Endocyte (United States)

ABSTRACTAcute chest syndrome (ACS) is a severe complication of sickle cell disease (SCD) occurring in ~50% of patients, some presenting frequent episodes. We lack tools to identify patients at high risk of ACS occurrence or frequent episodes. Epidemiological studies have found an association between asthma and ACS, but whether this link is causal is unclear. We used polygenic scores (PGS) to analyze whether the genetic propensity for asthma was associated with ACS and could be used to stratify the risk of ACS. We identified that PGS for asthma (PGSasthma) was associated with ACS rate (number of episodes per year), but not ACS occurrence, in both the CSSCD (n = 1278) and the GEN‐MOD (n = 406) prospective SCD cohorts, independently of fetal hemoglobin (HbF) (β = 0.17, standard error = 0.06, p = 0.006). This effect was most pronounced in patients with low HbF levels. Combining PGSasthma and HbF identified a population at high risk of frequent ACS after a first episode: individuals within the highest PGSasthma quintile and the lowest HbF quintile. Finally, we found that the genetic correlation between these two conditions only partially overlapped. This suggests that genetically determined asthma is not the unique contributor to the genetic propensity for ACS and that other genetic determinants may also play a role. In sum, our results suggest that patients with a high genetic propensity for asthma are prone to frequent ACS if not protected by high HbF levels. Combining PGSasthma and HbF may allow identifying patients at high risk of frequent ACS after a first episode for personalized management.

Foetal cells are detectable in women decades postpartum, a state termed foetal microchimerism. The interplay between these semi-allogeneic foetal cells and the mother could be affected by genetic mismatches in the HLA loci. Here, we relate HLA allele and molecular mismatch values to the presence and quantity of foetal microchimerism in the maternal circulation during pregnancy and postpartum. A total of 76 pregnant women were included, of which 59 were followed up 1-8 years postpartum. Maternal and foetal DNA was genotyped for HLA class I and II loci. Foetal cells in maternal buffy coat were detected by qPCR, targeting inherited paternal alleles. Antibody-verified eplet mismatch and Predicted Indirectly Recognisable HLA Epitopes (PIRCHE) scores were calculated to quantify foetal-maternal histocompatibility from the mother's perspective. Circulating foetal cells were detected in 50.0% (38/76) of women during pregnancy, and 25.4% (15/59) postpartum. During pregnancy, HLA class II antibody-verified eplet mismatch load and PIRCHE scores correlated negatively with the presence and quantity of foetal cells in the maternal circulation. Postpartum, HLA class I allele mismatches correlated negatively with foetal microchimerism presence, while HLA class II allele mismatches, HLA class I and II antibody-verified eplet mismatch load, and PIRCHE-I and PIRCHE-II scores correlated negatively with both microchimerism presence and quantity. The correlation between mismatch parameters aimed at evaluating the risk of humoral and T cell-mediated allorecognition and foetal microchimerism was more evident postpartum than during pregnancy. The observed predictive effect of foetal-maternal histocompatibility on foetal microchimerism suggests that circulating foetal cells are subject to clearance by the maternal immune system. We propose that allorecognition of foetal cells in the maternal circulation and tissues influences any long-term effect that foetal microchimerism may have on maternal health.

This Consensus Statement from an international, multidisciplinary workshop sponsored by the Pituitary Society offers evidence-based graded consensus recommendations and key summary points for clinical practice on the diagnosis and management of prolactinomas. Epidemiology and pathogenesis, clinical presentation of disordered pituitary hormone secretion, assessment of hyperprolactinaemia and biochemical evaluation, optimal use of imaging strategies and disease-related complications are addressed. In-depth discussions present the latest evidence on treatment of prolactinoma, including efficacy, adverse effects and options for withdrawal of dopamine agonist therapy, as well as indications for surgery, preoperative medical therapy and radiation therapy. Management of prolactinoma in special situations is discussed, including cystic lesions, mixed growth hormone-secreting and prolactin-secreting adenomas and giant andaggressive prolactinomas. Furthermore, considerations for pregnancy and fertility are outlined, as well as management of prolactinomas in children and adolescents, patients with an underlying psychiatric disorder, postmenopausal women, transgender individuals and patients with chronic kidney disease. The workshop concluded that, although treatment resistance is rare, there is a need for additional therapeutic options to address clinical challenges in treating these patients and a need to facilitate international registries to enable risk stratification and optimization of therapeutic strategies.

Abstract Introduction The vestibule is an external genital structure lying 90° to the vaginal canal opening. During the thrusting movements associated with penetrative sexual activity, the vestibule undergoes significant stretching during both invagination and evagination. The vestibule also undergoes extreme stretching during childbirth. The rete pegs in the non-keratinized squamous epithelium facilitate the ability of the vestibule to mechanically stretch. More importantly, however, the vestibular lamina propria, with nerves, blood vessels, glands, needs to contain a high density of extracellular matrix (ECM) proteins with stretching capabilities. In particular, the ECM protein elastin endows skin structures with the ability to stretch, recoil, and have elasticity. Elastin has previously been shown to influence mechanical stretching properties of tendons. Objective The objective of this study was to examine the density of elastin proteins in the vestibule in reproductive age patients as there are currently limited data. Methods A total of 7 reproductive age patients with normal sex steroid hormone values and complaints of entrance dyspareunia were diagnosed with neuroproliferative vestibulodynia and subsequently underwent complete vestibulectomy. Tissue blocks were created from the specimen, and 5 μm sections studied with immunohistochemical staining with primary antibody. Sections were then counterstained with Gill-modified hematoxylin. Results In all vestibular specimen, there was very high density of elastin underneath the epithelium in rete pegs and throughout the entire lamina propria (Fig 1). There is a density gradient with the highest density immediately deep to the basement membrane of the epithelium. Elastin fibers were arranged in all orientations: coronal (frontal), sagittal (longitudinal), and transverse (axial) planes. Conclusions The high density of elastin fibers in rete pegs and throughout the lamina propria in a multi-planar orientation is suggestive of extreme stretching capability in eugonadal vestibular tissues. The highest density of elastin staining was seen immediately below the basement membrane of the epithelium, which may provide tissue integrity and shear resilience to the epithelial layer. In sex steroid hormone insufficiency states such as menopause or with use of combined hormonal contraceptives, individuals may develop loss of introital elasticity and fissures due to a deficiency of the ECM protein elastin. More research is needed on elastic fiber production, organization, and integration with other cutaneous extracellular matrix proteins, and how they may be affected by sex steroid hormones. Disclosure Any of the authors act as a consultant, employee or shareholder of an industry for: Endoceutics, Inc.

Abstract Introduction The introitus has three embryologic tissue types: ectoderm (vulva), endoderm (vestibule), and mesoderm (vagina). The vulva and vagina have been shown previously to have estrogen receptors (ER), androgen receptors (AR) and progesterone receptors (PR). ER and AR have been demonstrated in vaginal epithelial cells, as well as in fibroblasts and smooth muscle of underlying stroma. Vulvar ER and AR were demonstrated in epidermal keratinocytes and dermal fibroblasts of hair and non-hair bearing skin. AR were also seen in vulvar sebaceous glands, sweat glands, and hair follicles. PR was seen in vaginal epithelium, fibroblasts, and smooth muscle but not in the vulva. Despite the importance of the vestibule to sexual health, there is limited information regarding vestibular hormone receptors. Objective To better understand vestibular hormonal physiology, immunohistochemical (IHC) studies of ER, AR, and PR was performed on excised vestibulectomy tissue specimen. Methods IHC was performed on 7 vestibular specimen from eugonadal individuals with confirmed diagnosis of neuroproliferative vestibulodynia based on positive CD117 and PGP9.5 staining, consistent with excess mast cells and nerve endings, respectively. Specimen were also studied with primary antibodies (Anti-ERα, Ab 32063; Anti-AR, MI 06680; Anti-PR, Dako PGR636). Finally, sections were lightly counterstained with Gill-modified hematoxylin. Results ER and AR were richly expressed in tissue samples from all 7 patients. Dense labeling of both receptor types (AR on left, on ER right) was identified in the germinal layer of the epithelium (Fig. 1) with comparatively sparse expression in the subepithelial stroma. AR, but not ER, were noted in mucinous vestibular glands (Fig. 2). While labeling of PR was highly variable (Fig. 3), all samples showed evidence of moderate PR labeling in the subepithelial stroma. Variability may be due to menstrual cycle phase during which samples were obtained, however this information was not collected. Conclusions This preliminary study provides evidence of dense ER and AR expression in the vestibule, consistent with it being in part an androgenic organ. This supports the concept that the vestibule may be more effectively treated with combined testosterone and estradiol cream than with topical estradiol alone in patients with entrance dyspareunia from hormonally mediated vestibulodynia or genitourinary syndrome of menopause. Also consistent with these findings is that treatment with intravaginal DHEA, which converts to testosterone and estradiol intracrinologically and spreads to the vestibule, successfully resolves entrance dyspareunia, as shown in a previous clinical trial. While the significance of variable vestibular progesterone receptor expression is unclear, the addition of topical progesterone in patients with entrance dyspareunia may be considered. Disclosure Any of the authors act as a consultant, employee or shareholder of an industry for: Endoceutics, Inc

While a 3-tier oral epithelial dysplasia grading system has been utilized for decades, it is widely recognized as a suboptimal risk indicator for transformation to cancer. A 2-tier grading system has been proposed, although not yet validated. In this study, the 3-tier and 2-tier dysplasia grading systems, and an S100A7 immunohistochemical signature-based grading system were compared to assess prediction of risk of transformation to oral cancer. Formalin-fixed, paraffin-embedded biopsy specimens with known clinical outcomes were obtained retrospectively from a cohort of 48 patients. Hematoxylin and eosin-stained slides were used for the 2- and 3-tier dysplasia grading, while S100A7 for biomarker signature-based assessment was based on immunohistochemistry. Inter-observer variability was determined using Cohen's kappa ( K ) statistic with Cox regression disease free survival analysis used to determine if any of the methods were a predictor of transformation to oral squamous cell carcinoma. Both the 2- and 3-tier dysplasia grading systems ranged from slight to substantial inter-observer agreement ( Kw between 0.093 to 0.624), with neither system a good predictor of transformation to cancer (at least P =0.231; ( P >>>0.05). In contrast, the S100A7 immunohistochemical signature-based grading system showed almost perfect inter-observer agreement ( Kw =0.892) and was a good indicator of transformation to cancer ( P =0.047 and 0.030). The inherent grading challenges with oral epithelial dysplasia grading systems and the lack of meaningful prediction of transformation to carcinoma highlights the significant need for a more objective, quantitative, and reproducible risk assessment tool such as the S100A7 immunohistochemical signature-based system.

BACKGROUND Chronic HBV infection is the leading cause of HCC, particularly in Asia and Africa. Despite demonstrating response rates ranging from 15% to 30% in patients with HCC, immune checkpoint inhibitor (ICI) immunotherapy has not achieved the optimal efficacy as observed in melanoma.1 Recent studies reveal that immunotherapies may be more effective in viral etiologies, particularly HBV, than nonviral HCC,2,3 while there is no difference related to the etiology of HCC in patients receiving tyrosine-kinase inhibitor therapy.2 These findings suggest the profound implication of the tumor microenvironment (TME) in response to immunotherapies. It is thus intriguing to ask why HBV+-HCC has a better response to ICI therapy, what are the roles of HBV-specific immune cells in modulating the debilitated immune in HCC, and whether it is possible to revive the HBV-related immune for anticancer therapy. Does HBV-specific immune sculpture the immune in TME of HCC? Multi-omics studies, including single-cell RNA sequencing, cytometry by time-of-flight, and multiplexed immunohistochemistry, have shown that the TME of HBV+-HCC is immune hot but highly immunosuppressive characterized by the enrichment of exhausted CD8+ T cells, regulatory T cells (Treg), and HBV-specific CD8+ T cells recognizing diverse HBV-derived epitopes, including tissue-resident memory T cells (TRM).4,5 In addition, Treg and TRM from HBV+-HCC express more PD-1 and are functionally immune suppressive and exhausted than nonviral HCC. Clinically, Treg cells in the TME were associated with a poor prognosis, while HBV-specific TRM cells were associated with a good prognosis in HCC.4,5 Together, these studies suggest that chronic HBV infection usually sculptures a hot but debilitated immune status favoring tumor development in the liver. How chronic HBV infection modulates immune profiles in the TME of HCC? You et al6 combined epigenomic and transcriptomic approaches to delineate the HBV-specific Treg, CD8+ exhausted T cells (Tex), and HBV-specific effector memory CD8 T cells and characterized their T-cell receptor repertoire clonality and specificity in a cohort of 43 patients with HBV+-HCC and 11 patients with HBV--HCC. They found a shared epigenomic program that regulates the differentiation of HBV-specific Treg and Tex and the HBV-related Treg-CTLA4 and CD8+Tex-TOX subsets in the TME of HCC. Meanwhile, a significant proportion of HBV-specific effector memory CD8 T cells and HBV-related FGFBP2+ CD8+ T cells were governed by shared transcription factor motifs of AP1, NFE2, and BACH1/2,6 suggesting the role of chronic HBV infection in sculpturing the immune profile of TME (Figure 1).FIGURE 1: The epigenomic programs leash and unleash the HBV-associated immune for tumor development and antitumor immunotherapy, respectively. Created with BioRender.com. Abbreviations: ICI, immune checkpoint inhibitor; TCR, T-cell receptor; TME, tumor microenvironment; Treg, regulatory T cells.Can we reinvigorate the HBV-induced immune from debilitation to empowered anti-tumor immunity? Clinically, Treg cells in the TME were associated with a poor prognosis, while HBV-specific TRM cells were associated with a good prognosis in HCC.4,5 Furthermore, HBV-specific CD8+T cells expressed FASLG, linked to the FASLG-FAS cell-to-cell communication network involving HBV-specific Treg cells and tumor cells.6 These findings hold substantial implications for targeting immunosuppressive cells like HBV-related Treg in TME to improve the efficacy of immunotherapy. Indeed, a recent study shows the treatment with a CCR4 antagonist blocked intratumoral Treg accumulation, overcame sorafenib resistance, rescued CD8+ T-cell function, and sensitized tumors to PD-1 checkpoint blockade in HBV+-HCC.7 In contrast, the anti-PD1 treatment led to an increase in the number and size of tumor nodules, which correlated with increased hepatic CD8+PD1+TOX+ T cells in NASH-HCC.3 These phenomena may partially explain why ICI therapy is more effective in patients with HBV+-HCC than in those with HBV−-HCC. Future challenges The discovery by You et al6 sheds light on the potential for virus-specific T cells to serve as elicitors of anti-tumor immunity in viral-associated HCC. However, the discrepancies in ICI responsiveness between HBV+-HCC and HCV+-HCC has not been fully explained. In recent years, more and more patients with chronic hepatitis B have achieved clinical HBV cured (HBsAg seroclearance), particularly after a finite nucleos(t)ide analog therapy.8 For these patients, it is unclear whether the "epigenetic scar" seen in patients cured of chronic hepatitis C9,10 is also present. As such, how are the potential impacts of the HBV-specific immune on immunotherapies of HCC in patients with HBsAg seroclearance? Nevertheless, reanimating the long-term infection-induced exhausted immune cells to treat virus-related cancers provides a promising paradigm for developing targeted immunotherapies.

Simple SummaryAge-related cardiovascular disease is the foremost cause of death globally. Despite medical advances, we have been unable to stop or reverse the underlying pathology. We offer a model that is consistent with all clinical and laboratory data, can explain the clinical heterogeneity of age-related disease both within (e.g., human patients) and between species, and is predictively valid for interventional clinical trials. Cell aging is not only the central mechanism of such disease, but studies of human cells, of human tissues, and in animals demonstrate that we can effectively reverse cell aging at the cellular, genetic, and epigenetic levels. Given current biotechnology techniques, the reversal of cell aging is now technically feasible in human trials. The implications for our ability to cure and prevent cardiovascular disease are without historical precedent. This approach may not only allow us to intervene effectively in age-related cardiovascular disease but will likely lower (rather than raise) the costs of global medicine.Despite progress in biomedical technologies, cardiovascular disease remains the main cause of mortality. This is at least in part because current clinical interventions do not adequately take into account aging as a driver and are hence aimed at suboptimal targets. To achieve progress, consideration needs to be given to the role of cell aging in disease pathogenesis. We propose a model unifying the fundamental processes underlying most age-associated cardiovascular pathologies. According to this model, cell aging, leading to cell senescence, is responsible for tissue changes leading to age-related cardiovascular disease. This process, occurring due to telomerase inactivation and telomere attrition, affects all components of the cardiovascular system, including cardiomyocytes, vascular endothelial cells, smooth muscle cells, cardiac fibroblasts, and immune cells. The unified model offers insights into the relationship between upstream risk factors and downstream clinical outcomes and explains why interventions aimed at either of these components have limited success. Potential therapeutic approaches are considered based on this model. Because telomerase activity can prevent and reverse cell senescence, telomerase gene therapy is discussed as a promising intervention. Telomerase gene therapy and similar systems interventions based on the unified model are expected to be transformational in cardiovascular medicine.

BackgroundMetastatic castration-resistant prostate cancer remains fatal despite recent advances. Prostate-specific membrane antigen (PSMA) is highly expressed in metastatic castration-resistant prostate cancer. Lutetium-177 (177Lu)–PSMA-617 is a radioligand therapy that delivers beta-particle radiation to PSMA-expressing cells and the surrounding microenvironment.MethodsWe conducted an international, open-label, phase 3 trial evaluating 177Lu-PSMA-617 in patients who had metastatic castration-resistant prostate cancer previously treated with at least one androgen-receptor–pathway inhibitor and one or two taxane regimens and who had PSMA-positive gallium-68 (68Ga)–labeled PSMA-11 positron-emission tomographic–computed tomographic scans. Patients were randomly assigned in a 2:1 ratio to receive either 177Lu-PSMA-617 (7.4 GBq every 6 weeks for four to six cycles) plus protocol-permitted standard care or standard care alone. Protocol-permitted standard care excluded chemotherapy, immunotherapy, radium-223 (223Ra), and investigational drugs. The alternate primary end points were imaging-based progression-free survival and overall survival, which were powered for hazard ratios of 0.67 and 0.73, respectively. Key secondary end points were objective response, disease control, and time to symptomatic skeletal events. Adverse events during treatment were those occurring no more than 30 days after the last dose and before subsequent anticancer treatment.ResultsFrom June 2018 to mid-October 2019, a total of 831 of 1179 screened patients underwent randomization. The baseline characteristics of the patients were balanced between the groups. The median follow-up was 20.9 months. 177Lu-PSMA-617 plus standard care significantly prolonged, as compared with standard care, both imaging-based progression-free survival (median, 8.7 vs. 3.4 months; hazard ratio for progression or death, 0.40; 99.2% confidence interval [CI], 0.29 to 0.57; P<0.001) and overall survival (median, 15.3 vs. 11.3 months; hazard ratio for death, 0.62; 95% CI, 0.52 to 0.74; P<0.001). All the key secondary end points significantly favored 177Lu-PSMA-617. The incidence of adverse events of grade 3 or above was higher with 177Lu-PSMA-617 than without (52.7% vs. 38.0%), but quality of life was not adversely affected.ConclusionsRadioligand therapy with 177Lu-PSMA-617 prolonged imaging-based progression-free survival and overall survival when added to standard care in patients with advanced PSMA-positive metastatic castration-resistant prostate cancer. (Funded by Endocyte, a Novartis company; VISION ClinicalTrials.gov number, NCT03511664.)