Cell Lymphoma Patients Research Articles

Can we truly define 'Fitness' for CAR-T therapy in large B cell lymphoma patients?

Can we truly define 'Fitness' for CAR-T therapy in large B cell lymphoma patients?

The Oncogenic Lipid Sphingosine-1-Phosphate Impedes the Phagocytosis of Tumor Cells by M1 Macrophages in Diffuse Large B Cell Lymphoma.

A total of 30-40% of diffuse large B cell lymphoma (DLBCL) patients will either not respond to the standard therapy or their disease will recur. The first-line treatment for DLBCL is rituximab and combination chemotherapy. This treatment involves the chemotherapy-induced recruitment of tumor-associated macrophages that recognize and kill rituximab-opsonized DLBCL cells. However, we lack insights into the factors responsible for the recruitment and functionality of macrophages in DLBCL tumors. We have studied the effects of the immunomodulatory lipid sphingosine-1-phosphate (S1P) on macrophage activity in DLBCL, both in vitro and in animal models. We show that tumor-derived S1P mediates the chemoattraction of both monocytes and macrophages in vitro and in animal models, an effect that is dependent upon the S1P receptor S1PR1. However, S1P inhibited M1 macrophage-mediated phagocytosis of DLBCL tumor cells opsonized with the CD20 monoclonal antibodies rituximab and ofatumumab, an effect that could be reversed by an S1PR1 inhibitor. Our data show that S1P signaling can modulate macrophage recruitment and tumor cell killing by anti-CD20 monoclonal antibodies in DLBCL. The administration of S1PR1 inhibitors could enhance the phagocytosis of tumor cells and improve outcomes for patients.

Clinical Features of Non-Hodgkin Lymphoma-Associated Hemophagocytic Syndrome: a Retrospective Study.

This study aims to improve the understanding of lymphoma-associated hemophagocytic syndrome, and find effective methods to identify and manage this fatal disease. Patients diagnosed with non-Hodgkin lymphoma-associated hemophagocytic syndrome from January 2008 to December 2022 in our center were included. Univariate and multivariate analyses were also conducted using the Cox proportional hazards model. Among 26 patients, 22 patients were diagnosed with T/NK cell lymphoma, while 4 patients were diagnosed with diffuse large B cell lymphoma. A total of 16 patients died with a median follow-up of 71 (26, 236) days. Compared with B cell lymphoma-associated hemophagocytic syndrome patients, T/NK cell lymphoma patients are younger, have lower platelet count, fibrinogen concentration, and serum albumin, have higher blood β2-mi-croglobulin levels and ferritin, are more likely to be infected with Epstein-Barr virus, are more inclined have a simultaneously occurrence of lymphoma and hemophagocytic syndrome. In multivariate analysis, fibrinogen, albumin, cholinesterase, uric acid, triglyceride, and ferritin are significantly associated with overall mortality. LAHS is a rare disease with poor prognosis. Early anti-inflammatory treatment combined with anti-lymphoma therapy can improve the overall survival time of patients. Prospective multi-center studies with larger sample sizes and longer follow-up periods are needed to further investigate optimal treatment and prognosis.

Application of Blood Lymphocyte Immunophenotype and TCR Gene Rearrangement in the Diagnosis of CTCL.

The immunophenotype of peripheral blood lymphocytes and T-cell receptor (TCR) gene rearrangement of cutaneous T cell lymphoma (CTCL) patients were retrospectively analyzed to explore their value in the diagnosis of CTCL. A total of fifty patients' results were enrolled from 2013 to 2021, including 29 malignant skin disorders and 21 benign skin disorders. The immunophenotype of peripheral blood lymphocytes were analyzed by flow cytometry and TCR gene rearrangement was detected by capillary electrophoresis. Lymphocyte subsets, CD4/CD8 ratio, the percentage of CD3+CD4+CD7- cells and CD45RA/CD45RO ratio was calculated between malignant and benign skin disorders. Peripheral blood lymphocyte immunophenotype and TCR gene rearrangement was compared with skin biopsy to evaluate their sensitivity and specificity. Lymphocyte subsets between malignant and benign groups have no significant difference in percentage of T cell (p > 0.05). The CD4/CD8 ratio is higher in patients with malignant lymphoma than the healthy range. The percentage of CD3+CD4+CD7- cells in malignant groups is higher than that in benign groups and CD45RA/ CD45RO ratio has significant difference between malignant and benign groups (p < 0.05). The sensitivity and specificity of TCR rearrangement for CTCL were 51.7% and 42.9%. The sensitivity and specificity of peripheral blood lymphocyte immunophenotype for CTCL were 44.8% and 33.3%. Combining the two methods, the sensitivity and specificity reached 69.0% and 38.1%, respectively. CD4/CD8 ratio of lymphocyte subsets, the proportion of CD4+CD7-T cells and CD45RA/CD45RO ratio can effectively distinguish benign and malignant dermatosis. TCR rearrangement method combined with lymphocyte immunophenotype can improve the sensitivity and specificity of CTCL diagnosis.

Modified BEAM as a conditioning regimen for mantle cell lymphoma patients undergoing autologous hematopoietic stem cell transplantation

Modified BEAM as a conditioning regimen for mantle cell lymphoma patients undergoing autologous hematopoietic stem cell transplantation

Immunoglobulin High Throughput Sequencing (Ig-HTS) Minimal Residual Disease (MRD) Analysis is an Effective Surveillance Tool in Patients With Mantle Cell Lymphoma

IntroductionMantle cell lymphoma (MCL) accounts for 4% to 6% of B-cell non-Hodgkin lymphoma with historically poor outcomes. With the advent of intensive first-line, targeted, and cellular therapies, outcomes have improved, and initial remission can be 8 to 10 years or longer. As patients experience longer remissions, this raises the question of the optimal surveillance modality. Peripheral blood minimal residual disease (MRD) analysis offers a potential alternative to surveillance imaging that is sensitive, less costly, and eliminates the risk of radiation exposure. Materials and MethodsThe clonoSEQ assay (Adaptive Biotechnologies) is an FDA-cleared commercially available Ig-HTS MRD assay with a sensitivity of 1 cell in 1,000,000. We performed a retrospective analysis of 34 patients from 2015 to 2021, who underwent MRD testing after achieving remission with first-line therapy. ResultsWith a median follow-up of 6.5 years, 10-year progression free survival (PFS) was 60% and 10-year overall survival was 92% of the entire cohort. Among 12 patients who sustained a radiographic relapse, peripheral blood became MRD+ either at or prior to the time of relapse in 11 patients (92%). The first MRD+ test had a lead time of 0 to 24 months (median 34 days) prior to radiographic relapse. Only 1 patient had a MRD− result while being found to have progressive disease on imaging. Among 22 patients who sustained continuous clinical remission, 21 have remained MRD−. Several patients were able to enjoy 2 to 4-year intervals without surveillance imaging. ConclusionsOur data suggest that the clonoSEQ MRD assay is an effective surveillance tool for MCL patients following first-line therapy and is predictive of relapse prior to imaging.

Mesenchymal Stem Cells as a Promising Cellular Anti-COVID-19 Therapy for Diffuse Large B Cell Lymphoma Patients affected by Severe SARS-CoV-2

Mesenchymal Stem Cells as a Promising Cellular Anti-COVID-19 Therapy for Diffuse Large B Cell Lymphoma Patients affected by Severe SARS-CoV-2

Signal regulatory protein beta 2 is a novel positive regulator of innate anticancer immunity.

In recent years, the therapeutic (re)activation of innate anticancer immunity has gained prominence, with therapeutic blocking of the interaction of Signal Regulatory Protein (SIRP)-α with its ligand CD47 yielding complete responses in refractory and relapsed B cell lymphoma patients. SIRP-α has as crucial inhibitory role on phagocytes, with e.g., its aberrant activation enabling the escape of cancer cells from immune surveillance. SIRP-α belongs to a family of paired receptors comprised of not only immune-inhibitory, but also putative immune-stimulatory receptors. Here, we report that an as yet uninvestigated SIRP family member, SIRP-beta 2 (SIRP-ß2), is strongly expressed under normal physiological conditions in macrophages and granulocytes at protein level. Endogenous expression of SIRP-ß2 on granulocytes correlated with trogocytosis of cancer cells. Further, ectopic expression of SIRP-ß2 stimulated macrophage adhesion, differentiation and cancer cell phagocytosis as well as potentiated macrophage-mediated activation of T cell Receptor-specific T cell activation. SIRP-ß2 recruited the immune activating adaptor protein DAP12 to positively regulate innate immunity, with the charged lysine 202 of SIRP-ß2 being responsible for interaction with DAP12. Mutation of lysine 202 to leucine lead to a complete loss of the increased adhesion and phagocytosis. In conclusion, SIRP-ß2 is a novel positive regulator of innate anticancer immunity and a potential costimulatory target for innate immunotherapy.

HSD39 Treatment Pathway and Unmet Needs of Mantle Cell Lymphoma Patients: An Exploratory Study from Portugal

HSD39 Treatment Pathway and Unmet Needs of Mantle Cell Lymphoma Patients: An Exploratory Study from Portugal

Long-term analysis of the RiBVD phase II trial reveals the unfavorable impact of TP53 mutations and hypoalbuminemia in older adults with mantle cell lymphoma; for the LYSA group.

Between 2011 and 2012, a phase II trial evaluated the use of the RiBVD (rituximab, bendamustine, velcade and dexamethasone) combination as first-line treatment for mantle cell lymphoma (MCL) patients over the age of 65. We have now re-examined the classic prognostic factors, adding an assessment of TP53 mutation status. Patients (N=74; median age 73 years) were treated with the RiBVD combination. Median progression-free survival (mPFS) was 79 months and median overall survival (mOS) was 111 months. TP53 mutation status was available for 54/74 (73%) patients. TP53 mutations (TP53mt) were found in 12 patients (22.2%). In multivariate analysis, among the prognostic factors (PF) evaluated, only TP53mt and an albumin level (Alb) 3.6 g/dL were independently associated with a shorter mPFS. A hazard ratio (HR) of 3.16 (1.3-9.9, P=0.014) was obtained for TP53mt versus TP53 wild-type (wt), and 3.6 (1.39-9.5, P=0.009) for Alb <3.6 g/dL versus Alb ≥3.6 g/dL. In terms of mOS, multivariate analysis identified three PF: TP53mt (HR: 5.9 [1.77-19.5, P=0.004]), Alb <3.6 g/dL (HR: 5.2 [1.46- 18.5, P=0.011]), and ECOG=2 (HR: 3.7 [1.31-10.6, P=0.014]). Finally, a score combining TP53 status and Alb distinguished three populations based on the presence of 0, 1, or 2 PF. For these populations, mPFS was 7.8 years, 28 months, and 2.5 months, respectively. Our prolonged follow-up confirmed the efficacy of the RiBVD regimen, comparing it favorably to other regimens. TP53mt and hypoalbuminemia emerge as strong PF that can be easily integrated into prognostic scores for older adult patients with MCL.

Dissemination feature based on PET/CT is a risk factor for diffuse large B cell lymphoma patients outcome

Background18F-FDG PET/CT provides precise information about dissemination of lymphoma lesions. Dmax, defined as distance between the two lesions that were farthest apart by PET/CT, was found to be a promising predictor of Diffuse large B-cell lymphoma (DLBCL) outcome in a small size of clinical trial data. We analyzed the impact of Dmax on the outcome of a large real-world DLBCL cohort.MethodsData of newly diagnosed DLBCL at the Third Affiliated Hospital of Soochow University were retrospectively collected. Baseline Dmax, clinical data and survival information were recorded. A metabolic parameter, metabolic bulk volume (MBV), was also measured to verify the independent impact of Dmax.ResultsOptimal cut-off values for Dmax and MBV were 45.34 cm and 21.65 cm3. With a median follow-up of 32 months, Dmax significantly impacted progression-free survival (PFS) and overall survival (OS) in 253 DLBCL patients. For Dmaxlow and Dmaxhigh groups, estimated 3-year OS were 87.0% and 53.8% (p < 0.001), while 3-year PFS were 77.3% and 37.3% (p < 0.001). And for MBVlow and MBVhighgroups, 3-year OS were 84.5% and 58.8% (p < 0.001), and 3-year PFS were 68.7% and 50.4% (p = 0.003). Multivariate analysis identified Dmax and Eastern Cooperative Oncology Group performance status (ECOG PS) independently associated with PFS and OS, while MBV only independently associated with OS. A Dmax revised prognostic index (DRPI) combining Dmax and ECOG PS identified an ultra-risk DLBCL population with 3-year PFS of 31.7% and 3-year OS of 38.5%. The area under the curve (AUC) showed that this model performed better than International prognostic Index (IPI).ConclusionDmax is a new and promising indicator to investigate dissemination of lymphoma lesions associated with the outcome of DLBCL. It significantly contributes to stratification of patients with disparate outcomes.Trial registrationThis research has been retrospectively registered in the Ethics Committee institutional of the Third Affiliated Hospital of Soochow University, and the registration number was approval No. 155 (approved date: 31 May 2022).

Outcomes of CAR-T Cell Therapy for Large B Cell Lymphoma in Patients of 70 Years and Older: Multicentric Experience on Behalf of Geth-TC/Geltamo

Background: Median age at diagnosis of diffuse large B cell lymphoma (DLBCL) patients is 66 years; 40% of patients are diagnosed at an age greater than 70 years. CAR-T cell therapy is approved for the treatment of relapsed/refractory DLBCL patients; however, a deeper understanding of outcomes in patients older than 70 years is needed. Our aim was to analyze outcomes of CAR-T cell therapy in this population and compare them to those obtained in younger patients in the real-world setting. Methods: A subgroup analysis of our real life experience report (Kwon, Iacoboni et al. Haematologica 2022) was performed. Data from consecutive patients treated in Spain with commercial CAR-T products were retrospectively collected on behalf of GETH-TC (Spanish Group of Stem Transplantation and Cell Therapy)-GELTAMO (Spanish Group of Lymphoma and Autologous Stem Cell Transplantation). Patients infused between November-2018 and August-2021 were included. Last update of the cohort was performed in June 2023. Cytokine release syndrome (CRS) and Immune effector cell-associated neurotoxicity syndrome (ICANS) were graded using the ASTCT consensus criteria. Response was assessed according to the Lugano criteria. Results: Characteristics of the patients are summarized in Table 1. A total of 307 patients underwent apheresis for CAR-T cell therapy as 3 rd or subsequent line of therapy. Fifty-four (18%) patients were 70 years or older. There were no differences between groups regarding product selection, proportion of patients infused, production failure and rate of bridging therapy. There were more patients with HCT-CI score ≥3 and a trend of higher proportion of patients with ECOG &amp;gt;2 at apheresis in the older group (31% vs. 19%, p=0.047; 9% vs. 5%, p=0.054); the remaining baseline characteristics did not differ. A similar proportion of patients were infused. Among the infused population (n=261), median time from apheresis to infusion was 49 days for patients ≥70 years (n=45) and 47 for younger patients (n=216) (p=0.824). A similar proportion of patients developed any grade of CRS and ICANS (88% vs. 80%, p=0.132 and 31% vs. 29%, p=0.843) and grade 3-4 events (11% vs. 6.5%, p=0.277 and 18% vs. 10%, p=0.146). Median duration of CRS (4 and 5 days, p=0.150), ICANS (4 and 5 days, p=0.540) and hospitalization length (20 days for both groups, p=0.995) did not differ between groups. There were not significant differences in the proportion of patients admitted to ICU (24% vs. 18%, p=0.284); however, the proportion of patients developing infection in the first 6 months after infusion showed a trend to be higher in the older group (42% vs. 29%, p=0.099). Non-relapse mortality was similar between groups (13% vs. 7%, 9=0.24). With a median follow-up of 19.2 months from infusion, both 12-m PFS (30% vs. 38%, p=0.281) and 12-m OS (49% vs. 54%, p= 0.473) were similar between older and younger patients (Figure 1). In the multivariate analysis of the whole population including CAR-T type, disease characteristics (primary refractory, progressive disease, disease stage, R-IPI and high LDH at apheresis), ECOG ≥2 and age ≥70, high LDH (HR 2.1, p=0.001), disease stage III-IV (HR 1.8, p=0.038) and ECOG ≥2 (HR 2, p=0.025) were risk factors for EFS. Age ≥70 (HR 1.5, p=0.054) and progressive disease at lymphodepletion (HR 1.8, p=0.055) were nearly significant risk factors. In the multivariate analysis for OS, high LDH (HR 1.7, p=0.013), high R-IPI (1.2, p=0.039), ECOG ≥2 (HR 2, p=0.014) and progressive disease at lymphodepletion (HR 1.9, p0.042) were risk factors for OS, while age had no impact (HR 1.1, p=0.674). Conclusions: In our real-life experience, CAR-T cell therapy showed a similar efficacy and safety in younger and older patients (&amp;gt;70 years). Consequently, this latter group should receive CAR-T cell therapy if treatment criteria are met.

A Novel Patient-Derived Xenograft Model IO-FIVE for Drug Discovery and Precision Medicine in Lymphoma and Leukemia Patients

Lymphoma and leukemia have been the top two hematopoietic cancers in US, leading to more than 40,000 deaths each year. Although the 5-year relative survival rates have been improved up to 60%, there is still a lot of patients showed not response to standard therapy. Patient-Derived Xenograft model (PDX model), as the classical mouse xenograft tumor model that best represents the genetic information characteristics of human tumors, can be used to predict the therapeutic effect, and develop individualized treatment for patients. Its greatest advantage is that the model retains the microenvironment of the original tumor cells, inherits all the molecular biological properties of the primary tumor, and preserves the heterogeneity of the tumor. However, due to the factors that the loss of tumor heterogeneity in hematopoietic cancers after ex vivo clonal expansion and selection, and this in vitro assay based system could not fully reflect the host response to the drugs, including a range of novel immunotherapeutic agents such as PD-1 monoclonal antibody, CD38 monoclonal antibody, etc. With the increasing clinical use of oncology immunological agents, recently we are developing a new assay, named IO-FIVE (Immuno-Oncology drugs Fast In Vivo Efficacy test) for research and therapy, by retaining an appropriate proportion of tumor-infiltrating immune cells (and stromal cells) mixed with tumor cells when digesting clinical tumor samples or patient-derived xenografts-preserved tumor samples into cell suspensions, and then injecting them into the specialized IO-FIVE device, a modified microencapsulation and hollow fiber culture system (OncoVee® MiniPDX), which is implanted subcutaneously into mice and administered systematically for immuno-drug susceptibility testing to screen for immuno-drugs or combinations suitable for individual treatment. The IO-FIVE testing cycle takes only 14 days and is performed using Celltiter Glo before and after the injection of cell suspensions into the specialized device. Celltiter Glo, flow cytometry and Omics (RNA-seq, DNA-seq) are used to measure the viability of total cell subpopulations, the relative ratio of tumor cells to immune cells and the alteration of cellular transcription levels in the device, in order to further explore the sensitivity of tumor tissues to the immunotherapeutic drugs and the potential molecular mechanism for the responders and non-responders. Immune regulatory CD38 antibody Daratumumab has been tested in more than 40 acute myeloid leukemia (AML) cases, with its overall in vivo efficacy around 25%, which is consistent with previous research. It is interesting to uncover novel pathways for drug discovery and clinic treatment since CD38 is well expressed in most AML patients, but at least half of those CD38 + AML patients are not sensitive or resistant to Daratumumab monotherapy. Expression of CD38 on AML is not enough to distinguish the responder vs non-responder of Daratumumab. IO-FIVE could potentially be a companion diagnosis on AML for the patient stratification of Daratumumab. PD1 antibody Sintilimab has been tested in all the patients (12 lymphoma, 52 leukemia), and the only two laboratory PD1-Ab non-responsive T cell lymphoma patients have been showed to be clinically non-responders as well. Immunotherapy and chemical or targeted therapy combination could enhance tumor killing ability and disease control. We will use the IO-FIVE data to further compare it with its source of real-world clinical patients to obtain more evidence-based evidence that can predict individualized treatment with immunotherapy.

Survival Impact of the Second Malignancies Among the Lymphoid Neoplasm

Background: The development of the lymphoma treatment increased the number of patients with long-term survival. Due to the nature of lymphoma heterogeneous group of hematological malignancies, lymphoma treatment varies based on histopathological features. Various cytotoxic chemotherapy or radiotherapy could increase patients' risk of secondary cancer. Few studies evaluated the impact on survival of co-occurring cancers in mature lymphoid malignancies. Methods: 3953 adult patients were diagnosed with lymphoma between January 2000 and April 2021 at Yeoido and Seoul St. Mary's Hospital. The cohorts compared cohorts Standardized incidence ratio applying the Korea Central Cancer Registry (KCCR). Results: Among the patients Initially diagnosed with Mature-B cell neoplasm were 3001 patients (76.3%) and 711(18%) with mature T and NK cell neoplasm (5.6%) for Hodgkin lymphoma. Among the B cell neoplasm, 57% diffuse large B cell lymphomas (DLBCL) were the most common form of B cell type, followed by follicular lymphoma (FL) and marginal zone lymphoma. PTCL NOS was the most dominant in the T cell lymphoma, accounting for 26%, followed by Extranodal NK/T cell lymphoma 23% and angioimmunoblastic T cell lymphoma. Among B cell lymphoma, 10% of solid cancer co-occurred, and 1% of hematological can be combined; in T cell types, 6% were solid ca and 12% were hematological ca. Two hundred one (4.8%) patients had historic or newly diagnosed solid cancer cases; the median diagnosed age was 64 (27-92) years. The median time to secondary cancer after diagnosis is 2 years, most common type of secondary were hung cancer, followed by thyroid and colorectal cancer. Comparing with the Korean register, the standardized Incidence Ratio of patients with lymphoma showed the highest in thyroid ca (hazard ratio (HR) = 2.65, [95% CI 1.45-4.45]), followed by lung (HR=2.14), stomach (HR=1.79), and colon (HR=1.59), In total lymphoma patients, secondary ca. doesn't offer a poor prognostic factor in total cohorts- In the case of MZL and FL (n=737), development of secondary cancer (n=131) acts as a poor prognostic factor 5 years in than count parts (n=565), 79.4 vs. 89.1%, respectively ( p &amp;lt;0.001). Multivariate analysis showed colorectal ca ( p &amp;lt;0.001, HR = 4.83, [95% CI 5.19-1103]) and transformation to DLBCL ( p &amp;lt;0.001, HR = 4.85, [95% CI 2.68-5.79]) affect worse outcomes on OS. In the case of CLL, multivariate analysis, treatment-related AML or MDS affects inferior survival outcomes ( p =0.02, HR = 3.99, [95% CI 1.25-12.79]). Conclusions: Secondary cancers could increase mortality in low-grade B cell lymphoma patients. There were different types of secondary cancer affected overall survival by each subtype. In the low-grade lymphoma, secondary hematologic malignancies or Richter syndrome poorer patient results. This study suggested awareness of various combined malignancies by subtypes of patients with lymphoma.

A Retrospective Analysis on the Efficacy and Saftey of Obinutuzumab in the Treatment of Non-Hodgkin B Cell Lymphoma

Backgroud: Obinutuzumab was approved by China's Drug Administration in June 2021, and there have been few real-world reports about efficacy and safety. Objective:Analysis the efficacy and safety of Obinutuzumab in the treatment of non-Hodgkin B cell lymphoma in our center. Methods ：We analysed the non-Hodgkin B cell lymphoma patients who received Obinutuzumab combined with chemotherapy or maintenance treatment during September 2021 and July 2023, median follow up 9 months（1-20）, to evaluate efficacy and safety. Results:In all 165 patients, male 88（53%), female 77（47%）, median 56 (26-83) years. FL 80 (48%), MZL 22 (13%), CLL/SLL 13 (8%), DLBCL 42 (25%), others 8 (5%). Newly diagnosed patients 114 (69%), relapsed/refractory 51 (31%). 124 patients received Obinutuzumab combined therapy, and 42 (25%) patients received Obinutuzumab maintenance treatment. Median treatment 4 cycles（1-11）. I/II 33 (20 %), III/IV 131 (79 %). Low-medium group 91（55%）, medium-high group 74（45%）. ORR 66.7%, CRR 40.6%. 3 patients died of progression of disease. 1 elder patient died of pneumonia. Further subgroup analysis revealed, ORR（78% vs 22%, p&amp;lt;0.001）and CRR（81% vs 19%, p=0.02）of newly diagnosed patients were better than relapsed/refractory patients. There was no difference in CRR among different stages patients（I/II vs III/IV, 24% vs 76%, p=0.424）, ORR（58% vs 42%, p=0.373）and CRR (63% vs 37%, p=0.481）between low-medium and medium-high patients. The patients received more than 4 cycles treatment with better ORR and CRR. The intensive treatment of Obinutuzumab in first cycle could improve ORR (87% vs 13%, p=0.036,）and decreased IgM ( p=0.0016）, but didn't not affect CRR （87% vs13%, p=0.343）, IgG ( p=0.13） and IgA ( p=0.41）. Maintenance treatment decreased IgM ( p=0.00064）but not for IgG or IgA. In treatment related side effects, any adverse events were 42（25%）, more than grade III were 18（11%）, tumor lysis syndrome 11 (7%), only one patient need blood purification treatment and recovered. Liver injury 8 (5%) . All the infusion-related response 23（14%）were grade I-II. Only 2 patients delayed treatment because of the side effects. Conclusion:Obinutuzumab is safe and effective in the treatment of non-Hodgkin B cell lymphoma, even for the III/IV or medium-high group patients will have similar results with I/II or low-medium patients. Further follow-up is still needed to determine the long-time efficacy and safety. Key words:retrospective analysis,Obinutuzumab, non-Hodgkin lymphoma.

Personal and Shared Tumor-Antigen Prioritization in Diffuse Large B Cell Lymphoma Patients Undergoing CD19 CAR T Cell Treatment

Backgroun d: Durable response to Axi-cel CD19 CAR T cell therapy is associated with activation and clonotypic expansion of the host T cell repertoire (Cheloni et al Blood, ASH, 2022). The characterization of the tumor microenvironment (TME) and the identification of tumor antigens associated with therapeutic efficacy is a critical area of investigation. Aim: We sought to determine the association between the host immune activation in response to tumor-associated and tumor-specific antigens and the clinical response to CAR-T cell treatment in diffuse large B cell lymphoma patients. Methods: RNAseq was performed on FFPE lymphoma samples collected before CAR T cell infusion (N=16). Patients were divided into 3 distinct cohorts: stable progressive disease at 1 month post-CAR T (non-responders, n=2), disease relapse within 6 months post-CAR T (relapsed, n=3), and on-going response lasting greater than 1 year (long-term responders, n=11). For neoantigens, mutations were retrieved following the GATK best practices for RNAseq data and annotated with Variant Effect Predictor (VEP). Tumor-specific antigens and tumor-associated antigens were detected by prioritizing 9-11mer amino acid sequences, preferentially presented by class I molecules, observed solely or primarily in tumor samples, respectively, as compared to a pool of healthy controls derived from 250 RNAseq samples, corresponding to 29 tissues (GTEx Portal). Antigen prioritization was performed following weighted scoring considering RNA expression, translation potential, peptide-MHC binding affinity, predicted immunogenicity, physicochemical properties, and clonality, in the case of neoantigens. Using deconvolution methods and canonical marker expression, we characterized the major immune cell types within the TME. Results: Higher cell proportions of lymphocytes and innate immune cells, including NK, and activated dendritic cells were observed in long-term responders, suggesting that a higher infiltration of these cell types in the tumor bed is associated with a better response to CAR T cell therapy. Notably, a positive correlation between the mutational burden and the a/b clonotypic expansion of peripheral blood cytotoxic T cell populations, detected from single-cell immunoprofiling (Cheloni et al Blood, ASH, 2022) was observed in eight long-term responders, consistent with prior studies highlighting the significance of neoantigens in eliciting robust and durable immune response against cancer cells. A high occurrence of neoantigens originating from the Titin ( TTN) gene in two long-term responders with the highest mutational burden was observed, and from ACTB and ACTG1, mutated genes associated with lymphoid cancers. A substantial enrichment of shared tumor antigens originating from retroelements was observed in long-term responders, suggesting the presence of common epitopes that drive an antigen-specific response. Conclusion: These findings shed light on the potential immunogenicity of specific neoantigens and shared antigens derived from repetitive elements, which could serve as promising targets for immunotherapeutic strategies in lymphoma patients. Understanding the tumor immune landscape and antigenic features associated with long-term response to CAR T cell therapy may pave the way for the development of personalized and effective combinatorial treatments for lymphoid cancers.

Long Term Durable Responses in Relapsed/Refractory (r/r) ALL, DLBCL, and FL Patients Treated with Tisagenlecleucel and Its Association with Persistence of CAR T-Cells

Introduction: Monitoring of chimeric antigen receptor (CAR) transgene levels in peripheral blood post tisagenlecleucel (tisa-cel) infusion provides information on expansion and persistence of CAR T-cells. In adult diffuse large B cell lymphoma (DLBCL) and follicular lymphoma (FL) patients (pts), the expansion of CAR T-cells was similar between responding and non-responding pts, while in pediatric and young adult acute lymphoblastic leukemia (ALL) pts, higher expansion in responding pts was observed. Based on long term follow up data from tisa-cel treated pts, analyses were performed to delineate the impact of CAR persistence and B cell aplasia on duration of remission (DOR). Methods: Transgene levels in blood measured by quantitative polymerase chain reaction were available from pivotal phase II studies in pts with r/r ALL (ELIANA [N=79], ENSIGN [N=64], NCT03123939 [N=69], NCT01626495 [N=60]), r/r DLBCL (JULIET [N=115]), and r/r FL (ELARA [N=97]), along with the long term follow up study (NCT02445222). The time related to loss of persistence of CAR T-cells (T loss) was defined as the time at which transgene levels first dropped below 50 transgene copies/µg DNA (approximates to lower limit of quantification (LLOQ) of the assay) after maximal expansion. Kaplan Meier analysis was performed to investigate the impact of duration of T loss and time corresponding to last quantifiable level (T last) on DOR/relapse. The impact of time to B cell recovery (&amp;gt;1% CD19+ B cells/WBCs or &amp;gt;3% CD19+ B cells/lymphocytes for ALL, and 80-616 cells/µL for DLBCL and FL), on DOR/relapse was also investigated. Results: Long term CAR persistence in tisa-cel treated pts has been observed for up to 9, 6, and 2.5 years for ALL, DLBCL, and FL pts, respectively, reflecting differing length of follow up based on initiation of trials in respective indications. Across indications, pts who lost transgene within 6 months or between 6-12 months had shorter DOR relative to pts with persistent transgene (Fig 1, ALL pts). In ALL, the median T loss was 27.4, 18.2, 9.7, and 18.0 months for ongoing complete response (CR) &amp;gt; 12 months, CR pts between 6-12 months, relapsed pts &amp;lt; 12 months, and relapsed pts &amp;gt; 12 months, respectively. However, of the pts who lost transgene within first 6 months, some pts maintained durable responses for ≥12 months (29%, 15%, and 50% in ALL, DLBCL, and FL, respectively). Among ALL relapsed pts, majority (17/26, 65%) of the pts with T loss &amp;lt; 6 months showed B cell recovery, however 6 pts had T loss at &amp;lt; 6 months as well as B-cell recovery but maintained response for ≥ 12 months. In ALL, Pulsipher et al.,(Blood Cancer Discov. 2022)have shown that NGS MRD within the first 6 months to 1 year post infusion may be a more reliable predictor of potential relapse than B cell recovery. The median time to B-cell recovery was 266 days in pts who relapsed/censored within 12 months but was not reached for pts with ongoing response at 12 months. On the contrary, B cell recovery seems to have no association with relapse in DLBCL or FL pts. Of the ALL pts with CAR persistence, pts with &amp;lt;50% morphologic blasts at any time prior to infusion of tisa-cel had longer DOR compared to pts with ≥50% blasts. This may also reflect a more resistant high risk ALL at the time of study entry or greater potential for stochastic loss of CD19 in pts with the higher disease burden. Conclusions: Long term sustained remission/response was observed in tisa-cel treated pts in the pivotal trials. The analyses demonstrate a positive association between CAR persistence and durable clinical responses across indications, however, some pts maintained durable responses despite early loss of transgene and/or early B cell recovery (&amp;lt; 6 months post infusion; Myers et al., J Clin Oncol. 2021). To note, in DLBCL and FL, the transgene levels in blood may not represent the levels at target sites including lymph nodes, moreover, T last and T loss are dependent on the duration of follow up, LLOQ etc. Of note, these analyses represent data from pivotal tisa-cel clinical studies with a heavily treated pt population post multiple lines of therapy including prior HSCT, and may not represent pts treated with commercial product, at earlier times post diagnosis in current studies with fewer lines of prior treatment, lower pre-infusion disease burden, and/or other CAR product. In pts with early loss of transgene, further research is needed to identify potential factors or patient characteristics resulting in durable responses.

Acalabrutinib with Rituximab As First-Line Therapy for Older Patients with Mantle Cell Lymphoma - a Phase II Clinical Trial

Introduction - Previously, our group reported results from ibrutinib with rituximab in older mantle cell lymphoma (MCL) patients (pts) with an atrial fibrillation rate of 22%. To develop a chemotherapy-free combination for elderly MCL pts, we designed this phase II clinical trial to investigate the efficacy and safety of acalabrutinib with rituximab (AR) as a first line approach in MCL pts (≥ 65 yrs). Methods - We enrolled 50 previously untreated pts in this single institution, single arm, phase II clinical trial - NCT05214183. Pts received acalabrutinib 100 mg orally twice a day and intravenous rituximab, weekly for first 4 weeks, followed by once a month for 12 months and subsequently once every 2 months totaling 24 months. Acalabrutinib was continued. The primary objective was to assess best overall response rate (ORR) rate after AR. Responses were assessed as per Lugano response criteria. Clonoseq based minimal residual disease (MRD) assessment was performed. Adverse events were coded as per CTCAE version 5. Genomic studies are ongoing. Results - Among 50 pts, the median age was 69 years (range: 65-81). Male/female were 36 and 14 respectively. Bone marrow was positive for MCL in 45/50 (90%) and GI tract in 35/47 (74%) pts. Forty-six pts had classic morphology while three were blastoid and one was pleomorphic. Ki-67% was available in 42 pts - 13/42 (31%) had high Ki-67% and 29/42 (69%) had a low Ki-67% (&amp;lt; 30%). Simplified MIPI risk stratification included: low (n=4), intermediate (n=35) and high risk (n=11). TP53 aberration status (mutations or deletion) was available in 43/50 pts and 12 pts had aberrant TP53 while 31/43 did not have aberrant TP53. Thirty-nine pts had baseline next generation sequencing (NGS) on tumor tissue biopsy and 25/39 (64%) had ≥ 3 mutations. Median number of AR cycles was 12 (range: 3-24). One pt was not evaluable for response at 12 weeks. Overall, the best response was 94% ORR and 90% CR, 4% PR; 6% were non-responders. Median number of cycles to get complete metabolic response was 3 (range 2-7). Fourteen pts were MRD negative at last follow-up of the 28 evaluable pts. With a median follow up of 17 months, the median PFS and OS were not reached (2 year 92% and 96% respectively). The median PFS and OS was not reached and not significantly different in pts with high and low Ki-67% or with/without TP53 aberrations or among pts with low, medium or high-risk categories. Nine pts (18%) came off study - 3 for adverse events (syncope, atrial fibrillation, intolerance), 3 for disease progression, one melanoma recurrence, one pt choice and one with an unusual monocytosis. Overall, 2 pts died (1 on trial with primary progression). The most common all-grade toxicities were fatigue (82%), myalgia (64%), headache (38%), bruising (28%) and &amp;lt;1% were grade 3 or higher. Among 50 pts, 1 pt had recurrence of grade 2 atrial fibrillation (2%) and one pt had recurrence of grade 3 unstable angina. Conclusions -Chemotherapy-free frontline therapy with AR is highly effective and safe and induced early CR in older pts with MCL.

Resistance Mechanisms Impacting Bispecific Antibody (BsAbs) and Chimeric Antigen Receptor (CAR) T-Cell Therapy Outcomes in Large B Cell Lymphoma (LBCL) Patients

Introduction: BsAbs and CAR T-cell therapy have revolutionized the treatment landscape of patients with relapsed/refractory (R/R) LBCL. Regrettably, less than 40% of patients will attain durable responses, and determinants of response remain unclear. Our aim was to identify tumoral characteristics that can predict resistance to CAR T-cell therapy and BsAbs in LBCL patients. Methods: We retrospectively collected clinical data and obtained pre-treatment formalin-fixed and paraffin-embedded specimens prior CAR T-cell and BsAb treatment from 56 LBCL patients from 4 centers. Targeted NGS was performed to identify variants and copy number variations (CNVs) across a custom gene panel of 200 genes. These include lymphoma related genes, and genes involved in tumor antigen presentation, B and T lymphocyte interaction, and cell death pathway. Variables assessing response where Overall Response Rate (ORR), Complete Response Rate (CRR), Progression-Free Survival (PFS), and Overall Survival (OS). To ascertain each gene's influence on these endpoints, a logistic model was used for ORR and CRR comparisons, while Kaplan-Meier and Cox methods were employed to analyze PFS and OS. Results: A total of 56 patients were included in the study, 36 patients treated with CAR T-cell and 36 with BsAbs (16 patients received both). Samples were obtained for 31 patients prior to CAR T-cell therapy (CAR T-cell cohort) and 34 cases prior to BsAbs (BsAbs cohort). Baseline characteristics of the patient population are shown in Table 1. In summary, median age was 62 years, 57% were male, 42% had a previous indolent lymphoma and median previous lines were 2 (range 1-4). Best response included CRR in 59%, with a median PFS and OS of 13.6 (CI95% 5.98 - Not reached [NR]) and 29.5 (CI95% 15.7 - NR) months, respectively. Median follow-up for the full cohort was 18.7 months (CI95% 15.1 - 24). As per the CAR T-cell cohort, ORR and CRR were 81% and 68%, respectively. Median PFS and OS for CAR-T-cell cohort was 29.4 months (CI95% 5.98 - NR) and NR, respectively. Regarding the BsAb cohort, ORR and CRR were 66% and 50%, respectively. Median PFS and OS was 10 (CI95% 5.16 - NR) and 22.5 (CI95% 14.4 - NR) months, respectively. For the full cohort, the most frequently altered genes, considering both mutations and CNVs, were KMT2D (66%), TP53 (60%), CREBBP (58%), IGLL5 (51%), REL (42%), EZH2 (38%), BCL2 (38%), TNFRSF14 (37%), SGK1 (32%) and KLHL6 (32%) (Figure 1). The most frequent CNVs were gains of 2p16.1 (34%), 3q28 (15%), 6p(14%), 11q (12%), 21q (12%), 3q (10%); and losses of 17p (32%), 6q (26%), 1p36.32 (25%), 13q14.2 (18%), 19p13.3 (18%), 4q35.1(16%), (10q23.31 (12%), 9p21.3 (10%). Focusing on the CAR T-cell cohort (n=31), mutations and/or deletions in EP300 (7%, n=2)( p=0.01) and KLHL6 (n=2, 7%) ( p=0.04) were associated with an inferior PFS. In addition, mutations and/or deletions in KLHL6 ( p=0.02) and EP300 (n=3, 10%) ( p=0.05), and amplifications in MYC (n=3,10%) ( p=0.031) were associated with shorter OS. No differences in PFS and OS were oberved in patients with mutations and/or deletions in TP53 (n=19, 63%). Regarding RHOA patients with mutations or deletions (n=2, 7%) did not present an inferior PFS ( p=0.88) or OS ( p=0.52). The analyzed alterations did not show an impact on response rate. In terms of patients who received BsAbs (n=34), we observed that mutations and/or deletions in TP53 (n=19, 58%) ( p=0.04) , RHOA (n=5, 15%) ( p=0.05) and GNAI2 (n=5,15%) ( p=0.02)were associated with aninferior PFS. Mutations and/or deletions in TP53 (p=0.04) , RHOA (p=0.009) , GNAI2 ( p=0.01) and CD274(PDL1)-PDCD1LG2(PDL2) (12%, n=3) ( p=0.01) were associated with an inferior OS. Regarding MYC, 8 cases were mutated (n=2, 6%), amplified (n=5, 15%) or both (n=1, 3%), but these alterations were not associated with an inferior PFS (p=0.46) or OS ( p=0.27). Again, no impact on response rate was observed for patients carrying alterations. Conclusions: In our study, a pattern of genetic alterations predicting worse OS was observed in patients treated with CAR T-cells ( KLHL6 EP300 and MYC) and BsAbs (TP53, RHOA, PDL1, PDL2, GNAI2). This important information will aid in the development of targeted strategies to overcome resistance and improve treatment outcomes.

Evaluation of the T-Cell Metabolic State of Starting Material and Manufactured CAR T Cell Products and Clinical Outcome of CAR T-Cell Therapy for Large B Cell Lymphoma

Background Although chimeric antigen receptor (CAR)-T cells are a viable treatment option for patients with relapsed or refractory (r/r) aggressive B-cell lymphomas, more than half of patients experience relapse within the first two years after product infusion. Successful autologous cell-based anti-cancer therapies require persistence of effector cells, features that highly depend on complex metabolic processes. We earlier demonstrated in chronic lymphocytic leukemia, that expansion and persistence of CD19-CAR T-cell infusion in vivo was correlated with increased mitochondrial mass and enhanced biogenesis(van Bruggen et al, Blood 2019). Metabolic features might therefore serve as early biomarker tools to predict efficacy and outcome of CAR T-cell therapy and might allow therapeutic targeting but did was never studied in a prospective clinical trial. Aims Perform extensive phenotyping and state-of-the-art metabolic state analyses of T cells prior and post CAR T cell production and investigate correlations with clinical outcomes and patient characteristics. Methods Samples were obtained from large B cell lymphoma (LBCL) patients that were part of the TRANSCEND-NHL-001 trial (NCT02631044, Abramson et al, Lancet 2020).Thirty-two patients with r/r B-cell lymphoma who received CAR-T cell treatment lisocabtagene maraleucel (16 responders (R) and 16 non-responders (NR)) were included in our analyses and (metabolically) analyzed by flow cytometry and extracellular flux analysis. Metabolic phenotyping was performed on 53 samples at baseline and 44 samples after an in vitro stimulation. Extracellular flux analysis was done on 14 DP samples. The cohort was split evenly between Responders and Non-responders, and balanced for a score calculated taking key clinical baseline characteristics that were found to be associated with response into account (NCT02631044, Abramson et al, Lancet 2020). Both T cells obtained at time of leukopheresis (starting material (SMAT)) and CAR-T drug products (DP), (following transduction of a CD19-targeted CAR T cell product with a defined CD4 and CD8 composition bearing a 4-1BB costimulatory domain and expansion) were analyzed directly after thawing and after stimulation with anti-CAR-antibody in case of CDP, or αCD3/αCD28 antibodies for SMAT samples. Patients consented to these studies and researchers were blinded for clinical outcomes at time of analysis. Results We found that the baseline clinical factors that associate with clinical response are related to specific metabolic cellular phenotypes, both in SMAT and DP, and with and without T cell stimulation. SMAT samples from patients with higher baseline scores show signs of an increased exhausted phenotype with regards to cellular ROS and exhaustion marker expression, whilst a more plentiful memory-like phenotype was seen in patients with a lower baseline score. When clustering by clinical response status, analysis of glucose and fatty-acid uptake, neutral lipid content, cellular ROS, mitochondrial mass and potential and flux data showed that responders, in both SMAT and DP samples, have an all-round increased metabolic activity upon αCD3/αCD28 or CAR T-receptor triggering, respectively, when compared to non-responders (Figure 1A). Furthermore, responding patients show enrichment of CD27 expression in the SMAT material, indicating a more abundant memory phenotype. Summary/Conclusions This is the first evidence of an association between a specific immune-metabolic phenotype of (CAR-) T cells and outcome of CAR-T cell therapy in patients with relapsed/refractory LBCL. More specifically, a higher abundance of memory phenotypes, metabolic activity and mitochondrial biogenesis of both harvested T cells and CAR-T product associates with improved clinical response. Combining metabolic, phenotypical and clinical characteristics and biomarkers of patient T cells (both pre- or post-production) shows the importance of the metabolic phenotype and fitness of T cell starting material, underlying the importance of early line CAR T therapy following diagnosis.