Citations

Background: Abortion is a spontaneous loss of pregnancy before 20 weeks. Approximately 42 million pregnancies end in abortion. The maternal infections that are transmissible from mother to fetus are caused by many pathogens, of which the TORCH complex contributes majorly to neonatal and infant deaths globally. The aim of this study is to detect the prevalence and types of infectious causes of abortion. One hundred aborted women admitted to King Faisal Medical Complex Maternity Hospital in Taif City between the period of 2018 and 2020 were enrolled in this study. The serological test reports (TORCH panel), as well as reports of hematological (CBC) and chemical parameters, were obtained from laboratory management system databases, reviewed, and then analyzed. The H&E-stained microscopic slides of their product of conception (POC) were examined under a microscope and compared with histopathological reports. The prevalence of TORCH infections was 8% in aborted women. Hepatitis B virus (HBV) and mixed TORCH infections constituted the highest percentage of TORCH pathogens in aborted women, constituting 6%. The most detected histopathological finding in seropositive cases (50%) was POC, with mixed inflammatory infiltrates and chronic endometritis, while in seronegative aborted women, POC was normal (64.1%). There is a statistically significant increase in the mean count of white blood cells in seropositive women. Therefore, it is important to provide health campaigns to bring awareness to the population about the risk factors of infectious agents to be avoided, especially during pregnancy.

Abstract The receptor tyrosine kinase AXL is a member of the TYRO3, AXL, and proto-oncogene tyrosine-protein kinase MER family and plays pleiotropic roles in cancer progression. AXL is expressed in immunosuppressive cells, which contributes to decreased efficacy of immunotherapy. Therefore, we hypothesized that AXL inhibition could serve as a strategy to overcome resistance to chimeric antigen receptor T (CAR T)–cell therapy. To test this, we determined the impact of AXL inhibition on CD19-targeted CAR T (CART19)–cell functions. Our results demonstrate that T cells and CAR T cells express high levels of AXL. Specifically, higher levels of AXL on activated Th2 CAR T cells and M2-polarized macrophages were observed. AXL inhibition with small molecules or via genetic disruption in T cells demonstrated selective inhibition of Th2 CAR T cells, reduction of Th2 cytokines, reversal of CAR T-cell inhibition, and promotion of CAR T-cell effector functions. AXL inhibition is a novel strategy to enhance CAR T-cell functions through two independent, but complementary, mechanisms: targeting Th2 cells and reversing myeloid-induced CAR T-cell inhibition through selective targeting of M2-polarized macrophages.

Introduction: Despite that trastuzumab (TZB) improves life expectancy and prognosis in patients with HER-2 positive breast cancer (BC), cardiotoxicity is a well-established complication. However, other potential cardiac complications of TZB such as pulmonary hypertension (PHTN) are poorly investigated. Aim: Evaluate the effect of TZB on pulmonary artery systolic pressure (PASP) and measure the predictors of developing elevated PASP. Methods: The medical records for all females who received TZB treatment for confirmed HER-2 positive BC at King Hussein Cancer Centre in Jordan between 2014 - 2019 who had a documented normal PASP of ≤35 mmHg before treatment were evaluated. Results: A total of 436 patients with a mean age of 53±11 years were included. The leading comorbidities were hypertension and diabetes mellitus affecting 26.2% and 15.4% of the patients, respectively. 15.6% of the study’s population were active smokers. Prior to initiating TZB, 22.3% had left ventricular hypertrophy (LVH), 14.3% had mild tricuspid valve regurgitation (TR), 11.5% had pericardial effusion, and 4.8% had mild aortic valve regurgitation (AR).Of alarming significance, 24.8% of the patients had their PASP increased after one year. Patients who developed elevated PASP tend to be older (56.9 vs 51.3, P<0.001), had baseline mild TR (26.3% vs 10.3%, P<0.001), LVH (39.4% vs 17.6%, P<0.001), and mild AR (9.1% vs 3.7%, P=0.031).Multivariate analysis showed that the only predictors of elevated PASP are TR at baseline (OR 3.9; 95% CI 1.58-6.17, P=0.001), and age (OR 1.04; 95% CI 1.01-1.06, P=0.005). However, baseline LVH (OR 1.79; 95% CI 0.98-3.26, P=0.057), number of the TZB cycles (OR 0.93; 95% 0.85-1.02, P=0.152), and the total dose of TZB (OR 1; 95% CI 1-1, P=0.284) were non-significant predictors. At 1 year follow up, the mean total cumulative dose of TZB was 6.8±1.8 grams that were given at a mean of 15±3 cycles. Moreover, major events (heart failure exacerbation, pulmonary embolism, and acute coronary syndrome) were recorded in 3.9% of the patients and death occurred in 4.1%. Conclusions: Trastuzumab demonstrated an increase in the PASP in a considerable portion of the patients at 1-year interval with this elevation being substantially concerning due to being highly indicative of PHTN.

The snATAC + snRNA platform allows epigenomic profiling of open chromatin and gene expression with single-cell resolution. The most critical assay step is to isolate high-quality nuclei to proceed with droplet-base single nuclei isolation and barcoding. With the increasing popularity of multiomic profiling in various fields, there is a need for optimized and reliable nuclei isolation methods, mainly for human tissue samples. Herein we compared different nuclei isolation methods for cell suspensions, such as peripheral blood mononuclear cells (PBMC, n = 18) and a solid tumor type, ovarian cancer (OC, n = 18), derived from debulking surgery. Nuclei morphology and sequencing output parameters were used to evaluate the quality of preparation. Our results show that NP-40 detergent-based nuclei isolation yields better sequencing results than collagenase tissue dissociation for OC, significantly impacting cell type identification and analysis. Given the utility of applying such techniques to frozen samples, we also tested frozen preparation and digestion (n = 6). A paired comparison between frozen and fresh samples validated the quality of both specimens. Finally, we demonstrate the reproducibility of scRNA and snATAC + snRNA platform, by comparing the gene expression profiling of PBMC. Our results highlight how the choice of nuclei isolation methods is critical for obtaining quality data in multiomic assays. It also shows that the measurement of expression between scRNA and snRNA is comparable and effective for cell type identification.

Abstract Glioblastoma multiforme (GBM) is the most aggressive and common form of brain tumor, and it is characterized by an immunosuppressive tumor microenvironment (TME) that supports tumor growth and pathology. An integral part of the GBM stroma is the mesenchymal stem cells (MSC), which promote GBM growth. Receptor tyrosine kinase EphA3, a member of the Eph family, is overexpressed in GBM tumors and in MSCs within TME, making it an attractive target for Chimeric antigen receptor T-cell (CART) therapy. CART has proven incredible promise in certain hematological malignancies therapy but remained short in solid tumors, particularly GBM. We hypothesize that concurrent targeting of cancers and TME by EphA3-CART can be a potential therapeutic option in GBM. We first showed the expression of the EphA3 receptor by Immunohistochemistry and western blot on GBM patient-derived xenograft (PDX), as well as on the SNB-19 and U87 human GBM cell lines and adipose-derived MSCs using flow cytometry. We then generated EphA3-CAR construct derived from a clinically validated anti-EphA3 scFv cloned in a lentiviral vector containing CD28 and CD3ζ, and generated EphA3 CART cells with over 90% efficiency. EphA3 CART cells demonstrated potent antigen specific effector functions. To demonstrate the antitumor activity in vivo, we subcutaneously grafted 1 × 106 GBM cells per NOD-SCID-γ-/- mice and at ~150-300 mm3 tumor volume, mice were then treated with 5 × 106 EphA3-CART or un-transduced (UTD) T cells control. Tumors were monitored by bioluminescence imaging (BLI) and caliper measurements, and mice were followed for survival. Mice treated with EphA3 CART cells exhibited a significant reduction in tumor volume (p&lt;0.005), and prolonged overall survival (p=0.0015). Finally, we tested the ability of EphA3-CART cells to overcome MSC induced CART cell suppression by co-culturing EphA3-CART cells with MSC and EphA3+ GBM target cells while using the CD19+ mantle cell lymphoma cell line JeKo-1 and CART19 as control. MSC conditioned coculture of GBM and CART showed significant suppression of CAR19 in the CART19 - JeKo-1 system (p=000.3), but no significant suppression of EphA3-CART cells. In summary, our results indicate that EphA3 CART cells exhibited potent and specific antitumor activity in vitro and in vivo and ameliorated MSC induced inhibition of CART cell functions, representing a potentially promising therapeutic option in GBM. Citation Format: Ekene J. Ogbodo, Michael W. Ruff, Reona L. Sakemura, Claudia ManriquezRoman, Truc Huynh, James H. Girsch, Olivia L. Sirpilla, Kun Yun, Carli M. Stewart, Ismail Can, Lionel K. Fonkoua, Mehrdad Hefazi, Elizabeth L. Siegler, Saad S. Kenderian. Simultaneous targeting of EphA3 on glioblastoma and tumor microenvironment to overcome resistance to CART cell therapy in brain cancer. [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 5017.

Abstract Objective To assess the relationship between ovarian hyperstimulation syndrome (OHSS) and adverse outcomes using population-based data in the United States. The hypothesis is that patients with OHSS were more likely to deliver preterm and more likely to have hypertensive disorders. Methods This retrospective cohort study identified 94 patients with OHSS and 183 matched referents in eight counties in Minnesota. Data were collected regarding pregnancy history, infertility treatment, and pregnancy outcomes. Using the Rochester Epidemiology Project, study subjects were identified from female patients, aged 18 to 49 years, who were diagnosed with infertility from January 2, 1995 to December 1, 2017, and had a pregnancy greater than 20 weeks' gestation. The primary outcome was preterm delivery or hypertensive disorder of pregnancy incidence in the OHSS group when compared with control patients. Chi-squared test, t test, and multivariate logistic models were used where appropriate. Results Patients with OHSS were more likely to deliver preterm (odds ratio, 2.14; 95% confidence interval, 1.26–3.65; P < 0.01), and their neonates were more likely to be small for gestational age (odds ratio, 4.78; 95% confidence interval, 1.61–14.19; P < 0.01). No significant differences between the groups were observed in any other outcome. Patients with OHSS are more likely to deliver preterm if they undergo fresh transfer compared with a freeze all and subsequent frozen transfer (odds ratio, 3.03, 95% confidence interval, 1.20–7.66, P = 0.02). Conclusion OHSS may lead to preterm birth and small-for-gestational-age neonates, which changes patient counseling and leads to arranging specialized obstetrical care for these patients with OHSS.

e17535 Background: High-grade serous ovarian carcinoma (HGSOC) is the most frequent and deadliest type of ovarian cancer (OC). With the recent advances in transcriptomic and epigenomic profiling of cancer at single cell resolution, it has become clear that intra-tumor heterogeneity driven by genetic and epigenetic factors may be the basis of drug resistance. Current treatments are not homogeneously effective against all the cancer cell subpopulations, thus enabling resistance. With single cell sequencing technology, characterization of cell signatures and regulatory mechanisms may translate to prognostic markers and novel drug targets. Methods: The transcriptomic and epigenomic landscape of HGSOC was mapped for five tumor samples from debulking surgeries and one tissue patient-derived xenograft (PDX) using the 10X genomics sequencing platform. This strategy permits the combination of single cell ATAC-seq and single cell RNA-seq within the same nucleus (Multiome). Isolation of nuclei, library preparation and sequencing were performed following 10X genomics protocols. Sample quality and library quality were assessed by nuclear morphology and fragment size analysis, respectively. Sequenced reads were mapped to the human genome and downstream analysis including copy number variant prediction, pathway enrichment, motif enrichment and gene regulatory network analysis were performed. The sampled patients continue to be followed for clinical outcomes, such as response to therapy. Clinical data including patient age, grade, and stage of cancer, debulking status, CA-125 levels, and neoadjuvant status, were used in conjunction with the genomics data to characterize patient specific molecular regulatory signatures. Results: Cells from the six samples (total N= 26,421) were projected via UMAP. Immune and stromal cells were shown to cluster by cell-type while cancer cells clustered by patient. Cancer cells were identified as cycling cells, ciliated cells, cells with enrichment of the JAK-STAT signaling pathway, and cells exhibiting cancer stemness signatures. Transcription factor motifs and binding-site enrichment in open chromatin regions reveal transcription regulation-based subpopulations. Using the combined gene expression and open chromatin information from these cells, there is potential to uncover the genetic regulatory network(s) that drives treatment resistance. Conclusions: With single cell technology, specific clusters of cancer and tumor microenvironment cells can be classified. Beyond characterizing patient specific signatures, multiome enables the discovery of genomic, transcriptomic and epigenomic signatures that provide insight in tumor progression and treatment resistance.

Accurate prediction of progression in subjects at risk of Alzheimer's disease is crucial for enrolling the right subjects in clinical trials. However, a prospective comparison of state-of-the-art algorithms for predicting disease onset and progression is currently lacking. We present the findings of "The Alzheimer's Disease Prediction Of Longitudinal Evolution" (TADPOLE) Challenge, which compared the performance of 92 algorithms from 33 international teams at predicting the future trajectory of 219 individuals at risk of Alzheimer's disease. Challenge participants were required to make a prediction, for each month of a 5-year future time period, of three key outcomes: clinical diagnosis, Alzheimer's Disease Assessment Scale Cognitive Subdomain (ADAS-Cog13), and total volume of the ventricles. The methods used by challenge participants included multivariate linear regression, machine learning methods such as support vector machines and deep neural networks, as well as disease progression models. No single submission was best at predicting all three outcomes. For clinical diagnosis and ventricle volume prediction, the best algorithms strongly outperform simple baselines in predictive ability. However, for ADAS-Cog13 no single submitted prediction method was significantly better than random guesswork. Two ensemble methods based on taking the mean and median over all predictions, obtained top scores on almost all tasks. Better than average performance at diagnosis prediction was generally associated with the additional inclusion of features from cerebrospinal fluid (CSF) samples and diffusion tensor imaging (DTI). On the other hand, better performance at ventricle volume prediction was associated with inclusion of summary statistics, such as the slope or maxima/minima of patient-specific biomarkers. On a limited, cross-sectional subset of the data emulating clinical trials, performance of the best algorithms at predicting clinical diagnosis decreased only slightly (2 percentage points) compared to the full longitudinal dataset. The submission system remains open via the website https://tadpole.grand-challenge.org, while TADPOLE SHARE (https://tadpole-share.github.io/) collates code for submissions. TADPOLE's unique results suggest that current prediction algorithms provide sufficient accuracy to exploit biomarkers related to clinical diagnosis and ventricle volume, for cohort refinement in clinical trials for Alzheimer's disease. However, results call into question the usage of cognitive test scores for patient selection and as a primary endpoint in clinical trials.

Background: Blastocystis hominis (B. hominis) is a ubiquitous parasite that has spread worldwide and is commonly present in human stool specimens. It was hypothesized that infection with B. hominis plays a role in the pathogenesis of autoimmune diseases in humans. The aim of this study is to test this hypothesis by investigating patients with autoimmune diseases. Patients with various types of autoimmune diseases with gastrointestinal symptoms were enrolled in this study as cases (n = 72) along with nongastrointestinal symptom patients as controls (n = 58). All participants in this study were subjected to history taking and were investigated for B. hominis infection via wet-mount microscopic stool examinations, staining with trichrome stain, and molecular-based tests applied to their fecal samples. Blood samples were also tested for complete blood counts. B. hominis were identified with specific PCR more in cases (12/72; 16.6%) than in controls (3/58; 5.2%), with a significant difference (p < 0.05). Significant decreases in white blood cell counts were demonstrated in systemic lupus erythematosus (SLE) and ulcerative colitis (UC) patients infected with B. hominis when compared to patients with nongastrointestinal symptoms (p-value < 0.05).