Treatment Of non-Hodgkin Lymphoma Research Articles

Recent Advances in Nanobiotechnology for the Treatment of Non-Hodgkin's Lymphoma.

Lymphoma is the eighth most common type of cancer worldwide. Currently, lymphoma is mainly classified into two main groups: Hodgkin's lymphoma (HL) and non-Hodgkin's lymphoma (NHL), with NHL accounting for 80% to 90% of the cases. NHL is primarily divided into B, T, and natural killer (NK) cell lymphoma. Nanotechnology is developing rapidly and has made significant contributions to the field of medicine. This review summarizes the advancements of nanobiotechnology in recent years and its applications in the treatment of NHL, especially in diffuse large B cell lymphoma (DLBCL), primary central nervous system lymphoma (PCNSL), and follicular lymphoma (FL). The technologies discussed include clinical imaging, targeted drug delivery, photodynamic therapy (PDT), and thermodynamic therapy (TDT) for lymphoma. This review aims to provide a better understanding of the use of nanotechnology in the treatment of non-Hodgkin's lymphoma.

E054 Rituximab dose minimisation in rheumatoid arthritis, a retrospective outcome analysis

Abstract Background/Aims Rituximab was originally developed as a non-Hodgkin lymphoma treatment and rheumatology adopted the same dosing for rheumatoid arthritis(RA); two 1000mg doses two weeks apart for each maintenance cycle. UHSussex utilises self-referral for maintenance rituximab dosing once disease control lessens. Evidence demonstrates patients treated with lower dose rituximab still enjoy good disease control with less immunosuppression, reduced hospital attendance and reduction in drug expenditure. Following one two-dose cycle, stable RA patients were prescribed a single 1000mg dose from 2020 onwards. A retrospective outcome analysis was completed in August 2022; however, results were highly impacted by Covid-19 and a repeat analysis was required. Aim: to assess impact of a single 1000mg dose rituximab regime on patient sustained outcomes, dose intervals, Disease Activity Scores (DAS) and drug expenditure. Methods The pharmacy database was used to identify RA patients receiving rituximab from March 2020 to August 2023. Patients received at least one standard two-dose cycle before switching to a single infusion. Patients were excluded if lost to follow up (under care of rheumatology for less than a year following dose switching) or considered clinically unstable. Patient letters and departmental records were accessed for disease scores and treatment intervals. Results Table1: Results from the two analysis periods From the original cohort (31/59)53% remained on single dosing. Switch back tended to occur within 18 months of switching. DAS scores were not collected reliably and were omitted. Conclusion Covid-19 impacted services and attitudes during the initial study period. Many patients stopped treatment, particularly rituximab, due to the prolonged duration of effect, excluding them from analysis. The dose-interval increase during the first analysis was considered a result of the pandemic. Patients and clinicians delayed infusions; minimising immunosuppression due to perceived increased risk of Covid-19. The continued interval increase in the second analysis may identify a patient cohort in deep remission eligible for therapy review. The study highlights poor practice in collecting and recording DAS scores and appropriate patient-clinician interaction opportunities. 53% of patients remained on single infusions in August 2023 resulting in resource savings, reduced hospital attendance, immunosuppression and significant cost savings demonstrating the rationale for this option. Disclosure S. Butler: Honoraria; Abbvie, Janssen. H. Smith: Honoraria; Novartis, Abbvie.

Establishment of cluster of differentiation 20 immobilized cell membrane chromatography for the screening of active antitumor components in traditional Chinese medicine

Non-Hodgkin lymphoma (NHL) is a heterogeneous group of malignant tumors occurring in B or T lymphocytes, and no small molecule-positive drugs to treat NHL have been marketed. Cluster of differentiation 20 (CD20) is an important molecule regulating signaling for the life and differentiation of B lymphocytes and possesses the characteristics of a drug target for treating NHL. 2-Methoxyestradiol induces apoptosis in lymphoma Raji cells and CD20 protein is highly expressed by Raji lymphoma cells. Therefore, in this study, a CD20-SNAP-tag/CMC model was developed to validate the interaction of 2-methoxyestradiol with CD20. 2-Methoxyestradiol was used as a small molecule control compound, and the system was validated for good applicability. The cell membrane chromatography model was combined with high-performance liquid chromatography ion trap time-of-flight mass spectroscopy (HPLC-IT-TOF-MS) in a two-dimensional system to successfully identify, analyze, and characterize the potential active compounds of Schisandra chinensis (Turcz.) Baill. extract and Lysionotus pauciflorus Maxim. extract, including Schisandrin A, Schizandrol A, Schizandrol B, Schisantherin B, and Nevadensin, which can act on CD20 receptors. The five potential active compounds were analyzed by non-linear chromatography. The thermodynamic and kinetic parameters of their interaction with CD20 were also analyzed, and the mode of interaction was simulated by molecular docking. Their inhibitory effects on lymphoma cell growth were assessed using a Cell Counting Kit-8 (CCK-8). Nevadensin and Schizandrin A were able to induce apoptosis in Raji cells within a certain concentration range. In conclusion, the present experiments provide some bases for improving NHL treatment and developing small molecule lead compounds targeting CD20 with low toxicity and high specificity.

Abstract 1865: HDP-102 - a CD37-targeting Amanitin-based-ADC for the treatment of NHL - non-clinical data package

Abstract Background: Antibody-drug conjugates (ADC) are gaining importance as anti-cancer therapy. Most ADCs are based on cytotoxic warheads targeting only proliferating cells. In contrast, HDP´s proprietary ATAC platform utilizes amatoxins as payload. Amatoxins are specific inhibitors of eukaryotic RNA polymerase II, suppressing a key function in the protein metabolism not limiting their cytotoxicity to proliferating cells. Furthermore, there are no known resistance mechanisms against amatoxins in mammalian cells (i.e. multi drug resistance transporters), making ATACs a promising new class of anti-cancer ADCs. Here we present pre-clinical data on the anti-CD37 ATAC HDP-102. CD37 is a transmembrane protein expressed exclusively on immune cells, mainly mature B-cells, and in many B-cell malignancies, including Non-Hodgkin’s Lymphoma (NHL). Material and Methods: HDP-102: anti-CD37 ADC based on cysteine-reactive, site-specific amatoxin-linker constructs produced by HDP Cytotoxicity assays: CellTiter Glo 2.0 (Promega) assays on CD37pos cell lines MEC-2, Raji-luc, Ramos and CD37neg Nalm-6 cells Efficacy studies: Disseminated MEC-2 and Raji-Luc mouse xenograft tumor models in single dose experiments Tolerability study in Cynomolgus monkeys: i.v. HDP-102 treatment on days 1 and 22; blood sampling for PK analysis; necropsy and histopathology on days 28 and 64 Results: HDP-102 showed full-blown cytotoxicity with EC50 values in the low nanomolar range in all CD37pos cell lines in vitro but no cytotoxicity on CD37neg cells. In vivo, the anti-tumor efficacy of HDP-102 was evaluated in disseminated CDX models of NHL. A single dose of 0.5 mg/kg HDP-102 already led to complete tumor remission in all animals in a Raji-luc model and 60% survival on day 100 in a MEC-2 model, demonstrating the high potency of HDP-102 in NHL models. In a tolerability study in Cynomolgus monkeys, i.v. administration of HDP-102 was well tolerated up to 2.5 mg/kg without clinical signs of toxicity. Microscopically, decreased cellularity in lymphoid tissue was the main finding, caused by the targeted effect of HDP-102 on CD37pos cells. The HNSTD of 2.5 mg/kg together with the potent anti-tumor efficacy of HDP-102 leads to a therapeutic window of ~200 for HDP-102. Conclusion: CD37 is an ideal target for ADCs due to its high prevalence in B-cell malignancies and limited expression on healthy cells. We have demonstrated that the anti-CD37 ATAC HDP-102 is a promising drug candidate for the treatment of NHL. HDP-102 showed excellent anti-tumor efficacy in CDX models even after single doses. Pharmacological activity of HDP-102 was also seen in Cynomolgus monkeys as evidenced by depletion of B-cells in lymphoid organs. Furthermore, HDP-102 was well tolerated in monkeys. Taken together, based on pre-clinical data, HDP-102 is a promising new treatment option for NHL patients combining a good safety profile with strong anti-tumor efficacy. Citation Format: Sarah-Jane Neuberth, Kristin Decker, Christian Orlik, Irina Dranova, Anikó Pálfi, Torsten Hechler, Andreas Pahl, Michael Kulke. HDP-102 - a CD37-targeting Amanitin-based-ADC for the treatment of NHL - non-clinical data package [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1865.

The Non-Hodgkin Lymphoma Treatment and Side Effects: A Systematic Review and Meta-Analysis.

This paper aims to review studies regarding side effects found during Non-Hodgkin Lymphoma treatment, to suggest the drug class most associated with these effects, as well as the most prevalent side effect grade. This review is registered in PROSPERO (IDCRD42022295774) and followed the PICOS strategy and PRISMA guidelines. The search was carried out in the databases PubMed/MEDLINE, Scientific Electronic Library Online, and DOAJ. Medical Subject Headings Terms were used and quantitative studies with conclusive results regarding side effects during the non-Hodgkin lymphoma treatment were selected. Patent information was obtained from google patents. Monoclonal antibodies were the main drug class associated with side effects during NHL therapy. The combination of Rituximab (Rituxan®; patent EP1616572B) and iInotuzumab (Besponsa®; patent EP1504035B3) was associated with a higher incidence of thrombocytopenia (p<0.05), while the combination of Rituximab and Venetoclax (Venclexta®; patent CN107089981A) was associated with a higher incidence of neutropenia (p<0.05) when compared to Bendamustine combinations (Treanda ™; patent US20130253025A1). Meta-analysis revealed a high prevalence of grade 3-4 neutropenia and thrombocytopenia in men. Finally, Americans and Canadians experienced a higher prevalence of these side effects, when compared to others nationalities (p<0.05). Patents regarding the use of monoclonal antibodies in NHL treatment were published in the last year. Monoclonal antibodies associated with neutropenia (grade 3-4) and thrombocytopenia, especially in North American men treated for NHL, and with an average age of 62 years demonstrated importance in this study.

Phase I Study of Duvelisib Study for Cytokine Release Syndrome Prophylaxis in Patients Receiving Chimeric Antigen Receptor T Cell Treatment for Non-Hodgkin Lymphoma

Phase I Study of Duvelisib Study for Cytokine Release Syndrome Prophylaxis in Patients Receiving Chimeric Antigen Receptor T Cell Treatment for Non-Hodgkin Lymphoma

Impact of Modern Low Dose Involved Site Radiation Therapy on Normal Tissue Toxicity in Cervicothoracic Non-Hodgkin Lymphomas: A Biophysical Study.

This biophysical study aimed to determine fitting parameters for the Lyman-Kutcher-Burman (LKB) dose-response model for normal tissue complication probability (NTCP) calculations of acute side effects and to investigate the impact of reduced radiation doses on the probability of their occurrence in supradiaphragmatic non-Hodgkin lymphoma (NHL) irradiation. A cohort of 114 patients with NHL in the cervicothoracic region, treated between 2015 and 2021 at the University Hospitals of Münster, Hamburg, and Essen, with involved site radiation therapy (ISRT), were included. Among them, 68 patients with aggressive NHL (a-NHL) received consolidative radiation therapy with 24-54 Gy following (R-)CHOP chemotherapy. Additionally, 46 patients with indolent NHL (i-NHL) underwent radiotherapy with 22.5-45.0 Gy. Two treatment plans were prospectively created for each patient (a-NHL: 30.0/40.0 Gy; i-NHL: 24.0/30.0 Gy). NTCP were then calculated using the optimized LKB model. The adapted dose-response models properly predicted the patient's probability of developing acute side effects when receiving doses ≤ 50 Gy. In addition, it was shown that reduced radiation doses can influence the NTCP of acute side effects depending on the aggressiveness of NHL significantly. This study provided a foundation to prospectively assess the probability of adverse side effects among today's reduced radiation doses in the treatment of NHL.

Key Findings from a Transformative Quality Improvement Initiative on Advancing CAR T-Cell Treatment in Non-Hodgkin Lymphoma

Background Chimeric antigen receptor (CAR) T-cell therapy can transform outcomes for eligible patients with relapsed/refractory non-Hodgkin lymphoma (NHL). However, community-based care teams face challenges, including identifying eligible patients and coordinating referral and care, hindering integration into clinical practice. PRIME conducted a quality improvement (QI) initiative to better understand the challenges community oncology teams encounter in coordinating CAR T-cell therapy for patients with NHL and to bridge the gap between tertiary referral centers and large CAR T-cell centers. Methods This QI initiative included baseline provider surveys (n = 60), pre- and post-surveys of providers (n = 55) participating in team-based audit-feedback (AF) sessions (Table 1) and 90-day follow-up surveys. Survey questions were designed to evaluate practice patterns, challenges, and barriers to providing CAR T cell therapy to NHL patients. Care teams from each clinic, along with a CAR T-cell expert from the regional academic center and nationally renowned expert NHL CAR T-cell physician, participated in AF sessions to (a) assess system-specific practice gaps identified via the provider surveys, (b) prioritize areas for improvement, and (c) develop action plans for addressing root causes. Results Provider Surveys: Providers' top reported challenges managing patients with NHL were individualizing treatment plans (33%), patient nonadherence/lack of follow-up (23%), and patient health literacy (20%). Specific barriers to using CAR T-cell therapy included identifying patients eligible for treatment (45%), gaining approval from the insurance company (40%), and transferring the patient to a CAR T-cell center for infusion (27%). When asked about clinical practices, 48% of providers reported discussing novel therapies for NHL in detail with their patients, and 51% of providers often or always refer eligible patients for CAR T-cell therapy. While 45% of providers rated the level of collaboration/communication between CAR T-cell centers and oncology clinics as very good/excellent, more than half felt it could be better, citing insufficient sharing of patient information (38%), and lack of expertise in CAR T-cell administration among clinic staff (18%) as common challenges. Small Group AF Sessions: HCPs participating in the A/F sessions reported key challenges of treatment with CAR T-cell therapy as gaining approval from insurance companies (33%), managing patients post-treatment (26%), and identifying patients eligible for treatment (22%) (Figure 1).Improvement in very high/high confidence were noted in identifying eligible patients for CAR T-cell therapy (51% to 84%) and managing adverse/late effects of CAR T-cell therapy in patients with NHL (46% to 85%). Action plans outlined to achieve CAR T-Cell therapy goals included: increase efforts to facilitate timely patient referrals for CAR T-cell therapy, improve communication between referral centers and CAR T-cell therapy performing centers, and enhance patient knowledge and expand supportive care discussions for CAR T-cell therapy. Follow-up Surveys: In 90-day follow up surveys (n = 12), 92% of respondents reported good/very good/excellent communication and collaboration between CAR T-cell centers and oncology clinics. Additionally, 75% report improved identification of eligible patients for CAR T-cell therapy. Despite these improvements, clinics top challenge of gaining approval from the insurance company for CAR T-cell therapy (33%) persists. Conclusions Through this QI initiative, clinical teams created and implemented action plans to close gaps in identifying eligible patients, coordinating CAR T-cell therapy referral and approval, and providing counseling and obtaining consent among hematology/oncology team members. Following the initiative, meaningful confidence gains were noted in identifying eligible patients for CAR T-cell therapy and managing adverse/late effects of treatment. Action plans developed by HCPs following this QI initiative, will be addressed in future initiatives to support evidence-based integration of CAR T-cell therapies into NHL care to optimize patient outcomes. Study Sponsor Statement The study reported in this abstract was funded by an independent educational grant from Bristol Myers Squibb, who had no role in the study design, execution, analysis, or reporting.

A Multicenter, Prospective Clinical Study of the R-CDOP Regimen in Previously Untreated Non-Hodgkin Lymphoma with High Tumor Burden

Background: The treatment of non-Hodgkin lymphoma (NHL) with high tumor burden is facing severe challenges. The response rate and survival of R-CHOP treatment still need to be further explored in these patients. Pegylated liposomal doxorubicin (PLD) is a promising alternative drug to the current regimen, with a safety and efficacy profile through the enhanced permeability and retention effect and longer circulation1-3, and especially less cardiotoxic compared with traditional anthracyclines4. We conducted this prospective study to evaluate the efficacy and safety of PLD combined with R-COP (R-CDOP) regimen in treatment for previously untreated (TN) NHL with high tumor burden. Methods: This study was a single-arm, multicenter, prospective clinical trial (NCT05040555). TN Patients with CD20+ diffuse large B-cell lymphoma or grade 3b follicular lymphoma with high tumor burden were recruited. Patients are considered to have high tumor burden if they meet at least one of the following criteria: (1) presence of at least three lymph nodes with a diameter of 3 cm or more; (2) presence of any lymph nodes or extranodal masses with a diameter of 7 cm or more; (3) presence of hepatomegaly and splenomegaly confirmed by PET-CT; Spleen: female &amp;gt; 15cm, male &amp;gt; 16cm); (4) presence of pleural and peritoneal effusions determined through cytological examination, flow cytometry, or diagnosed by PET-CT, (5) LDH levels exceeding 3 times the upper limit of normal value, and (6) PET-CT TMTV exceeding 220cm3. The patients received PLD (30-35mg/ m2, d1) plus Rituximab (375mg/m2, d0), Cyclophosphamide (750mg/ m2, d1), Vindesine (3mg/ m2, d1) / Vincristine (1.4mg/m2, d1) and Prednisone (60mg/ m2, d1-5) Q3W for 4 (Limited-stage lymphoma) or 6 (Advanced lymphoma) cycles with an additional 2 cycles of Rituximab therapy. The primary endpoints was objective response rate (ORR). The secondary endpoint were complete response rate (CRR), disease control rate (DCR), progression-free survival (PFS), overall survival (OS) and safety. Results: At the data cut-off of 5 June 2023, 44 eligible pts were enrolled (Table 1). The ORR and CRR was 84.2% (32/38) [95%CI, 68.8%-94.0%] and 60.5% (23/38) [95%CI, 43.4-76.0%], respectively. The DCR was 89.5% (34/38) [95%CI, 75.2%-97.1%]. The median PFS was 14.4 months [95%CI, 4.7%-24.2%], and the median OS was not reached. The median PFS and OS will be updated after long-term follow-up. The most common grade 3/4 AEs were neutropenia (29.5%), leucopenia (25.0%) and anemia (11.4%). Left ventricular ejection fraction (LVEF) declined in 1(1/44, 2.3%) pt, but did not fall below 50%. Congestive heart failure was not occurred. Dose adjustment and treatment delay due to AE occurred in only one pt, and no treatment termination due to AE occurred except for 1 patient death. Conclusions: The preliminary results of the study indicate that the R-CDOP regimen is safe and shows promising efficacy for treating TN NHL with a high tumor burden. References： 1. Hashizume H, Baluk P, Morikawa S, et al. Openings between defective endothelial cells explain tumor vessel leakiness. Am J Pathol. 2000;156 (4):1363-1380 2. Gabizon A, Shmeeda H, Grenader T. Pharmacological basis of pegylated liposomal doxorubicin: impact on cancer therapy. Eur J Pharm Sci. 2012;45(4):388-398. 3. Sadzuka Y, Sugiyama I, Tsuruda T, et al. Characterization and cytotoxicity of mixed polyethyleneglycol modified liposomes containing doxorubicin. Int J Pharm. 2006;312(1-2):83-89. 4. Lu B, Shen L, Ma Y, et al. Cardiovascular adverse events associated with cyclophosphamide, pegylated liposomal doxorubicin, vincristine, and prednisone with or without rituximab ((R)-CDOP) in non-Hodgkin's lymphoma: A systematic review and meta-analysis. Front Pharmacol. 2022;13:1060668.

Infectious Complications in Patients with Relapsed or Refractory (R/R) Non-Hodgkin's Lymphoma (NHL) Treated with the Anti-CD20xCD3 Bispecific Antibody (BsAb) Mosunetuzumab

Introduction:NHL treatment with anti-CD20 antibodies is associated with an increased risk of infections. Mosunetuzumab (mosun) is a novel anti-CD20xCD3 BsAb that has been examined in clinical trials for different NHL subtypes and was recently approved for treating R/R follicular lymphoma. While the open-label clinical trial of mosun reported that 17% of the patients experienced infectious complications, a detailed analysis of the incidence and types of infections in patients receiving this treatment has not been reported. Methods: This is a single-center, IRB-approved, retrospective analysis of patients with R/R NHL receiving mosun, either on a study protocol (NCT02500407) at the recommended phase 2 dose or as commercial use. Patients who completed at least 1 treatment cycle and had at least 30 days of follow-up were included in this analysis. Patient demographics, information on the lymphoma, and infection variables were collected by chart review. All infection-specific variables were collected from the start of mosun therapy until the initiation of the next line of therapy or the last follow-up, whichever came first. The data cut-off was on 7/22/23. Infectious events were graded according to common terminology criteria for adverse events (CTCAE) version 5.0. Baseline characteristics were analyzed for infection risk by performing a Cox regression analysis. Results: Between October 2016 and July 2023, 48 patients were treated with mosun at our center, and 44 met the study inclusion criteria. Four patients had only received one dose or did not have sufficient follow at the time of the analysis. Baseline characteristics are summarized in Table 1. After a median follow-up of 6.8 months (range 0.7-59.7), we recorded 37 infection episodes in 16/44 patients (36.4%). Amongst these, the frequencies of viral, bacterial, and fungal infections were 54.1% (20/37), 43.2% (16/37), and 2.7% (1/37), respectively. The most common infection types were upper respiratory tract infections 42.1% (20/37), skin and soft tissue infections16.2% (6/37), including 3 shingles episodes), genitourinary infections 10.8% (4/37), and lower respiratory tract infections 10.8% (4/37). Of all the infections, 9/37 (24.3%) were Grade 3 or higher; of these, 2/37 (5.4%) were Grade 5 (1 biliary sepsis, 1 COVID pneumonia). The cumulative incidence of infection was 25% over the follow-up period (figure 1). The median time to the first infection was 144 days (range 29-1821). Among the patients who experienced infections, the infection rate was 1.54 every 100 days at risk within the first three months. The infection rate was 1.69 every 100 days at risk between three and six months and 0.37 every 100 days at risk beyond six months for patients with infection. We evaluated the type of lymphoma (i.e., aggressive vs. indolent), patients' age, number of prior therapies before BsAb administration, prior CAR-T therapy, prior autologous SCT, baseline hypogammaglobulinemia, and baseline cytopenia's as risk factors of infection with a univariate Cox regression analysis. We found that none of these characteristics were associated with the risk of infection with statistical significance. However, in a multivariate Cox regression analysis, patients who received CAR-T therapy before starting a BsAb were at a slightly higher risk of developing an infection (OR = 6.13; 95% CI: 0.90, 41.74, p = 0.063). Conclusion: Infection risk in patients with R/R NHL treated with mosun either on protocol or for commercial use appears to be higher than in the clinical trial experience reported in the literature. Viral infections were especially common, including respiratory infections and shingles episodes. Our findings highlight the need for careful monitoring and implementation of preventive strategies to decrease the infection burden in these vulnerable patients.

XPO1 Inhibitor Triggers Autophagy of TP53-Mutated Burkitt's Lymphoma Cells

INTRODUCTION Burkitt lymphoma, a highly aggressive B-cell non-Hodgkin's lymphoma (NHL), has distinct pathological and clinical characteristics. Intensive combination chemotherapy has been widely used in the treatment of Burkitt lymphoma and survival has improved for Burkitt lymphoma patients. TP53 mutation frequency is not age related, and TP53 mutations are significantly associated with higher incidence of relapse. XPO1 inhibitors, novel targeted drugs, reversibly inhibit the nuclear export of tumor-associated proteins and mRNAs by blocking nuclear export protein 1 (XPO1). Preclinical and clinical studies of XPO1 inhibitors have been successfully used in treatment of AML, MM and NHL. METHODS We used CCK8 assays to detect growth inhibition in 4 lymphoma cell lines (Raji, Jeko-1, Daudi, and CA46) under the treatment of different concentrations of KPT330 (XPO1 inhibitor). The 4D-Label-free quantitative proteomics technology was used to determine the changes of protein distribution in the nucleoplasm and cytoplasm of CA46 cells before and after KPT330 treatment (50nM, 48h). The proteomic results were validated by QPCR, westernblotting assay and fluorescent marker analysis. RESULTS The lymphoma cell lines we choosed, Raji, Jeko-1, Daudi, and CA46, all have MYC rearrangement and TP53 mutations. CCK8 assay showed Jeko-1, Daudi and CA46 were relatively sensitive to KPT330 (IC50=86.87nM, 48.34nM and 87.85nM, respectively). The results of the quantitative proteomics suggested proteins with increased nucleo-cytoplasmic ratio after KPT330 treatment were mainly focused on autophagy regulation and p53 signal pathways, including TFEB, USP10, DAP, NRBF2, SQSTM1, etc ( Fig1, a). To confirm the effect of KPT330-triggered autophagy, first, we assessed the apoptosis rates of CA46 after exposure to KPT330 for 24 hours, all of which were below 10%, ruling out apoptosis-induced autophagy ( Fig1, b). Then, westernblotting results showed that the cytoplasm load of autophagy flux related proteins, LC3B-1 and LC3B-II, both increased after KPT330 treatment ( Fig1, c). To confirm whether KPT330 effects the generation or degradation of LC3B-II, we used the late autophagy inhibitor chloroquine (CQ) to cells and observed the p62 protein level reduced and LC3B-II protein level increased with additional use of CQ compared with only KPT330 treatment ( Fig1, d). Furthermore, the ratio of LC3B-II-CQ-presence to LC3B-II-CQ-absence in the KPT330-treated group was higher than that in the control group ( Fig1, d), suggesting that KPT330 may enhance the generation of LC3B-II. We further utilized the pLenti-CMV-mCherry-GFP-LC3B plasmid to establish an LC3B stable-overexpressing CA46 cell line. Flow cytometry analysis showed that the median fluorescence intensity of both GFP and mCherry signals increased after KPT330 treatment( Fig1, e), further confirming the overall elevation of LC3B levels. To investigate the mechanism behind the increased autophagy flux, we utilized westernblotting analysis to validate the results from quantitative proteomics. TFEB protein level showed an increase in nuclear-cytoplasmic ratio upon KPT330 treatment ( Fig2, a). TFEB is known as a transcriptional activation protein, and its nuclear translocation is associated with the upregulated expression of genes involved in autophagosome formation, autolysosome fusion and lysosome biogenesis. QPCR analysis validated that genes associated with the aforementioned processes, such as LC3B, VPS11, CTSD, and CLCN7, exhibited increasing expression levels from 6 to 24 hours after KPT330 treatment ( Fig2, b). More interestingly, USP10, a component of a regulatory loop that controls autophagy and p53/TP53 levels, significantly increased in cell nucleus after drug action ( Fig2, c), which suggested that KPT330 may stabilize p53 function. Finally, we also attempted to discover the potential synergistic therapeutic effect of KPT330 with commonly used chemotherapy drugs in clinical lymphoma treatment. However, none synergistic effect was observed according to KPT330 combined with Cytarabine, Doxorubicin, Azacitidine, Decitabine, Rapamycin, Bortezomib, Duvelisib or Venetoclax, respectively ( Fig2, d). CONCLUSION XPO1 Inhibitor, KPT330 can restrict nuclear export of autophagy-related proteins thus regulate autophagy and stabilize p53 function in TP53-mutated Burkitt's lymphoma cells, which revealed a promising treatment.

Evaluation of CD70 Chimeric Antigen Receptor T (CAR-T) Cells As a Potential Treatment for Non-Hodgkin's Lymphoma: A Preclinical Study

Background ：CD70 is a member of the tumor necrosis factor (TNF) superfamily. While its expression is limited in healthy tissues, CD70 highly constitutive expressed on cell surface of various malignancies. This expression profile make CD70 a promising target for cancer immunotherapy. We described a CD70 CAR-T based on a potent VHH nanobody named 11C9 in treatment of renal cell carcinoma (RCC). Recently, we received approval to initiate a phase I clinical study by Center for Drug Evaluation, National Medical Products Administration (NMPA) in China. Motivated by promising evidence that CD70 aberrantly expressed on Non-Hodgkin's lymphoma (NHL), including diffuse large B-cell lymphomas, follicular lymphomas, Burkitt and mantle cell lymphomas, we conducted a preclinical evaluation of CD70 CAR-T cells as a potential treatment by preclinical model. Method: We successfully generated CD70 CAR-T cells based on a VHH nanobody (11C9) and CD19 CAR-T cells based on murine scFv (FMC63) following the exact manufacturing protocols used for phase I and phase II clinical trial products respectively. Then, we evaluated CAR-T cell cytotoxicity, phenotype, proliferation, and cytokine production in vitro. Furthermore, we established NHL tumor cell-xenograft NOG mice models to compare their anti-tumor activity in vivo. Result: In vitro test demonstrated two types of CAR-T cells have comparable proliferation, and cytokine production. We observed more potency for CD70 CAR-T to kill CD70 and CD19 double positive Raji cells than CD19 CAR-T cells. In vivo test showed that both CD70 CAR-T cells and CD19 CAR-T cells eradiated Raji cells xenografted NOG mice models at high dose. However, after treated at lower dose CD19 CAR-T, tumor relapsed in one week when the same lower dose CD70 CAR-T treated mice keeping tumor free statue. Conclusion: Our results indicate that CD70 CAR-T cell is potential treatment for NHL. We will conduct an investigation of single and dual CD70 CAR-T on treatment of lymphoma especially for CD19 negative NHL.

Novel CD20 Mutations As a Mechanism of Resistance to CD20-CD3 Targeted Therapies in Non-Hodgkin's Lymphoma

Non-Hodgkin's Lymphoma (NHL) is one of the most common blood cancer types in the world. CD20 is a B cell restricted lineage marker that is a well-established target for the treatment of NHL and anti-CD20 agents are included as in standard of care regimens. Given the success of these targeted therapies, strategies to further enhance therapeutic activity are being explored, including the CD20-CD3 bispecific molecule, mosunetuzumab, which is approved for treatment of relapsed and refractory Follicular Lymphoma in adults who have received &amp;gt;2 lines of treatment. Mosunetuzumab acts by redirecting CD3+ T cells to engage and eliminate CD20-expressing B cells. As with other targeted therapies, mutations and loss of target expression have been described following mosunetuzumab therapy [1]. Here we describe the impact of novel mutations identified in patient biopsy samples from the mosunetuzumab monotherapy trial, GO29781 that were located within the extracellular and transmembrane domains of CD20 on protein expression, localization and T-cell mediated killing. NHL cell lines, SU-DHL-16 and MAVER-1, were engineered using CRISPR-Cas9 system to create CD20 knock out (KO) cell lines. Wild-type (WT) or mutant CD20 were exogenously re-expressed in the CD20 KO cell lines and in the CD20 negative acute lymphoblastic leukemia cell line, REH, to create isogenic cell lines for further analysis. Complete CD20 loss was observed in cells expressing CD20 P160fs (frameshift) and Q187* (truncating) mutations. CD20 missense mutations located in the extracellular domain (C167G, K175E) expressed CD20 protein levels comparable to CD20 WT cells and membrane localization was confirmed by flow cytometry and immunofluorescence microscopy. To test the functional effect of CD20 mutations, in vitro co-culture cell killing assays were carried out using healthy donor CD8+ T cells, isogenic cell lines (3:1 effector to tumor cell ratio) and a proof-of-concept CD20-CD3 bispecific antibody. Differences in T-cell mediated killing were observed across mutation types. CD20 WT cell lines demonstrated 95-99% cell killing following 48 hour treatment while the CD20 KO, frameshift and truncating mutations all showed negligible T-cell mediated killing (~10-17% cell death). Cells expressing the missense mutations in the extracellular domain showed resistance to T-cell mediated killing (~30-45% cell death) compared to CD20 WT cells. T-cell activation was assessed by CD69 (early) and CD25 (late) surface markers. In all co-cultures, the level of CD69+ T cells was equivalent, while the CD69+CD25+ CD8 T cells were only significantly increased in the CD20 WT co-cultures, consistent with the high level of T-cell mediating killing. We have confirmed that these mutations can play a role in the development of resistance through both loss of target protein expression and interference within the anti-CD20 binding site. These studies help to characterize mechanisms of resistance to mosunetuzumab that could be extended to other anti-CD20 targeted agents. [1] Schuster et al. Journal of Clinical Oncology 2022, 40 (16), 7526.

The Impact of Pre-Existing Autoimmune Disease on the Safety and Efficacy of CAR T-Cell Therapy for Non-Hodgkin Lymphoma

Introduction: Chimeric antigen receptor T-cell therapy (CAR-T) has revolutionized the treatment of non-Hodgkin lymphoma (NHL). Despite having an increased risk of NHL, patients with pre-existing autoimmune disease (AID) were excluded from pivotal CAR-T trials due to concerns that CAR-T may exacerbate AID. As a result, there are limited data on the safety and efficacy of CAR-T for NHL in patients with AID. Interestingly, there is also growing evidence that CAR-T can be therapeutically effective for AID, such as in the treatment of systemic lupus erythematosus (SLE). As CAR-T utilization for NHL increases, it is essential to characterize the safety and efficacy of this potentially curative therapy in patients with AID. This study aimed to evaluate the impact of AID on outcomes in patients treated with commercial CD19-targeted CAR-T for NHL. Methods: Following Institutional Review Board approval, we conducted a retrospective analysis of all patients who were treated with commercial CD19-targeted CAR-T for NHL and had a pre-existing AID across the Mayo Clinic Enterprise (Arizona, Florida, and Minnesota) between January 2018 and May 2023. Variables collected included age, gender, NHL type, AID type, and CAR-T product. Efficacy outcomes, including response rates at 6 and 12 months after CAR-T, were collected. Toxicities, including cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS), were collected and analyzed. Our chart review for AID included SLE, sarcoidosis, ulcerative colitis, celiac disease, pernicious anemia, Crohn's disease, psoriasis, Sjogren's syndrome, hypothyroidism (Hashimoto's disease), hyperthyroidism (Graves' disease), rheumatoid arthritis (RA), vasculitis, scleroderma, and mixed connective tissue disease (MCTD). Results: A total of 235 patients who received commercial CD19-targeted CAR-T for NHL across the Mayo Clinic Enterprise were identified (Table 1). Of these 235 patients, 27 had a pre-existing AID and 208 did not. The median age was 66 years (range 39-81) for patients with AID and 59 years (range 18-88) for patients without AID. Over half of the patients were male (59% of patients with AID and 51% of patients without AID). The most common NHL type was diffuse large B cell lymphoma (41% of patients with AID and 56% of patients without AID). Most patients received axicabtagene ciloleucel (85% of patients with AID and 84% of patients without AID). The most common AID was hypothyroidism (81%, n=22), although other conditions were also identified, including RA (11%, n=3), SLE (4%, n=1), pulmonary sarcoidosis (4%, n=1), psoriasis (4%, n=1), and MCTD (4%, n=1). Two patients had more than one AID. Median follow-up was 18 months (range 1-36) for patients with AID and 14 months (range 2-49) for patients without AID. Six months after CAR-T, the complete response (CR) rate was 77.2% and 45.1% in patients with AID and without AID, respectively (p=0.0042). Twelve months after CAR-T, the CR rate was 50% and 31.4% in patients with AID and without AID, respectively (p=0.0801). CRS of any grade occurred in 78% and 88% of patients with and without AID, respectively. ICANS of any grade occurred in 41% and 54% of patients with and without AID, respectively. Conclusion: AID did not appear to adversely impact the safety or efficacy of CAR-T for NHL. Limitations of our study include the small number of patients with AID and the fact that the most common AID was hypothyroidism. Larger studies are warranted to confirm these findings. Additional data are needed to characterize the impact of CAR-T for NHL on AID outcomes and to describe the effect of disease-modifying antirheumatic drugs on NHL outcomes after CAR-T.

Changing Trends in B-Cell Non-Hodgkin Lymphoma Treatment: The Role of Novel Monoclonal Antibodies in Clinical Practice.

We are currently witnessing a dramatic shift in our approach to the treatment of B-cell non-Hodgkin lymphoma (B-NHL). In the evolving clinical landscape, novel treatments for this clinically heterogeneous disease span a wide range of interventions, encompassing targeted agents, cell therapy approaches, and novel monoclonal antibodies (NMABs). Among these, the latter are likely to exert the most profound impact due to their distinctive high efficacy and versatile applicability. NMABs represent a heterogeneous group of agents, including naked antibodies, immunotoxins, and T-cell-engaging molecules. In recent times, several NMABs have either gained regulatory approval or are on the verge of introduction into clinical practice, addressing multiple therapeutic indications and treatment regimens. Their anticipated impact is expected to be broad, initially in the context of relapsed/refractory (R/R) disease and subsequently extending to early treatment lines. The scope of this review is to provide a comprehensive overview of the biological characteristics, clinical properties, efficacy, and toxicity profiles of NMABs that have recently been introduced or are nearing integration into clinical practice.

CAR-T Cell Therapy in the Treatment of Pediatric Non-Hodgkin Lymphoma.

Non-Hodgkin lymphomas (NHL) are a group of cancers that originate in the lymphatic system, especially from progenitor or mature B-cells, T-cells, or natural killer (NK) cells. NHL is the most common hematological malignancy worldwide and also the fourth most frequent type of cancer among pediatric patients. This cancer can occur in children of any age, but it is quite rare under the age of 5 years. In recent decades, available medicines and therapies have significantly improved the prognosis of patients with this cancer. However, some cases of NHL are treatment resistant. For this reason, immunotherapy, as a more targeted and personalized treatment strategy, is becoming increasingly important in the treatment of NHL in pediatric patients. The objective of the following review is to gather the latest available research results, conducted among pediatric and/or adult patients with NHL, regarding one immunotherapy method, i.e., chimeric antigen receptor (CAR) T cell therapy. We focus on assessing the effectiveness of CAR-T cell therapy, which mainly targets B cell markers, CD19, CD20, and CD22, their connections with one another, sequential treatment, or connections with co-stimulatory molecules. In addition, we also evaluate the safety, aftermath (especially neurotoxicities) and limitations of CAR-T cell therapy.

The academic point-of-care anti-CD19 chimeric antigen receptor T-cell product varnimcabtagene autoleucel (ARI-0001 cells) shows efficacy and safety in the treatment of relapsed/refractory B-cell non-Hodgkin lymphoma.

Varnimcabtagene autoleucel (var-cel) is an academic anti-CD19 chimeric antigen receptor (CAR) product used for the treatment of non-Hodgkin lymphoma (NHL) in the CART19-BE-01 trial. Here we report updated outcomes of patients with NHL treated with var-cel. B-cell recovery was compared with patients with acute lymphoblastic leukaemia (ALL). Forty-five patients with NHL were treated. Cytokine release syndrome (any grade) occurred in 84% of patients (4% grade ≥3) and neurotoxicity in 7% (2% grade ≥3). The objective response rate was 73% at Day +100, and the 3-year duration of response was 56%. The 3-year progression-free and overall survival were 40% and 52% respectively. High lactate dehydrogenase was the only covariate with an impact on progression-free survival. The 3-year incidence of B-cell recovery was lower in patients with NHL compared to ALL (25% vs. 60%). In conclusion, in patients with NHL, the toxicity of var-cel was manageable, while B-cell recovery was significantly prolonged compared to ALL. This trial was registered as NCT03144583.

Radiotherapy dose de-escalation in patients with high grade non-Hodgkin lymphoma in a real-world clinical practice.

The standard treatment of non-Hodgkin lymphoma (NHL) comprises combined modality treatment, radiotherapy (RT), and chemotherapy with rituximab which has significantly improved both disease-free survival (DFS) and overall survival (OS). However, there is no uniformity in radiation dose usage in these patients. In this retrospective study, we compared lower radiation dose with higher in patients with aggressive NHL. From 2007 to 2017, treatment records of all high-grade NHL or diffuse large B-cell lymphoma and non-central nervous system NHL were included. We compared response rates, OS and DFS of patients who received ≤30 Gy RT to those with >30 Gy. Univariate and multivariate analyses were done to determine factors affecting prognosis, i.e., age, sex, stage, International Prognostic Index (IPI), adding rituximab, and radiation dose. A total of 184 NHL patients treated with combined modality or radiation alone having complete follow-up details were analyzed. At median follow-up of 66.8 months, 5-year OS was 72.8% in high-dose group versus 69.9% in low-dose group (p = 0.772) and 5-year DFS 64.7% versus 64.1% (p = 0.871). Patients having early-stage disease receiving low dose and those with advanced disease treated with >30 Gy had better OS and DFS though not statistically significant. Adding rituximab was associated with significantly better OS and DFS irrespective of radiation dose delivered. High IPI score and omitting rituximab were the only factors that significantly worsened both OS and DFS. Acute radiation toxicities were comparable in both groups (p = 0.82). Among late toxicities, no patient developed a second malignancy and 5% died due to cardiovascular complications (p = 0.595) though only two patients (1.1%) had received thoracic radiation. The two groups had comparable response rates, acute toxicities, DFS and OS. This study suggests that RT dose reduction may be possible in high-grade NHL without compromising the DFS and OS.

POSTER: HL-122 Microbiota Changes in Hodgkin and Non-Hodgkin Lymphoma Treatment: Assessment With Il-17 and Il-12 Levels

POSTER: HL-122 Microbiota Changes in Hodgkin and Non-Hodgkin Lymphoma Treatment: Assessment With Il-17 and Il-12 Levels

HL-122 Microbiota Changes in Hodgkin and Non-Hodgkin Lymphoma Treatment: Assessment With Il-17 and Il-12 Levels

HL-122 Microbiota Changes in Hodgkin and Non-Hodgkin Lymphoma Treatment: Assessment With Il-17 and Il-12 Levels