Disease Process Research Articles

CircRNAs as a novel class of potential diagnostic biomarkers in bipolar disorders.

Bipolar disorder (BD) is a severe mood disorder with uncertain causes and debilitating signs and symptoms. Gene expression is crucial to the pathophysiology of BD and could be influenced by genetic or epigenetic factors, by either direct modification of mRNA templates or by regulation of post-transcriptional translation. Recent evidence has shown that several critical processes in psychiatric diseases, such as neuronal activity or plasticity, synaptic transmission, and neuronal depolarization, have all been linked to circular RNAs (circRNAs). The circRNA profile of neuronal cells, which may be easily ascertained by a liquid biopsy, may shed light on the molecular pathophysiology of psychiatric disorders, including BD. This approach could aid in future development in diagnosis and treatment. In this review, we provide an in-depth understanding of the roles of circRNAs in the pathophysiology of BD and offer new insight into their potential as emerging diagnostic tools and therapeutic targets.

Concomitant Wilms tumor and autosomal dominant polycystic kidney disease.

Concomitant Wilms tumor (WT) and autosomal dominant polycystic kidney disease (ADPKD) is exceedingly rare, presenting a diagnostic and technical challenge to pediatric surgical oncologists. The simultaneous workup and management of these disease processes are incompletely described. We performed a retrospective analysis of patients treated at our institution with concomitant diagnoses of WT and ADPKD. We also review the literature on the underlying biology and management principles of these conditions. We present three diverse cases of concomitant unilateral WT and ADPKD who underwent nephrectomy. One patient had preoperative imaging consistent with ADPKD with confirmatory testing postoperatively, one was found to have contralateral renal cysts intraoperatively with confirmatory imaging post nephrectomy, and one was diagnosed in childhood post nephrectomy. All patients are alive at last follow-up, and the patient with longest follow-up has progressed to end-stage kidney failure requiring transplantation and dialysis in adulthood. All patients underwent germline testing and were found to have no cancer predisposition syndrome or pathogenic or likely pathogenic variants for WT. Concomitant inheritance of ADPKD and development of WT are extremely rare, and manifestations of ADPKD may not present until late childhood or adulthood. ADPKD is not a known predisposing condition for WT. When ADPKD diagnosis is made by family history, imaging, and/or genetic testing before WT diagnosis and treatment, the need for extensive preoperative characterization of cystic kidney lesions in children and increased risk of post-nephrectomy kidney failure warrant further discussion of surgical approach and perioperative management strategies.

Predictive effect of lipopolysaccharide-stimulated inflammatory cytokines on symptoms of generalized anxiety disorder

BACKGROUND Generalized anxiety disorder (GAD) is a relatively common mental disorder. Recently, inflammation, an important factor for the development of depression, has attracted increasing attention. Several studies have shown that inflammatory cytokines can affect the pathophysiological processes of several nervous system diseases. We hypothesized that there is a correlation between the levels of lipopolysaccharide (LPS)-stimulated inflammatory cytokines and the clinical symptoms of GAD. AIM To investigate the predictive effect of LPS-stimulated inflammatory cytokines on symptoms of GAD. METHODS This was a cross-sectional study in which 89 patients with GAD diagnosed at The First Hospital of Hebei Medical University from January 2022 to December 2022 and 70 individuals without anxiety and depression (controls) during the same period were included. Fasting venous blood was collected from all the subjects in heparin tubes, and another 3 ml of blood was supplemented with LPS (10 ng/ml). The plasma levels of 12 cytokines [Interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, tumor necrosis factor (TNF)-α, interferon (IFN)-γ, IL-17A, IL-12p70, and IFN-α] were detected. RESULTS Post-LPS stimulation, the levels of IL-1β, IL-6, IL-8, IL-10, and TNF-α in both the control and GAD groups were significantly elevated above those in the nonstimulated groups, with IL-6 and IL-8 showing marked increases. Increases in IL-8 and TNF-α were statistically significant in the GAD group (P < 0.05). IL-1β, IL-6, IL-8, IL-10, and TNF-α were found to be significantly correlated with Hamilton Anxiety Rating Scale (HAMA) scores (P < 0.05). A negative correlation was observed between IL-10 levels and HAMA scores. Further analysis revealed that TNF-α was associated with mental anxiety, whereas IL-1β, IL-8, and IL-10 were associated with physical anxiety symptoms, with IL-10 showing a negative correlation with physical anxiety. IL-6 was associated with both mental and physical aspects of anxiety. CONCLUSION The physical symptoms of GAD are related to inflammatory factors. IL-1β, IL-8, IL-10, and TNF-a can be used as predictors of physical or mental anxiety in patients with GAD.

Hair cortisol concentrations in clipped and combed hair and associations with characteristics, health status and stress in domestic cats

Hair cortisol concentrations (HCC) are measured to assess long-term HPA-axis activity and may represent a valuable non-invasive tool to evaluate chronic stress in cats. This study investigated combing as a novel, low-stress method for HCC assessment, as well as possible associations between HCC and cat characteristics in 167 owned cats. Hair was sampled at veterinary clinics through clipping and/or combing the cat, or at home by the owner combing the cat. A questionnaire was sent to cat owners, including inquiries about the cat’s sex, health status, and exposure to stress. HCC was quantified using a commercial cortisol assay kit. Despite variations within and between sampling methods, Spearman’s correlation and Bland–Altman plots revealed a moderate correlation between clipped and combed samples (rs = 0.61, LOA -5.51 ± 22.54). In multiple linear regression, variations in HCC were observed based on sex, health status and cat group size. No associations were found between HCC and stress as assessed by owners. Despite study limitations and remaining uncertainty regarding factors influencing HCC, combing presents a convenient approach for evaluating long-term HPA-axis activity in clinical settings. The association between health and HCC suggests alterations in cortisol levels that are related to disease processes and stress-inducing events associated with the disease.

Early life epigenetics and childhood outcomes: a scoping review.

Epigenetics is the study of changes in gene expression, without a change in the DNA sequence that are potentially heritable. Epigenetic mechanisms such as DNA methylation, histone modifications, and small non-coding RNA (sncRNA) changes have been studied in various childhood disorders. Causal links to maternal health and toxin exposures can introduce epigenetic modifications to the fetal DNA, which can be detected in the cord blood. Cord blood epigenetic modifications provide evidence of in-utero stressors and immediate postnatal changes, which can impact both short and long-term outcomes in children. The mechanisms of these epigenetic changes can be leveraged for prevention, early detection, and intervention, and to discover novel therapeutic modalities in childhood diseases. We report a scoping review of early life epigenetics, the influence of maternal health, maternal toxin, and drug exposures on the fetus, and its impact on perinatal, neonatal, and childhood outcomes. IMPACT STATEMENT: Epigenetic changes such as DNA methylation, histone modification, and non-coding RNA have been implicated in the pathophysiology of various disease processes. The fundamental changes to an offspring's epigenome can begin in utero, impacting the immediate postnatal period, childhood, adolescence, and adulthood. This scoping review summarizes current literature on the impact of early life epigenetics, especially DNA methylation on childhood health outcomes.

Childhood Multiple Endocrine Neoplasia (MEN) Syndromes: Genetics, Clinical Heterogeneity and Modifying Genes

Multiple endocrine neoplasia (MEN) syndromes are part of a spectrum of clinically well-defined tumor syndromes ultimately characterized by histologically similar tumors arising in patients and families with mutations in one of the following four genes: MEN1, RET, CDKN1B, and MAX. The high level of genetic and phenotypic heterogeneity has been linked to phenocopies and modifying genes, as well as unknown mechanisms that might be investigated in the future based on preclinical and translational considerations. MEN1, also known as Wermer’s syndrome (OMIM *131100), is an autosomal dominant syndrome codifying for the most frequent MEN syndrome showing high penetrance due to mutations in the MEN1 gene; nevertheless, clinical manifestations vary among patients in terms of tumor localization, age of onset, and clinical aggressiveness/severity, even within the same families. This has been linked to the effect of modifying genes, as described in the review. MEN 2-2b-4 and 5 also show remarkable clinical heterogeneity. The traditional view of genetically predisposing monogenic or multifactorial disorders is no longer valid, and mandates a change in scientific focus. Phenotypes are indeed rarely consistent across genetic backgrounds and environments. In the future, understanding factors and genetic variants that control cellular functions and the expression of disease genes should provide insights into fundamental disease processes, providing implications for counseling and therapeutic and prophylactic possibilities.

Correlation between gut microbiota dysbiosis and colorectal cancer: review

Introduction and Purpose: The human gut microbiota, comprising a diverse consortium of approximately 100 trillion microorganisms, is integral to maintaining health and modulating disease processes. Its development begins at birth, influenced by maternal microbiota and environmental factors. Dysbiosis, defined as an imbalance in the gut microbial composition, has been implicated in a range of gastrointestinal pathologies, including colorectal cancer (CRC). This review endeavors to elucidate the relationship between gut microbiota and CRC, examining the impact of specific bacterial taxa on the pathogenesis CRC. Description of the State of Knowledge: Gut microbiota encompasses a multitude of microbial species, with their composition differing along the gastrointestinal tract. Healthy gut microbiota perform essential functions such as pathogen protection, metabolic processes, and immune system modulation. Factors like diet and genetics significantly influence microbial composition. Dysbiosis contributes to CRC through inflammation, genotoxin production, and immune modulation. Specific bacteria, such as Fusobacterium nucleatum and Bacteroides fragilis, are associated with CRC. Mechanisms of carcinogenesis include bacterial adherence, invasion of epithelial cells, and activation of pro-inflammatory pathways. Summary:Understanding the gut microbiota's role in CRC highlights the importance of maintaining a balanced microbiome for cancer prevention. Dietary interventions promoting beneficial bacteria and reducing harmful species could mitigate CRC risk. Further research should prioritize the identification of microbial biomarkers for early CRC detection and the development of therapeutic strategies aimed at modulating the gut microbiota. These efforts will enhance CRC prevention and treatment modalities.

Personalized Nutrition for Obesity Management: A Mini-Review

Abstract Obesity is a pro-inflammatory state and a chronic health condition, which falls within the spectrum of immune-mediated inflammatory diseases (IMIDs). It is caused mainly by the consumption of excess calories and egregious food substances. A Personalized Food Avoidance Dietary Approach for Management of IMIDs (PFA-DAMI) addresses primary and secondary immune intolerances of different individuals to dietary constituents. Exposure to toxic food substances is responsible for inflammatory immune dysfunction that mediates obesity. Hence, a PFA-DAMI is promising for the management of obesity. T cell dysfunction disease mediating models describing inflammatory disease processes underlying IMIDs exist. However, they do not highlight inflammatory processes underlying the diseases concerning toxin-mediated epigenetic T cell dysfunction. Online searches were conducted on databases, such as Google Scholar, PubMed, Biomed Central, and SciELO. Articles were reviewed using keywords, such as obesity, personalized nutrition, immune optimization/dysfunction, T lymphocyte activation/dysfunction, cytokines, and adipokines. There is a putative T cell toxin-mediated dysfunction disease model for IMIDs, which may apply to other diseases. The putative disease model may highlight the actual inflammatory immune dysfunctional processes underlying T cell disease mediation in inflammatory diseases, which may be validated by multiomic studies. Validation of the putative disease model using obesity as an example should pave the way for a better understanding of the role of personalized nutrition in obesity management.

The Effects of Diet, Dietary Supplements, Drugs and Exercise on Physical, Diagnostic Values of Urine Characteristics

Background/Objectives: This review summarizes the current knowledge about factors that affect the physical characteristics of urine. It highlights proper urine sample collection and displays factors like diet, hydration status, and medications that can alter urine color, odor, clarity, specific gravity and pH. Results: Urinalysis is a minimally invasive examination of a patient’s health, especially concerning nephrological and endocrinological abnormalities, as well as dietary habits and stimulants used. Certain deviations in appearance, composition or frequency/pain during urination may indicate an ongoing disease process in the body. Based on laboratory results, further medical treatment is determined. The reason for a change in the color of the urine, for its clouding or intense odor may be a disease, as well as the consumption of food, medication, intensive physical exercise or inadequate hydration of the body. Well-standardized procedures for collecting, transporting, preparing and analyzing samples should become the basis for an effective diagnostic strategy in urinalysis. It is worth noting that pharmacists in pharmaceutical care are often the first people to whom a patient turns for health advice and for the interpretation of simple laboratory tests. Acquiring the ability to interpret the results of laboratory tests and the principles of proper sampling for laboratory tests is indispensable in the process of possible counseling and providing reliable answers to patients’ questions. Conclusions: Although urinalysis is not recommended as a routine screening tool for the general population, it can prove to be a valuable source of patient health data in some cases as the data will be useful to physicians and pharmacists to more effectively diagnose and better care for patients.

Development of a General Composite Scale (GENCOMS) for Progressive Neurodegenerative Diseases and Implications for the Assessment of Disease-Modifying Therapies.

The reliable assessment of treatment outcomes for disease-modifying therapies (DMT) in neurodegenerative disease is challenging. The objective of this paper is to describe a generalized framework for developing composite scales that can be applied in diverse, degenerative conditions, termed "GENCOMS." Composite scales optimize the sensitivity for detecting clinically meaningful effects that slow disease progression. The GENCOMS method relies on robust natural history data and/or placebo arm data from DMT trials. Validated scales that are core to the disease process have been identified, and item level data obtained to standardize the response outcomes from 0 (best possible score) to 1 (worst possible score). A partial least squares regression analysis was conducted with temporal change as the dependent variable and change scores in standardized items as the explanatory variables. The derived model coefficients constitute a weighted sum of items that most effectively measure disease progression. The resultant composite scale was optimized to detect disease progression and can be examined in a range of slow or fast progressing populations. The scale can be used in studies with comparable patient populations as an endpoint optimized to measure disease progression and therefore ideally suited to assess treatment effects in DMTs. The methodology presented here provides a generalizable framework for developing composite scales in the assessment of neurodegenerative disease progression and evaluation of DMT effects. By objectively selecting and weighting items from previously validated measures based solely on their sensitivity to disease progression, this methodology allows for the creation of a more responsive measurement of clinical decline. This heightened sensitivity to clinical decline can be utilized to detect modest yet meaningful treatment effects in the early stages of neurogenerative diseases, when it is optimal to begin a DMT.

The Cervical Intervertebral Foramen: Microanatomy, Pathology, and Clinical Implications.

This is an evidence-based narrative review article. We hope to provide a primer on cervical intervertebral foramen (cIVF) anatomy for spine surgeons, interventionalists, and physiatrists who regularly treat cervical spine pathology, and encourage further exploration of this topic. This corridor for exiting cervical nerve roots is characterized by its intricate microanatomy involving ligamentous, nervous, and vascular structures. Degenerative changes such as facet hypertrophy and disc herniations alter these relationships, potentially leading to nerve root compression and cervical radiculopathy. This review synthesizes existing knowledge on the cIVF. Key imaging, cadaveric, and clinical studies serve as a foundation for this anatomic review. We explore topics such as dynamic changes that affect foraminal size and their implications for nerve root compression, the relationship of the dorsal root ganglion to the cervical foramen, and the function and clinical significance of foraminal ligaments, arteries, and veins. Changes in the cIVF are frequently the basis of cervical degenerative pathologies. A comprehensive understanding of its microanatomical structure will allow the practitioner to better treat the underlying disease process causing their symptoms and signs.

Tissue-aware interpretation of genetic variants advances the etiology of rare diseases.

Pathogenic variants underlying Mendelian diseases often disrupt the normal physiology of a few tissues and organs. However, variant effect prediction tools that aim to identify pathogenic variants are typically oblivious to tissue contexts. Here we report a machine-learning framework, denoted "Tissue Risk Assessment of Causality by Expression for variants" (TRACEvar, https://netbio.bgu.ac.il/TRACEvar/ ), that offers two advancements. First, TRACEvar predicts pathogenic variants that disrupt the normal physiology of specific tissues. This was achieved by creating 14 tissue-specific models that were trained on over 14,000 variants and combined 84 attributes of genetic variants with 495 attributes derived from tissue omics. TRACEvar outperformed 10 well-established and tissue-oblivious variant effect prediction tools. Second, the resulting models are interpretable, thereby illuminating variants' mode of action. Application of TRACEvar to variants of 52 rare-disease patients highlighted pathogenicity mechanisms and relevant disease processes. Lastly, the interpretation of alltissue models revealed that top-ranking determinants of pathogenicity included attributes of disease-affected tissues, particularly cellular process activities. Collectively, these results show thattissue contexts and interpretable machine-learning models can greatly enhance the etiology of rare diseases.

Abstract B011: Predictors of survival and cachexia in pancreatic cancer: results from the CAMPP study

Abstract Introduction: Pancreatic Ductal Adenocarcinoma (PDAC) is a major cause of cancer-related death in the US and has an overall 5-year survival rate of only 13%. Cachexia, or cancer associated muscle wasting, is common and negatively affects treatment outcomes and quality of life. Despite these poor outcomes, our understanding of PDAC disease processes and improvements in treatments are limited. Here, we present the first results of the Cancer Associations in MUSC PDAC Patients (CAMPP) study. Methods: This retrospective analysis of 133 surgically resectable PDAC patients at the Medical University of South Carolina (MUSC) between May 2016 to 2021 includes chart review data on survival, treatment, and clinical outcomes. Cancer cachexia was evaluated via total psoas area at the L3 vertebral level, which was standardized by height to produce a Total Psoas Index (TPI). Survival was plotted via unadjusted Kaplan-Meier analysis and TPI and cachexic status was examined for 124 patients with both linear and logistic regressions for unadjusted and adjusted data. Results: When analyzing population survival, we observed a mean overall survival (OS) of 23.6 months and mean progression free survival (PFS) of 17.3 months. Notable survival differences include perioperative vs adjuvant only chemotherapy groups in OS at 2 years (p=0.069) and 5 years (p=0.13). Within the perioperative population, there were differences in PFS (p=0.017) and OS (p=0.056) for patients who responded to therapy at 5 years. To explore potential relationships with cachexia, we performed simple bivariate linear regressions for TPI at diagnosis. We found preliminary associations with sex (p<0.001) and creatinine (p=0.062). When adjusting the model for the confounding effects of race, resection status, and perioperative status, sex (p<0.001) and age (p=0.02) were significantly associated. Importantly, creatinine was not significantly associated (p=0.16). Regardless, an adjusted logistic regression using sex-dependent cachexia cutoffs revealed a clinically relevant odds ratio of 0.9 (95% CI [0.72, 1.09]). This indicates a 10% reduction in the odds of cachexia with every 0.1 mg/dL increase in creatinine, all else held constant. Stratifying with sex-dependent cutoffs for both TPI and creatinine revealed significant differences in 5-year OS (p=0.1) between all four groups. Conclusions: Observing survival trends for patients receiving perioperative care is notable as there are currently no clear guidelines for perioperative chemotherapy despite growing evidence for its efficacy. Further, while sex and age are known predictors of cachexia, creatinine is much less studied. Our associations between creatinine and TPI agree with observations in lung cancer, supporting its use as a clinically relevant marker. Ongoing work is exploring longitudinal measures of cachexia and correlating the CAMPP dataset with tumor genomics to reveal individualized factors impacting patient outcomes. Citation Format: Julie A Disharoon, Arash Velayati, Chloe Matila, Andrew Disharoon, Joseph A Karam, John Redhead, Rachel Burge, Elizabeth G Hill, David T Long, Toros A Dincman. Predictors of survival and cachexia in pancreatic cancer: results from the CAMPP study [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr B011.

Are the Effects of Bioactive Components on Human Health a Myth?: Black Elderberry (Sambucus nigra L.) from Exotic Fruits.

Black elderberry has come to the fore in recent years due to its health benefits. Black elderberry fruit (Sambucus nigra L.), collected from natural sources, has a rich content of protein, vitamins, antioxidants, unsaturated fatty acids, and minerals as it contains conjugated and free forms of amino acids. Black elderberry can prevent oxidative stress and reduce blood pressure and prevent cardiovascular diseases, diabetes mellitus, neurodegenerative diseases thanks to the polyphenols it contains. It can prevent diseases, stimulate the immune system, show an antitumor effect, and be effective in the course of disease processes by rising the activity of antioxidant enzymes, including glutathione. Since black elderberry is a promising food in terms of bioactive components, it is considered as promising to treat and prevent many diseases. However, it is not correct to prove its beneficial effects on the prevention of chronic diseases and to make generalisations. Therefore, there is a need of more comprehensive evidence-based clinical studies and data. This review examined current evidence and discussions about the health-related effects of black elderberry, which contains many biologically active components such as polyphenols, phenolic acids, flavonols, proanthocyanidins and anthocyanins, as well as terpenes and lectins, and offered some suggestions on its use in the future.

The clinical significance of pyogenic liver abscess after transarterial chemoembolization or microwave ablation on malignant liver tumors: A retrospective study.

Pyogenic liver abscess (PLA) is a rare but severe complication of interventional therapy that has been little studied. We aimed to find the risk factors for PLA after transarterial chemoembolization (TACE) or microwave ablation (MWA), further explore its clinical significance and summarize our experience with its treatment. Twenty-two patients with PLA and 118 randomly selected patients without PLA after TACE/MWA were enrolled. Logistic regression was used to analyze risk factors, a nonparametric test was used to compare recovery duration, the log-rank test was used to compare progression-free survival, and Spearman correlation coefficient was calculated between the time from fever to drainage and the total duration of fever. The disease process and treatment were summarized. Sphincter of Oddi manipulation increased the risk of PLA by 70.781-fold. The PLA group took longer to recover (36.56 ± 16.42 days) than the control group (5.54 ± 4.33 days), and had a shorter progression-free survival. Escherichia coli was the major pathogenic bacterium, and multidrug resistance was found in 8 patients with E coli or Enterococcus faecium. The time from fever to drainage was 15.89 ± 13.78 days, which was positively correlated with the total duration of fever (24.29 ± 18.24 days). Overall, 18 patients recovered, and 4 patients died of PLA, for a mortality rate of 18.18%. The fever of 10 patients (45.45%) was controlled by cefoperazone sodium and sulbactam sodium or piperacillin sodium and tazobactam sodium; the fever of 7 patients (31.81%) was controlled by imipenem and cilastatin sodium; and the fever of 3 patients (13.63%) was controlled by tigecycline. Sphincter of Oddi manipulation is a high-risk factor for PLA after TACE or MWA. PLA can accelerate cancer progression and even lead to death. E coli was the major pathogenic bacterium, and multidrug resistance was most common in E coli and E faecium. Timely drainage and appropriate antibiotics are the key primary measures for treating PLA. Cefoperazone sodium and sulbactam sodium or piperacillin sodium and tazobactam sodium is a good choice for the first treatment of PLA, especially before pathogenic bacteria are identified. With the emergence of drug resistance, imipenem and cilastatin sodium, and tigecycline can be used for posterior treatment.

Unfolding Mechanism and Fibril Formation Propensity of Human Prion Protein in the Presence of Molecular Crowding Agents

The pathological process of prion diseases implicates that the normal physiological cellular prion protein (PrPC) converts into misfolded abnormal scrapie prion (PrPSc) through post-translational modifications that increase β-sheet conformation. We recently demonstrated that HuPrP(90–231) thermal unfolding is partially irreversible and characterized by an intermediate state (β-PrPI), which has been revealed to be involved in the initial stages of PrPC fibrillation, with a seeding activity comparable to that of human infectious prions. In this study, we report the thermal unfolding characterization, in cell-mimicking conditions, of the truncated (HuPrP(90–231)) and full-length (HuPrP(23–231)) human prion protein by means of CD and NMR spectroscopy, revealing that HuPrP(90–231) thermal unfolding is characterized by two successive transitions, as in buffer solution. The amyloidogenic propensity of HuPrP(90–231) under crowded conditions has also been investigated. Our findings show that although the prion intermediate, structurally very similar to β-PrPI, forms at a lower temperature compared to when it is dissolved in buffer solution, in cell-mimicking conditions, the formation of prion fibrils requires a longer incubation time, outlining how molecular crowding influences both the equilibrium states of PrP and its kinetic pathways of folding and aggregation.

The conceptualisation of cardiometabolic disease policy model in the UK

BackgroundDecision models are increasingly used to inform policy-making processes, and there is a need to improve their credibility. The estimation of health and economic outcomes generated from decision models is influenced by the development process itself. This paper aims to present the conceptual model development process of cardiometabolic disease (CMD) policy models in the UK setting.MethodsThis conceptual model followed the International Society of Pharmacoeconomics and Outcomes Research-Society of Medical Decision Making (ISPOR-SMDM) Modelling Good Research Practices Task Force-2.ResultsFirst, for the conceptualisation of the problem, the CMD disease staging, progression and current clinical guidelines were summarised, followed by a systematic review of published policy models. We critically appraised policy models such as cardiovascular disease and type 2 diabetes. Key messages from the review emphasised the importance of understanding various determinants influencing model development, including risk factors, model structure, models’ parameters, data utilisation, economic perspective, equality/equity consideration, transparency and validation process. Second, as a sequential process, is model conceptualisation, to determine which modelling types and their attributes best represent the defined problem. Expert opinions, including a clinician and experienced modellers, provided input on the state transition model to ensure the structure is clinically relevant. From this stage, the consideration and agreement to establish a disease state in a state transition model was discussed.ConclusionThis conceptual model serves as a basis for representing the systematic process for structuring a CMD policy model to enhance its transparency and credibility.

PH-sensing GPR68 inhibits vascular smooth muscle cell proliferation through Rap1A.

Phenotypic transformation of vascular smooth muscle (VSM) from a contractile state to a synthetic, proliferative state is a hallmark of cardiovascular disease (CVD). In CVD, diseased tissue often becomes acidic from altered cellular metabolism secondary to compromised blood flow, yet the contribution of local acid/base imbalance to the disease process has been historically overlooked. In this study, we examined the regulatory impact of the pH-sensing G protein-coupled receptor GPR68 on vascular smooth muscle (VSM) proliferation in vivo and in vitro in wild-type (WT) and GPR68 knockout (KO) male and female mice. Arterial injury reduced GPR68 expression in WT vessels and exaggerated medial wall remodeling in GPR68 KO vessels. In vitro, KO VSM cells showed increased cell cycle progression and proliferation compared to WT VSM cells, and GPR68-inducing acidic exposure reduced proliferation in WT cells. mRNA and protein expression analyses revealed increased Rap1A in KO cells compared to WT cells, and RNA silencing of Rap1A reduced KO VSM cell proliferation. In sum, these findings support a growth-inhibitory capacity of pH-sensing GPR68 and suggest a mechanistic role for the small GTPase Rap1A in GPR68-mediated VSM growth control. These results shed light on GPR68 and its effector Rap1A as potential targets to combat pathologic phenotypic switching and proliferation in VSM.

Sodium thiosulfate treatment rescues hyperglycaemia-induced pronephros damage in zebrafish by upregulating nitric oxide signalling.

Sodium thiosulfate (STS) is gaining increasing attention in research for its potential therapeutic applications across a spectrum of disease processes beyond its current uses. However, the precise mechanisms of action remain incompletely understood. We investigated the efficacy of STS in treating hyperglycaemia-induced pronephros damage in zebrafish to gain further insight into the underlying mechanisms. Hyperglycaemia was induced in zebrafish by suppressing the pdx1 transcription factor, which plays a crucial role in maintaining physiological pancreatic function. STS was administered by introducing it into the medium of zebrafish larvae. The pronephros structure was analysed at 48h post-fertilization. Metabolomic profiling and RNA sequencing were conducted on groups exposed to various experimental conditions. Our findings reveal a downregulation of nitric oxide (NO) signalling in zebrafish with a knocked-down pdx1 gene, both metabolomically and transcriptionally. Notably, treatment with STS led to a compensatory upregulation of the NO signalling, ultimately resulting in the rescue of the pronephros structure. Our study provides compelling evidence that targeting NO metabolism by the administration of STS offers a promising strategy for addressing hyperglycaemia-induced organ damage. These findings underscore the potential of STS as a promising therapeutic agent for diabetic complications and warrant further investigation of its clinical applications. KEY POINTS: Sodium thiosulfate (STS) is increasingly drawing attention in research for its potential therapeutic applications across a spectrum of disease processes. Here, we demonstrate that STS treatment rescues hyperglycaemia-induced pronephros damage in zebrafish. We identified upregulation of nitric oxide signalling as the major driver behind STS-mediated rescue. Our data suggest that STS offers a promising strategy for addressing hyperglycaemia-induced organ damage, including diabetic nephropathy.

Discovery of Covalent MLKL PROTAC Degraders via Optimization of a Theophylline Derivative Ligand for Treating Necroptosis.

Mixed lineage kinase domain-like pseudokinase (MLKL) initiates necroptosis and could serve as a therapeutic target related to a series of human diseases. Proteolysis-targeting chimeras (PROTACs) are useful tools for degrading pathological proteins and blocking disease processes. Using computer-aided modeling and molecular dynamics simulations, we developed a series of covalent MLKL PROTACs by linking and optimizing a theophylline derivative that covalently targets MLKL. Via structure-activity relationship studies, MP-11 was identified as a potent MLKL PROTAC degrader. Furthermore, MP-11 showed lower toxicity than the original MLKL ligand, exhibiting nanomolar-scale antinecroptotic activity on human cell lines. Xenograft model studies showed that MP-11 effectively degraded MLKL in vivo. Importantly, our study demonstrates that the covalent binding strategy is an effective approach for designing MLKL-targeting PROTACs, serving as a model for developing PROTACs to treat future necroptosis-related human diseases.