Allopurinol and Febuxostat Equally Attenuate Methotrexate-Induced Testicular Injury In Rats Via Activation Of EGF/ ERK1/2/HO‐1 Signaling Pathway.

Methotrexate (MTX) is commonly used in the management of several malignancies and autoimmune disorders; however, testicular damage is one of the most detrimental effects of MTX administration. In the current study, we evaluated the possible protective effect of xanthine oxidase inhibitors either purine analogue; allopurinol (ALL) or non-purine analogue; febuxostat (FEB) in testicular injury induced by MTX in rats. Gonadotoxicity was induced by a single dose of MTX (20 mg/kg, i.p.). ALL and FEB were administered orally in the following daily doses (100, 10 mg/kg, respectively) for 15 days. Total and free testosterone were measured in serum. In addition, total antioxidant capacity (TAC), epidermal growth factor (EGF), malondialdehyde (MDA), tumor necrosis factor-α (TNF-α), extracellular signal-regulating kinase1/2 (ERK1/2), and nitric oxide (NO) end products were measured in testicular tissues. At the same time, immunoexpression of HO-1in testicular tissue was measured. Histopathological examination was done. ALL and FEB increased total and free serum testosterone. Both drugs showed a significant reduction in testicular MDA, NO, TNF-α levels with an increase in TAC, EGF, and ERK1/2 levels in testicular tissue. Furthermore, both drugs enhanced HO-1 immunoexpression in testicular tissue. All these findings were parallel to the preservation of normal testicular architecture in rats treated with ALL and FEB. In conclusion, All and FEB were protective against testicular damage induced by MTX through anti-inflammatory, anti-apoptotic, and antioxidant actions which might be through activation of the EGF/ERK1/2/HO-1 pathway. At the same time, no significant difference between ALL and FEB was noticed in this protective effect.

Profound Lymphopenia at the Time of ATG Administration Is Not Predictive of Survivals after Allotransplant Using Purine Analogue/Busulfan-Based Conditioning Regimen

Introduction: Prophylactic T cell depletion with antithymocyte globulin (ATG) remains a standard of care for GVHD prophylaxis during allotransplant (ASCT). Although the optimal ATG dosing strategy is still unknown, recent studies have reported that recipient absolute lymphocyte counts (ALC) at the time of ATG administration may predict survivals in ASCT with unrelated donors, suggesting that the dose (especially at the cut off of <0.1x109/L) and timing of ATG administration must be taken into account (Soiffer et al, JCO 2017; Kennedy et al, BBMT 2018). Our experience on the impact of lymphopenia at the time of ATG administration during allotransplant is reported here. Materials & Methods: All adults transplanted in our department between 01/2009 and 03/2019 with a Purine analogue/Busulfan/ATG based conditioning regimen and PBSC as source of graft from a matched or 9/10 mismatched donor were eligible. Reduced-intensity conditioning (RIC) regimen consisted of fludarabine 30mg/m²/day (d) from d-6 to d-2, busulfan 3,4 mg/kg/d from d-4 to d-3 and ATG (Thymoglobuline, Sanofi, Lyon, France) 2,5 mg/Kg/d, d-2 and d-1 (FB2A2) or the same but with clofarabine 30mg/m²/d in replacement of fludarabine with 1 or 2 d of ATG (CloB2A2/CloB2A1). Reduced-toxicity myeloablative conditioning regimens (RT-MAC) consisted of the same as FB2A2 but with 3 or 4 d of busulfan instead of 2 (FB3A2/FB4A2). All grafts were administered freshly on the day of the collection while patients (pts) had already received ATG. GVHD prophylaxis was ciclosporine alone for pts with a sibling donor while pts grafted with a matched (MUD) or a 9/10 mismatch (mmUD) unrelated donor received ciclosporine+MMF. We exhaustively looked at pts for whom a blood differential was available at the time of ATG administration in order to study the impact on OS, DFS and GRFS (no grade 3-4 acute GVHD, no moderate/severe chronic GVHD and no relapse) of a profound lymphopenia vs not. Results: Of 395 eligible pts, 116 (median follow-up for alive pts: 49 months) were documented with a differential at time of ATG administration, confirming that this analysis is not a routine practice in our department and probably in many centers. RIC was administered in 80 (69%) of the pts including 39 FB2A2, 12 CLOB2A2 and 29 CLOB2A1. RT-MAC was administered in 36 (31%) pts, including 27 FB3A2 and 9 FB4A2. Seventy-six pts had a myeloid disease while 40 had a lymphoid disease. Donor types were siblings (n=33), MUD (n=70) or 9/10 mmUD (n=13). For the entire cohort, 4y OS, DFS and GRFS were 56.2% (47-66), 40.9% (32-51) and 34.5% (26-45), respectively. No difference in survivals was observed between lymphoid vs myeloid pts, pts transplanted with sibling vs other donors, pts receiving a RIC vs a MAC or a CloB2 vs a FB2 RIC regimen. Median ALC at time of start of conditioning was .915x109/L (range: .010-15.780). No difference in terms of survivals was observed when considering pts under this threshold vs others. ROC curve analysis failed to identify a cut-off allowing to predict better survivals according to ALC at the time of ATG administration (ALC/ATG). Median ALC/ATG was .070x109/L (range: 0-2.300). No difference in terms of survivals was observed when considering pts under this threshold vs others. The same was true when considering .100x109/L as ALC/ATG cut-off. Regarding MAC, the median ALC/ATG was .100x109/L with no difference in survivals between pts under or above this value. The same was true for RIC with ALC/ATG cut-offs < median (.055x109/L) or <.100 x109/L. Interestingly, considering pts with ALC/ATG <.100 x109/L within the RIC setting, survivals were similar between those who received 1d (n=25) vs 2d (n=28) of ATG. This analysis was not performed for pts with ALC/ATG >.100 x109/L as only 4 of them received 1d of ATG vs 23 2d. The dose of CD34+ and CD3+ T cells infused had no impact also on survivals. Conclusion: This study demonstrates that profound lymphopenia at the time of ATG administration as part of a RIC as well as a RT-MAC Purine analogue/Busulfan/ATG based conditioning regimen has no impact on outcomes. Moreover, a reduced dose of ATG in RIC pts with profound lymphopenia at the time of ATG administration did not translate into better survivals. Other unknown factors rather than recipient lymphopenia remain to be discovered to optimize individualized dosing of ATG. Disclosures Peterlin: AbbVie Inc: Consultancy; Astellas: Consultancy; Jazz Pharma: Consultancy; Daiichi-Sankyo: Consultancy. Moreau:Celgene: Consultancy, Honoraria; Janssen: Consultancy, Honoraria; Takeda: Consultancy, Honoraria; AbbVie: Consultancy, Honoraria; Amgen: Consultancy, Honoraria. Le Gouill:Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Travel support; Roche-Genentech: Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Travel support. Chevallier:Daiichi Sankyo: Honoraria; Incyte: Consultancy, Honoraria; Jazz Pharmaceuticals: Honoraria.

Soluble Interleukin-2 Receptor and C-Reactive Protein Levels Are Associated with the Efficacy of Bendamustine As a Salvage Treatment for Indolent Lymphoma

Background: Bendamustine has demonstrated a favorable efficacy and toxicity profile in relapsed or refractory indolent lymphoma. Limited information is available regarding the prognostic factors (particularly laboratory parameters) of bendamustine treatment, although studies have shown a relationship between the hampered effect of bendamustine and an increase in lactate dehydrogenase (LDH) levels. Therefore, we retrospectively evaluated clinical and laboratory factors immediately prior to initiating bendamustine treatment and analyzed its correlation with the clinical outcome among patients with relapsed or refractory indolent lymphoma. Patients and methods: We analyzed 55 relapsed or refractory indolent lymphoma patients who had been treated with bendamustine alone (n = 19) or rituximab plus bendamustine (n = 36) at our hospital from 2000 to 2015. The median age at diagnosis was 65 years and the median follow-up period was 552.5 days. Histological analysis revealed follicular lymphoma (n = 21), mantle cell lymphoma (n = 10), MALT lymphoma (n = 7), lymphoplasmacytoid lymphoma (n = 5), and other low-grade B cell lymphomas (n = 4). We further analyzed the relationships between the clinical outcome [overall response rate (ORR), complete response (CR), progression-free survival (PFS), and overall survival (OS)] and clinical data, including sex, patient age (≥65 years), performance status (≥2), IPI (≥HI), FLIPI (≥HI), no. of previous therapies (≥2), prior purine analogue treatment, response to last treatment, and laboratory parameters [white blood cell count (≥5000/µl), lymphocyte count (≥1000/µl), platelet count (≥10000/µl), LDH (elevated or normal), sIL-2R (elevated or normal), and CRP (elevated or normal)], before starting bendamustine treatment. Results: The median number of cycles of bendamustine was 4 (range: 1-8). The ORR was 80.3% with a CR of 39.2%. Moreover, the CR rate was significantly worse in patients who had elevated sIL-2R and CRP, high or high-intermediate IPI scores, and a high WBC count (P = 0.002, P < 0.001, P = 0.072, and P < 0.046, respectively). Among follicular lymphoma patients, elevated CRP was only associated with a low CR rate (P = 0.028). In multivariate analysis, sIL-2R, CRP, and a high WBC count were all significantly associated with a worse CR rate (P = 0.044, P = 0.002, and P = 0.032, respectively). The 1- and 2- year OS rates were 80.3% and 76.1%, respectively. The OS was significantly higher in a group of the patients who obtained CR after bendamustine, were treated with rituximab plus bendamustine, and did not receive prior purine analogue treatment (P = 0.007, P = 0.008, and P < 0.001, respectively). An elevated CRP was associated with worse OS (P = 0.055). In multivariate analysis, only CR after bendamustine was significantly associated with improved OS (P = 0.023). The 1- and 2-year PFS rates were 69.2% and 60.5%, respectively. PFS was significantly better in patients who obtained CR after bendamustine treatment, had a normal CRP, had not received more than 2 previous therapies, and had a good performance status (P < 0.001, P = 0.007, P = 0.031, and P = 0.033, respectively). In multivariate analysis, any prognosis factors were significantly associated with PFS. Conclusion: This is the first study to demonstrate a correlation between laboratory parameters (i.e., CRP and sIL-2R) and the clinical outcome in patients with relapsed or refractory indolent lymphoma who were treated with bendamustine. In this study, CRP and sIL-2R levels as well as the WBC count were associated with the CR rate. In addition, the CRP levels were associated with OS and PFS, but LDH levels were not associated with any clinical outcomes. Previous studies have reported that the elevation of serum CRP and sIL-2R levels were associated with poor OS or PFS in patients who were treated with rituximab combined with CHOP in diffuse large B-cell lymphoma or follicular lymphoma. Our study indicates that serum CRP and sIL-2R levels are also important laboratory parameters for patients with indolent lymphoma who underwent bendamustine treatment. Disclosures No relevant conflicts of interest to declare.

Radioiodination and biological evaluation of Cladribine as potential agent for tumor imaging and therapy

Cladribine, a purine analogue antimetabolite, was radioiodinated with