FASEB bioAdvances最新文献

A “quiet revolution” in medicine has been taking place over the past two decades. There are two converging dynamic forces that have propelled precision medicine to the limelight, garnering wide public attention. The first driver is the realization that populations within a disease area can be stratified, thus developing therapies tailored to their specific needs, and the capability to identify these populations by analyzing large, diverse datasets. The second driver is technology advances in multi-omics approaches and applications (i.e., molecularly informed medicine) enabling a more comprehensive portrait of disease biology. This promises to not only accelerate the development of precision medicine processes but also presents challenges for healthcare professionals and health systems that are struggling to interconnect and integrate disparate data sources into a cohesive clinical strategy to the benefit of their patients. We coin here the term next-generation precision medicine (ngPM), which is bound to become conventional in the clinics sooner or later. Artificial intelligence (AI) and machine learning (ML) in healthcare have transformative potential and are a strategic response to today's challenges and tomorrow's opportunities. The chief challenges here are how well precision medicine (PM) permeates primary care to become a standard of care and drive toward precision wellness or precision lifestyle (ngPM), while ensuring access to care is feasible, streamlined, and routine. We present here a perspective that would harness the power of ngPM for precision wellness.

In vivo cell type-specific genetic recombination based on the Cre-loxP system has contributed to the understanding of biological processes and diseases. Neuronal nuclei (NeuN)/RBFOX3 is a widely used mature neuron marker in developmental biology and neuroscience. Here, we generated Rbfox3-improved Cre (iCre) knock-in mouse model and investigated the effect of iCre knock-in into the Rbfox3 gene and Cre recombination activity in the central nervous system (CNS) and peripheral tissues. The knock-in of internal ribosome entry site (IRES)-iCre cassette into the Rbfox3 3′ UTR did not affect birth rate, growth, and brain weight. In the adult brain, iCre protein expression was confirmed, whereas RBFOX3 protein expression was partially reduced in the knock-in mice. Cre recombination analysis using R26GRR fluorescent reporter strain revealed that Rbfox3-driven iCre-induced gene recombination in the CNS and heart during embryonic development. In the adult brain, gene recombination was observed in neurons, however, not in other glial cells. In the peripheral tissues, iCre activity was found in the sciatic nerve and in other peripheral tissues, including the heart, bladder, and testis. We validated gene recombination rate in the germline and found that 100% recombination occurred in male germ cells and approximately 50% in female germ cells. Concludingly, Rbfox3-iCre mice induce genetic recombination in neurons within CNS as well as in some peripheral tissues and germ cells. In addition to establishing a novel Cre mouse line, the findings of this study offer valuable insights into the development and application of mouse tools that utilize the Rbfox3 gene locus.

Acute kidney injury (AKI) is a complex clinical syndrome associated with increased incidence and mortality rates among critically ill patients, often leading to multiple organ dysfunction, which underscores the need to better understand its molecular mechanisms. In this study, common differentially expressed genes (DEGs) between various AKI models and control groups were extracted using the Gene Expression Omnibus (GEO) database, followed by an exploration of potential signaling pathways involved in AKI. Key genes in the development of AKI were identified through Weighted Gene Co-expression Network Analysis (WGCNA) and protein–protein interaction (PPI) networks, and the expression of hub genes was validated using quantitative PCR (qPCR), Western blotting, immunohistochemistry (IHC) and flow cytometry. A total of 1265 DEGs significantly associated with AKI were identified, with GO and KEGG analyses revealing significant enrichment in pathways related to kidney development, muscle regulation, and amino acid biosynthesis. WGCNA further screened AKI-related modules, identifying 290 DEGs significantly linked to the disease state. PPI network analysis revealed Fosb as a significantly upregulated hub gene in AKI, with experimental validation demonstrating its substantial upregulation in patients with acute tubular necrosis (ATN), HR-induced HK-2 cells injury and ischemia–reperfusion injury (IRI) mice. Inhibition of Fosb alleviated hypoxia-reoxygenation (HR)-induced apoptosis and inflammation in HK-2 cells by suppressing the AP-1 complex (Fosb/C-Jun) signaling pathway. Therefore, Fosb is significantly upregulated in AKI and associated with inflammation mediated by the AP-1 signaling pathway, suggesting its potential as a diagnostic biomarker and therapeutic target for AKI.

Hair follicle neural crest stem cells reside in the bulge region of the outer root sheath of hair follicles, originate from the ectoderm, and have multidirectional differentiation potential, making them ideal candidates for tissue engineering applications. These cells mainly reside in a hypoxic microenvironment that favors the maintenance of stemness. Recently, many studies have elucidated the involvement of the Hippo pathway in the regulation of stem cell fate. However, few studies have investigated whether the Hippo signaling pathway regulates the growth of hair follicle neural crest stem cells in hypoxic environments. In the present study, we investigated the role of the Hippo pathway in the regulation of hair follicle neural crest stem cells under hypoxic conditions. We identified neural crest-derived stem cells from single-cell RNA-seq data of skin organoids in a public database, and reported that the Hippo pathway was activated in the cell population. Hair follicle neural crest stem cells were isolated from rat hair follicles and cultured under hypoxic (3% oxygen) and normoxic (20% oxygen) conditions. Cell viability was assessed via the CCK8 assay. The expression levels of several key genes, including Hif2α, Nestin, Sox10, Oct4, Nanog, Sox2, and Klf4, were evaluated via quantitative real-time PCR, after which we treated the cells with verteporfin, a small molecule inhibitor of the Hippo pathway. Changes in the subcellular localization of the hair follicle neural crest stem cell-specific marker SOX10 were assessed via immunofluorescence. Western blotting was used to analyze the expression levels of proteins associated with stemness and hypoxia responses, including HIF2α, SOX10, OCT4, NANOG, SOX2, and KLF4. The results showed that hypoxic conditions facilitated the maintenance of stemness in hair follicle neural crest stem cells, including the promotion of proliferation and the expression of multipotential markers. Inhibition of the Hippo pathway results in a significant decrease in cell proliferation. The protein expression of HIF2α, SOX10, OCT4, NANOG, SOX2, and KLF4 was also reduced under hypoxic conditions.

G protein-coupled receptors (GPCRs), which play crucial roles in various physiological functions, often assembled into dimers and higher-order oligomers. This oligomerization phenomenon has been observed in diverse physiological and pathological contexts, presenting promising opportunities for drug discovery targeting vital systems such as the cardiovascular, nervous, endocrine, and renal systems. This review offers a concise understanding of GPCR dimerization, its signaling mechanisms, and its implications. Furthermore, we explored therapeutic strategies aimed at modulating receptors involved in dimer/oligomer formation within the renin-angiotensin system.

Sarcopenia refers to the decline in muscle mass and function that occurs with advancing age. It is driven by alterations in multiple cellular processes. AMP-activated protein kinase (AMPK) is a cellular energy sensor that opposes many age-related changes, making it an attractive target for the treatment of sarcopenia. This study aimed to test the effect of chronic treatment of old mice with the AMPK-activating prodrug, AICAR, on treadmill running capacity and muscle mass, force production, gene expression, and intracellular markers relevant to sarcopenia. Old (23 months) mice were tested for treadmill running capacity, then randomly assigned to receive daily treatment with AICAR (OA; 300 to 500 mg/kg, delivered via subcutaneous injection) or an equivalent volume of saline vehicle (OS) for 31 days. Young (5 months) saline-treated mice (YS) served as controls. Treadmill posttesting was performed after 24 days, and the mice were euthanized after 31 days of treatment. Extensor digitorum longus (EDL) muscles were tested for force generation and RNA sequencing, RT-PCR, and western blotting were performed on quadricep muscles. Treadmill running capacity declined from pre- to posttesting by 24.5% in OS mice. This decline was not observed in YS or OA mice. Quadricep weight was ~8% higher, and tetanic force production by the EDL muscle increased by 26.4% in OA versus OS. These phenotypic improvements with AICAR treatment were accompanied by changes in gene expression in OA/YS versus OS muscles consistent with the “rejuvenation” of gene ontologies associated with connective tissue, neurodegenerative disease, Akt signaling, and mitochondrial function, among others. AICAR increased the mitochondrial markers cytochrome C by ~33%, and citrate synthase by ~22%. Serum insulin-like growth factor-1 levels increased, and Akt phosphorylation tended (p = 0.07) to increase with AICAR treatment. Although protein levels of the mTORC1 signaling pathway intermediate, rpS6, were higher in OA versus OS muscles, the phosphorylation of mTORC1 pathway intermediates was unaffected. On the other hand, gene expression of the muscle-specific ubiquitin ligases Mafbx and Murf1 were reduced with AICAR treatment. AICAR treatment mildly increased/preserved muscle mass and force production and prevented a decline in treadmill running performance in old mice. These effects were associated with altered skeletal muscle gene and protein expression, suggesting improved mitochondrial content and metabolic signaling (particularly through Akt) as contributing factors to the observed phenotypic effects. Our findings support further development of AMPK-activating drugs as a therapeutic strategy for improving age-related organismal dysfunction and sarcopenia.

Maturation represents a process characterized by adaptive changes, particularly in the circulatory system. However, it is not known whether, in neonates, potassium channels contribute to NO-induced vasorelaxation at all and, if so, which potassium channels these are. Therefore, this study tested the hypothesis that potassium channels mediate NO-induced vasorelaxation in newborn rats. Young (10- to 15-day-old) and adult (2- to 3-month-old) male rats were studied using real-time PCR, isometric myography, and the sharp microelectrode technique on saphenous arteries. We observed prominent mRNA expression of several distinct isoforms of potassium channel families known to potentially mediate SNP-induced vasodilation. Further, in both adult and young rats, SNP can relax vessels independently of potassium channels. A solely potassium channel-independent anticontractile effect of SNP was observed also when either Kir6, or Kir2, or Kv2 channels, respectively, were available in both adult and young rats. However, when Kv1 channels were available, a Kv1 channel-dependent component contributed to the anticontractile effect of SNP in young rats. When BK_Ca channels were available, a BK_Ca channel-dependent component contributed to the anticontractile effect of SNP in adult rats. A considerable Kv7 channel-dependent component contributed to the anticontractile effect of SNP in both adult and young rats. Thus, the data of the present study show for the first time that potassium channels, even multiple ones, contribute to SNP-induced vasorelaxation in newborn rats and that the potassium channels involved in SNP-induced vasorelaxation change from Kv1/Kv7 channels to BK_Ca/Kv7 channels during postnatal development.

Amylin, also known as islet amyloid polypeptide (IAPP), is a pancreatic β-cell peptide hormone involved in satiation and control food intake. It is also produced in smaller quantities by neurons, the gastrointestinal tract, and spinal ganglia. Numerous studies have revealed that patients with type 2 diabetes mellitus (T2DM) and cognitive deficits exhibit IAPP deposits in the pancreas, brain, and blood vessels. IAPP has also been shown to exert neuroprotective effects against Alzheimer's disease (AD) and cognitive impairments. The objective of this review paper is to provide recent information about the pathophysiological roles of IAPP in metabolic and in neurological disorders, and its potential as a druggable target. We have reviewed preclinical and clinical human and animal research studies of IAPP. We discuss the IAPP structure, its receptors, and its physiological functions in metabolism, satiation, adiposity, obesity, and in the brain. Then we discuss its role in metabolic and neurological disorders like diabetes, obesity, bone disorder, neurodegeneration, cerebrovascular disorders, depression, alcohol use disorder, epilepsy, and in ovarian cysts. Overall, this review provides information on the progress of research into the roles of IAPP and its receptor in food intake, energy homeostasis, glucose regulation, satiation, and its role in metabolic and neurological disorders making it a potential target for therapeutic approaches. This review also suggests that the utilization of rodents overexpressing human IAPP in neurodegeneration models may unearth some significant therapeutic potentials for neurological disorders.

Transplanted adipose stem cells (ASC) have a low survival rate in the body, and there are not many ASC that can be effectively used, which weakens their tissue repair function. Based on this status quo, a new type of copper-based metal–organic network (Cu-MON) was used to pretreat cells to regulate cell activity in order to improve the efficacy of cell therapy or reduce the number of cells used, thus reducing the cost of clinical treatment. Gene expression changes before and after Cu-MON treatment of normal donor adipose stem cells (ND-ASC) and type 2 diabetes mellitus adipose stem cells (T2DM-ASC) were evaluated through RNA sequencing, KEGG and GO enrichment analysis. The results showed that Cu-MON improved ASC cell quality by regulating immune response and promoting paracrine secretion. IL-17 signaling pathway and IL-6, CXCL8, and MMP-9 were key pathways and necessary genes that affected the ability of stem cells. In addition, Cu-MON also improved stem cell antiviral ability through Type I interferon signaling pathway. Our research showed that Cu-MON had improved the cell quality of ASC by regulating immune response, promoting paracrine secretion, and improving antiviral capabilities. This approach to biomaterial pretreatment is fast, convenient, and relatively safe, and provides new strategies for improving the efficiency of cell therapies.

6-Nitrodopamine (6-ND) is the main catecholamine released from human isolated vas deferens and the adrenergic nervous system is known to play a major role in the contractions of the epididymal portion of the vas deferens. Here it was investigated the interactions of 6-ND on the contractions of the human isolated vas deferens induced by either classical catecholamines or electric-field stimulation (EFS). The vas deferens obtained from 106 patients who underwent vasectomy surgery were mounted in a 10-mL glass chamber filled with warmed (37°C) and oxygenated Krebs–Henseleit's solution. The strips were pretreated (30 min) with 6-ND (0.1–100 nM) and exposed to increasing concentrations of noradrenaline (0.01–300 M), dopamine (0.00001–10 mM), or adrenaline (0.01–300 M). The strips were also submitted to EFS in tissues pre-incubated or not with 6-ND (1–100 nM), noradrenaline (100 nM), adrenaline (100 nM), or dopamine (100 nM). Catecholamine basal release was evaluated by LC–MS/MS and expression of tyrosine hydroxylase by both immunohistochemistry (IC) and fluorescence in-situ hybridization (FISH). Pre-incubation of the vas deferens with 6-ND caused marked potentiation of the contractions induced by noradrenaline, adrenaline, and dopamine, as characterized by significant increases in E_max, without changes in pEC₅₀ values. 6-nitrodopamine also caused significant increases in the EFS-induced contractions. The basal release of 6-ND was not affected by pre-treatment of the tissues with tetrodotoxin. Tyrosine hydroxylase was detected in epithelial cells of human vas deferens samples by both IC and FISH. The results clearly demonstrate that epithelium-derived 6-ND is a major modulator of human vas deferens contractility.