Neuroendocrine Status Research Articles

Metabolic Syndrome in Reproductive Age Women of Various Ethnic Groups. Neuroendocrine Status and Lipid Peroxidation System.

We analyzed the state of neuroendocrine regulation and LPO-antioxidant defense systems in reproductive age women with metabolic syndrome (MetS), representatives of the Russian and Buryat ethnic groups. Compared to the corresponding control groups, women from the Russian ethnic group with MetS had elevated levels of thyroid-stimulating hormone and free androgen index (FAI) and reduced levels of sex hormone-binding globulin, while women from the Buryat ethnic group had increased levels of prolactin and FAI. Changes in the LPO system in women of the Russian ethnic group with MetS consisted in an increase in the levels of substrates with double bonds, TBA-reactive substances, and fat-soluble vitamins. Buryat women with MetS had a higher content of primary oxidation products and reduced levels of glutathione. The results of the study indicate a hyperandrogenic shift in the neuroendocrine regulation system, as well as compensatory influences from different parts of the antioxidant defense system in women of reproductive age with MetS, depending on their ethnicity. These findings indicate the need for assessing and monitoring the levels of these metabolites in women with MetS, considering their ethnicity.

SMARCA4 controls state plasticity in small cell lung cancer through regulation of neuroendocrine transcription factors and REST splicing

IntroductionSmall Cell Lung Cancer (SCLC) can be classified into transcriptional subtypes with distinct degrees of neuroendocrine (NE) differentiation. Recent evidence supports plasticity among subtypes with a bias toward adoption of low-NE states during disease progression or upon acquired chemotherapy resistance. Here, we identify a role for SMARCA4, the catalytic subunit of the SWI/SNF complex, as a regulator of subtype shift in SCLC.MethodsATACseq and RNAseq experiments were performed in SCLC cells after pharmacological inhibition of SMARCA4. DNA binding of SMARCA4 was characterized by ChIPseq in high-NE SCLC patient derived xenografts (PDXs). Enrichment analyses were applied to transcriptomic data. Combination of FHD-286 and afatinib was tested in vitro and in a set of chemo-resistant SCLC PDXs in vivo.ResultsSMARCA4 expression positively correlates with that of NE genes in both SCLC cell lines and patient tumors. Pharmacological inhibition of SMARCA4 with FHD-286 induces the loss of NE features and downregulates neuroendocrine and neuronal signaling pathways while activating non-NE factors. SMARCA4 binds to gene loci encoding NE-lineage transcription factors ASCL1 and NEUROD1 and alters chromatin accessibility, enhancing NE programs. Enrichment analysis applied to high-confidence SMARCA4 targets confirmed neuron related pathways as the top GO Biological processes regulated by SMARCA4 in SCLC. In parallel, SMARCA4 also controls REST, a known suppressor of the NE phenotype, by regulating SRRM4-dependent REST transcript splicing. Furthermore, SMARCA4 inhibition drives ERBB pathway activation in SCLC, rendering SCLC tumors sensitive to afatinib.ConclusionsThis study nominates SMARCA4 as a key regulator of the NE state plasticity and defines a novel therapeutic strategy for SCLC.

Microenvironment shapes small-cell lung cancer neuroendocrine states and presents therapeutic opportunities

Small-cell lung cancer (SCLC) is the most fatal form of lung cancer. Intratumoral heterogeneity, marked by neuroendocrine (NE) and non-neuroendocrine (non-NE) cell states, defines SCLC, but the cell-extrinsic drivers of SCLC plasticity are poorly understood. To map the landscape of SCLC tumor microenvironment (TME), we apply spatially resolved transcriptomics and quantitative mass spectrometry-based proteomics to metastatic SCLC tumors obtained via rapid autopsy. The phenotype and overall composition of non-malignant cells in the TME exhibit substantial variability, closely mirroring the tumor phenotype, suggesting TME-driven reprogramming of NE cell states. We identify cancer-associated fibroblasts (CAFs) as a crucial element of SCLC TME heterogeneity, contributing to immune exclusion, and predicting exceptionally poor prognosis. Our work provides a comprehensive map of SCLC tumor and TME ecosystems, emphasizing their pivotal role in SCLC's adaptable nature, opening possibilities for reprogramming the TME-tumor communications that shape SCLC tumor states.

The influence of a course of complex application of supravenous laser irradiation of blood and phototherapy on indicators of the neurohormonal status of patients with true eczema

BACKGROUND: In the pathogenesis of true eczema, an important place is occupied by stress, under the influence of which stimulation of the hypothalamus and brain stem occurs, as well as activation of the hypothalamic-pituitary-adrenal axis and the autonomic nervous system. Stress-induced secretion of neurotransmitters impairs the barrier function of the epidermis, slows wound healing, increases itching and inflammation, and suppresses immunity by changing the immune response of T-helper cells. AIM: Assessment of the severity of the corrective effect of a course of application of supravenous laser irradiation of blood and broadband medium-wave phototherapy on the blood levels of adrenocorticotropin, cortisol and beta-endorphin in patients with true eczema with a moderate form of the course. MATERIALS AND METHODS: The study was carried out with the participation of 120 patients with a moderate form of true eczema. Using simple fixed randomization, all patients were divided into 4 groups. The first group (control, n=30) received only basic drug therapy, including the use of detoxification, hyposensitizing, antihistamines and topical glucocorticosteroids. Patients of the second group (comparison group 1, n=30) additionally underwent a course of supravenous laser irradiation of blood. In the third group (comparison group 2, n=30), broadband medium-wave phototherapy was performed against the background of basic therapy. Patients of the fourth group (main group, n=30) in addition to basic therapy received a course of combined exposure to supravenous laser blood irradiation and phototherapy. The concentrations of adrenocorticotropin, cortisol and beta-endorphin were determined in the blood of patients twice (before the start of treatment and 1 month after the start of therapy) using ELISA. The clinical condition of patients in the groups was assessed by changes in the EASI index. Additionally, we studied the dynamics of the level of reactive and personal anxiety using the Spielberger–Khanin questionnaire. RESULTS: Positive dynamics of adrenocorticotropin, cortisol and beta-endorphin were noted only in the comparison groups and the main group, which proves the high efficiency of correction of the neurohumoral status of patients through a course of complex use of supravenous laser blood irradiation and broadband mid-wave phototherapy. Quantitative changes in the assessed parameters during complex therapy with therapeutic physical factors indicate a supra-additive version of their interaction, realized through different modalities, points of application and mechanisms for realizing the biological potential of physiotherapeutic effects. CONCLUSION: The results obtained convincingly prove the effectiveness of an additional course of use of supravenous laser irradiation of blood in combination with broadband medium-wave phototherapy when correcting the neuroendocrine status in patients with true eczema. The close correlation between neurohumoral indicators and clinical and psychological patterns of the disease proves the pathogenetic significance of the studied neuroendocrine indicators, which allows us to consider their dynamics as informative criteria for the effectiveness of the treatment.

Роль определения хромогранина А в лечении больных кастрационно-резистентным раком предстательной железы.

Introduction. Chromogranin A has the greatest diagnostic value in detecting neuroendocrine differentiation (NED) of a tumor. This work is devoted to the study of the therapy of castration-resistant prostate cancer (CRPC) using somatostatin (AS) analogues based on the assessment of the neuroendocrine status of the tumor. Material and methods. The study included 89 patients with CRPC aged 72.2±1.4 years. Localized prostate cancer was diagnosed in 6 (6.7%), locally distributed prostate cancer (T3-4N0M0) in 12 (13.5%) patients, metastatic (T3-4N0-1M1)–in 71 (79.8%). An increase in CgA was observed in 31.5% (n=28) of CRPC patients. Combination therapy was performed with Octreotide depot (Pharm–Sintez, Russia) according to the scheme: 20 mg n/a once every 28 days in combination with dexamethasone at a dose of 4 mg per day for 1 month. Then the dose of dexamethasone was reduced. Patients were monitored every 28 days. The effectiveness was evaluated by the dynamics of PSA and CgA, as well as (ultrasound, abdominal MSCT, pelvic MRI and osteoscintigraphy). According to PSA, the answer was considered to be a decrease in the median PSA, or a lack of growth of more than 10%. Results. In patients with normal levels of CgA after 3 months of treatment, the median PSA significantly increased: from 57.0 [29.8; 94] ng/ml initially to 89.0 [47.4; 131.9] ng/ml, which was 56.1%, the decrease in CgA was 9.5% (from 2.1 to 1.9 nmol/l). In patients with elevated levels of CgA, there was a decrease in the median PSA value by 35.7%, from 169.5 [66.3; 235.3] ng/ml to 109.0 [44.8; 236.9] and CgA by 25% – from 5.2 [3.6; 7.1] to 3.9 [2.3; 5.2]. The median values and the proportion of PSA reduction in the group of «responders» stratified by CgA levels before and after 3 months of treatment were, respectively: in the group with normal CgA values – 43.0 [16.5; 154.3] nmol/l and 15.2 [7.2; 48.8] nmol/l or 64.7%; with CgA 3-7 nmol/l – 160.5 [35.1; 278.6] nmol/l and 42.4 [14.1; 88.4] nmol/l or 73.6%; in patients with CgA more than 7 nmol/L – 166.5 [28.0; 222.0] nmol/l and 81.7 [16.4; 151.0] nmol/l or 50.9%. Conclusion. Early detection of NED in CRPC with the use of the CgA marker makes it possible to stratify patients according to its severity and off r combination therapy with the inclusion of AS. An analysis of the immediate results of treatment with octreotide depot in combination with GnRH agonists and dexamethasone showed significantly higher efficacy in patients with initially increased levels of CgA, and the results achieved look preferable in patients with a moderate increase in this marker (within 3-7 nmol/l).

EVALUATION OF THE EFFECT OF NEUROPEPTIDES OF THE MELANOCORTIN FAMILY ON STRESS-ASSOCIATED CHANGES IN THE LEVEL OF CORTICOTROPIN-RELEASING HORMONE AND CORTICOSTERONE

Introduction. To date, the problem of studying the neuropeptide system of regulation of adaptive processes is relevant, since the stress reaction is a trigger for the occurrence of various diseases, in particular depression. Corticoliberin is considered as the main regulator of the hypothalamic-pituitary-adrenal axis, released in response to stress from the paraventricular nucleus of the hypothalamus, modulating the synthesis of adrenal hormones and further the processes of integration of neuroendocrine, autonomic and behavioral responses to stress. The aim of the study was to evaluate the effect of neuropeptides on stress–induced changes in corticotropin-releasing hormone and corticosterone. Material and methods. The experiment was carried out on male rats in the amount of 70 individuals, which were divided into several groups: a group of intact animals receiving an equiobject of water solution for intraperitoneally injection for 20 days (10 individuals), a group of animals receiving an equiobject of water solution for intraperitoneally injection in an experimentally created model of a depressive-like state for 20 days (20 individuals); experimental groups of animals who received intraperitoneally studied neuropeptide compounds Met-Glu-His-Phe-Pro-Gly-Pro and His-Phe-Arg-Trp-Pro-Gly-Pro at a dose of 100 mcg/kg/day for 20 days from day 1 of an experimentally created model of a depressive-like state. A depressive-like state was modeled by daily intermale confrontations for 20 days, as a result of which submissive and aggressive types of behavior were formed in rats. Neuroendocrine status was assessed by determining the level of key hormones of the hypothalamic-pituitary-adrenal axis (corticotropin-releasing hormone (corticoliberin) and corticosterone) in blood serum using enzyme immunoassay (ELISA). Results. In conditions of stress-induced depression, a statistically significant increase in corticoliberin and corticosteroid in the blood of rats of both types of behavior was found, the difference in comparative aspect with the indicators of the control group, while exposure to neuropeptides against the background of stressful exposure contributed to a decrease in corticosterone levels by an average of 30% (p<0.05). Conclusion. Thus, it was found that stress-induced depression is accompanied by hyperactivation of the hypothalamic-pituitary-adrenal axis, presumably due to a violation of the reverse inhibitory effects of endogenous glucocorticoids due to dysfunction of glucocorticoid receptors, which contributes to the development of a complex of manifestations of the main reactions, in particular, a decrease in the level of neurotrophic factors and the development of neurogenic inflammation. Neuropeptides of the glyproline series, in turn, showed the ability to reduce the activity of the hypothalamic-pituitary-adrenal axis by inhibiting the level of corticoliberin and corticosterone in conditions of stress-induced depression.

Intravenous infusion of lidocaine as a component of opioid-free anesthesia in thoracic surgery: prospective randomized single-center observational study

BACKGROUND: The possibility of reducing doses of narcotic analgesics during the perioperative period is widely discussed in modern scientific literature. This direction is called opioid-free anesthesia and is used in abdominal surgery, oncology, gynecology, and other areas. Publications on the use of opioid-free anesthesia in thoracic surgery are scarce.
 OBJECTIVE: To evaluate the analgesic effectiveness of intravenous lidocaine infusion as a component of anesthesia in thoracic surgery.
 MATERIALS AND METHODS: Ninety patients who underwent open lobectomy or pneumonectomy were examined. Depending on the analgesic component, patients were divided into three groups: group 1 (n=30) received intravenous infusion of lidocaine, group 2 (n=30) received epidural block, and group 3 (n=30) had fentanyl infusion. Blood pressure, heart rate, cortisol, and blood serum glycemia levels were measured. The intensity of postoperative pain syndrome and the need for promedol, tramadol, and sanitation fibrobronchoscopy were assessed. Arterial hypotension and frequency of postoperative nausea and vomiting were recorded. The length of stay of the patients in the intensive care unit and hospital was recorded.
 RESULTS: Cortisol levels did not differ between groups (p=0.26). The glucose level in the epidural block group was significantly lower than that in other groups (p=0.011). A significant increase in mean blood pressure and heart rate was observed in the opioid analgesia group (p 0.001). The lowest severity of pain 6 h after surgery was observed in the epidural blockade and lidocaine groups (p 0.001). Perioperative hypotension was more common in the epidural analgesia group (p=0.045). The incidence of postoperative nausea and vomiting was higher in the opioid anesthesia group (p=0.004). In the fentanyl infusion group, sanitary fibrobronchoscopy was more often required to eliminate atelectasis (p=0.039). The number of bed days spent in the ICU was significantly higher in the opioid analgesic group (p=0.002); however, no significant differences were noted between the groups regarding the number of days spent in the hospital (p=0.228).
 CONCLUSION: Opioid-free anesthesia based on intravenous lidocaine infusion, as a component of anesthetic management in thoracic surgery, provides sufficient perioperative stability of hemodynamic and neuroendocrine status, has a pronounced analgesic effect, with less opioid consumption on the first day after surgery, and helps reduce the length of stay in the intensive care unit and hospital.

Temporal evolution reveals bifurcated lineages in aggressive neuroendocrine small cell prostate cancer trans-differentiation

Trans-differentiation from an adenocarcinoma to a small cell neuroendocrine state is associated with therapy resistance in multiple cancer types. To gain insight into the underlying molecular events of the trans-differentiation, we perform a multi-omics time course analysis of a pan-small cell neuroendocrine cancer model (termed PARCB), a forward genetic transformation using human prostate basal cells and identify a shared developmental, arc-like, and entropy-high trajectory among all transformation model replicates. Further mapping with single cell resolution reveals two distinct lineages defined by mutually exclusive expression of ASCL1 or ASCL2. Temporal regulation by groups of transcription factors across developmental stages reveals that cellular reprogramming precedes the induction of neuronal programs. TFAP4 and ASCL1/2 feedback are identified as potential regulators of ASCL1 and ASCL2 expression. Our study provides temporal transcriptional patterns and uncovers pan-tissue parallels between prostate and lung cancers, as well as connections to normal neuroendocrine cell states.

BIOM-01. PROGNOSTIC IMPLICATIONS OF SMALL CELL LUNG CANCER (SCLC) TRANSCRIPTIONAL SUBTYPING FOR CENTRAL NERVOUS SYSTEM (CNS) METASTASES

Abstract BACKGROUND Small cell lung cancer (SCLC) is characterized by a high propensity for brain metastases (BM) and is associated with a poor prognosis. Subtypes characterized by high versus low neuroendocrine states (expressing ASCL1 or NEUROD1) have been identified. This study aims to evaluate the influence of transcription factor-defined SCLC subtypes on the incidence and outcomes of BM. METHODS We identified 164 patients with SCLC based on their ASCL1 and NEUROD1 expression status by immunohistochemistry. Four transcriptional subtypes were defined: ASCL1+/NEUROD1- (A+/N-), ASCL1+/NEUROD1+ (A+/N+), ASCL1-/NEUROD1+ (A-/N+), and ASCL1-/NEUROD1- (A-/N-). Cumulative incidence competing risk analyses examined the incidence of central nervous system (CNS) progression. Cox proportional hazards models were utilized to assess both overall survival (OS) and CNS progression-free survival (CNS-PFS). RESULTS Out of 164 patients, the majority fell into either A+/N- or A+/N+ subtypes (n = 62, n = 63, respectively). BM were detected at the time of diagnosis in 12% (20/164) of patients, while 18% (29/164) developed BM later. In two distinct cohorts of patients (initially free from BM and with de novo BM at diagnosis), the 12-month cumulative incidence of CNS progression was numerically highest for the A+/N- subtype (HR = 11.8, 95%CI: 4.7-22.4, and HR = 44, 95%CI: 10.5-74.7, respectively). OS in both A+/N- and A-/N+ subtypes was worse compared to the A+/N+ subtype (HR = 1.62, 95%CI: 1.03-2.56, and HR = 1.63, 95%CI: 1.06-2.60, respectively). Similarly, CNS-PFS was poorer in same A+/N- and A-/N+ subtypes (HR = 3.10, 95%CI: 1.39-6.93, and HR = 3.38. 95% CI: 1.49-7.67) when compared to A+/N+. CONCLUSIONS This study is the first to explore CNS-specific outcomes based on transcription factor SCLC subtypes. Among patients without BM at SCLC diagnosis, those exclusively expressing ASCL1 or NEUROD1 demonstrated poorer outcomes than those with co-expression. Further investigation is warranted to elucidate the implications of SCLC subtyping, ultimately facilitating tailored disease management.

Leptin excites basolateral amygdala principal neurons and reduces food intake by LepRb-JAK2-PI3K-dependent depression of GIRK channels.

Leptin is an adipocyte-derived hormone that modulates food intake, energy balance, neuroendocrine status, thermogenesis, and cognition. Whereas a high density of leptin receptors has been detected in the basolateral amygdala (BLA) neurons, the physiological functions of leptin in the BLA have not been determined yet. We found that application of leptin excited BLA principal neurons by activation of the long form leptin receptor, LepRb. The LepRb-elicited excitation of BLA neurons was mediated by depression of the G protein-activated inwardly rectifying potassium (GIRK) channels. Janus Kinase 2 (JAK2) and phosphoinositide 3-kinase (PI3K) were required for leptin-induced excitation of BLA neurons and depression of GIRK channels. Microinjection of leptin into the BLA reduced food intake via activation of LepRb, JAK2, and PI3K. Our results may provide a cellular and molecular mechanism to explain the physiological roles of leptin in vivo.

Varied impacts of social relationships on neuroendocrine state

Social relationships, affiliative social attachments, are important for many species. The best studied types of relationships are monogamous pair bonds. However, it remains unclear how generalizable models of pair bonding are across types of social attachments. Zebra finches are a fascinating system to explore the neurobiology of social relationships because they form various adult bonds with both same- and opposite-sex partners. To test whether different bonds are supported by a single brain network, we quantified individuals' neuroendocrine state after either 24 h or 2 weeks of co-housing with a novel same- or opposite-sex partner. We defined neuroendocrine state by the expression of 22 genes related to 4 major signaling pathways (dopamine, steroid, nonapeptide, and opioid) in six brain regions associated with affiliation or communication [nucleus accumbens (NAc), nucleus taeniae of the amygdala (TnA), medial preoptic area (POM), and periaqueductal gray (PAG), ventral tegmental area, and auditory cortex]. Overall, we found dissociable effects of social contexts (same- or opposite-sex partnerships) and duration of co-housing. Social bonding impacted the neuroendocrine state of four regions in males (NAc, TnA, POM, and PAG) and three regions in females (NAc, TnA, and POM). Monogamous pair bonding specifically appeared to impact male NAc. However, the patterns of gene expression in zebra finches were different than has previously been reported in mammals. Together, our results support the view that there are numerous mechanisms regulating social relationships and highlight the need to further our understanding of how social interactions shape social bonds.

Multimodal approach to perioperative protection in children in abdominal surgery: A prospective comparative study

OBJECTIVE: To increase the effectiveness of perioperative protection by using combined multimodal analgesia (CMA) with epidural blockade (EA) with bupivacaine in combination with low-flow anesthesia with sevoflurane in traumatic abdominal surgery in children.
 MATERIALS AND METHODS: A prospective comparative study was conducted (n=102, from 1 to 17 years). Surgical planned pathology: Hirschsprung disease, Ledd and Payr syndrome, liver echinococcosis, enterocystoma, pancreatic cyst. Study period: 20182022. Patients were divided into two groups depending on the technique of anesthesia and management of the postoperative period. Group 1 (n=55) received CMA + EA with bupivacaine 0.5%. In the postoperative period prolonged EA with bupivacaine 0.25% 0.51 mg/kg. Comparison group 2 (n=47) traditional general anesthesia based on opioids. For the treatment of postoperative pain syndrome (PPS) promedol 0.3 mg/kg. The effectiveness of perioperative analgesia in pediatric abdominal surgery, the time of development and intensity of postoperative pain syndrome were evaluated.
 RESULTS: The data obtained as a result of the study clearly showed that the advantages lie behind the optimized method of combined multimodal analgesia CMA + EA with bupivacaine 0.5%. In general, the effectiveness of CMA in combination with epidural analgesia in the intraoperative period was quite high. Additional intraoperative administration of fentanyl was registered in 8 (14.5%) of cases. The stability of hemodynamic parameters and neuroendocrine status was noted in almost all patients of the main group. Analyzing the time of development and intensity of PPS, it can be argued that patients of group 1 had better indicators: the duration of the pain-free period was 2 times longer and the intensity of PPS was lower than in children of group 2. Motor blockade gradually resolved in patients on average, after 20946 minutes. By this time, there was a recovery of the sensation of a pinprick and motor activity in 32 (58.1%) of patients according to the Bromage 2 scale. Analysis of the results of an oral survey of patient satisfaction with pain relief conducted 24 hours after surgery showed that in group 1 the number of cases 43 (78.1%) satisfied with the level of pain relief was 1.5 times higher compared to the control group 24 (51.1%).
 CONCLUSION: Combined multimodal analgesia in combination with epidural blockade with bupivacaine in traumatic abdominal surgery in children provides sufficient perioperative stability of the hemodynamic and neuroendocrine status, significantly reduces the pharmacological burden, reduces complications, early activation of patients and rapid postoperative rehabilitation compared to the traditional method.

Abstract B025: MicroRNA regulators of neuroendocrine differentiation of prostate cancer

Abstract Background: Therapy-induced neuroendocrine prostate cancer (NEPC) is an extremely aggressive variant of castration-resistant prostate cancer (CRPC) that is increasing in incidence with the widespread use of second generation of androgen receptor (AR)-pathway inhibitors (APIs) such as Enzalutamide (ENZ) and Abiraterone. This aggressive variant arises from CRPC-Adenocarcinomas (CRPC-Adeno) via a reversible trans-differentiation process, referred to as neuroendocrine differentiation (NED) wherein cells undergo a lineage switch and exhibit neuroendocrine (NE) features, characterized by expression of neuronal markers such as enolase 2 (ENO2), chromogranin A (CHGA) and synaptophysin (SYP). There is an urgent need of identifying novel molecular markers to assess emergence of NED in CRPC patients. We demonstrated that progression of advanced CRPC with adenocarcinoma characteristics (CRPC-Adeno) to therapy-induced, androgen-independent NE (CRPC-NE) states is associated with a characteristic set of miRNA alterations that promote plasticity of advanced prostate adenocarcinomas to NEPC (Bhagirath et al., Oncogene, 2020). Importantly, we could develop a ‘novel miRNA classifier’ to robustly stratify CRPC-NE tumors from CRPC-Adenocarcinomas. Here we further validate the classifier in independent clinical cohorts and deduce the optimal miRNA genes required for NEPC diagnosis. We further study the functional significance of miR-28-3p, a miRNA identified by sequencing, in NEPC. Methods: Human clinical samples with corresponding clinical information were procured from two independent sites of Prostate Cancer Biorepository Network (PCBN). Samples included human CRPC-adeno vs CRPC-NE. FFPE sections from these clinical samples were microdissected, RNA were extracted and small RNA sequencing was performed using an Illumina NextSeq 500 platform. Sequencing data were analyzed and machine learning algorithms were applied (random forest machine learning technique with leave-pair-out cross validation (LPOCV)). The performance of classifier was measured using receiver operating characteristic (ROC) analysis with area under the curve (AUC) as the primary evaluation metric. The functional significance of miR-28-3p was studied by its overexpression and knockdown in PCa cell lines followed by functional assays. Results: Unsupervised analysis of sequencing data by principal component analyses (PCA) revealed distinct clustering of the CRPC-NE tumors from CRPC-Adenocarcinomas based on miRNA profiles suggesting that miRNA profiles can be used to stratify these tumor types. Our analyses showed that a set of 5 miRNAs of the classifier are important in distinguishing between CRPC-Adeno vs CRPC-NE with an AUC=0.8318. Further, miR 28-3p potentially has a biphasic role in PCa, where it is oncogenic in early stages and acts as a tumor suppressor in late stage PCa. Conclusions: A ‘5- miRNA’ classifier was validated to be of significance in two independent validation cohorts. We propose this miRNA classifier as an important tool for diagnosing NED in CRPC patients. Citation Format: Amritha Sreekumar, Sharanjot Saini. MicroRNA regulators of neuroendocrine differentiation of prostate cancer [abstract]. In: Proceedings of the AACR Special Conference: Advances in Prostate Cancer Research; 2023 Mar 15-18; Denver, Colorado. Philadelphia (PA): AACR; Cancer Res 2023;83(11 Suppl):Abstract nr B025.

Abstract 3141: Epiregulon infers single-cell transcription factor activity to dissect mechanisms of lineage plasticity and drug response

Abstract Single-cell multiomic technologies enable holistic reconstruction of cell states and hold the potential to identify drivers of tumor evolution and drug resistance. However, the data sparsity of single-cell assays can preclude accurate estimation of transcription factor activity. In addition, gene expression alone cannot capture transcription factor activity, especially in the context of drug treatment which alters transcription factor function without suppressing gene expression. To circumvent these challenges, we have developed epiregulon, a R package that constructs gene regulatory networks and infers transcription factor (TF) activity in single cells by integrating single cell gene expression, chromatin accessibility and bulk TF occupancy data. Epiregulon applies tests of independence to identify likely transcription factor - regulatory element - target gene relationships occurring at joint probabilities exceeding the expected probabilities of independent events. With epiregulon, we are able to detect lineage factor activity at enhanced sensitivity and predict perturbations more accurately than gene expression could. We further applied this tool to understand transcription factor activity to enzalutamide treatment and identify potential drivers of enzalutamide resistance. Finally, we generated a ground truth dataset using reprogram-seq, a technology that captures multi-omic profiles of single cells upon expression of defined transcription factors. Epiregulon correctly predicts NKX2-1 targets and demonstrates that NKX2-1 expression reprograms the epigenome towards a neuroendocrine state in LNCaP cells. Epiregulon is a useful tool that constructs gene regulatory networks to model the underlying gene regulation hierarchies that drive gene expression and cell states. Citation Format: Tomasz Wlodarczyk, Jenille Tan, Kerstin Seidel, Diana Wu, Shang-Yang Chen, Aleksander Chlebowski, Timothy Keyes, Yu Guo, Aaron Lun, Christopher Siebel, Shiqi Xie, Xiaosai Yao. Epiregulon infers single-cell transcription factor activity to dissect mechanisms of lineage plasticity and drug response [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 3141.

Clinical and morphological features of large-cell neuroendocrine carcinomas and small-cell lung carcinomas expressing the DLL3 and ASCL1 oncoproteins.

Intratumoral similarities and differences between large-cell neuroendocrine carcinomas (LCNECs) and small-cell lung carcinomas (SCLCs) are determined partially by the Notch signaling pathway, which controls the switch from neuroendocrine to slight/non-neuroendocrine cell fate. LCNECs are divided into two subgroups according to genomic alterations: type I LCNECs exhibit a neuroendocrine profile characterized by achaete-scute homolog 1 (ASCL1)high/delta-like protein 3 (DLL3)high/NOTCHlow and type II LCNECs show the pattern ASCL1low/DLL3low/NOTCHhigh. Here, we used immunohistochemistry, transmission electron microscopy, and digital analysis to examine the role of the Notch ligand DLL3 as an immunomarker of the neuroendocrine state and ASCL1 as a regulator of cell-cell interactions in SCLCs and LCNECs. High DLL3 and ASCL1 expression was associated with atypical submicroscopic characteristics involving nuclear size, chromatin arrangement, Golgi apparatus, and endoplasmic reticulum, and was characteristic of type I LCNECs with similarity to SCLCs, whereas low DLL3 and ASCL1 expression was found in both SCLCs and type II LCNECs. In patients diagnosed at an early stage who did not have metastasis and who underwent chemotherapy, DLL3high and ASCL1high SCLCs and type I LCNECs were associated with a better prognosis and a lower risk of death. The present findings suggested that DLL3/ASCL1 are potential therapeutic targets and prognostic indicators in patients with SCLCs or LCNECs.

Sexual dimophism in plasma nitrogenous metabolites levels and some psycho-neuro-endocrine parameters

Background. We previously showed that the factor structure of the relationships between the plasma levels of nitrogenous metabolites (uric acid, bilirubin, urea, and creatinine), on the one hand, and the parameters of anxiety and neuro-endocrine status, on the other hand, is significantly different in men and women of reproductive age and postmenopausal. The purpose of this study is to identify the metabolic and psycho-neuro-endocrine features of these three cohorts of people. Materials and methods. The object of observation were 31 males (24÷69 y) and 30 females, from among them 18 postmenopausal (45÷76 y) and 12 of reproductive age (30÷42 y), with dysfunction of neuro-endocrine-immune complex. In basal conditions we determined plasma levels of nitrogenous metabolites and adaptation hormones, estimated the severity of the trait and reactive anxiety, recorded the ongoing HRV and EEG. After 4 or 7 days, repeated testing was performed. Results. To achieve the goal, the registered parameters were subjected to discriminant analysis. Along with the quite expected nitrogenous metabolites, age, testosterone and calcitonin (both raw and sex- and age-standardized values), trait anxiety, Kerdö's Vegetative Index, one of the vagal markers (SDNN), as well as 10 EEG parameters emerged as characteristic features of the cohorts. In the information space of two discriminant roots, three cohorts are clearly demarcated. Conclusion. We interpret this as another proof of the existence of connections between nitrogenous metabolites and psycho-neuro-endocrine parameters in line with the concept of the functional-metabolic continuum.

Pathology of the Thyroid Gland in Women Rheumatoid Arthritis

Endocrine disorders are of primary importance in the occurrence and progression of rheumatic diseases. Often, endocrine restructuring that occurs during puberty, menopause, pregnancy, childbirth, and abortion serves as a background for the development of connective tissue diseases. At the same time, the change in neuroendocrine status is an integral component of the progression of rheumatic diseases. Especially distinct hormonal shifts were found on the part of the thyroid gland in rheumatoid arthritis (RA). The prevalence of thyroid pathology in general among patients with rheumatoid arthritis, according to literature data, reaches 34%, and Hashimoto's thyroiditis, according to various authors, from 4 to 13.5%. In addition, the greatest susceptibility to RA in women of reproductive age indicates a certain role of sex hormones in the pathogenesis of the disease. Metabolic and degenerative changes in connective tissue largely determine the pathogenesis and clinical picture of various diseases. Rheumatic diseases, the most common of which is rheumatoid arthritis (RA), make up a significant proportion of the pathology of the joints, musculoskeletal system, and connective tissue. It affects 0.5—1% of the world's population, which is about 63 million patients.

DLL3 regulates Notch signaling in small cell lung cancer.

Tumor heterogeneity plays a critical role in tumor development and response to treatment. In small-cell lung cancer (SCLC), intratumoral heterogeneity is driven in part by the Notch signaling pathway, which reprograms neuroendocrine cancer cells to a less/non-neuroendocrine state. Here we investigated the atypical Notch ligand DLL3 as a biomarker of the neuroendocrine state and a regulator of cell-cell interactions in SCLC. We first built a mathematical model to predict the impact of DLL3 expression on SCLC cell populations. We next tested this model using a single-chain variable fragment (scFv) to track DLL3 expression invivo and a new mouse model of SCLC with inducible expression of DLL3 in SCLC tumors. We found that high levels of DLL3 promote the expansion of a SCLC cell population with lower expression levels of both neuroendocrine and non-neuroendocrine markers. This work may influence how DLL3-targeting therapies are used in SCLC patients.

Characteristics of the Immune and Neuroendocrine Status of Children Exposed to Non-Ionizing Radiation in the School Environment

Introduction: The modern educational environment involves the use of various electronic devices generating physical hazards. In addition to the widespread use of these devices, other physical factors of the school environment also affect schoolchildren’s health. However, the impact of the combined exposure to physical factors of various nature on the regulatory systems of the body has been studied insufficiently so far. However, the influence of a complex of physical factors of various nature on the regulatory systems of the body has not been sufficiently studied. Objective: To evaluate the features of the immune and neuroendocrine status of schoolchildren exposed to heterogeneous physical factors of a non-ionizing nature at school and to analyze the relationship between the studied physical factors and changes in the parameters of regulatory systems. Materials and methods: The exposed group included 144 students of a gymnasium from the city of Perm and the unexposed (reference) group consisted of 114 schoolchildren from the town of Kungur, Perm Region. We measured physical factors of the school environment, including noise, artificial illumination, electromagnetic radiation, and air ion concentrations affecting elementary, secondary, and high school students from both groups and compared the results with respective permissible levels. The indicators of immune (CD-phenotyping, phagocytic activity of leukocytes, concentrations of immunoglobulins and cytokines) and neuroendocrine (TSH, free T4, and cortisol) systems were established and compared. Mathematical models were calculated and analyzed in order to find the links between the exposure to physical factors under study and the parameters of the immune and neuroendocrine systems. Results: The indoor air study showed differences in the number and charge of air ions between the gymnasium with its numerous e-learning tools and the school of comparison. The study of the immune and neuroendocrine systems in the exposed group revealed age-related changes in the expression of lymphocyte subpopulations, moderate changes in humoral and phagocytic parameters, an imbalance in the expression of pro- and anti-inflammatory cytokines and hormones. The construction of logistic regression models allowed us to establish the link between the parameters of electromagnetic radiation in the school environment and changes in the immune status of students (CD19+ and CD3+CD8+ lymphocytes). Conclusion: The study demonstrated a number of changes in the regulatory systems of schoolchildren associated with the impact of physical factors of the school environment.

Molecular clusters reveal opportunities for personalised small cell lung cancer immunotherapy

Molecular clusters reveal opportunities for personalised small cell lung cancer immunotherapy