Regulation of necroptotic-apoptotic signaling in vitro and in vivo

Programmed cell death (PCD) is an evolutionarily conserved process observable in all metazoans. In mammals this process is requisite for organ and structural development of the fetus, pathogen clearance, tissue remodeling and homeostasis, responses to cellular stress, and suppression of oncogenesis. Several major forms of PCD have been characterized to date, including apoptosis, necroptosis and autophagy-dependent cell death (ADCD). Each of these forms of PCD demonstrate their own morphologic and molecular features. In particular, apoptotic and necroptotic cell death have been identified in many circumstances to be competitive in nature, often displaying a ‘winner take all’ dynamic where necroptosis provides a backup role in many tissues upon failure of apoptosis. To better understand the regulation of PCD and more specifically the interactions between apoptosis and necroptosis, I have examined these forms of cell death in two models: (1) cisplatin-mediated cell death of homogeneous populations of embryonic stem (ES) cells in vitro and (2) endothelin-1 mediated ischemic/reperfusion injury in the murine cortex in vivo. In addition to homologous gene recombination, I have also utilized pharmacologic inhibitors to interrogate the interaction of these pathways in detail for both models. Using these methods, I demonstrate that ES cells exposed to cisplatin demonstrate a hitherto uncharacterized novel form of cooperative PCD between apoptotic caspase-3 and necroptotic RIPK1 and RIPK3. Further analyses suggest that RIPK1 and RIPK3 signaling lie upstream of caspase-3 in this cooperative pathway and that the resulting cell morphology and molecular details of dead/dying cells reflect neither traditional apoptosis nor necroptosis, instead demonstrating a hybrid of both. Characterization of the neuronal cell death involved in endothelin-1-mediated cortical ischemic/reperfusion injury in vivo was similarly observed to be protected through inhibition of either RIPK1 or caspase-3. Interestingly, protection induced by inhibition of RIPK1 seem to occur upstream of caspase-3-mediated apoptosis similar to features observed in ES cells. This identification of a novel co-operative apoptotic/necroptotic signaling in turn have important implications for cell death regulation in primary tissues – in terms of both pathologic processes and pharmacologic interventions.Ph.D.2021-11-30 00:00:0

Impact of Haemophilus influenzae type B (Hib) and viral influenza vaccinations in pregnancy for improving maternal, neonatal and infant health outcomes

Background: Infections during pregnancy confers increased risk of maternal and perinatal morbidity and mortality. However, the case for advocating Haemophilus influenzae type B (Hib) and viral Influenza vaccinations in pregnancy is still debatable.Objectives: To assess the impact of Hib and viral Influenza vaccinations during pregnancy on maternal, neonatal and infant health outcomes compared to placebo/control.SEARCH Methods: We searched the Cochrane Pregnancy and Childbirth Group\u27s Trials Register (29 January 2015) and reference lists of retrieved studies.SELECTION CRITERIA: All randomised controlled clinical trials (including cluster-randomised trials) and quasi-randomised trials evaluating Hib or viral influenza vaccination during pregnancy compared with no vaccination or placebo.DATA COLLECTION AND ANALYSIS: Two review authors independently assessed trials for inclusion, risk of bias and extracted data. Data were checked for accuracy.MAIN Results: Two trials were included this review. One (involving 213 women and 213 neonates) evaluated the impact of Hib vaccination during pregnancy and the other study (involving 2116 women and 2049 neonates) evaluated the impact of viral influenza vaccination during pregnancy. Overall, the HiB vaccination trial was judged to be at \u27high risk of bias\u27 due to inadequate randomisation while the other trial was judged to be at \u27low risk of bias\u27. Hib vaccination during pregnancy versus placeboOne trial involving 213 women and 213 neonates evaluating the impact of Hib vaccination during pregnancy was included under this comparison. The study did not report on any of this review\u27s prespecified primary outcomes (including mortality, respiratory tract infection and sepsis) or secondary outcomes (including adverse events) except preterm delivery. There was no clear difference between the Hib vaccination and placebo control groups in terms of preterm delivery (risk ratio (RR) 1.28, 95% confidence interval (CI) 0.12 to 13.86, one study, 213 participants), fetal distress (RR 1.23, 95% CI 0.67 to 2.26, one study, 213 infants), intubation (RR 1.03, 95% CI 0.55 to 1.95, one study, 213 infants) and neonatal jaundice (RR 1.01, 95% CI 0.52 to 1.97, one study, 213 infants). We could not grade the evidence for quality due to lack of outcome data. Viral influenza vaccination during pregnancy versus placeboOne trial involving 2116 women and 2049 infants evaluating the impact of trivalent inactivated influenza vaccine (IIV3) during pregnancy was included under this comparison.There was no clear difference between the viral influenza and placebo control group in terms of most of this review\u27s primary outcomes: maternal death (RR 4.96, 95% CI 0.24 to 103.24, moderate quality evidence), infant death up to 175 days after birth (RR 0.71, 95% CI 0.37 to 1.37, moderate quality evidence), perinatal death (stillbirth and death in the first week of life) (RR 1.32, 95% CI 0.73 to 2.38, moderate quality evidence), influenza-like illness in women (RR 0.96, 95% CI 0.79 to 1.16) or their babies (RR 1.02, 95% CI 0.94 to 1.09), any respiratory illness in women (RR 0.97, 95% CI 0.91 to 1.04, high quality evidence) or their babies (RR 1.01, 95% CI 0.95 to 1.07, high quality evidence). There were also no clear differences between vaccination and placebo control groups in terms of maternal hospitalisation for any infection (RR 2.27, 95% CI 0.94 to 5.49; 2116 women, moderate quality evidence), and neonatal hospitalisation for sepsis (RR 1.60, 95% CI 0.73 to 3.50; 2049 infants, moderate quality evidence). However, viral influenza vaccination during pregnancy was associated with a reduction in reverse-transcriptase-polymerase-chain-reaction (RT-PCR) confirmed influenza among infants (RR 0.51, 95% CI 0.30 to 0.88, one study, 2049 infants) and women (RR 0.50, 95% CI 0.29 to 0.86, one study, 2116 women).In terms of this review\u27s secondary outcomes, there were no clear differences in terms of the impact on pregnancy outcomes (miscarriage, preterm labour and stillbirth), hospitalisation for respiratory infection among women and infants. Similarly, there was no difference between the viral influenza vaccine and placebo control groups in terms of any adverse systemic reactions.AUTHORS\u27 CONCLUSIONS: There is limited evidence (from one small trial at a high risk of bias) on the effectiveness on Hib during pregnancy for improving maternal, neonatal and infant health outcomes.Evidence from one large high quality trial on the effectiveness of viral influenza vaccine during pregnancy suggests reduced RT-PCR confirmed influenza among women and their babies, suggesting the potential of this strategy for scale up but further evidence from varying contexts is required.Further trials for both Hib and viral influenza vaccines with appropriate study designs and suitable comparison groups are required. There are currently two \u27ongoing\u27 studies - these will be incorporated into the review in future updates