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  • 1
    In: Blood, American Society of Hematology, Vol. 132, No. Supplement 1 ( 2018-11-29), p. 2158-2158
    Abstract: Introduction: Sarcopenia, defined as low muscle mass and low muscle function and/or reduced physical performance, has been identified as a potential risk factor for adverse outcomes in a variety of conditions like elderly patients and patients with solid tumors. However, fewer studies assessed this association in patients undergoing autologous hematopoietic stem cell transplantation (autoHSCT). With this aim, we performed a retrospective study to evaluate the effect of sarcopenia on complications (such as toxicities, infections and intensive care unit - ICU - admission) and survival after autoHSCT for adults with lymphoma. Methods: We enrolled 135 patients with lymphoma who underwent autoHSCT between January, 2014 and December, 2017, in a single center. Computed tomography (CT) imaging scans were used for muscle mass analysis. Skeletal muscle area (SMA, cm2) was measured on axial CT images at the level of the third lumbar vertebra (L3) using appropriate software (Slice O´Matic, Tomovision, CA). SMA was normalized for height to obtain the skeletal muscle index (SMI, cm2/m2). We set our own cut-off values at lowest specific tertiles of SMI based on sex and body mass index (BMI, Kg/m2) for diagnosing sarcopenia. Sarcopenia was defined in this cohort as SMI 〈 46 cm²/m² if BMI 〈 25 kg/m² or 〈 51 cm²/m² if BMI ≥25 kg/m² in men, and ≤41.5 cm²/m² in women. Toxicities (overall, renal, hepatic and gastrointestinal) after autoHSCT were graded according to the National Cancer Institute Common Toxicity Criteria (version 4.0). In addition, infections and ICU admission were also analyzed. Patients with previous autoHSCT CT scans more than 120 days were excluded. Probabilities of overall survival (OS), event-free survival (EFS) and treatment-related mortality (TRM) were calculated according to the Kaplan-Meier method. Unadjusted and adjusted analyses were conducted by using Logistic Regression models to determine the association of covariates with outcome. The significance level was adopted at p 〈 0.05. Results: A total of 110 patients met the eligibility criteria (39 sarcopenic and 71 non-sarcopenic). Mean age at transplant was similar in both groups (43 years, p=0.96), with 18% of elderly. The majority of patients was male (65%) and had an aggressive disease at diagnosis (77%). Sarcopenics had a higher Charlson comorbidity index (≥3) at transplant than non-sarcopenics (p=0.02). The conditioning regimen used was different between groups, with a higher proportion of bussulfan-based conditioning in the sarcopenic group (p=0.04). Table 1 list characteristics between sarcopenics and non-sarcopenics patients. Neutrophil recovery was achieved at a median time of 11 days after transplant in both groups (p=0.55). We found association between grade 3-4 gastrointestinal toxicity and sarcopenia (44% vs. 24%; OR 2.46, CI95% 1.07-5.66, p=0.03), although, after adjustment by conditioning regimen, no significant difference was detected (OR 2.09, CI95% 0.89-4.97, p=0.10). No statistically differences were detected in ICU admission, occurrence of infection, hepatic, renal or overall toxicities. Median follow-up of survivors was 20 months (range, 1-50). The 2-years OS and EFS were 72% (CI95% 61-81; sarcopenic 70% [50-84] vs. non-sarcopenic 74% [59-84] , p=0.99) and 62% (CI95% 51-71; sarcopenic 68% [49-81] vs. non-sarcopenic 59% [46-70] , p=0.37), respectively. The cumulative incidence of TRM was 6.5% at day 100 (CI95% 2.5-11.9; sarcopenic 5.1% [1.3-19.0] vs. non-sarcopenic 5.7% [2.2-14.5] , p=0.70). Conclusion: In our study, sarcopenia was not statistically associated with severe toxicity and higher mortality. To the best of our knowledge, this is the first study to assess CT images and compare sarcopenia with toxicity in aHSCT for lymphoma. Prospective studies are needed to investigate the real impact of sarcopenia on outcomes after autologous transplantation. Disclosures No relevant conflicts of interest to declare.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
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    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2018
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  • 2
    In: Cancer Immunology Research, American Association for Cancer Research (AACR), Vol. 8, No. 4_Supplement ( 2020-04-01), p. A22-A22
    Abstract: Chronic signaling by molecules generated in the tumor microenvironment has local and systemic effects on the immune system. We have previously shown that cervical cancer cells express cytokines as IL-6 and G-CSF, among other factors. These cytokines have been associated with cancer progression in several types of cancer, either by promoting chronic inflammation and/or tolerance. Trying to better understand how cells from the immune system and tumor integrate signals generated by the tumor microenvironment, we have investigated the activation status of signaling pathways involved in cell proliferation and survival, inflammation and immune responses. We observed a cohort of cervical cancer patients, all of whom had cervical biopsies positive for high oncogenic risk human papillomavirus (HPV). In the cervical cancer biopsies, we found constitutive expression of phosphorylated STAT3, Akt, p65 NFκB and CREB, in both the tumor and leukocyte compartments. In the peripheral blood, however, we found constitutive expression of phosphorylated STAT3 and Akt, but significantly lower expression of phosphorylated p65 NFκB than in controls. Data previously published by our laboratory using a HPV16 experimental model showed high constitutive expression of phospho-STAT3 and low negative expression of phospho-p65 NFκB in splenocytes of tumor-bearing mice compared to controls. Given the data from our patients, we hypothesized that the high STAT3 expression and low NFκB expression could have a correlation, with a biologic effect on tumor-bearing subjects. Again, using an HPV16 experimental model, we neutralized IL-6 and G-CSF expression or pharmacologically inhibited STAT3 with the NSC74859 compound. In both cases, we observed significant reduction in tumor growth, decrease in phospho-STAT3 expression (when using the neutralizing antibodies), and importantly, significant increase in phospho-p65 NFκB expression. We also observed a significant increase in T-cell infiltration in tumors from treated animals and increase in antigen-specific T-cell proliferation. Our data indicate that cytokines that signal chronically through STAT3 can inhibit antitumor immune responses through NFκB inhibition. We also suggest that inhibition of STAT3 may be used as therapeutic tool against cervical cancer and other HPV-associated cancers. Financial support: FAPESP 2014/19326-6; 2008/57889-1; CNPq 573799/2008-3. Citation Format: Renata Ariza Marques Rossetti, Gretel Rodriguez Rodriguez, Ildelfonso Alves Da Silva, Jr. Jesus De Paula Carvalho, Maria Beatriz Sartor De Faria Rosa, Luisa Lina Villa, Ana Paula Lepique. Cervical cancer immunomodulation through stat3 and p65 NFκb: Effects beyond the tumor microenvironment [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr A22.
    Type of Medium: Online Resource
    ISSN: 2326-6066 , 2326-6074
    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2020
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  • 3
    In: Journal of Leukocyte Biology, Oxford University Press (OUP), Vol. 105, No. 5 ( 2019-04-19), p. 1041-1054
    Abstract: Cervical cancer continues to be a public health problem in developing countries. Previous studies have shown that cervical cancer cells display markers of aerobic glycolysis, indicating that these tumors are likely to secrete lactate. Mostly, lactate is recognized as a molecule capable of suppressing immune responses, through inhibition of T cells, Mϕs, and dendritic cells. We and others have previously shown that Mϕs are frequent cells infiltrating cervical cancers with the ability to inhibit antitumor immune responses and promote tumor growth through angiogenesis. Here, we have tested the hypothesis that lactate, secreted by cervical cancer cells, can modulate Mϕ phenotype. First, we showed higher lactate plasma concentrations in patients with increasing cervical lesion grades, with maximum concentration in the plasma of cancer patients, which supported our hypothesis. We then inhibited lactate production in tumor cell spheroids established from cervical cancer derived cell lines, using the lactate dehydrogenase inhibitor, oxamate, prior to co-culture with monocytes. Lactate mediated part of the crosstalk between tumor cells and Mϕs, promoting secretion of IL-1β, IL-10, IL-6, and up-regulation of hypoxia induced factor-1α expression, and down-regulation of p65-NFκB phosphorylation in Mϕs. We also showed that Mϕs from co-cultures treated with oxamate were better inducers of T cell activation. Of note, experiments performed with inhibition of the monocarboxylate transporters rendered similar results. Our data confirms the hypothesis that lactate, secreted by cervical tumor cells, influences the phenotype of tumor Mϕs, promoting a suppressive phenotype.
    Type of Medium: Online Resource
    ISSN: 1938-3673 , 0741-5400
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    Language: English
    Publisher: Oxford University Press (OUP)
    Publication Date: 2019
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  • 4
  • 5
    In: Journal of Leukocyte Biology, Oxford University Press (OUP), Vol. 96, No. 4 ( 2014-06-26), p. 619-631
    Abstract: Tumors are complex structures containing different types of cells and molecules. The importance of the tumor microenvironment in tumor progression, growth, and maintenance is well-established. However, tumor effects are not restricted to the tumor microenvironment. Molecules secreted by, as well as cells that migrate from tumors, may circulate and reach other tissues. This may cause a series of systemic effects, including modulation of immune responses, and in some cases, leukocytosis and metastasis promotion. Leukocytosis has been described as a poor prognostic factor in patients with cervical cancer. The main etiological factor for cervical cancer development is persistent infection with high oncogenic risk HPV. Our laboratory has been exploring the effects of high oncogenic risk, HPV-associated tumors on lymphoid organs of the host. In the present study, we observed an increase in myeloid cell proliferation and alteration in cell signaling in APCs in the spleen of tumor-bearing mice. In parallel, we characterized the cytokines secreted in the inflammatory and tumor cell compartments in the tumor microenvironment and in the spleen of tumor-bearing mice. We show evidence of constitutive activation of the IL-6/STAT3 signaling pathway in the tumor, including TAMs, and in APCs in the spleen. We also observed that IL-10 is a central molecule in the tolerance toward tumor antigens through control of NF-κB activation, costimulatory molecule expression, and T cell proliferation. These systemic effects over myeloid cells are robust and likely an important problem to be addressed when considering strategies to improve anti-tumor T cell responses.
    Type of Medium: Online Resource
    ISSN: 0741-5400 , 1938-3673
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    Language: English
    Publisher: Oxford University Press (OUP)
    Publication Date: 2014
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  • 6
    In: Scientific Reports, Springer Science and Business Media LLC, Vol. 7, No. 1 ( 2017-08-21)
    Abstract: Cervical cancer is the last stage of a series of molecular and cellular alterations initiated with Human Papillomavirus (HPV) infection. The process involves immune responses and evasion mechanisms, which culminates with tolerance toward tumor antigens. Our objective was to understand local and systemic changes in the interactions between HPV associated cervical lesions and the immune system as lesions progress to cancer. Locally, we observed higher cervical leukocyte infiltrate, reflected by the increase in the frequency of T lymphocytes, neutrophils and M2 macrophages, in cancer patients. We observed a strong negative correlation between the frequency of neutrophils and T cells in precursor and cancer samples, but not cervicitis. In 3D tumor cell cultures, neutrophils inhibited T cell activity, displayed longer viability and longer CD16 expression half-life than neat neutrophil cultures. Systemically, we observed higher plasma G-CSF concentration, higher frequency of immature low density neutrophils, and tolerogenic monocyte derived dendritic cells, MoDCs, also in cancer patients. Interestingly, there was a negative correlation between T cell activation by MoDCs and G-CSF concentration in the plasma. Our results indicate that neutrophils and G-CSF may be part of the immune escape mechanisms triggered by cervical cancer cells, locally and systemically, respectively.
    Type of Medium: Online Resource
    ISSN: 2045-2322
    Language: English
    Publisher: Springer Science and Business Media LLC
    Publication Date: 2017
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  • 7
    Online Resource
    Online Resource
    Wiley ; 2014
    In:  Immunity, Inflammation and Disease Vol. 2, No. 2 ( 2014-08), p. 63-75
    In: Immunity, Inflammation and Disease, Wiley, Vol. 2, No. 2 ( 2014-08), p. 63-75
    Type of Medium: Online Resource
    ISSN: 2050-4527 , 2050-4527
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2014
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  • 8
    Online Resource
    Online Resource
    American Association for Cancer Research (AACR) ; 2023
    In:  Cancer Research Vol. 83, No. 7_Supplement ( 2023-04-04), p. 4055-4055
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 83, No. 7_Supplement ( 2023-04-04), p. 4055-4055
    Abstract: Adoptive transfer of tumor infiltrating lymphocytes (TIL) is a feasible and effective therapy for melanoma and lung cancer[1,2]. Multiple factors may determine the quality of the TIL product including components of the tumor microenvironment. In this work, we analyzed the role of melanoma infiltrating B cells in the context of TIL expansion based on their documented association with response to other types of immunotherapies[3] . We stimulated melanoma infiltrating B cells using human recombinant CD40L on the first day of ex-vivo TIL expansion. Samples were expanded from cryopreserved melanoma tumor single cell suspensions, in high dose IL-2 alone (standard protocol), or in high dose IL-2 plus CD40L. After 48h, analysis of activation markers on the CD40-expressing cells by flow cytometry was performed. For further investigation of the changes induced by CD40L stimulation, TIL expansion cultures (+/- CD40L) were analyzed using scRNA-seq (10X Genomics Chromium NextGEM Single Cell 5’ v2 and V(D)J Reagent kits; Illumina NovaSeq 6000 instrument with S4 sequencing flow cell) at 48h of culture (n=7 patients). The TIL expansion success rate was 68% with the CD40L treatment condition compared to 36% with the standard protocol. TILs cultured in the presence of CD40L expanded to on average three times more than with the standard protocol (P ≤ 0.01). Treatment with CD40L increased the percentage of CD39- CD69- T cells (P ≤ 0.05). Within the tumor digests, a higher percentage of B cells, including switched memory B cells (CD27+ IgD− ), was associated with successful TIL expansion (P=0.04). scRNA-seq analysis demonstrated different clustering patterns within the B cell compartment based on culture conditions. No clear partition was observed for other cell types, including the myeloid compartment. B cells displayed 126 DEGs associated to CD40L addition, CCL22, CD83, EBI3 and CD58 were among the upregulated genes in the CD40L-treated B cells. Other cell types experienced minimal to no change in transcriptomic profiles. B cell clusters were sub-classified based on CD27 and IgD expression[4], showing a predominance of naïve and switched memory B cells. Our results show that higher presence of B cells within tumors is associated with better TIL expansion, suggesting an interplay between T and B cells, and providing rationale for the design of improved TIL expansion protocols based on B cell stimulation with CD40L. This work has been supported in part by the Flow Cytometry, Genomics and Biostatistics and Bioinformatics Core Facilities at Moffitt Cancer Center, an NCI designated Comprehensive Cancer Center (P30-CA076292). We acknowledge Moffitt’s Melanoma Center of Excellence and the Mark Foundation for the financial support. [1] Sarnaik, A.A. et al. JCO 39, 2656-2666 (2021). [2] Creelan, B. et al. Nat Med 27, 1410-1418 (2021). [3] Cabrita, R. et al. Nature 577, 561-565 (2020). [4] Sanz, I. et al. Front Immunol 10, 2458 (2019). Citation Format: Renata Ariza Marques Rossetti, Leticia Tordesillas, Matthew Beatty, Dongliang Du, Yian Ann Chen, Amod Sarnaik, Shari Pilon-Thomas, Daniel Abate-Daga. CD40L stimulates melanoma infiltrating B cells and enhances ex vivo TIL expansion. [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 4055.
    Type of Medium: Online Resource
    ISSN: 1538-7445
    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2023
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  • 9
    In: Transplant Infectious Disease, Wiley, Vol. 22, No. 2 ( 2020-04)
    Abstract: Hepatitis B virus (HBV) infection is a worldwide concern with a broad distribution. In immunosuppressed populations, such as solid organ and hematopoietic stem cell transplant (HSCT) recipients, it can reactivate leading to acute hepatic failure. Different risk factors are known for higher rates of reactivation, and entecavir, tenofovir, and lamivudine are often used for prophylaxis and treatment. However, data regarding the impact of antiviral drugs in neutrophil and platelet engraftment are still unknown and concern the management of viral hepatitis post‐HSCT. Methods We performed a single‐center, retrospective, observational study reviewing medical records of patients referred for hematopoietic stem cell transplant from 2010 to 2017, which were also HBV infected, aiming to describe outcomes related to antiviral treatment and also the impact on platelet and neutrophil recovery after transplant. A secondary goal consisted of analyzing the impact of HBV infection in early and late mortality post‐HSCT. The study included patients with positive blood bank screening for hepatitis B infection (HBsAg, Anti‐HBc or HBV‐NAT), confirmed later on by a laboratory routine serology. Results A total of 1132 hematopoietic stem cell recipients were assessed between 2010 and 2017. Eighty‐six patients were confirmed to have HBV infection, of which six were HBsAg‐positive, 20 were isolated anti‐HBc‐positive, and 60 had resolved infection (anti‐HBc‐positive and anti‐HBs‐positive). With regard to prophylaxis, 19 patients underwent HSCT on HBV antiviral therapy or prophylaxis: two were HBeAg‐positive, three were HBeAg‐negative and HBV‐DNA was only detectable in three of them. Moreover, one patient had an occult HBV infection. Regarding therapy, 9 patients were on entecavir, 6 patients on lamivudine, two on tenofovir, and two of them on a combination of tenofovir + lamivudine due to HIV co‐infection. Reverse seroconversion was not identified in any patients receiving antiviral therapy or prophylaxis, but it was detected in one patient with occult hepatitis B and another with resolved infection. No severe side effects led to therapy discontinuation in the treated group, which also did not have any significant delay in neutrophil or platelet engraftment when compared to patients without antiviral therapy. In addition, the only factors associated with increased mortality were transplant onset after 50 years, allogeneic transplant and myeloablative conditioning regimens. Interestingly, the presence of HBsAg or detectable HBV‐DNA was not related to worse outcomes, neither the use of rituximab. In multivariate analysis, the use of antiviral therapy, the occurrence of graft‐versus‐host disease or CMV reactivation also was not linked to increased mortality. Conclusions To sum up, HBV serology, ALT, and HBV‐DNA monitoring are essential to detect hepatic flares earlier, even in populations with chronic inactive hepatitis, due to the possibility of later seroconversion. HBV infection was not related to increased 2‐year mortality post‐transplant. Antiviral prophylaxis did not cause any important clinical or laboratory side effects that could demand discontinuation, and its use was not associated with later neutrophil and platelet engraftments.
    Type of Medium: Online Resource
    ISSN: 1398-2273 , 1399-3062
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2020
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