Beta 2-microglobulin Levels Research Articles

BSBM-16 HLA CLASS-I ANTIGEN PRESENTATION MACHINERY AND IFN-γ PATHWAY ALTERATIONS IN LUNG CANCER BRAIN METASTASES

Abstract BACKGROUND Immune checkpoint inhibitors (ICI) can have activity in lung cancer brain metastases (LBM). However, the duration of responses is limited and most patients with LBM do not benefit from these therapies. Alterations in HLA class-I antigen presentation machinery via beta-2 microglobulin (B2M) downregulation and disruption of IFN-γ sensing were identified as mechanisms of immune evasion and ICI resistance in solid tumors. However, the expression and clinical significance of these pathways in LBM are poorly understood. METHODS Using multiplex quantitative immunofluorescence we measured the localized expression of B2M protein; and the IFN-γ pathway markers phospho-STAT1 (pSTAT1), and Interferon Regulatory Factor 1 (IRF-1) in 67 immunotherapy naïve LBM and 45 primary lung tumors (PLT), including 15 patients with paired samples represented in a tissue microarray. The markers were selectively measured in cytokeratin + tumor cells and in cytokeratin- stromal cells. Associations between the markers, PD-L1, TILs and clinicopathologic variables were compared between LBM and PLT. RESULTS LBM showed lower levels of B2M in both tumor and stroma compartments compared to PLT. In contrast, high tumor and stromal levels of pSTAT1 were observed in LBM and comparable levels of IRF-1 were found between both tumor sites. Consistently across LBM and PLT samples, pSTAT1 and IRF1 were strongly correlated, and a positive but weaker association was observed with B2M and TILs. Only in PLT high tumor-cell PD-L1 expression was associated with high levels of IRF-1. No statistically significant association was observed between the markers and clinicopathologic variables. Tumor B2M downregulation in PLT was associated with worse survival. CONCLUSION LBM show distinct immunomodulatory properties relative to PLT, characterized by reduced HLA class-I antigen presentation machinery, elevated IFN-γ signaling and reduced local adaptive immune responses. These results support differences in immune evasion mechanisms in LBM compared with the PLT, which may have biological and clinical implications.

Role of Selected Circulating Tumor Biomarkers in Patients with Skeletal Metastatic Pancreatic Neuroendocrine Neoplasms.

We investigated the diagnostic capacity of selected circulating biomarkers (CBMs) for the early detection of bone metastasis (BMets) in patients with pancreatic neuroendocrine neoplasms (PanNENs). A total of 115 patients with PanNENs and 40 controls were enrolled. We measured the serum levels of ferritin, cytokeratin 18 (CY18), CA19-9, CA125, AFP, CEA, and beta-2 microglobulin (B2M). A total of eight PanNEN patients developed BMets, and one hundred seven remained BMets-free. We observed a significantly higher level of CA125 and CY18 in BMets patients vs. non-BMets patients (p = 0.01 and p = 0.04, respectively). CA125, CY18, and B2M area under receiver operator characteristic (AUROC) analyses differentiated both patients groups; CA125 area under the curve (AUC) 0.77, p < 0.01; CY18 AUC data were 0.72, p = 0.03, and B2M AUC 0.67, p = 0.02. On the basis of CBM metrics in both subgroups, we reached a sensitivity/specificity for CA125 of 75/76%; for CY18 of 75/69%, for B2M of 100/50%, for CA125, and the CY18 combination of 93/90%, respectively. According to current results, CA125 and CY18 seem to have the potential capacity as fair biomarkers for BMets detection, despite the small number of cases. Further studies are warranted in the larger PanNEN patient group.

Design of Beta-2 Microglobulin Adsorbent Protein Nanoparticles.

Beta-2 microglobulin (B2M) is an immune system protein that is found on the surface of all nucleated human cells. B2M is naturally shed from cell surfaces into the plasma, followed by renal excretion. In patients with impaired renal function, B2M will accumulate in organs and tissues leading to significantly reduced life expectancy and quality of life. While current hemodialysis methods have been successful in managing electrolyte as well as small and large molecule disturbances arising in chronic renal failure, they have shown only modest success in managing plasma levels of B2M and similar sized proteins, while sparing important proteins such as albumin. We describe a systematic protein design effort aimed at adding the ability to selectively remove specific, undesired waste proteins such as B2M from the plasma of chronic renal failure patients. A novel nanoparticle built using a tetrahedral protein assembly as a scaffold that presents 12 copies of a B2M-binding nanobody is described. The designed nanoparticle binds specifically to B2M through protein-protein interactions with nanomolar binding affinity (~4.2 nM). Notably, binding to the nanoparticle increases the effective size of B2M by over 50-fold, offering a potential selective avenue for separation based on size. We present data to support the potential utility of such a nanoparticle for removing B2M from plasma by either size-based filtration or by polyvalent binding to a stationary matrix under blood flow conditions. Such applications could address current shortcomings in the management of problematic mid-sized proteins in chronic renal failure patients.

Characterization of Urate Metabolism and Complications of Patients with Renal Hypouricemia.

Background Both renal hypouricemia (RHU) and gout are associated with renal dysfunction and urolithiasis. The difference in renal complications associated with RHU and gout, however, has not been studied. We characterized the urate metabolism and complications of patients with RHU and compared them with patients with gout. Methods Eighteen patients with RHU who had a serum uric acid (SUA) level <2 mg/dL (10 men and 8 women), 44 patients with gout (44 men) and 16 normouricemic patients (4 men and 12 women) were included. The blood and urinary biochemical data were evaluated. A genetic analysis of uric acid transporter 1 (URAT1) was also conducted in 15 cases with RHU. Results The SUA level of RHU was 0.9±0.5/mg/dl, and the Uur/Ucr and Cur/Ccr were 0.56%±0.14% and 45.7%±18.0%, respectively. A genetic analysis of URAT1 in 15 RHU patients showed that 13 harbored a URAT1 gene mutation, whereas 2 harbored the wild-type gene. The SUA level was significantly lower in RHU patients (n=11) than in either gout patients (n=44) or normouricemic patients (n=16). This reduction was accompanied by the elevation of Cua/Ccr. Urinary beta 2-microglobulin levels were higher in RHU patients than in gout or normouricemia patients. Cua/Ccr correlated with normalized urinary beta 2-microglobulin levels. The prevalence of urolithiasis was 18.2% in RHU cases and 6.8% in gout cases. A homozygous URAT1 mutation was associated with urolithiasis. Conclusion Besides urolithiasis, RHU can be associated with tubular dysfunction, such as elevated urinary beta 2-microglobulin levels.

Comparison of International Prognostic Indices and Validation Study for Patients with Diffuse Large B-Cell Lymphoma in the Rituximab Era

ABSTRACT In diffuse large B-cell lymphoma (DLBCL), patients needing new alternative regimens in first-line treatment should be selected with better risk classification. After International Prognostic Index (IPI), Revised-IPI (R-IPI), National Comprehensive Cancer Network (NCCN)-IPI, and Grupo Español de Linfomas y Trasplante Autólogo de Médula Ósea (GELTAMO)-IPI, have been developed to improve risk predictions. This study compared performances of four prognostic indices concerning differentiation of overall survival (OS), the most critical endpoint. The study was conducted on 116 patients diagnosed with DLBCL. Patients with primary nervous system and testicular DLBCL, and post-transplant lymphoproliferative disorders, were excluded. The fitting of prognostic indices for database and the prediction of patient discrimination were compared using Akaike’s information criterion and concordance index. Of the study cohort, 63.8% were male, the median age was 56 (18-88), and median follow-up term was 45.6 (0.3-75.2) months. All factors, constituting IPI and R-IPI scores, demonstrated a significant difference in OS. Involvements of the extranodal regions specified in NCCN-IPI and elevated serum beta-2 microglobulin levels in GELTAMO-IPI had prognostic significance (p= 0.005 and p= 0.040, respectively). Each of the four prognostic indices, resulted in risk groups with significantly different OS. R-IPI provided the best fit for database, while NCCN-IPI provided the best discrimination between patients with high and low OS. Although NCCN-IPI provides the best discrimination between patients with short and long OS, using original IPI still seems acceptable in the rituximab era. Integration of tumor molecular characteristics into NCCN-IPI may better characterize high-risk group in new treatment approaches. Keywords: International Prognostic Index, National Comprehensive Cancer Network, Validation, Diffuse Large B-Cell Lymphoma, Rituximab

#3255 BETA2 MICROGLOBULIN IS AN INDEPENDENT PARAMETER RELATED WITH DYSFUNCTION OF NATURAL KILLER CELL ACTIVITY IN HEMODIALYSIS PATIENTS

Abstract Background and Aims End-stage renal disease patients are characterized by immune dysfunction. Natural killer (NK) cells area lymphocytes of innate immune system that play a key role in an immune response towards viral infections and tumors. We aimed to analyze clinical factors related to the NK cell activity in chronic hemodialysis (HD) patients. Method Clinically stable 196 patients who were treated by HD for more than 3 months were enrolled from 4 outpatient HD clinics. NK cell activity was assessed using NK Vue assay (NKMAX, Sungnam, Korea) that uses serum of ex-vivo stimulated whole blood to detect interferon (IFN)-γ secreted from NK cells as an indicator of NK cell activity. All patients were stratified as abnormal (&amp;lt;250 pg/ml) and normal (≥250 pg/ml) groups according to NK cell activity. Results Mean age of the HD patients was 62.7 years [range 31 to 95 years] and mean HD duration was 49.2 months [range 3 to 221 months]. In total, 68 (35%) of HD patients showed abnormal NK cell activity. The abnormal NK cell activity group showed significantly increased age and shorter HD duration compared to the normal NK cell activity group (66.7 ± 11.9 vs. 60.6 ± 13.4 years, p = 0.001; 33.8 ± 33.0 vs. 57.5 ± 48.5 months, p&amp;lt;0.001, respectively). The serum albumin and parathyroid hormone levels were significantly lower in the abnormal NK cell activity group (3.7 ± 0.4 vs. 3.9 ± 0.3 g/dL, p = 0.005 and 176 ± 153 vs. 240 ± 260 pg/mL, respectively). In contrast, hemoglobin, blood urea nitrogen, urea reduction ratio, and C-reactive protein levels were comparable between the two groups. In multivariate regression analysis, old age, short dialysis duration, low serum albumin and high beta2 microglobulin levels were independent risk factors of abnormal NK cell activity in HD patients (adjusted odd ratio [AOR], 1.033; 95% confidence interval [CI], 1.005-1.062, p = 0.02; AOR, 0.981; 95% CI, 0.971-0.992; p&amp;lt;0.001, AOR, 0.395; 95% CI, 0.157-0.995; p = 0.049, and AOR, 1.042; 95% CI, 1.011-1.075; p = 0.008, respectively). Conclusion Our results suggest that young age and good nutrition as well as lower burden of middle molecule uremic toxin is associated with greater NK cell activity in HD patients.

COMBINING BASELINE TOTAL METABOLIC TUMOR VOLUME AND BETA‐2‐MICROGLOBULIN LEVELS PREDICTS OUTCOMES IN HIGH BURDEN FOLLICULAR LYMPHOMA

Introduction: Follicular lymphoma (FL) usually presents an indolent course with repeated events of disease progression but treatment-free time intervals and patients’ survival are reduced at each relapse. Many prognostic scores exist but few are used in practice due to their complexity. Total metabolic tumor volume (TMTV) has demonstrated a strong prognostic value in FL and its routine use is increasing in academic centers. We aim to explore the predictive value of a simple prognostic score based on TMTV and beta-2-microglobulin (B2M) levels at baseline in high tumor burden FL to identify patients with the highest unmet medical need. Methods: In this monocentric retrospective analysis, we enrolled adult patients admitted to our hospital between 2006 and 2018 and fulfilling the following inclusion criteria: grade 1 to 3A FL with at least one high tumor burden criteria defined by the Groupe d'Etude des Lymphomes Folliculaires (GELF), available PET-CT images collected before first-line standard immunochemotherapy start and suitable for TMTV assessment. Baseline TMTV was determined using the 41% SUVmax threshold method. B2M levels were routinely measured at diagnosis. Primary endpoints were progression-free survival (PFS) and overall survival (OS) according to a score combining TMTV and B2M. Results: We included 126 FL patients in this analysis. Median age at time of treatment was 61 years (55; 57). Patients displayed advanced-stage disease (n = 112, 88.9%), intermediate (n = 51, 40.8%) or high (n = 62, 49.6%) FLIPI and intermediate (n = 40, 33.3%) or high (n = 31, 25.8%) PRIMA-PI scores. Median baseline SUVmax and TMTV were 12.1 (10; 17) and 606 mm3 (268;1219), respectively. Baseline B2M levels were measured for 125 (97%) patients and >3 mg/L in 30 (24%). Treatment regimens used were mostly R-CHOP like (n = 109, 86.5%) and R-lenalidomide (n = 9, 7.1%). 80 patients (63.5%) received a maintenance therapy during 24 months. We defined three risk categories : (i) Low risk (n = 45, 36%): TMTV ≤510 mm3 and B2M ≤3 mg/L; (ii) Intermediate risk (n = 57, 45.6%) : TMTV >510 mm3 or B2M >3 mg/L ; (iii) High risk (n = 23, 18.4%) : TMTV >510 mm3 and B2M >3 mg/L. With a median follow-up of 83 (52; 116) months, the 5-year PFS was 81.7% (95% CI: 71–94), 53.7% (95% CI: 41.8–69) and 38.6% (95% CI: 23–65) in the low, intermediate and high risk groups, respectively (p < 0.0001). Five-year OS was based on 14 deaths and estimated at 96.1% (95% CI: 91–100), 89.1% (95% CI: 81.2–97.7) and 73.7% (95% CI: 57.6–94.2) in the low, intermediate and high risk groups, respectively (p = 0.003). Keywords: aggressive B-cell non-Hodgkin lymphoma, diagnostic and prognostic biomarkers, PET-CT No conflicts of interests pertinent to the abstract.

Attempt to assess direct interactions between tumor burden, myeloid-derived suppressor cells and PD-1- and TIM-3-expressing T cells in multiple myeloma patients

The avoidance of immune surveillance by malignant plasma cells (PCs) in multiple myeloma (MM) is mediated by different mechanisms, among which an induction of T cell exhaustion and expansion of myeloid-derived suppressor cells (MDSCs) appear to play substantial roles, but it is still a lack of data on possible MDSC-mediated induction of T cell exhaustion. The aim of the present work was to evaluate possible relationship between frequencies of MM PCs, MDSCs and phenotypically exhausted PD-1+ and TIM-3+ T cells in bone marrow (BM) samples and peripheral blood (PB) of MM patients at various disease stages. Peripheral blood (n = 88) and BM samples (n = 56) were obtained from MM patients (newly diagnosed (n = 6), patients in remission (n = 71) and with progressive disease (n = 11)). Frequencies of T cells expressing checkpoint receptors PD-1 and TIM-3, polymorphonuclear MDSCs (PMN-MDSCs, Lin-CD14-HLA-DR- CD33+CD15+/CD66b+), monocyte MDSCs (M-MDSCs, CD14+HLA-DRlow/-), early MDSCs (E-MDSCs, Lin-HLA-DR-CD33+CD15-/CD66b-), and MM PCs (CD45dimCD38+CD138+CD56+CD19-CD117+CD27- CD81-) were assessed with flow cytometry. Circulating and BM-resident PD-1+/TIM-3+T cell subsets, BM E-MDSCs, as soon as MM PCs and serum beta2-microglobulin (B2-M) levels were gradually increased in patients at different stages. Despite that, there were no associations between the markers of tumor load and the studied cell subsets. In patients in remission, BM PMN-MDSCs negatively correlated with CD4+T cells, CD4+PD-1+ and CD8+TIM-3+T cell subsets; there were positive correlations between BM E-MDSCs and CD4+PD-1+TIM-3+ cells and PB M-MDSCs and CD8+PD-1+ and (as a trend) CD8+TIM-3+T cells. We found no associations for the samples of patients at diagnosis and with progression. We can conclude that a possible mutual influence of malignant PCs, MDSCs and PD-1+/TIM-3+T cells is nonlinear, especially during a manifest tumor growth at diagnosis and progression. The detected negative correlations between resident PMN- MDSCs and T cell subsets might be associated with MDSC suppressive function, affecting both predominantly activated PD-1+ cells and exhausted TIM-3+ subsets. The positive correlations between BM E-MDSCs and CD4+PD-1+TIM-3+ cell subset and circulating M-MDSCs and PD-1+ and TIM-3+ CD8+T cells might confirm an ability of MDSCs to induce T cell exhaustion.

A classifier based on 273 urinary peptides predicts early renal damage in primary hypertension.

Renal diseases caused by primary hypertension (HTN) are often asymptomatic without sensitive markers for early diagnosis and prediction, easily progressing to severe and irreversible renal damage in patients with clinical manifestations. This study explored whether a classifier developed based on 273 urinary peptides (CKD273) could serve as a potential biomarker for early prediction of renal damage in HTN. Urinary CKD273 level of healthy individuals, HTN + normoalbuminuric and HTN + albuminuria patients were compared, and 22 baseline data including sex, age, renal function, and hypertensive fundus lesions were collected. Patients diagnosed with HTN, albuminuria, and normal renal function were followed up. According to the follow-up results, the cut-off value of CKD273 in predicting hypertensive renal injury was calculated and analyzed in the high-risk and low-risk groups of HTN patients for its performance in detecting early hypertensive renal injury. Among a sum of 319 participants, average urinary CKD273 level was significantly higher in patients with HTN than in normal individuals. A total of 147 HTN patients with normal albuminuria were followed up for a mean of 3.8 years. Thirty-five patients showed urinary albumin-to-creatinine ratio (uACR) at least 30 mg/g for three consecutive times. The receiver-operating characteristic (ROC) curve showed that the urinary CKD273 cut-off value for evaluating new-onset proteinuria in patients with HTN was 0.097. Based on this cut-off value, 39 and 108 patients were included in the high-risk and low-risk groups, respectively. Specifically, compared with patients in the low-risk group, those in the high-risk group showed significantly longer duration of HTN, higher proportions of hypertensive fundus lesions and at least 30 mg/g uACR, and higher levels of homocysteine (Hcy), cystatin C (CysC), beta-2 microglobulin (β2-MG), and uACR. 76.9% of high-risk patients had significantly higher new-onset proteinuria than the low-risk group. Correlation analysis demonstrated a positive correlation between urinary CKD273 and UACR ( r = 0.494, P = 0.000). The incidence of new-onset albuminuria was significantly higher in the high-risk group than in the low-risk group, as shown by Cox regression analysis. The areas under the curve of CKD273, Hcy, β2-MG, and CysC were 0.925, 0.753, 0.796, and 0.769, respectively. Urinary CKD273 is a predictor of new-onset proteinuria in patients with HTN, therefore, it can be used for diagnosing patients with early renal injury in patients with HTN, contributing to early prevention and treatment of hypertensive nephropathy.

Abstract 6635: Lipid and glycolipid antigen presentation faults as a mechanism of resistance to immune checkpoint therapy in poorly immunogenic melanomas

Abstract Introduction: Cold tumors are characterized by lacking infiltration of antitumor T cells. These tumors are universally refractory to immune checkpoint therapies (ICT). While poor T cell infiltration has been partly ascribed to low tumor mutational burden (TMB), TMB doesn’t necessarily exclude immunogenic responses nor ensure a response to ICT. Mechanisms of T cell exclusion in tumors with sufficient TMB may be caused by molecular failure of the antigen processing and presentation (APP), which impacts T cell generation. It was previously described that loss of heterozygosity (LOH) of an essential component of the MHC class I molecule, the beta-2-microglobulin (B2M), is associated with low antigen presentation, tumor T cell infiltration, and ICT resistance in melanoma. Here we provide further insight into APP and ICT response in melanoma. Methods: Transcriptomic analysis and visualization of genomic data from the GDC-TCGA skin cutaneous melanoma (SKCM) study were performed using the Xena platform. Four anti-PD1 metastatic melanoma cohorts were normalized with transcripts per million units. STRING protein-protein interaction analysis was performed in the human database with 11 nodes and 39 edges. Nanostring transcriptomic data from metastatic uveal melanoma (mUM) was revisited from our previous publications for Spearman’s correlational and group analysis of APP signatures amongst ICT responders and non-responders. Results: In an expanded analysis of 4 normalized anti-PD1 treated metastatic melanoma cohorts (n=272), we confirmed that transcriptomic levels of B2M are significantly reduced in non-responders. Moreover, STRING protein-protein interaction analysis of B2M revealed CD1D as a potential molecular partner of B2M. We, therefore, observed that CD1D transcriptomic expression is not only reduced in anti-PD1 poor responders but is a survival prognosticator in primary tumors from the GDC-TCGA study. Interestingly, CD1D expression is also significantly reduced in poorly immunogenic mUM tumors, a cancer type regarded as universally refractory to ICT. Conclusions: These findings suggest that APP faults in ICT-resistant tumors are not restricted to faults in peptide presentation to T lymphocytes (via B2M) but may also impact the presentation of tumor-associated lipid and glycolipid antigens via CD1d, which can impact antitumor responses of natural killer T cells (NKT). Adjuvant therapies that restore MHC-I and CD1d-mediated antigen presentation may unleash the response to ICT via NKT. Understanding CD1d expression faults locally and systemically may provide valuable prognostication information likely to impact how cancer is treated by optimally selecting patients for ICT and validating novel therapeutic targets for immunotherapy development. Citation Format: Carlos Rogerio Figueiredo. Lipid and glycolipid antigen presentation faults as a mechanism of resistance to immune checkpoint therapy in poorly immunogenic melanomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6635.

Association of Urinary Biomarkers of Renal Tubular Injury with Cognitive Dysfunction in Older Patients with Chronic Kidney Disease: A Cross-Sectional Observational Study

Epidemiological data suggest that individuals in all stages of chronic kidney disease (CKD) have higher risks of developing cognitive impairment. The relationship between CKD and cognition has been assessed exclusively using glomerular function markers; however, kidney tubule injury has not been assessed. We assessed the association between urinary biomarkers of renal tubular injury and cognitive dysfunction in older patients with CKD Stages 3-4. According to the Montreal Cognitive Assessment, participants were divided into cognitive dysfunction and control groups. Compared with the control group, the cognitive dysfunction group had significantly higher percentages of smokers, noticeably lower average education, and higher mitochondrial DNA (mtDNA) levels in the peripheral blood. Spearman correlation analysis showed that higher urine neutrophil gelatinase-associated lipocalin, kidney injury molecule-1, and beta-2 microglobulin (β2M) levels were significantly associated with lower cognitive scores. Multivariate logistic regression analysis showed that only increased urinary β2M levels were independently associated with cognitive worsening in CKD after adjusting for confounders. Logistic regression identified a promising role of urinary β2M combined with smoking and education for predicting cognitive impairment in CKD. Urinary β2M and cognitive function negatively correlated with mtDNA content, suggesting that mitochondrial dysfunction is a common pathophysiological mechanism linking CKD and cognitive dysfunction.

The association between βeta 2-microglobulin and bronchopulmonary dysplasia

Abstract Objectives Previous studies showed that increased urinary Beta 2-microglobulin (β2-M) level is associated with fetal inflammatory response and successfully predict bronchopulmonary dysplasia (BPD). We aimed to investigate the clinical utility of serum β2-M levels to predict BPD in preterm infants. Method Infants born between May and November 2018 and whose gestational age (GA) was &lt;32 weeks were included into the study. During routine blood work in the first couple of hours of life an extra 0.5 mL blood was drawn to study β2-M levels later on. β2-M levels were compared between infants who developed BPD or not. Results Data analysis of 111 infants was performed. Out of 111 infants, 37 died before BPD diagnosis and out of the rest 74 infants, 38 (34.2%) were diagnosed with BPD. Mean GA was 28 ± 1.8 and 29.9 ± 1.4 weeks (p &lt; 0.01) and mean birth weights (BW) were 1,086 ± 316 and 1,395 ± 348 g (p &lt; 0.01) in BPD group and without BPD respectively. Demographic characteristics of the two groups were similar. While the white blood cell count, CRP and IL-6 levels were similar in both groups, β2-M levels were significantly higher in BPD group (4.84 ± 1.0 and 3.79 ± 0.95 mg/L, p = 0.01). Furthermore a weak correlation between β2-M level and BPD was observed (r = 0.23, p = 0.04). Conclusion Serum β2-M levels which obtained in the early postnatal life could predict developing BPD. Monitoring β2-M levels in infants who have high clinical risk factors for BPD development may provide additional benefit in predicting BPD.

Evaluation of serum interleukin 2 receptor and beta-2-microglobulin as prognostic factors for canine lymphoma: A pilot study.

Interleukin 2 receptor (IL-2R) is released from activated T cell lymphocytes and related to proliferation of B cells and T cells. Beta-2-microglobulin (B2M) is synthesized from all nucleated cells and constitutes a major histocompatibility complex class I antigen. In human medicine, high concentrations of these two factors have been found to be related to prognosis in aggressive non-Hodgkin's lymphoma. In this pilot study, we aimed to assess the correlation between the serum concentration of IL-2R and B2M and the diagnosis and prognosis of canine lymphoma. This study included 8 healthy dogs and 17 dogs with lymphoma. To measure the serum concentration of IL-2R and B2M, a commercial enzyme-linked immunosorbent assay was used. In dogs with lymphoma, IL-2R concentrations were significantly high at the time of diagnosis, but B2M concentrations were not. In relapsed dogs, both IL-2R and B2M concentrations were significantly higher than those in the control and chemotherapy response groups. When the serum concentrations of IL-2R and B2M during chemotherapy were monitored in four relapsed dogs, B2M levels were more closely related with relapse. This study demonstrated that serum IL-2R and B2M concentration can be a diagnostic or prognostic tool for canine lymphoma. Monitoring of serum B2M concentration seems to be useful for predicting relapse.

Prognostic Value of the Expression of Receptor Tyrosine Kinase-Like Orphan Receptor 1 (ROR-1) in Chronic Lymphocytic Leukemia.

Background: The transmembrane receptor tyrosine kinase-like orphan receptor 1 (ROR1) has acted on the causation and sustentation of mature B-cell lymphomagenesis for chronic lymphocytic leukemia (CLL) cells.The study attempted to show whether there is a relationship between the level of ROR1 surface expression in CLL cells and disease findings. Materials and Methods: The level of ROR1 cell surface expression was determined in accordance with the flow cytometric analysis of CLL patients at the first diagnosis time. Two groups were formed according to the high and low ROR1 levels. The cut-off point for the ROR1 level was calculated for advanced-stage disease using receiver operating characteristic (ROC) curves. A two-sided p-value <0,05 was considered statistically significant. Results: 108 CLL cases with a median age of 60 were enrolled. The median percentage of ROR1 cell surface marker positivity in the CD5/CD19 positive leukemic cell was 62%.The CLL cases with high ROR1 levels have thrombocytopenia (p=0.042), anemia (p=0.028), and high beta-2 microglobulin value ≥3 mg/dL (p=0.002) and the need for first-line treatment (p=0.043). Conclusion: The poor prognostic parameters such as splenomegaly, anemia, higher beta-2 microglobulin levels, intermediate/advanced RAİ stage disease, and need for first-line treatment had associated high-level ROR 1 expression of our CLL patients. It needs to be investigated for its effect on predicting disease burden and aggressiveness with more comprehensive studies on ROR1 expression levels in CLL cases.

The Short-Term Efficacy and Safety of Obinutuzumab Plus Chemotherapy in B-NHL with High Tumor Burden: A Real-World, Retrospective Study

Introduction: The prognosis of patients with B-cell non-Hodgkin's lymphoma (B-NHL) in high tumor burden is significant poor and patients frequently occur tumor lysis syndrome (TLS) during chemotherapy. There are no standard treatment regimens at present, so it is very crucial to explore effective treatment options to improve prognosis. The aim of present study is to analysis the short-term efficacy and safety of obinutuzumab plus chemotherapy in B-NHL with high tumor burden. Methods: We retrospectively investigated the records of 19 patients of B-NHL with high tumor burden and received obinutuzumab treatment at the First Hospital of Jilin University from November 2021 to July 2022, including 9 marginal zone lymphoma (MZL), 7 chronic lymphocytic leukemia /small lymphocytic lymphoma (CLL/SLL), 2 mantle cell lymphoma (MCL), 1 B-prolymphocytic leukemia (B-PLL). All patients received obinutuzumab at a dose of 1000 mg (on days 1, 8, and 15 of cycle 1 and on day 1 of subsequent cycles). The primary endpoint was objective response rates (ORR), and secondary endpoints was incidence of adverse events (AEs). Results: This study enrolled 19 patients completed ≥ 1cycles obinutuzumab plus chemotherapy, including 8 patients with initial treatment and 11 with relapsed and refractory (R/R). Among them, 15 received obinutuzumab with bendamustine (BG), 3 with Bruton tyrosine kinase inhibitors (G-BTKi), 1 with COP (cyclophosphamide, vincristine, and prednisone; G-COP). The median age was 65 years (range, 41-83 years); 12 males and 7 females. In addition, 94.7% patients (18/19) were identified with advanced stage; 73.7% (14/19) with B symptoms; 52.6% (10/19) with ECOG ≥ 2; 57.9% (11/19) with elevated lactate dehydrogenase (LDH) levels; 100% with elevated beta-2 microglobulin (β2MG) levels. 94.7% patients could quickly relieve the high tumor burden status after 1cycle obinutuzumab treatment (Figure 1A). Furthermore, there are 6 patients with hyperleukocytosis (blood cell count greater than 100,000/mL), all reduce to normal level on day 4 of obinutuzumab treatment (Figure 1B); 12 patients with splenomegaly, including 11 massive splenomegaly (long axis >20 cm), all significantly decreased after obinutuzumab treatment, and 7 patients has returned to normal level, and patients with enlarged lymph nodes also significantly reduced after obinutuzumab treatment (Figure 1C). Besides, 8 patients measured minimal residual disease (MRD) in peripheral blood and 6 (75%) achievement of undetectable MRD. Significantly, the ORR was 94.7% (18/19), and 57.9% (11/19) achieved complete remission by evaluation of images. (Figure 1D). The most common AE of any grade were infusion-related reactions 7/19 (36.8%); expiratory dyspnea 2/19 (10.5%); nausea 1/19 (5.3%). In addition, 9/19(47.4%) neutropenia, which may also associate with chemotherapy. There was no incidence of AEs leading to treatment withdrawal or death under obinutuzumab plus chemotherapy treatment. Conclusions: Obinutuzumab could quickly relieve the high tumor burden status and decrease the occurrence of TLS. Thus, obinutuzumab is a promising treatment in patients of B-NHL with high tumor burden. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Obinutuzumab Versus Rituximab Based First-Line Chemoimmunotherapy for Follicular Lymphoma - a Real-World Multicenter Retrospective Cohort Study

Introduction - The recent GALLIUM study showed an advantage of 7% in progression free survival in favor of obinutuzumab (G) vs. rituximab (R) based immunochemotherapies as first line therapy in follicular lymphoma (FL) patients. Yet, the toxicity appears to be increased with obinutuzumab based therapy. Methods - To assess for toxicity we compared treatment for FL in the era before the approval of obinutuzumab by the Israeli health authorities on 1/1/2018 (rituximab-based therapy) to the era post approval (obinutuzumab-based therapy). This is a multicenter retrospective cohort study comprising five medical centers in Israel. All adult FL patients treated with first line chemo-immunotherapy were included. Comparison of G vs. R based chemo-immunotherapy was performed in order to assess real life toxicity profile between the two anti-CD20 monoclonal antibodies (R- and G-groups). Primary outcome was any infection recorded during the induction and 6 months post induction period. Secondary outcomes included rates of febrile neutropenia, severe infections defined as grade 3-4 infections, infections requiring intravenous (IV) antibiotics, life threatening infections or fatal infections, as well as rates of other non-hematological and hematological toxicities and treatment discontinuation. Response rates at the end of induction therapy were also recorded. Results - A total of 156 patients, treated between 1/1/2016-1/1/2019, were included in the analysis - 78 patients in each group. Most patients received bendamustine (n= 92, 59%) or CHOP (n=49, 31.4%) as the adjacent chemotherapy regimen. Baseline characteristics of patients were comparable between groups, apart from a higher incidence of diabetes mellitus and hypertension in the R-group. Any infection occurred in 69 patients (44.2%) of the cohort, and a total of 106 infectious episodes were recorded. Patients in the R- and G-groups had similar rates of any infection (44.8% and 43.5%, p=1), severe infections (43.3% vs. 47.8%, p=0.844), febrile neutropenia (15% vs. 19.6%, p=0.606) and treatment discontinuation (14.1% vs. 19.2%). Characteristics of the 106 infection episodes according to the R- versus G- groups are described in Table 1. Most other adverse events recorded, including hematological and non-hematological toxicities, were also comparable between groups, apart from higher rates of nausea and vomiting (30.7% vs. 11.5%, p=0.029) and fever from a non-infectious origin (32% vs. 10%, p=0.032) in the R-group. Overall, no statistically significant difference was evident in adverse events of grades 3 to 5 (hematological and non-hematological) between R and G-based treatments (76.9% vs. 82%, p=0.427) nor in complete remission rates at the end of induction therapy (79.5% vs. 84.6%, p=0.147). Baseline factors significantly associated with any infection in univariate analysis included female gender, chronic obstructive pulmonary disease, use of GCSF prophylaxis, elevated LDH, beta2 microglobulin and lower levels of hemoglobin before `treatment commencement. None of the parameters was found to be significantly associated with any infection in multivariable analysis, albeit gender and LDH values had a trend towards significance. Conclusions - In this first and largest real-life study of first line treated patients with follicular lymphoma, we did not observe excess toxicity with G- compared to R- based therapy during the induction and 6 months post induction period. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

The Prognostic Significance of Serum Beta-2 Microglobulin (sβ2m) Levels in Patients with Hodgkin Lymphoma (HL): Final Analysis on 915 Patients Treated with ABVD or Equivalent (ABVDeq) Chemotherapy or Combined Modality Therapy (CT/CMT) Focusing to the Determination of Optimal Cut-Offs

Objective/Aim: Sβ2m is a well- established prognostic factor for several hematologic malignancies, but its role in HL is still controversial. We aimed to investigate the prognostic significance of sβ2m levels in a large series of homogenously treated HL patients. Methodology: We analyzed 915 patients with HL treated with ABVDeq (1990-2019) and selected solely based on the availability of pretreatment sβ2m levels. Sβ2m levels (upper normal limit 2.4mg/L) were analyzed in relation to other baseline features and the outcome. Freedom From Progression (FFP) was defined as time between treatment initiation and treatment failure (toxic death, primary refractoriness, PR with switch to alternative chemotherapy or relapse); deaths of unrelated causes were censored. Overall Survival (OS) and Disease-Specific Survival (DSS) were measured from treatment initiation to death of any cause or HL-related causes respectively. Sequential cut-offs (1.8-3.5 by 0.1mg/L increments) were used to explore the potential impact of sβ2m on outcomes. Results: The median serum β2m levels were 2.20mg/L (IQR 1.80-3.00, range 0.50-14.40) and 383/915 patients (42%) had elevated levels (>2.4mg/L). Sβ2m correlated strongly with all baseline features. Univariate Analysis: FFP was significantly inferior in patients with higher β2m at all tested cut-offs. At 2.4 mg/L ("normal versus high") the 10-year FFP was 80% vs 70% (p=0.001) after a median follow-up of 81.1 months. However, the best cut-off was 2.0mg/L [10-year FFP 83% vs 70% (p=0.001)]. A dose-response relationship was seen across quartiles Q1-4 with 10-year FFP 84%,78%,73% and 68% (p=0.001). In early stages (IA/IIA), statistically significant results were obtained at cut-offs between 1.8 and 2.1mg/L. The best cut-off was 1.9mg/L [10-year FFP 88% vs 78% (p=0.003)]. In advanced stages, none of the cut-offs yielded statistically significant results (borderline at 2.0mg/L; 10-year FFP 74% vs 64%, p=0.09). Multivariate Analysis: Sβ2m levels remained significant for FFP after adjustment for IPS factors, ESR and B-symptoms at 2.0mg/L [hazard ratio (HR) 1.55, p=0.01) in the whole series of 915 patients; it was not significant at the "normal versus high" comparison at 2.4mg/L. In early stages, sβ2m was a significant predictor of FFP at both cut-offs of 2.0 mg/L and 2.4 mg/L (HR 1.65, p=0.034 and 1.67, p=0.038). In advanced stages, sβ2m was an independent prognostic factor for FFP at 2.0mg/L (HR 1.44, p=0.098 despite the lack of univariate significance), but not at 2.4mg/L. The 10-year OS and DSS was lower in patients with elevated β2m levels (10-year rates 90% vs 77%, p<0.001 and 93% vs 86%, p=0.002). Similar results to FFP were obtained by multivariate analysis of DSS for all 915 and early-stage patients. Conclusion: Higher sβ2m was a significant independent predictor of FFP in HL but the optimal cut-off appears to lie within the normal limits, performing better than a "normal versus high" evaluation (cut-off 2.4 mg/L) and being 2.0mg/L for the whole series and 1.9 mg/L for early-stage patients. The prognostic significance in advanced stages was weaker (best cut-off 2.0 mg/L). Serum β2m was also highly predictive of OS and DSS. An analysis of corrected serum β2m according to renal function is ongoing.

Idecabtagene Vicleucel Chimeric Antigen Receptor T-Cell Therapy for Relapsed/Refractory Multiple Myeloma with Renal Insufficiency: Real World Experience

Introduction: Idecabtagene vicleucel (ide-cel) is an autologous BCMA directed chimeric antigen receptor therapy T (CAR-T) cell therapy approved for relapsed/refractory multiple myeloma (RRMM). However, patients with renal insufficiency (RI) were excluded from the registrational KarMMa clinical trial. RI impacts a significant proportion of patients with multiple myeloma and the safety and efficacy profile of CAR-T remains unknown in this patient population. We evaluated the real-world outcomes of patients with RI treated with standard of care ide-cel. Methods: RI was defined as creatinine clearance (CrCl) < 50 ml/min at the time of CAR-T therapy. CrCl of < 30 ml/min or being on dialysis was defined as severe RI. Dosing of lymphodepletion chemotherapy and grading/management of toxicities was per institutional guidelines, while responses were graded based on the IMWG response criteria. Results: The study cohort includes 211 patients receiving ide-cel from 11 medical centers, of which 28 (13%) patients had RI at the time of CAR-T therapy with CrCl < 50 ml/min. Among these, 11 (39%) patients had severe RI including one patient on dialysis. Table 1 describes baseline and treatment characteristics of patients with and without RI. Patients with RI were older (69 vs 63 years, p=0.003) and more likely to be female (68% vs 35%, p=0.001). They had lower albumin, higher beta-2-microglobulin (B2M) and higher likelihood of having revised ISS stage 3 disease, driven primarily by high B2M levels. Median prior lines of therapy in patients with and without RI was 8 vs 6, respectively. Fludarabine was dose reduced in 82% patients with RI and amongst this group 61% of patients had > 20% dose reduction. There was no difference in CAR-T cell dose infused (median: 416 vs 408 million cells). Cytokine release syndrome (CRS) was seen in 89% vs 84%, p=0.7 of patients with and without RI, including grade ≥ 2 CRS in 14% vs 20%, p=0.6 of patients, respectively. Neurotoxicity was observed in 21% vs 19%, p=0.7 of patients, respectively. Patients with RI had a longer hospital stay (median 13 vs 9 days, p=0.05), although ICU admission rates were similar (14% vs 7%, p=0.3). Infections were seen in 43% vs 32%, p=0.2 of patients, respectively. At one month, any ≥ grade 3 cytopenia was more common in patients with RI (86% vs 55%, p=0.001), including grade ≥ 3 neutropenia (54% vs 33%, p=0.04) and grade ≥ 3 thrombocytopenia (75% vs 41%, p<0.001). By 90 days post CAR-T, there was no difference in > grade 3 cytopenias amongst the 2 groups. Renal function did not significantly change after CAR-T in most patients. Amongst patients with paired baseline and day 30 data, no patient with CrCl 30-49 ml/min experienced worsening of CrCl to < 30 ml/min at day 30. Amongst 10 patients with CrCl < 30 ml/min, 3 patients experienced improvement in CrCl to 30-49 ml/min, while 7 patients had similar CrCl at day 30. New need for dialysis post CAR-T was observed in two patients, both with baseline CrCl > 50 ml/min; in one patient at 1 week post CAR-T in the setting of critical illness/severe CRS and in another patient 8 months post CAR-T in the setting of salvage therapy for relapse and critical illness. 34 patients have died by last follow-up, of which 8 were due to CAR-T related toxicities including infections (1 patient with RI, 7 without RI, p=0.9). 189 patients were evaluable for response. Patients with RI had an overall response rate of 96% comparted to 83% in patients without RI, p=0.045. Complete response (58% vs 65%, p=0.1) and very good partial response rates (77% vs 65%, p=0.2) were similar. At a median follow-up of 6 months, the median progression-free survival (PFS) in the two groups was 6.5 vs 8.1 months, p=0.6 (Fig 1). On multivariable analysis incorporating RI, age, cytogenetics and prior BCMA-directed therapy, RI was not an independent predictor for PFS (HR 1.4, 95% CI: 0.7-2.7, p=0.4), while high-risk disease, prior BCMA therapy, and age < 65 years were independent adverse prognostic factors. Conclusions: This multicenter retrospective study supports the feasibility of ide-cel CAR-T cell therapy in RRMM patients with RI. CRS, neurotoxicity and non-relapse mortality were comparable in patients with or without RI, although patients with RI had a longer hospital stay and were more likely to experience short-term high-grade cytopenias. Importantly, patients with RI experienced similar benefit in terms of response rates and PFS. S.S & L.P: co-first authors. L.S, K.K.P & D.K.H: joint senior authors Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

IgD Multiple Myeloma, Does It Still Have a Poor Prognosis?

IgD Multiple myeloma is considered unusual myeloma, with just 2% of cases of MM being diagnosed as IgD subtype. The infrequency of its presentation and the poor prognosis make the disease challenging to treat, with many patients being selected for clinical trials. A 63-year-old female presented to the ER after falling on ice. After undergoing a CT Head, multiple lytic lesions in the occipital region were found, confirmed by MRI. 2 months later, a bone marrow biopsy found 90% of marrow replaced by CD138 plasma cells, positive for kappa & negative for lambda, consistent with myeloma. On evaluation, she was found to have pan-hypo-gammglobulinemia. However, the skeletal survey did not find any other lytic lesions. Seeing this, IgD levels were measured and found to be >100 times over the upper normal limit with normal beta2-microglobulin levels, confirming monoclonal gammopathy. A FISH study found the patient positive for t(11;14)/CCND1IGH translocation (85% of cells) and negative for TP53. Palliative radiotherapy for the skull base was performed before the patient started chemotherapy. The patient was deemed a suitable candidate, selected for a clinical trial, and placed on RVd [revlimid + velcade + dexamethasone] with delayed bone marrow transplant. As the patient developed a rash and neuropathy from velcade [bortezomib], it was paused and restarted after three months. The patient was doing well on this chemotherapy regimen for two years and was placed on lenalidomide-based maintenance therapy. Two years later, her disease started progressing with an increased frequency of bone pains and opioid dosing. At this juncture, it was decided to change course and start her on pomalidomide/dexamethasone. This did not help the patient, and the neurological side effects of dexamethasone made us change course and initiate a daratumumab, bortezomib, cyclophosphamide and dexamethasone. This proved unfruitful, too, as the patient developed adverse effects of polyneuropathy, pancytopenia requiring transfusion, and worsening GERD. Seeing the disease relapse and intolerance to the previous regimen, we decided to initiate a once-per-week regimen of selinexor, bortezomib, and dexamethasone. The patient has improved on this regimen and is back to her baseline for physical activity, mental status and pain, and an improved appetite. In this case, we wanted to highlight how incidental findings on a CT lead us to diagnose a rare type of multiple myeloma and how, in the era of novel agents, the survival of patients can be prolonged significantly. Considering the rarity of this subtype, international collaborative studies are suggested to further elucidate the underlying mechanisms for developing potent therapeutic approaches. Most of the data reported on how IgD multiple myeloma progresses was written before the arrival of novel agents. More evaluation of survival times and progression of the disease needs to be done as it's possible that IgD MM is not as poor a prognosis as it once was.

Serum beta2-microglobulin level in systemic lupus erythematosus patients: Relation to disease activity

Aim of the workTo assess the level of β2-microglubulin (β2M) in systemic lupus erythematosus (SLE) patients and its association with disease activity and other disease parameters. Patients and methods40 SLE patients and 22 matched controls were studied. Serum β2M was assessed using enzyme-linked immunosorbent assay (ELISA). SLE Disease Activity Index (SLEDAI) and the damage index were assessed. ResultsThe patients were 36 females and 4 males (F:M 9:1) with a mean age of 28.5 ± 7.9 years and disease duration of 6.7 ± 3.3 years. The SLEDAI was 9.3 ± 5.2 and the damage index 1.83 ± 1.84. The mean level of serum β2M was significantly higher in SLE patients (6.42 ± 2.46 mg/L) than control (2.47 ± 0.4 mg/L) (p < 0.01).The serum level of β2M was significantly higher in patients with nephritis (n = 22) (7.45 ± 2.47 mg/L) compared to those without (n = 18) (5.17 ± 1.82 mg/L)(p = 0.002), And it was similar in those with and without arthritis (7.24 ± 2.3 mg/L vs 5.88 ± 2.4 mg/L (p0.07).The β2M significantly correlated with disease activity (r = 0.86, p 0.001), serum creatinine (r = 0.52, p > 0.001), urea (r = 0.63, p < 0.001), 24 h urinary protein (r = 0.56, p < 0.001), hematuria (r = 0.4, p < 0.01) and pyuria (r = 0.41; p < 0.01), ESR (r = 0.48; p < 0.01) and inversely with hemoglobin level (r = −0.34; p = 0.03). No significant correlation was found with C-reactive protein or with disease damage. Serum (β2M) significantly predicted nephritis and disease activity (sensitivity 63.6 %, specificity 77.8 %; p < 0.001 and 95 %CI: 0.25–0.41; p < 0.001 respectively). ConclusionSerum β2M is significantly associated with disease activity and lupus nephritis, suggesting that serum β2M may serve as a potential biomarker to monitor the disease activity and predicting lupus nephritis. However its association to disease severity needs further longitudinal studies.