Retinoic acid-responsive CD8 effector T-cells are selectively increased in IL-23-rich tissue in gastrointestinal GvHD.

Gastrointestinal (GI) graft-versus-host disease (GvHD) is a major barrier in allogeneic hematopoietic stem-cell transplantation (AHST). The metabolite retinoic acid (RA) potentiates GI-GvHD in mice via alloreactive T-cells expressing the RA-receptor-alpha (RARα), but the role of RA-responsive cells in human GI-GvHD remains undefined. We therefore used conventional and novel sequential immunostaining and flow cytometry to scrutinize RA-responsive T-cells in tissues and blood of AHST patients and characterize the impact of RA on human T-cell alloresponses. Expression of RARα by human mononuclear cells was increased after RA exposure. RARαhi mononuclear cells were increased in GI-GvHD tissue, contained more cellular RA-binding proteins, localized with tissue damage and correlated with GvHD severity and mortality. Using a targeted candidate protein approach we predicted the phenotype of RA-responsive T-cells in the context of increased microenvironmental IL-23. Sequential immunostaining confirmed the presence of a population of RARahi CD8 T-cells with the predicted phenotype, co-expressing the effector T-cell transcription factor T-bet and the IL-23-specific receptor. These cells were increased in GI- but not skin-GvHD tissues and were also selectively expanded in GI-GvHD patient blood. Finally, functional approaches demonstrated RA predominantly increased alloreactive GI-tropic RARahi CD8 effector T-cells, including cells with the phenotype identified in vivo. IL-23-rich conditions potentiated this effect by selectively increasing b7 integrin expression on CD8 effector T-cells and reducing CD4 T-cells with a regulatory cell phenotype. In conclusion we have identified a population of RA-responsive effector T-cells with a distinctive phenotype which are selectively expanded in human GI-GvHD and represent a potential new therapeutic target

Low HER2 expression in normal breast epithelium enables dedifferentiation and malignant transformation via chromatin opening.

Overexpression of the HER2 protein in breast cancer patients is a predictor of poor prognosis and resistance to therapies. We used an inducible breast cancer transformation system that allows investigation of early molecular changes. HER2 overexpression to similar levels as those observed in a subtype of HER2-positive breast cancer patients induced transformation of MCF10A cells and resulted in gross morphological changes, increased anchorage-independent growth of cells, and altered the transcriptional programme of genes associated with oncogenic transformation. Global phosphoproteomic analysis during HER2 induction predominantly detected an increase in protein phosphorylation. Intriguingly, this correlated with chromatin opening, as measured by ATAC-seq on acini isolated from 3D cell culture. HER2 overexpression resulted in opening of many distal regulatory regions and promoted reprogramming-associated heterogeneity. We found that a subset of cells acquired a dedifferentiated breast stem-like phenotype, making them likely candidates for malignant transformation. Our data show that this population of cells, which counterintuitively enriches for relatively low HER2 protein abundance and increased chromatin accessibility, possesses transformational drive, resulting in increased anchorage-independent growth in vitro compared to cells not displaying a stem-like phenotype

Surrogacy tourism: the ethical and legal challenges

Although surrogacy seemed to have been practised since ancient times, its resurgence in the contemporary era has been nothing short of phenomenal. With advances made in reproductive technology, it is now possible to fertilise eggs and sperms in laboratories and have the embryo transferred into the womb of a surrogate mother for gestation. Through a combination of push and pull factors, this possibility of gestational surrogacy has led to the meteoric rise of cross-border surrogacy. This paper seeks to highlight the ethical and legal challenges associated with the practice, and calls for better legal oversight at international level

Investigating Mechanisms of Immune Escape leading to Relapse of Acute Myeloid Leukaemia after Allogeneic Haematopoietic Stem Cell Transplantation

Acute myeloid leukaemia (AML) is a haematological malignancy with an incidence of around 3000 cases per year in the UK. Despite recent advances in treatment, allogeneic haematopoietic stem cell transplantation (AHSCT) remains the only route to a potential cure for many patients. Unfortunately, 20-50% patients relapse following AHSCT, with extremely limited treatment options. Relapse in this scenario is commonly held to be due to immune evasion of the allogeneic graft versus leukaemia effect but the precise mechanisms underlying this are not fully understood. Furthering our understanding of these mechanisms will allow us to develop better treatments for this group of patients. This project utilised a cohort of eleven patients for a genome wide multi-omic approach to interrogate AML blasts at diagnosis and relapse post AHSCT to identify cell intrinsic changes prompting relapse and identify candidate pathways for pharmacological targeting in an in vitro model of alloreactivity. Firstly, a supervised analysis was conducted to investigate known mechanisms of immune evasion targeting the antigen presenting capacity of AML blasts. In three out of four cases, AML blasts were less able to provoke an in vitro allogeneic T cell response at post AHSCT relapse than at diagnosis, and it was shown that the ability to provoke T cell proliferation responses at diagnosis correlated with the surface expression of HLA-DR. Gene expression analysis showed a significant downregulation between diagnosis and post-transplant relapse of both MHC class I and class II genes, as well as upregulation of MUC1, known to contribute to myeloid derived suppressor cell activation in AML. Secondly, an unsupervised genome wide analysis was carried out examining the genetic and epigenetic landscapes of AML blasts. Mutational analysis demonstrated evidence of clonal evolution in each of the samples tested and identified a novel mutation in TXNIP which was both recurrent and relapse specific. Bioinformatic identification of potential neoantigens showed that the number of HLA-DR restricted neoantigens correlated with the change in HLA-DR expression between diagnosis and post AHSCT relapse. Gene expression analysis identified 25 genes which were significantly differentially expressed between diagnosis and post AHSCT relapse, including the chemokines CXCL2, CXCL3 and CXCL8, and the cell survival molecule BCL2A1 and gene set enrichment analysis identified significantly enriched pathways including immune pathways and NF-kB targets, as well as cell cycle pathways. Finally, in vitro functional testing into the role of the differentially expressed chemokines in alloreactivity showed that the microenvironment was vital in modulating the response to these chemokines, and pharmacological targeting of AML relapse post AHSCT highlighted both BCL2 inhibition and LSD1 inhibition as promising therapeutic pathways

Very Long-Term Complete Remission Can Be Achieved in Men With High-Risk Localized Prostate Cancer and a Very High PSA Value: An Analysis of the GETUG 12 Phase 3 Trial.

Serum prostate specific antigen (PSA) is a well-known prognostic parameter in men with prostate cancer. The treatment of men with very high PSA values and apparently no detectable metastases is not fully established. Ancillary analysis from the GETUG 12 phase 3 trial. Patients with non-metastatic high-risk prostate cancer by bone and computerized tomography (CT) scan were randomly assigned to receive androgen deprivation therapy (ADT) and docetaxel plus estramustine or ADT alone. Relapse-free survival (RFS), clinical RFS, metastases-free survival (MFS), overall survival (OS), and prostate cancer-specific survival (PCSS) were estimated using the Kaplan-Meier method for different levels of PSA (50 ng/mL, 75 ng/mL, and 100 ng/mL). The relationship between PSA and outcomes was studied using residual-based approaches and spline functions. The median follow-up was 12 years (range: 0-15.3). Baseline PSA (<50 ng/mL, n = 328; ≥50ng/mL, n = 85) was associated with improved RFS (P = .0005), cRFS (P = .0024), and MFS (P = .0068). The 12-year RFS rate was 46.33% (CI 40.59-51.86), 33.59% (CI 22.55-44.97), and 11.76% (1.96-31.20) in men with PSA values <50 ng/mL (n = 328), 50-100 ng/mL (n = 68), and ≥100 ng/mL (n = 17), respectively. Exploratory analyses revealed no deviation from the linear relationship assumption between PSA and the log hazard of events. Men with apparently localized prostate cancer and a high baseline PSA value have a reasonable chance of being long-term disease-free when treated with curative intent combining systemic and local therapy