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Introduction: DLBCL presents clinical, biological, and prognostic heterogeneity. The composition of the tumor TME strongly contributes for its biological diversity, being able to predict treatment response and determine prognosis. DLBCL with TME enriched in CD8+T-lymphocytes and NK cells demonstrate increased OS compared to those with TME enriched in histiocytes. Data from patients with solid tumors have established associations between absolute quantification of lymphocyte subpopulations (LS) in peripheral blood (PB) and prognosis. Therefore, it is hypothesized that the cellular constituents of PB may mirror the TME, and their quantification may constitute an easy and fast tool to access biological biomarkers predictive of therapeutic response and prognosis in malignancies. This study aims to determine the absolute quantification of LS in the PB of DLBCL patients, seeking associations with outcomes and specific clinical phenotypes. Methods: This study involved 54 newly-diagnosed DLBCL patients. PB collection was performed before starting any therapy, for blood count and immunophenotyping on a BD Facs Calibur flow cytometer. End points included OS and EFS. Survival curves were constructed using the KM method. The Mann-Whitney test and the Kruskal-Wallis test were used to establish the relationship between the lymphocyte profile, clinical variables, therapeutic response and outcomes. The Contal O'Quingley test was used to determine cut-offs for each LS capable of discriminating outcomes. Univariate analysis was performed using the Cox method and a p-value ≤ 0.05 was considered statistically significant. Results: The median age at diagnosis was 54 years and 55.6% were female. Clinical stage III/IV was observed in 77.8%, 57.4% had bulky ≥ 7 cm, 63.0% had B-symptoms, and 53.7% presented IPI ≥ 3. The ORR was 80.8%. With a median follow-up of 21.3 months, the estimated 2-year OS and EFS were 73.3% and 64.7%. The median absolute values obtained for the PB lymphocyte subpopulations were: 1026 cells for CD3+ T-lymphocytes, 591 cells for CD4+ T-helper lymphocytes, 412 cells for CD8+ T-cytotoxic lymphocytes, 1.6 for CD4/CD8 ratio, 240 cells for T-reg lymphocytes, 112 cells for LG T-lymphocytes, 173 cells for NK cells, and 70 cells for B-lymphocytes. In univariate analysis, using a cut-off value = 367 CD8+T-cells/mm3 and 141 NK-cells/mm3, cytotoxic T-lymphodepletion was associated with decreased OS (p = 0.005) and EFS (p < 0.001), as well as NK-lymphodepletion was associated with decreased EFS (p = 0.033), but not with shortened OS (p = 0.240). DLBCL patients presenting higher NK-cell counts had lower rates of chemoresistance to the R-CHOP (p = 0.024), and those with higher NK-cell counts (p = 0.033) and higher CD8+T-lymphocyte counts (p = 0.039) had lower probability to develop relapse, progression or death, confirming the central role of these PB cellular constituents in anti-tumor immunity. We observed that B-cell lymphodepletion was associated with an adverse clinical phenotype, including higher frequency of bulky disease (p = 0.009), B-symptoms (p < 0.001) and advanced clinical stage (p = 0.033). Conclusion: In newly-diagnosed DLBCL, the lymphodepletion of CD8+T-cells and NK cells quantified in PB were associated with decreased OS and EFS. Similarly, peripheral T-cytotoxic and NK lymphodepletion were associated with absence of response to the R-CHOP regimen, as well as higher probability of relapse, progression or death. Therefore, the absolute quantification of these LS in PB may be considered potential biomarkers capable of predicting response and prognosis in DLBCL. Also, we believe it may mirror the cellular composition of the immune TME.

Introduction: DLBCL commonly affects the older population. Elderly patients often present high-risk molecular profile, lower tolerability to chemotherapy, and poor outcomes. In this setting, attenuated therapeutic regimens, such as R-miniCHOP and R-miniCHOP of the elderly, have emerged for this fragile population. Albeit its relevance, response rates, clinical outcomes, and toxicities of these regimens remain poorly understood, because these individuals are usually excluded from controlled trials. Therefore, this study aims to assess the outcomes, determine survival predictors, and compare responses and toxicities between R-CHOP, R-miniCHOP, and R-miniCHOP of the elderly protocols in the management of elderly DLBCL patients. Methods: This retrospective, observational, and single-center study involved 185 DLBCL patients older than 70 years, treated at USP, from 2009 to 2020. Endpoints included ORR, OS, PFS. Survival curves were constructed using the KM method and the Log-Rank test was used to assess the relationship between variables and outcomes. The chi-square test and the Kruskal-Wallis test were applied to assess statistically significant differences in clinical characteristics, adverse event profile, and responses between different treatment modalities. Univariate analysis was performed using the Cox test and multivariate analysis by Cox regression method. The results were presented in HR and 95% CI, and a p-value ≤0.05 was considered significant. Results: The median age at diagnosis was 75 years, and 58.9% were female. Comorbidities were prevalent, including 19.5% with immobility, 28.1% with malnutrition and 24.8% with polypharmacy. Advanced clinical stage (III/IV) was observed in 72.4%, 48.6% had bulky ≥ 7 cm, 63.2% had B-symptoms, and 67.0% presented IPI ≥ 3. Among the 182 (98.4%) effectively treated cases, 57.1% received R-CHOP, 18.0% R-miniCHOP, 13.2% R-miniCHOP of the elderly, and 1.7% were palliated using R-CVP. The ORR for the whole cohort was 68.1%, with CR achieved in 65.9%. ORR was 72.1% for R-CHOP, 70.6% for R-miniCHOP, and 45.6% for R-miniCHOP of the elderly, p = 0.040. Although R-miniCHOP of the elderly regimen promoted lower ORR, patients in this group had higher rates of unfavorable clinical-laboratory findings, including hypoalbuminemia (p = 0.001), IPI ≥ 3 (p = 0.013), and NCCN-IPI ≥ 3 (p = 0.002). With a median follow-up of 6.3 years, the estimated 5-year OS and PFS were 50.2% and 44.6%, respectively. The estimated 2-year OS was 82.0% for R-CHOP, 67.5% for R-miniCHOP, and 48.1% for R-miniCHOP of the elderly, p = 0.003. The estimated 2-year PFS was 61.1% for R-CHOP, 56.4% for R-miniCHOP, and 20.5% for R-miniCHOP of the elderly, p = 0.005. Although correlated with increased OS and PFS in comparison to attenuated protocols, R-CHOP regimen was associated with higher rates of severe neutropenia (p = 0.003), but not translated into febrile neutropenia (p = 0.907), therapy interruption (p = 0.671), or higher early mortality rates (p = 0.681). In multivariate analysis, age≥75 years (HR: 2.08, p = 0.001), neutrophilia (HR: 2.18, p = 0.007), low lymphocyte/monocyte ratio (HR: 1.98, p = 0.010), and clinical stage III/IV (HR: 2.36, p = 0.003) were predictors of decreased OS. Conclusion: In this large and real-life LATAM cohort, we demonstrated that DLBCL patients older than 70 years still do not have satisfactory outcomes, with half of cases not reaching 5 years of life expectancy after diagnosis. Although a significant portion of older DLBCL patients is highly fragile and ineligible for enhanced regimens, attenuated anthracycle-based protocols promoted remarkably inferior outcomes compared to those achieved by the R-CHOP.

Introduction: FL has heterogeneous biological behavior, variable clinical outcomes, and a natural history marked by histological transformation (HT). The management of FL and its outcomes are highly dependent on staging (CS) and tumor burden (TB). The identification of new molecular biomarkers is crucial, not only to understand FL's biology, but also, can guide better prognostic stratification, and determine new therapeutic targets. This study aims to outline the GEP in tissue samples from FL categorized by CS and TB using a large-scale gene microarray platform with subsequent expansion by RT-qPCR. Methods: This study involved 219 FL cases. FL patients were categorized into early-stage disease (I/II) (ES), advanced-stage (AS) (III/IV) with low tumor burden (LTB), and AS with high tumor burden (HTB) according to GELF criteria. The experimental phase involved 126 FFPE samples. Firstly, 42 samples (12 ES, 11 AS-LTV, 12 AS-HTV, 6 control lymph nodes and 1 control HELA) were submitted to the array using the Clariom D Array Human platform. The identification of differentially expressed genes (DEGs) between the 4 groups used the Transcriptome Analysis Console by the eBay ANOVA method. Bidimensional PCA sought to identify discriminatory DEGs clusters. A comparison between pairs was subsequently performed. The 7 most discriminatory DEGs were selected for a cohort expansion by RT-qPCR. Results: According to CS and TB, 14.6% of FL cases were categorized as ES, 17.8% as AS-LTB, and 67.6% as AS-HTB. With a median follow-up of 80.7 months, the median OS was 208.2, 130.5, and 126.3 months for ES, AS-LTB, and AS-HTB,p = 0.249. The median PFS was 113.3, 110.7, and 59.5 months, for the same groups, p = 0.018. The array identified 135,750 genes comparisons between the control, ES, AS-LTB, and AS-HTB. PCA analysis was not able to identify discriminatory DEGs clusters among the 4 groups. Pairwise comparative analysis identified 1913 DEGs among the 4 groups. Of these, 1759 (91.9%) were DEGs between FL and healthy controls, but only 218 (11.3%) were DEGs among the 3 different FL subgroups, evidencing a low molecular diversity in FL when categorized by CS and TB. In the cohort expansion by qRT-PCR, 7 pre-selected DEGs based on the microarray were tested, including the genes GNG7, ITGAM, AZN1, TIMD4, IL7R, PTGS2, and MTTL10. The comparison between the median expression of these genes among the 3 FL's groups showed that the expression of the ILR7 was downregulated with the progression of the disease, with marked hypoexpression in AS-HTB patients. These data may suggest downregulation of IL7R gene in the FLs natural history. We found that hypoexpression of the GNG7 (p = 0.024) and PTGS2 (p = 0.090) were associated with higher rates of disease progression/relapse, and hypoexpression of the GNG7 (p = 0.006) was associated with a higher probability of HT, highlighting the potential prognostic impact of these molecular biomarkers. Finally, we established phenotypic-genotypic associations, as hypoexpression of ITGAM was associated with higher rates of B-symptoms (p = 0.017) and bulky disease (p = 0.026), as well as hypoexpression of IL7R was correlated with a higher probability of bulky (p = 0.025) and extranodal involvement (p = 0.016). Conclusion: Although CS and TB are strong determinants of the prognostic heterogeneity of FL, this study did not find high molecular diversity by this categorization. We observed a relatively homogeneous GEP with low biological diversity in FL presenting different disease evolution phases. Moreover, new molecular biomarkers potentially implicated in FLs natural history, prognostication and specific clinical phenotypes were revealed.

Unsupervised machine learning techniques are employed to understand patterns and behaviors of variables in databases. Multiple Correspondence Analysis (MCA), an extension of correspondence analysis, can be used to verify the association of categorical variables and their categories. In this study, we evaluated the relationship between clinical-demographic variables of 154 patients with nodal peripheral T-cell lymphoma (PTCL) to understand how these variables relate to the unfavorable clinical outcome, death. Methodology: MCA was used to reduce the dimensionality of the real world database by creating two dimensions, 1 and 2, which were subsequently categorized into group 1 and group 2. Dimension 1, comprising ECOG, IPI, treatment, overall response, remission, and bone marrow transplant, represented about 30% of the total observed variance. Survival analysis was conducted to assess the association between these groups and overall survival (OS) and mortality rate. Results: The categories of dimension 1, group 1 and group 2, were associated with OS and mortality rate, with group 1 being associated with 5.5 months (95% CI: 2.70 – 8.40) of OS, while group 2 had a survival time of 277.20 months (95% CI: 91.21 – 463.12). Moreover, the mortality rate in group 1 was 87% (n = 67) and in group 2 it was 36% (n = 28) (p < 0.001). The risk of death for group 1 was 11.11 times (95% CI: 9.75-18.30; β= 2.41; p < 0.001). Discussion: These findings indicate a significant disparity in survival outcomes based on the clinical-demographic profiles of the patients. Group 1, characterized by poorer clinical indicators, exhibited substantially shorter OS and higher mortality rates. The ability of MCA to effectively reduce dimensionality while preserving the clinical relevance of the variables underscores its utility in identifying key prognostic factors. Conclusion: The MCA technique was effective in reducing the dimensionality of the database, maintaining the clinical characteristics of the variables robustly for use in Cox regression. This approach can aid in the identification of high-risk patient groups and inform treatment strategies aimed at improving clinical outcomes.

Introduction: Peripheral T-cell lymphomas (PTCL) are a rare group of malignancies derived from mature T-cells or natural killer cells, characterized by significant clinical-biological heterogeneity despite being grouped together. These entities have unique clinical, morphological, phenotypic, and molecular signatures. The predominantly nodal presentation group includes the histological variants ALK-positive anaplastic large cell lymphoma (ALK1+ ALCL), ALK-negative anaplastic large cell lymphoma (ALK1- ALCL), angioimmunoblastic T-cell lymphoma (AITL), and PTCL, not otherwise specified (PTCL-NOS). The central pillar of treatment for this subgroup of lymphomas is based on anthracycline-containing polychemotherapy regimens and upfront consolidation with autologous hematopoietic stem cell transplantation, although unfavorable outcomes such as primary refractory disease and early relapses are still common, except for the ALK1+ ALCL subgroup. Methodology: Hence, a multicenter, real-world, ambispective study was conducted, involving reference institutions in Brazil and two in the USA, to evaluate the main clinical-demographic characteristics associated with the outcomes of 179 patients diagnosed with nodal PTCL. Results and discussion: Individuals with nodal PTCL often presented at diagnosis with unfavorable clinical features, including a high frequency of advanced disease (stages III and IV), high tumor burden, poor performance status (ECOG ≥ 2 points), and frequent allocation in high-risk categories of the IPI and PIT prognostic indices. Patients with nodal PTCL included in this study globally exhibited low Complete Response rates, as well as shortened Overall Survival (OS) and Progression-Free Survival (PFS). In this cohort, we did not identify a robust survival benefit from the intensification of the CHOP regimen with the addition of etoposide (CHOEP protocol) or with BV-CHP. Age ≥ 60 years, ECOG ≥ 2, and lack of complete remission after primary therapy centered on CHOP-like chemotherapy were identified as independent prognostic factors associated with worse survival in this cohort represented by the four main histopathological subtypes of nodal PTCL. Conclusion: This analysis demonstrated a high frequency of unfavorable clinical features and poor OS and PFS outcomes in patients with nodal PTCL, regardless of the frontline chemotherapy regimen.

Introduction: Circulating free DNA (cfDNA), particularly circulating tumor DNA (ctDNA), has emerged as a pivotal tool for diagnosis, response assessment, and monitoring of various malignancies. However, in most cancers, it has yet to be incorporated into clinical practice due to the lack of reproducible and widely accessible methodological standardization. In this context, numerous studies have been conducted to achieve such standardization. The purpose of this study was to evaluate the quantification of cfDNA and the identification of plasma ctDNA using the Z-scan technique in patients with diffuse large B-cell lymphoma (DLBCL). Methods: Peripheral blood samples were prospectively collected from DLBCL patients at diagnosis, after the fourth treatment cycle, and at the end of R-CHOP-like treatment. Simultaneously, samples from healthy individuals matched by sex and age were collected. After obtaining cfDNA, its concentration was assessed using Nanodrop, Qubit, Bioanalyzer, and Z-scan equipment. The results were compared and correlated with the clinical characteristics of the patients. Results: From January 2018 to December 2022, 54 DLBCL patients were recruited, with a median age of 58 years (ranging from 21 to 91 years), 33 (62.9%) of whom were female. The overall response rate of the cohort was 68.52%, with a median follow-up of 24.6 months, showing an overall survival of 32 months and event-free survival of six months. The concentrations of cfDNA varied significantly among the different reading methodologies used. The peak-to-valley distance (Theta values - Θ) of cfDNA from patients at diagnosis (mean of 0.438) obtained in the Z-scan was statistically distinct between the control group (background) and patients at different collection points. The peak-to-valley distance (Θ) of cfDNA samples collected at diagnosis was higher than that of the control group (background), p = 0.068, those collected after cycle 4 (mean 0.33), p = 0.016, and after treatment completion (mean 0.33), p = 0.0005. Several cut-off points for “Θ” were tested to identify the best discriminatory value between DLBCL and healthy control, with a sensitivity of 64.71% (95% CI: 50.99% to 76.37%) and specificity of 86.61% (95% CI: 62.86% to 93.02%) for Θ > 0.406. No correlations were found between “Θ” values and the clinical and demographic characteristics of the patients. Conclusion: The Z-Scan technique was able to distinguish DLBCL patients from healthy controls, as well as samples obtained during and after treatment from those collected before the start of anti-lymphoma therapy. We hypothesize that this discrimination was possible because the Z-scan technique may have been capable of distinguishing ctDNA from cfDNA.

Introduction: Infection with Human T-Lymphotropic Virus Type 1 (HTLV-1) is endemic in various regions worldwide. Brazil has an estimated 800,000 to 2.5 million infected individuals. This virus is associated with a broad spectrum of clinical manifestations, ranging from asymptomatic infection to severe diseases such as tropical spastic paraparesis/HTLV-1-associated myelopathy (HAM/TSP) and adult T-cell leukemia/lymphoma (ATLL). Both conditions are linked to high morbidity and mortality. HTLV-1 can infect and alter the function of immune system cells, particularly CD4+ T lymphocytes, causing abnormalities in genes involved in cellular senescence, proliferation, and apoptosis. This study aimed to evaluate the differential expression of genes regulating senescence and cellular proliferation in asymptomatic HTLV-1 carriers. Methods: Peripheral blood samples were collected from healthy individuals (controls), asymptomatic HTLV-1 carriers, and ATLL patients. Gene expression analysis was performed using RNA extracted from CD4+ T lymphocytes separated by magnetic microbeads. The extracted RNA was converted into cDNA for quantitative real-time polymerase chain reaction (qRT-PCR). Results: A total of 32 asymptomatic HTLV-1 carriers, 30 healthy controls, and 7 ATLL cases were analyzed (N = 69). Of the 32 asymptomatic HTLV-1 carriers, 22 (68.8%) were female; in the control group, 16 (53.3%) were female, and in the ATLL group, 5 (71.4%) were female. The age of asymptomatic HTLV-1 carriers ranged from 20 to 87 years (median 58 years), from 31 to 66 years (median 52 years) in the control group, and from 43 to 79 years (median 64 years) in the ATLL group. Three (43%) patients were classified as having smouldering ATLL, and 1 (14.3%) as acute ATLL. The genes CDKN1A, PIK3CA, and RBL2 were underexpressed in asymptomatic HTLV-1 carriers. ATLL patients exhibited underexpression of CDKN1A and overexpression of COL3A1. Conclusion: Asymptomatic HTLV-1 carriers showed underexpression of CDKN1A, PIK3CA, and RBL2 in CD4+ T lymphocytes. CDKN1A was also significantly underexpressed in ATLL. Asymptomatic carriers and ATLL patients did not show differential expression of the genes AKT1, ID1, IFNG, NFKB1, PTEN, TERT, TWIST1, and SPARC. COL3A1 was overexpressed in ATLL patients.

ABCs and LRP transport proteins are involved in several metabolic processes, among which the transport of lipids and xenobiotics can be highlighted. Thus, the overexpression of some of these proteins, such as ABC-G2 and the LRP protein, are associated with the phenotype of resistance to multiple drugs, one of the main causes of relapses and relapses in patients with acute myeloid leukemia. Mesenchymal stem cells (MSC) are multipotent cells present in the bone marrow and are part of the hematopoietic niche. Phenotypic changes in MSCs are related to AML progression. The oxysterols, 7-KC and Triol, are derived from the auto-oxidation of the cholesterol molecule; and have functions in cell signaling, survival and death. Therefore, the objective of this work was to evaluate the action of oxysterols, in subtoxic concentrations, of 7-KC (30 μM, 50 μM and 70 μM) and Triol (20 μM, 30 μM and 40 μM) on the ABCs and LRP transport proteins of MSCs derived from bone marrow from patients with AML (MSC-D), being used as a comparator MSC of bone marrow from healthy individuals (MSC-S). Using the indirect immunofluorescence technique, it was observed that the amount of ABC-A1 protein was higher in MSC-S strains and that treatment with 30 μM of 7-KC increased ABC-A1 only in MSC-S strains. On the other hand, treatment with Triol decreased the concentration of ABC-A1 in MSC-S strains, and this decrease in ABC-A1 was associated with a decrease in the amount of the LXR-β receptor. Furthermore, treatment with 70 μM of 7-KC decreased LRP protein expression in MSC-S strains, while in MSC-D strains there was no difference. Treatment with Triol showed no significant effect on LRP protein expression in MSC-S, only in MSC-D strains, where there was an increase in the concentration of 20 μM, followed by a decrease in the concentrations of 30 μM and 40 μM of Triol. Both 7-KC and Triol treatment increased the concentration of SMO protein in the cytoplasm and its translocation to the nucleus in MSC-S cell lines. This effect was not observed in MSC-D strains. Treatment with Triol increased the concentration of ABC-D4 protein in the cytoplasm. Furthermore, the presence of ABC-D4 protein was observed in the nucleus of MSC-S and MSC-D strains. Our results demonstrated that in the MSCs lineages the effect of 7-KC and Triol is mainly metabolic, in which an increase in the oxidized glutathione content, alteration in the processes of mitochondrial dynamics with induction of genes related to the fission and fusion process in both 7-KC and Triol-treated MSC lines. Increased expression of the BCL2 gene and cytoplasmic survivin was also observed in MSCs treated with 7-KC. In conclusion, it is possible to observe differences in the expression of ABC transporters and LRP between CTM-S and CTM-D strains. Furthermore, at the same concentrations of 7-KC and Triol used, the observed effect was different between MSC-S and MSC-D, which demonstrates that there are different mechanisms between mesenchymal stem cell lines. In summary, our results are characteristic of the metabolic action of the oxysterols 7-KC and Triol on MSCs.

Introduction: Mantle cell lymphoma (MCL) is a rare malignancy with heterogeneous behavior. Despite the therapeutic advances recently achieved, MCL remains incurable. Currently, the standard of care for young and fit patients involves induction immunochemotherapy followed by up-front ASCT. However, the role of more intensive induction regimens, such as those based on high-doses of cytarabine (HDAC) remains controversial in the management of ASCT-eligible patients. Based on this premise, the current study aims to describe clinical and laboratory characteristics, assess outcomes, determine survival predictors, and compare responses between different primary therapeutic strategies, with focus on assessing the impact of HDAC-based regimens on outcomes in ASCT-eligible patients. Methods: This retrospective, observational, and single-center study involved 165 MCL patients, diagnosed and treated at the University of São Paulo, Brazil, from 2010 to 2022. Endpoints included overall OS, EFS, and POD-24 from up-front treatment. Survival curves were constructed using the Kaplan-Meier method and the Log-Rank test was used to assess the relationship between variables and outcomes. Univariate analysis was performed using the semi-parametric Cox test and multivariate analysis by Cox regression method or proportional ratios model. The results were presented in HR and 95% CI, and a p-value ≤ 0.05 was considered statistically significant. Results: The median age at diagnosis was 65 years and 73.9% were male. More than 90% of cases had a classic nodal variant (cnMCL), 76.4% had bone marrow infiltration, and 56.4% presented splenomegaly. Bulky ≥ 7 cm, B-symptoms, ECOG ≥ 2, and advanced-stage III/IV were observed in 32.7%, 64.8%, 32.1%, and 95.8%, respectively. Sixty-four percent of patients were categorized as high-risk according to MIPI score. With a median follow-up of 71.1 months (95% CI: 61.8-80.4) the estimated 2-year OS and EFS were 64.1% (95% CI: 56.2-71.9%) and 31.8% (95% CI: 23.0-40.6%), respectively. The ORR for the whole cohort was 64.7% (95% CI: 57.1-71.7%), with CR achieved in 41.5% (95% CI: 34.1-49.1%). POD-24 was observed in 46.7% (95% CI: 39.1-54.3%), and the overall mortality rate during all follow-up was 58.2% (95% CI: 50.6-65.5%), with disease progression being the main cause of death. Patients treated with R-HDAC-based regimens (43.4%-66/152) had higher ORR (85.9% vs 65.7%, p = 0.007) compared to those receiving R-CHOP (46.0%-70/152), as well as lower POD-24 rates (61.9% vs 80.4%, p = 0.043) and lower mortality (43.9% vs 68.6%, p = 0.004). However, intensified induction regimens using R-HDAC were not associated with a real OS benefit in MCL patients undergoing up-front consolidation with ASCT (2-year OS: 88.7% for R-HDAC plus ASCT vs 78.8% for R-CHOP plus ASCT, p = 0.289). Furthermore, up-front ASCT was independently associated with increased OS (p < 0.001), EFS (p = 0.005), and lower POD-24 rates (p < 0.001) in MCL. Additionally, CNS involvement (HR: 3.12, p = 0.004), tumor lysis syndrome (HR: 2.79, p = 0.026), albumin < 3.5 g/dL (HR: 2.69, p < 0.001), and failure to achieve remission after induction therapy (HR: 3.71, p < 0.001) were predictors of poor OS. Conclusion: In the largest Latin American cohort of MCL reported to date, we confirmed the OS benefit promoted by up-front consolidation with ASCT in young anf fit patients, regardless of the intensity of the immunochemotherapy regimen used in the pre-ASCT induction. Although HDAC-based regimens were not associated with an unequivocal increase in OS, it was associated with higher ORR and lower rates of early-relapses.

Introduction: FL is the most common low-grade B-cell NHL. Although indolent, it has heterogeneous biological behavior and variable clinical outcomes. Histologic transformation (HT) to high-grade B-cell NHL is associated with dismal survival. The therapeutic management of FL and its prognosis are highly dependent on its staging and tumor burden. Here we aimed to describe clinical characteristics, assess outcomes, determine predictors of survival and HT, and compare responses between different therapies applied in a large cohort of FL patients. Methods: This retrospective study involved 223 patients with FL G1-3A, diagnosed at USP, Brazil, from 2006 to 2022. FL patients were categorized into early-stage (I/II) (ES), advanced-stage (AS) (III/IV) with low tumor burden (LTV), and AS with high tumor burden (HTV) according to the GELF criteria. Endpoints were OS, PFS, early-relapse (< 24 months) and HT rates. Survival curves were constructed using the KM method and the Log-Rank test was used to assess the relationship between variables and outcomes. Univariate and multivariate analysis were performed. The results were presented in HR and 95% CI and p ≤ 0.05 was considered significant. Results: The median age at diagnosis was 60 years and 54.8% were female. 15% of cases had ES, 18.4% had AS-LTV, and 66.8% had AS-HTV. BM involvement, leukemic presentation, splenomegaly, and serous effusions occurred in 48.9%, 11.7%, 12.6%, and 21.1%, respectively. Bulky ≥ 7 cm, B-symptoms, ECOG ≥ 2, and involvement of ≥ 4 nodal areas were observed in 44.4%, 51.6%, 15.2%, and 56%, respectively. 49% of patients were categorized as high-risk according to the FLIPI score. With a median follow-up of 79.5 months (95% CI 67.0-92.0) the OS medians were 18 years (95% CI 14.9-21.1), 11 years (95% CI 9.2-12.9), and 10.5 years (95% CI 9.3-11.8) for ES, AS-LTV, and AS-HTV, respectively, p = 0.138. Similarly, the PFS medians were 6.7 years (95% CI 3.6-9.8), 8.4 years (95% CI 2.0-14.8), and 3.5 years (95 CI 1.8-5.3) for ES, AS-LTV, and AS-HTV, respectively, p = 0.171. Early-relapses occurred in 36.3% of cases, being 18.2%, 31.7%, and 41.0% for ES, AS-LTV, and AS-HTV, respectively, p = 0.032. The overall mortality rate was 29.1% for the whole cohort. HT was documented in 16.7% of cases, being 3.0%, 14.6% and 20.1% for ES, AS-LTV, and AS-HTV, respectively, p = 0.053. Among patients with AS-HTV, the R-CHOP regimen did not promote increased OS compared to the R-CVP regimen (p = 0.415), however it was associated with a substantial increase in PFS (p = 0.005). Age ≥ 60 years (HR 1.06, p < 0.001), ≥ 2 comorbidities (HR 3.85, p = 0.014), B-symptoms (HR 2.35, p = 0.015), HT (HR 2.77, p = 0.002) and thrombocytopenia (HR 2.96, p = 0.028) were predictors of poor OS. Similarly, age ≥ 60 years (HR 1.08, p < 0.001), involvelment of ≥ 4 nodal areas (HR 1.27, p < 0.001), and high LDH (HR 1.65, p = 0.002) predicted decreased PFS. Serous effusions (HR 2.09, p = 0.05), albumin < 3.5 g/dL (HR 2.82, p = 0.05), B-symptoms (HR 2.00, p = 0.006), involvement of ≥ 4 nodal areas (HR 1.21, p = 0.014), and BM infiltration (HR 2.11, p = 0.05) were the main predictors for HT. Conclusion: Although it is characteristically an indolent disease, we demonstrated that a significant portion of FL patients have shortened survival. We confirmed this prognostic heterogeneity, particularly considering the clinical staging and tumor burden. Therefore, FL patients with AS-HTV had higher HT rates and early-relapses, both classic adverse prognostic markers, as well as a tendency to higher mortality. Additionally, we identified clinical and laboratory predictors for HT, which may in a near future direct adapted-risk therapeutic strategies.

Introduction: DLBCL is a malignancy with marked clinical, biological and prognostic heterogeneity. Its different outcomes may be predicted by the identification of molecular biomarkers. Therefore, quantification of circulating cell-free DNA (ccfDNA) and circulating tumor DNA (ctDNA) have emerged as minimally invasive methods able to predict its prognosis. However, such measurements are based on the use of costly technologies and with restricted access in clinical practice. Here, we aimed to quantify ccfDNA in DLBCL using capillary electrophoresis (CEF), a low-cost and widely available methodology, in order to establish prognostic associations between its serum concentrations, clinical characteristics and outcomes. Methods: This prospective study involved 54 DLBCL patients, diagnosed at USP, from 2019 to 2020, and 23 healthy-controls. All cases were treated with 6-8 cycles of R-CHOP. Plasma samples were collected from DLBCL at three moments: at diagnosis, after the 4th CT cycle and at the end of treatment. ccfDNA was extracted using the QIAamp MinElute ccfDNA kit and quantified by CEF (Agilent 2100 Bioanalyzer). Comparison between two groups was performed using the Mann-Whitney test, and between three groups using the Kruskall-Wallis test. The cut-off for [ccfDNA] (1147 ng/μl) was determined by ROC methodology. Survival curves were constructed using the KM method and Log-Rank test was used to assess the relationship between variables and outcomes, and p≤0.05 was considered significant. Results: The median age at diagnosis was 57 years and 61.1% were female. Advanced-stage was observed in 59.3%, 29.6% had ECOG ≥ 2, 52.6% had IPI ≥ 3, and 44.4% had bulky ≥ 10 cm. BM infiltration and involvement of ≥ 2 EN sites occurred in 5.6% and 16.8%, respectively. With a median follow-up of 32 months, the medians OS and EFS were 32 months (95% CI 28-35) and 31 months (95% CI 28-34), respectively. The ORR was 76% (95% CI 63.5-86). The overall mortality rate was 20.4% (95% CI 11.1-32.3). ccfDNA concentrations were significantly higher in DLBCL compared to controls (p < 0.001), however, it did not differ significantly between samples from DLBCL collected at the 3 different time points (p = 0.602). We observed positive associations between ccfDNA [ ] in diagnostic DLBCL samples and biomarkers of high tumor burden, including higher LDH levels (p = 0.003), bulky (p = 0.011), ECOG ≥ 2 (p < 0.001), and IPI ≥ 3 (p = 0.069). However, ccfDNA[ ] at diagnosis were not statistically associated with ORR (p = 0.264), nor with clinical outcomes, including OS (1-year OS: 92% for [ccfDNA] < 1147 ng/μl vs 76% for [ccfDNA] ≥ 1147 ng/μl, p = 0.217) and EFS (1-year EFS: 92% for [ccfDNA] < 1147 ng/μl vs 69% for [ccfDNA] ≥ 1147 ng/μl, p = 0.270). We believe that the short follow-up time and sample number restriction may have contributed to the lack of statistically significant association between ccfDNA [ ] and responses/outcomes observed. In this sense, cohort expansion and NGS techniques for ctDNA identification are ongoing to assess its potential impact as a prognostic biomarker in our cohort. Conclusion: We demonstrated that ccfDNA [ ] measured by CEF were able to accurately discriminate healthy individuals from DLBCL, highlighting its potential role as a minimally invasive diagnostic biomarker. Furthermore, ccfDNA [ ] at diagnosis in DLBCL patients were positively associated with markers of high tumor burden and adverse prognosis. Our results suggest that measurement of ccfDNA by CEF may be a highly cost-effective methodology to identify high-grade B-cell NHLs and recognize DLBCL subgroups with adverse clinical-prognostic characteristics.