31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival

Does Lin28 antagonize miRNA-mediated repression by displacing miRISC from target mRNAs?

Lin28 is a developmentally regulated RNA-binding protein that plays important roles in diverse physiological and pathological processes including oncogenesis and brain synaptic function. These pleiotropic roles of Lin28 are mechanistically linked both to its ability to directly stimulate translation of genes involved primarily in cell growth and metabolism and to its ability to block biogenesis of a subset of miRNAs including the let-7 family of miRNAs. In the case of direct stimulation of gene expression, Lin28 binds to targeted mRNAs through recognition of Lin28-responsive elements (LREs) within mRNAs and recruits RNA helicase A (RHA) to promote translation. RHA belongs to the DEAD-box protein family of RNA helicases, which generally catalyze ATP-dependent unwinding of RNA duplexes or remodeling of ribonucleoprotein complexes (RNPs). Since any given mRNA can potentially be inhibited by miRNAs bearing complementary sequences, we hypothesize that binding of Lin28 to LREs not only nucleates the binding of multiple Lin28 molecules to the same mRNA, but also leads to remodeling of RNPs through recruitment of RHA and causes release of inhibitory miRNA-induced silencing complexes (miRISCs) bound to the mRNA. This mode of action may contribute to Lin28-mediated stimulation of translation in both tumor and neuronal cells

The relationship between H19 and parameters of ovarian reserve

Context The H19 long noncoding RNA (lncRNA) belongs to a highly conserved, imprinted gene cluster involved in embryonic development and growth control. We previously described a novel mechanism whereby the Anti-mullerian hormone (Amh) appears to be regulated by H19. However, the relationship between circulating H19 and markers of ovarian reserve including AMH not been investigated. Objective To determine whether H19 expression is altered in women with decreased ovarian reserve. Design Experimental study. Setting Yale School of Medicine (New Haven, USA) and Gazi University School of Medicine (Ankara, Turkey). Patients or other participants A total of 141 women undergoing infertility evaluation and treatment. Intervention Collection of discarded blood samples and cumulus cells at the time of baseline infertility evaluation and transvaginal oocyte retrieval, respectively. Main outcome measure Serum and cumulus cell H19 expression. Results Women with diminished ovarian reserve (as determined by AMH) had significantly lower serum H19 expression levels as compared to controls (p < 0.01). Serum H19 was moderately positively correlated with serum AMH. H19 expression was increased 3.7-fold in cumulus cells of IVF patients who demonstrated a high response to gonadotropins, compared to low responders (p < 0.05). Conclusion In this study, we show that downregulation of H19 in serum and cumulus cells is closely associated with decreased ovarian reserve, as measured by decreased AMH levels and reduced oocyte yield at oocyte retrieval. Further study with expanded sample sizes is necessary to determine whether H19 may be of use as a novel biomarker for diminished ovarian reserve

hMG addition affects the change in progesterone level during IVF stimulation and LBR: a retrospective cohort study

Abstract Background Premature progesterone (P) rise during IVF stimulation reduces endometrial receptivity and is associated with lower pregnancy rates following embryo transfer (ET), which can influence provider recommendation for fresh or frozen ET. This study aimed to determine whether change in P level between in IVF baseline and trigger (P) is predictive of pregnancy outcome following fresh ET, and whether the ratio of gonadotropins influences P rise and, as a result, clinical pregnancy outcomes: clinical pregnancy rate (CPR) and live birth rates (LBR). Methods Retrospective cohort study at a single fertility center at an academic institution. The peak P level and P were modeled in relation to prediction of CPR and LBR, and the ratios of hMG:rFSH were also modeled in relation to prediction of peak P level on day of trigger, P, and CPR/LBR in a total of 291 patients undergoing fresh embryo transfer after controlled ovarian hyperstimulation-IVF (COH-IVF). Results P correlates with CPR, with the most predictive range for success as P 0.7–0.85 ng/mL (p = 0.005, 95% CI 0.635, 3.636; predicting CPR of 88.9%). The optimal range for peak P in regard to pregnancy outcome was 0.15–1.349 ng/mL (p = 0.01; 95% CI for coefficient in model 0.48–3.570). A multivariable logistic model for prediction of CPR and LBR using either peak or P supported a stronger association between P and CPR/LBR as compared to peak P. Furthermore, an hMG:rFSH ratio of > 0.6 was predictive of lowest peak P (p = 0.010, 95% CI 0.035, 0.256) and smallest P (p = 0.012, 95% CI 0.030, 0.243) during COH-IVF cycles. Highest CPRs were observed within hMG:rFSH ratios of 0.3–0.4 [75.6% vs. 62.5% within and outside of the range, respectively, (p = 0.023, 95% CI 0.119, 1.618)]. Highest LBRs were seen within the range of 0.3–0.6 hMG:rFSH, [LBR of 55.4% vs. 41.4% (p = 0.010, 95% CI 0.176, 1.311)]. Conclusions Our data supports use of P to best predict pregnancy rates and therefore can improve clinical decision making as to when fresh ET is most appropriate. Furthermore, we found optimal gonadotropin ratios can be considered to minimize P rise and to optimize CPR/LBR, emphasizing the importance of luteinizing hormone (LH) activity in COH-IVF cycles

Poor ovarian response in women undergoing in vitro fertilization is associated with altered microRNA expression in cumulus cells

WOS: 000355552600019PubMed ID: 25910568Objective: To analyze the association of micro-ribonucleic acid (miRNA) expression with the number of oocytes retrieved, in women undergoing in vitro fertilization (IVF). Design: Experimental study. Setting: Academic medical center. Patient(s): A total of 189 women undergoing IVF-intracytoplasmic sperm injection (ICSI). Intervention(s): Pooled cumulus cells were collected. Main Outcome Measure(s): Poor responders were identified as patients who produced fewer oocytes than the 25th percentile of their respective age group. MicroRNAs were extracted from cumulus cells, and an miRNA microarray was performed, comparing poor responders (n = 3) to non-poor responders (n = 3). Expression of miR-21-5p (active strand of miR-21) and miR-21-3p was tested in poor responders (n - 21) and non-poor responders (n = 29), using reverse transcription real-time polymerase chain reaction (qRT-PCR). Regulation of miR-21-5p and miR-21-3p, in human granulosa-like tumor (KGN) cells, by estradiol (E-2), was tested in vitro. Result(s): MicroRNA microarray analysis showed up-regulation of 16 miRNAs and down-regulation of 88 miRNAs in poor responders. Notably, miR-21 was significantly up-regulated 5-fold in poor-responder samples. Analysis using qRT-PCR confirmed that miR-21-5p expression was significantly up-regulated in poor responders, whereas miR-21-3p expression was significantly lower, suggesting that elevated miR-21-5p expression in cumulus cells is not regulated at the pre-miR-21 level in poor responders. Both miR-21-5p and miR-21-3p were increased in KGN cells in response to higher doses of E2; their expression was not affected at lower E2 concentrations. Conclusion(s): We found that poor response to IVF is associated miR-21-5p, and that this elevated expression is independent of lower serum E2 levels in poor responders. (C) 2015 by American Society for Reproductive Medicine.NICHD NIH HHS [L50 HD081739, K12 HD000849

Poor ovarian response in women undergoing in vitro fertilization is associated with altered microRNA expression in cumulus cells

Objective: To analyze the association of micro-ribonucleic acid (miRNA) expression with the number of oocytes retrieved, in women undergoing in vitro fertilization (IVF). Design: Experimental study. Setting: Academic medical center. Patient(s): A total of 189 women undergoing IVF-intracytoplasmic sperm injection (ICSI). Intervention(s): Pooled cumulus cells were collected. Main Outcome Measure(s): Poor responders were identified as patients who produced fewer oocytes than the 25th percentile of their respective age group. MicroRNAs were extracted from cumulus cells, and an miRNA microarray was performed, comparing poor responders (n = 3) to non-poor responders (n = 3). Expression of miR-21-5p (active strand of miR-21) and miR-21-3p was tested in poor responders (n - 21) and non-poor responders (n = 29), using reverse transcription real-time polymerase chain reaction (qRT-PCR). Regulation of miR-21-5p and miR-21-3p, in human granulosa-like tumor (KGN) cells, by estradiol (E-2), was tested in vitro. Result(s): MicroRNA microarray analysis showed up-regulation of 16 miRNAs and down-regulation of 88 miRNAs in poor responders. Notably, miR-21 was significantly up-regulated 5-fold in poor-responder samples. Analysis using qRT-PCR confirmed that miR-21-5p expression was significantly up-regulated in poor responders, whereas miR-21-3p expression was significantly lower, suggesting that elevated miR-21-5p expression in cumulus cells is not regulated at the pre-miR-21 level in poor responders. Both miR-21-5p and miR-21-3p were increased in KGN cells in response to higher doses of E2; their expression was not affected at lower E2 concentrations. Conclusion(s): We found that poor response to IVF is associated miR-21-5p, and that this elevated expression is independent of lower serum E2 levels in poor responders. (C) 2015 by American Society for Reproductive Medicine

Efficacy of intrauterine perfusion of peripheral blood mononuclear cells (PBMC) for infertile women before embryo transfer: meta-analysis

This meta-analysis was intended to evaluate the effects of intrauterine perfusion of peripheral blood mononuclear cells (PBMC) on the pregnancy outcomes including clinical pregnancy rates, embryo implantation rates, live birth rates and miscarriage rates of infertile women who were undergoing in vitro fertilisation (IVF) treatment. By searching Pubmed, Embase database, five articles meeting the inclusion criteria were included, and 1173 women were enrolled (intrauterine PBMC group: n = 514; NO-PBMC group: n = 659). For the entire IVF/ICSI population and one or two embryo transfer failure patients, there was no significant difference in endometrial thickness, embryo implantation rates, live birth rates, and miscarriage rates between the PBMC group and NO-PBMC group. Although the clinical pregnancy rates of the PBMC group were higher than that of the NO-PBMC group, the confidence interval was close to the line of unity. As for the patients with three or more implantation failures, the clinical pregnancy rates, embryo implantation rates and live birth rates were much higher in the PBMC group than that of the NO-PBMC group. In summary, current evidence suggests that intrauterine perfusion of PBMC can significantly improve pregnancy outcomes in patients who have three or more implantation failures.Impact statement What is already known on this subject? An increasing number of studies have shown that immune cells play an important role in embryo transfer. There is no reliable evidence to confirm the clinical efficacy of intrauterine perfusion of PBMC. What do the results of this study add? The current evidence suggests that intrauterine perfusion of PBMC can significantly improve pregnancy outcomes in patients who have three or more implantation failures. What are the implications of these findings for clinical practice and/or further research? To the best of our knowledge, this meta-analysis is the first to evaluate the effect of intrauterine perfusion of PBMC on pregnancy outcomes before embryo transfer. Our study indicated that intrauterine perfusion of PBMC significantly increased clinical pregnancy rates, embryo implantation rates, and live birth rates in patients who failed more than three implants