting that PSB202 might be potentially efficacious against multiple types of B-cell malignancies. Synergy was demonstrated for PSB202 in combination with lenalidomide. Toxicokinetic assessment in cynomolgus monkeys showed a dose proportional exposure for both antibody components, with mean terminal elimination half-life (T1/2) of ~60 and ~68 hours for the anti-CD20 and anti-CD37 component, respectively. PBS202 was well tolerated, with observed toxicities mostly related to its activity as a B-cell depletion agent. As expected, PSB202 when administered at ≥ 10 mg/kg resulted in nearly complete depletion of B lymphocytes 24 hours following the 1st dose. B-cells returned to near baseline within 2 months after the last dose at 10 mg/kg. PSB202 can deplete normal and malignant B-cells by targeting two clinically validated targets through multiple distinct mechanisms including direct B-cell killing, enhanced ADCC activity, and CDC activity. PSB202 offers the potential to improve clinical efficacy, decrease drug resistance, provide administration convenience especially when combining with additional therapeutics, lower the cost for treatment, and reduce the unwanted side effects from chemotherapy. PSB202 is currently being evaluated in patients with previously treated, relapsed, indolent B-cell malignancies (NCT05003141). Citation Format: Paul Algate, Zhi Liu, Saying Yuan, Hua Liu, Cristina Domeier, Haiming Jiang, Yuanzhi Lao, Yanling Xu, William C. Fanslow, Ron Schoner, Wei Yan. Development of PSB202, a bifunctional antibody pair that target CD20 and CD37, for the treatment of B-cell malignancies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2902.