Genes&Diseases

语种：英文出版周期：双月刊

E-ISSN：2352-3042P-ISSN：2352-4820

主管单位：重庆市教育委员会主办单位：重庆医科大学

Genes and Diseases是本由重庆医科大学于2014年创办的双月刊，也是国内第一本分子医学与转化医学相结合的全英文综合期刊，并入选“中国科技期刊卓越行动计划”高起点新刊项目。

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High-intensity interval training (HIIT) has been found to be more effective in relieving heart failure (HF) symptoms, than moderate-intensity continuous aerobic training (MICT). Additionally, higher meteorin-like protein (Metrnl) levels are seen after HIIT versus MICT. We investigated whether Metrnl contributed to post-HF cardiac functional improvements, and the signaling pathways involved. 50 HF patients underwent MICT, and another 50, HIIT, which was followed by cardiac function and serum Metrnl measurements. Metrnl was also measured in both blood and skeletal muscle samples of mice with transverse aortic constriction-induced HF after undergoing HIIT. Afterward, shRNA-containing adenovectors were injected into mice, yielding five groups: control, HF, HF + HIIT + scrambled shRNA, HF + HIIT + shMetrnl, and HF + Metrnl (HF + exogenous Metrnl). Mass spectrometry identified specific signaling pathways associated with increased Metrnl, which was confirmed with biochemical analyses. Glucose metabolism and mitochondrial functioning were evaluated in cardiomyocytes from the five groups. Both HF patients and mice had higher circulating Metrnl levels post-HIIT. Metrnl activated AMPK in cardiomyocytes, subsequently increasing histone deacetylase 4 (HDAC4) phosphorylation, leading to its cytosolic sequestration and inactivation via binding with chaperone protein 14-3-3. HDAC4 inactivation removed its repression on glucose transporter type 4, which, along with increased mitochondrial complex I-V expression, yielded improved aerobic glucose respiration and alleviation of mitochondrial dysfunction. All these changes ultimately result in improved post-HF cardiac functioning. HIIT increased skeletal muscle Metrnl production, which then operated on HF hearts to alleviate their functional defects, via increasing aerobic glucose metabolism through AMPK-HDAC4 signaling.

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FK506-binding protein 9 (FKBP9) is involved in tumor malignancy by resistance to endoplasmic reticulum (ER) stress, and the up-regulation of FKBP9 is associated with patients' poor prognosis. The current knowledge of the molecular mechanisms is still limited. One previous study showed that FKBP9 could confer glioblastoma cell resistance to ER stress through ASK1-p38 signaling. However, the upstream regulatory mechanism of FKBP9 expression is still indistinct. In this study, we identified the FKBP9 binding proteins using co-immunoprecipitation followed by mass spectrometry. Results showed that FKBP9 interacted with the binding immunoglobulin protein (BiP). BiP bound directly to FKBP9 with high affinity. BiP prolonged the half-life of the FKBP9 protein and stabilized the FKBP9 protein. BiP and FKBP9 protein levels were positively correlated in patients with glioma, and patients with high expression of BiP and FKBP9 showed a worse prognosis. Further studies showed that FKBP9 knockout in genetically engineered mice inhibited intracranial glioblastoma formation and prolonged survival by decreasing cellular proliferation and ER stress-induced CHOP-related apoptosis. Moreover, normal cells may depend less on FKBP9, as shown by the absence of apoptosis upon FKBP9 knockdown in a non-transformed human cell line and overall normal development in homozygous knockout mice. These findings suggest an important role of BiP-regulated FKBP9-associated signaling in glioma progression and the BiP–FKBP9 axis may be a potential therapeutic target for glioma.

Abnormal mitochondrial dynamics can lead to seizures, and improved mitochondrial dynamics can alleviate seizures. Vacuolar protein sorting 13D (VPS13D) is closely associated with regulating mitochondrial homeostasis and autophagy. However, further investigation is required to determine whether VPS13D affects seizures by influencing mitochondrial dynamics and autophagy. We aimed to investigate the influence of VPS13D on behavior in a rat model of acute epileptic seizures. Hence, we established an acute epileptic seizure rat model and employed the CRISPR/CAS9 technology to construct a lentivirus to silence the Vps13d gene. Furthermore, we used the HT22 mouse hippocampal neuron cell line to establish a stable strain with suppressed expression of Vps13d in vitro. Then, we performed quantitative proteomic and bioinformatics analyses to confirm the mechanism by which VPS13D influences mitochondrial dynamics and autophagy, both in vitro and in vivo using the experimental acute epileptic seizure model. We found that knockdown of Vps13d resulted in reduced seizure latency and increased seizure frequency in the experimental rats. Immunofluorescence staining and western blot analysis revealed a significant increase in mitochondrial dynamin-related protein 1 expression following Vps13d knockdown. Moreover, we observed a significant reduction in LC3II protein expression levels and the LC3II/LC3I ratio (indicators for autophagy) accompanied by a significant increase in P62 expression (an autophagy adaptor protein). The proteomic analysis confirmed the up-regulation of P62 protein expression. Therefore, we propose that VPS13D plays a role in modulating seizures by influencing mitochondrial dynamics and autophagy.

Resistance to sorafenib, an effective first-line treatment for advanced hepatocellular carcinoma (HCC), greatly compromised the prognosis of patients. The extracellular matrix is one of the most abundant components of the tumor microenvironment. Beyond acting as a physical barrier, it remains unclear whether cell interactions and signal transduction mediated by the extracellular matrix contribute to sorafenib resistance. With the analysis of primary HCC organoid RNA-seq data combined with in vivo and in vitro experiments validation, we discovered that fibronectin extra domain A (FN-EDA) derived from cancer-associated fibroblasts played a critical role in sorafenib resistance. Mechanistically, FN-EDA stimulates the up-regulation of the key one-carbon metabolism enzyme SHMT1 in HCC cells via the TLR4/NF-κB signaling pathway, thereby countering the oxidative stress induced by sorafenib. Moreover, we reinforced the clinical significance of our discoveries by conducting in vivo assays with an immunodeficiency subcutaneous xenograft tumor model, which was established using primary cancer-associated fibroblasts derived from clinical HCC tissues, and through the analysis of HCC samples obtained from The Cancer Genome Atlas (TCGA) database. Our findings suggest that targeting the FN-EDA/SHMT1 pathway could be a potential strategy to improve sorafenib responsiveness in HCC patients.

Recombinant adenovirus (rAdV) is a commonly used vector system for gene transfer. Efficient initial packaging and subsequent production of rAdV remains time-consuming and labor-intensive, possibly attributable to rAdV infection-associated oxidative stress and reactive oxygen species (ROS) production. Here, we show that exogenous GAPDH expression mitigates adenovirus-induced ROS-associated apoptosis in HEK293 cells, and expedites adenovirus production. By stably overexpressing GAPDH in HEK293 (293G) and 293pTP (293GP) cells, respectively, we demonstrated that rAdV-induced ROS production and cell apoptosis were significantly suppressed in 293G and 293GP cells. Transfection of 293G cells with adenoviral plasmid pAd-G2Luc yielded much higher titers of Ad-G2Luc at day 7 than that in HEK293 cells. Similarly, Ad-G2Luc was amplified more efficiently in 293G than in HEK293 cells. We further showed that transfection of 293GP cells with pAd-G2Luc produced much higher titers of Ad-G2Luc at day 5 than that of 293pTP cells. 293GP cells amplified the Ad-G2Luc much more efficiently than 293pTP cells, indicating that exogenous GAPDH can further augment pTP-enhanced adenovirus production. These results demonstrate that exogenous GAPDH can effectively suppress adenovirus-induced ROS and thus accelerate adenovirus production. Therefore, the engineered 293GP cells represent a superfast rAdV production system for adenovirus-based gene transfer and gene therapy.

Recent studies have explored the spatial transcriptomics patterns of Alzheimer's disease (AD) brain by spatial sequencing in mouse models, enabling the identification of unique genome-wide transcriptomic features associated with different spatial regions and pathological status. However, the dynamics of gene interactions that occur during amyloid-β accumulation remain largely unknown. In this study, we performed analyses on ligand-receptor communication, transcription factor regulatory network, and spot-specific network to reveal the dependence and the dynamics of gene associations/interactions on spatial regions and pathological status with mouse and human brains. We first used a spatial transcriptomics dataset of the AppNL-G-F knock-in AD and wild-type mouse model. We revealed 17 ligand-receptor pairs with opposite tendencies throughout the amyloid-β accumulation process and showed the specific ligand-receptor interactions across the hippocampus layers at different extents of pathological changes. We then identified nerve function related transcription factors in the hippocampus and entorhinal cortex, as well as genes with different transcriptomic association degrees in AD versus wild-type mice. Finally, another independent spatial transcriptomics dataset from different AD mouse models and human single-nuclei RNA-seq data/AlzData database were used for validation. This is the first study to identify various gene associations throughout amyloid-β accumulation based on spatial transcriptomics, establishing the foundations to reveal advanced and in-depth AD etiology from a novel perspective based on the comprehensive analyses of gene interactions that are spatio-temporal dependent.

Head and neck squamous cell carcinoma (HNSC) represents nearly 90% of all head and neck tumors. The current treatment modality for HNSC patients primarily involves surgical intervention and radiotherapy, but its therapeutic efficacy remains limited. The mRNA vaccine based on tumor antigens seems promising for cancer treatment. Ferroptosis, a novel form of cell death, is linked to tumor progression and cancer immunotherapy. Nevertheless, the effectiveness of ferroptosis-associated tumor antigens in treating HNSC remains uncertain. In this study, we identified three ferroptosis-associated tumor antigens, namely caveolin1 (CAV1), ferritin heavy chain (FTH1), and solute carrier 3A2 (SLC3A2), as being overexpressed and mutated based on data obtained from The Cancer Genome Atlas and Gene Expression Omnibus databases. These antigens were strongly associated with poor prognosis and infiltration of antigen-presenting cells in HNSC. We further identified two ferroptosis subtypes (FS1 and FS2) with distinct molecular, cellular, and clinical properties to identify antigen-sensitive individuals. Our findings indicate that FS1 exhibits an immune “hot” phenotype, whereas FS2 displays an immune “cold” phenotype. Additionally, differential expression of immunogenic cell death modulators and immune checkpoints was observed between these two immune subtypes. Further exploration of the HNSC's immune landscape revealed significant heterogeneity among individual patients. Our findings suggest that CAV1, FTH1, and SLC3A2 are potential targets to prevent HNSC in FS2 patients. Overall, our research reveals the potential of ferroptosis-associated mRNA vaccines for HNSC and identifies an effective patient population for vaccine treatment.

Programmed cell death contributes to neurological damage in ischemic stroke, especially during the reperfusion stage. Several cell death pathways have been tested preclinically and clinically, including ferroptosis, necroptosis, and apoptosis. However, the sequence and complex interplay between cell death pathways during ischemia/reperfusion remains under investigation. Here, we unbiasedly investigated cell death pathways during ischemia/reperfusion by utilizing RNA sequencing analysis and immunoblot assays and revealed that ferroptosis and necroptosis occurred early post-reperfusion, followed by apoptosis. Ferroptosis inhibitor Liproxstatin-1 effectively inhibited necroptosis during reperfusion, while the necroptosis inhibitor Necrostatin-1 suppressed protein expression consistent with ferroptosis activation. Protein–protein interaction analysis and iron chelation therapy by deferoxamine mesylate indicate that iron is capable of promoting both ferroptosis and necroptosis in middle cerebral artery occlusion/repression modeled mice. Treatment of cells with iron led to a disruption in redox balance with activated necroptosis and increased susceptibility to ferroptosis. Collectively, these data uncovered a complex interplay between ferroptosis and necroptosis during ischemic stroke and indicated that multiple programmed cell death pathways may be targeted co-currently.

Stimulator of interferon genes (STING) has recently been found to play a crucial role in cardiac sterile inflammation and dysfunction. The role of stimulator of interferon genes (STING) in cardiac sterile inflammation and dysfunction has been recently discovered. This study aims to examine the involvement of STING in pathological cardiac remodeling and the mechanisms that govern the activation of the STING pathway. To investigate this, transverse aortic constriction (TAC) was performed on STING knockout mice to induce pressure overload-induced cardiac remodeling. Subsequently, cardiac function, remodeling, and inflammation levels were evaluated. The STING pathway was found to be activated in the pressure overload-stressed heart and angiotensin II (Ang II)-stimulated cardiac fibroblasts. Loss of STING expression led to a significant reduction in inflammatory responses, mitochondrial fragmentation, and oxidative stress in the heart, resulting in attenuated cardiac remodeling and dysfunction. Furthermore, the exacerbation of pressure overload-induced STING-mediated inflammation and pathological cardiac remodeling was observed when mitophagy was suppressed through the silencing of Parkin, an E3 ubiquitin ligase. Taken together, these findings indicate that STING represents a newly identified and significant molecule implicated in the process of pathological cardiac remodeling and that mitophagy is an upstream mechanism that regulates STING activation. Targeting STING may therefore provide a novel therapeutic strategy for pathological cardiac remodeling and heart failure.

The pathogenic effects of type 2 diabetes on bone tissue are gaining attention, but the cellular and molecular mechanisms underlying osteoimmunology are still unclear in diabetes-related bone diseases. We delineated the single-cell transcriptome of bone marrow cells from both wide type and type 2 diabetes mice, which provided the first detailed global profile of bone marrow cells and revealed a distinct bone immune microenvironment at the genetic level under type 2 diabetic condition. It was observed that osteoclast activity was inhibited due to a dysregulated cytokine network, which ultimately led to decreased osteoclast formation and differentiation. In type 2 diabetes mice, a specific Cd36+ cluster (cluster 18, monocytes/macrophages 2) was identified as the precursor of osteoclasts with diminished differentiation potential. AP-1 was demonstrated to be the key transcription factor in the underlying mechanism.

Dysregulated calcium (Ca2+) signaling pathways are associated with tumor cell death and drug resistance. In non-excitable cells, such as hepatocellular carcinoma (HCC) cells, the primary pathway for Ca2+ influx is through stromal interaction molecule 1 (STIM1)-mediated store-operated calcium entry (SOCE). Previous studies have demonstrated the involvement of STIM1-mediated SOCE in processes such as genesis, metastasis, and stem cell self-renewal of HCC. However, it remains unclear whether STIM1-mediated SOCE plays a role in developing acquired resistance to sorafenib in HCC patients. In this study, we established acquired sorafenib-resistant (SR) HCC cell lines by intermittently exposing them to increasing concentrations of sorafenib. Our results showed higher levels of STIM1 and stronger SOCE in SR cells compared with parental cells. Deleting STIM1 significantly enhanced sensitivity to sorafenib in SR cells, while overexpressing STIM1 promoted SR by activating SOCE. Mechanistically, STIM1 increased the transcription of SLC7A11 through the SOCE-CaN-NFAT pathway. Subsequently, up-regulated SLC7A11 increased glutathione synthesis, resulting in ferroptosis insensitivity and SR. Furthermore, combining the SOCE inhibitor SKF96365 with sorafenib significantly improved the sensitivity of SR cells to sorafenib both in vitro and in vivo. These findings suggest a potential strategy to overcome acquired resistance to sorafenib in HCC cells.

Chronic hepatitis B virus (HBV) infection is a leading cause of liver cirrhosis and hepatocellular carcinoma, representing a global health problem for which a functional cure is difficult to achieve. The HBV core protein (HBc) is essential for multiple steps in the viral life cycle. It is the building block of the nucleocapsid in which viral DNA reverse transcription occurs, and its mediation role in viral-host cell interactions is critical to HBV infection persistence. However, systematic studies targeting HBc-interacting proteins remain lacking. Here, we combined HBc with the APEX2 to systematically identify HBc-related host proteins in living cells. Using functional screening, we confirmed that proteasome activator subunit 1 (PSME1) is a potent HBV-associated host factor. PSME1 expression was up-regulated upon HBV infection, and the protein level of HBc decreased after PSME1 knockdown. Mechanistically, the interaction between PSME1 and HBc inhibited the degradation of HBc by the 26S proteasome, thereby improving the stability of the HBc protein. Furthermore, PSME1 silencing inhibits HBV transcription in the HBV infection system. Our findings reveal an important mechanism by which PSME1 regulates HBc proteins and may facilitate the development of new antiviral therapies targeting PSME1 function.

The recent advances in high throughput sequencing technology have drastically changed the practice of medical diagnosis, allowing for rapid identification of hundreds of genes causing human diseases. This unprecedented progress has made clear that most forms of intellectual disability that affect more than 3% of individuals worldwide are monogenic diseases. Strikingly, a substantial fraction of the mendelian forms of intellectual disability is associated with genes related to the ubiquitin-proteasome system, a highly conserved pathway made up of approximately 1200 genes involved in the regulation of protein homeostasis. Within this group is currently emerging a new class of neurodevelopmental disorders specifically caused by proteasome pathogenic variants which we propose to designate “neurodevelopmental proteasomopathies”. Besides cognitive impairment, these diseases are typically associated with a series of syndromic clinical manifestations, among which facial dysmorphism, motor delay, and failure to thrive are the most prominent ones. While recent efforts have been made to uncover the effects exerted by proteasome variants on cell and tissue landscapes, the molecular pathogenesis of neurodevelopmental proteasomopathies remains ill-defined. In this review, we discuss the cellular changes typically induced by genomic alterations in proteasome genes and explore their relevance as biomarkers for the diagnosis, management, and potential treatment of these new rare disease entities.

Cancer cachexia is a multifactorial syndrome characterized by progressive weight loss and a disease process that nutritional support cannot reverse. Although progress has been made in preclinical research, there is still a long way to go in translating research findings into clinical practice. One of the main reasons for this is that existing preclinical models do not fully replicate the conditions seen in clinical patients. Therefore, it is important to understand the characteristics of existing preclinical models of cancer cachexia and pay close attention to the latest developments in preclinical models. The main models of cancer cachexia used in current research are allogeneic and xenograft models, genetically engineered mouse models, chemotherapy drug-induced models, Chinese medicine spleen deficiency models, zebrafish and Drosophila models, and cellular models. This review aims to revisit and summarize the commonly used animal models of cancer cachexia by evaluating existing preclinical models, to provide tools and support for translational medicine research.

Pancreatic cancer, a highly fatal malignancy, is predicted to rank as the second leading cause of cancer-related death in the next decade. This highlights the urgent need for new insights into personalized diagnosis and treatment. Although molecular subtypes of pancreatic cancer were well established in genomics and transcriptomics, few known molecular classifications are translated to guide clinical strategies and require a paradigm shift. Notably, chronically developing and continuously improving high-throughput technologies and systems serve as an important driving force to further portray the molecular landscape of pancreatic cancer in terms of epigenomics, proteomics, metabonomics, and metagenomics. Therefore, a more comprehensive understanding of molecular classifications at multiple levels using an integrated multi-omics approach holds great promise to exploit more potential therapeutic options. In this review, we recapitulated the molecular spectrum from different omics levels, discussed various subtypes on multi-omics means to move one step forward towards bench-to-beside translation of pancreatic cancer with clinical impact, and proposed some methodological and scientific challenges in store.

Renal fibrosis is a complex and multifactorial process that involves inflammation, cell proliferation, collagen, and fibronectin deposition in the kidney, ultimately leading to chronic kidney disease and even end-stage renal disease. The main goal of treatment is to slow down or halt the progression of fibrosis and to improve or preserve kidney function. Despite significant progress made in understanding the underlying mechanisms of renal fibrosis, current therapies have limited renal protection as the disease progresses. Exosomes derived from stem cells are a newer area of research for the treatment of renal fibrosis. Exosomes as nano-sized extracellular vesicles carry proteins, lipids, and nucleic acids, which can be taken up by local or distant cells, serving as mediators of intercellular communication and as drug delivery vehicles. Exosomes deliver molecules that reduce inflammation, renal fibrosis and extracellular matrix protein production, and promote tissue regeneration in animal models of kidney disease. Additionally, they have several advantages over stem cells, such as being non-immunogenic, having low risk of tumor formation, and being easier to produce and store. This review describes the use of natural and engineered exosomes containing therapeutic agents capable of mediating anti-inflammatory and anti-fibrotic processes during both acute kidney injury and chronic kidney disease. Exosome-based therapies will be compared with stem cell-based treatments for tissue regeneration, with a focus on renal protection. Finally, future directions and strategies for improving the therapeutic efficacy of exosomes are discussed.

The cardiac troponin complex (cTn) is a regulatory component of sarcomere. cTn consists of three subunits: cardiac troponin C (cTnC), which confers Ca2+ sensitivity to muscle; cTnI, which inhibits the interaction of cross-bridge of myosin with thin filament during diastole; and cTnT, which has multiple roles in sarcomere, such as promoting the link between the cTnI-cTnC complex and tropomyosin within the thin filament and influencing Ca2+ sensitivity of cTn and force development during contraction. Conditions that interfere with interactions within cTn and/or other thin filament proteins can be key factors in the regulation of cardiac contraction. These conditions include alterations in myofilament Ca2+ sensitivity, direct changes in cTn function, and triggering downstream events that lead to adverse cardiac remodeling and impairment of heart function. This review describes gene expression and post-translational modifications of cTn as well as the conditions that can adversely affect the delicate balance among the components of cTn, thereby promoting contractile dysfunction.

Histone deacetylases (HDACs) are proteases that play a key role in chromosome structural modification and gene expression regulation, and the involvement of HDACs in cancer, the nervous system, and the metabolic and immune system has been well reviewed. Our understanding of the function of HDACs in the vascular system has recently progressed, and a significant variety of HDAC inhibitors have been shown to be effective in the treatment of vascular diseases. However, few reviews have focused on the role of HDACs in the vascular system. In this study, the role of HDACs in the regulation of the vascular system mainly involving endothelial cells and vascular smooth muscle cells was discussed based on recent updates, and the role of HDACs in different vascular pathogenesis was summarized as well. Furthermore, the therapeutic effects and prospects of HDAC inhibitors were also addressed in this review.

Murine double minute 2 (MDM2) plays an essential role in the cell cycle, apoptosis, DNA repair, and oncogene activation through p53-dependent and p53-independent signaling pathways. Several preclinical studies have shown that MDM2 is involved in tumor immune evasion. Therefore, MDM2-based regulation of tumor cell-intrinsic immunoregulation and the immune microenvironment has attracted increasing research attention. In recent years, immune checkpoint inhibitors targeting PD-1/PD-L1 have been widely used in the clinic. However, the effectiveness of a single agent is only approximately 20%–40%, which may be related to primary and secondary drug resistance caused by the dysregulation of oncoproteins. Here, we reviewed the role of MDM2 in regulating the immune microenvironment, tumor immune evasion, and hyperprogression during immunotherapy. In addition, we summarized preclinical and clinical findings on the use of MDM2 inhibitors in combination with immunotherapy in tumors with MDM2 overexpression or amplification. The results reveal that the inhibition of MDM2 could be a promising strategy for enhancing immunotherapy.

Ovarian cancer is one of the most common malignant tumors of the female reproductive system. The majority of patients with advanced ovarian cancer are mainly treated with cisplatin-based chemotherapy. As the most widely used first-line anti-neoplastic drug, cisplatin produces therapeutic effects through multiple mechanisms. However, during clinical treatment, cisplatin resistance has gradually emerged, representing a challenge for patient outcome improvement. The mechanism of cisplatin resistance, while known to be complex and involve many processes, remains unclear. We hope to provide a new direction for pre-clinical and clinical studies through this review on the mechanism of ovarian cancer cisplatin resistance and methods to overcome drug resistance.

Sodium-glucose co-transporter inhibitors (SGLTis) are the latest class of anti-hyperglycemic agents. In addition to inhibiting the absorption of glucose by the kidney causing glycosuria, these drugs also demonstrate cardio-renal benefits in diabetic subjects. miR-30 family, one of the most abundant microRNAs in the heart, has recently been linked to a setting of cardiovascular diseases and has been proposed as novel biomarkers in kidney dysfunctions as well; their expression is consistently dysregulated in a variety of cardio-renal dysfunctions. The mechanistic involvement and the potential interplay between miR-30 and SGLT2i effects have yet to be thoroughly elucidated. Recent research has stressed the relevance of this cluster of microRNAs as modulators of several pathological processes in the heart and kidneys, raising the possibility of these small ncRNAs playing a central role in various cardiovascular complications, notably, endothelial dysfunction and pathological remodeling. Here, we review current evidence supporting the pleiotropic effects of SGLT2is in cardiovascular and renal outcomes and investigate the link and the coordinated implication of the miR-30 family in endothelial dysfunction and cardiac remodeling. We also discuss the emerging role of circulating miR-30 as non-invasive biomarkers and attractive therapeutic targets for cardiovascular diseases and kidney diseases. Clinical evidence, as well as metabolic, cellular, and molecular aspects, are comprehensively covered.

Osteoarthritis (OA) is a common chronic joint disease characterized by articular cartilage degeneration, subchondral sclerosis, synovitis, and osteophyte formation. OA is associated with disability and impaired quality of life, particularly among the elderly. Leptin, a 16-kD non-glycosylated protein encoded by the obese gene, is produced on a systemic and local basis in adipose tissue and the infrapatellar fat pad located in the knee. The metabolic mechanisms employed by leptin in OA development have been widely studied, with attention being paid to aging as a corroborative risk factor for OA. Hence, in this review, we have attempted to establish a potential link between leptin and OA, by focusing on aging-associated mechanisms and proposing leptin as a potential diagnostic and therapeutic target in aging-related mechanisms of OA that may provide fruitful guidance and emphasis for future research.

The NRF2 pathway is a metabolic- and redox-sensitive signaling axis in which the transcription factor controls the expression of a multitude of genes that enable cells to survive environmental stressors, such as oxidative stress, mainly by inducing the expression of cytoprotective genes. Basal NRF2 levels are maintained under normal physiological conditions, but when exposed to oxidative stress, cells activate the NRF2 pathway, which is crucial for supporting cell survival. Recently, the NRF2 pathway has been found to have novel functions in metabolic regulation and interplay with other signaling pathways, offering novel insights into the treatment of various diseases. Numerous studies have shown that targeting its pathway can effectively investigate the development and progression of age-related musculoskeletal diseases, such as sarcopenia, osteoporosis, osteoarthritis, and intervertebral disc degeneration. Appropriate regulation of the NRF2 pathway flux holds promise as a means to improve musculoskeletal function, thereby providing a new avenue for drug treatment of age-related musculoskeletal diseases in clinical settings. The review summarized an overview of the relationship between NRF2 and cellular processes such as oxidative stress, apoptosis, inflammation, mitochondrial dysfunction, ferroptosis, and autophagy, and explores the potential of targeted NRF2 regulation in the treatment of age-related musculoskeletal diseases.

Ovarian cancer is a common cancer for females, and the incidence and mortality rates are on the rise. Many treatment strategies have been developed for ovarian cancer, including chemotherapy and immunotherapy, but they are often ineffective and prone to drug resistance. Protein ubiquitination is an important class of post-translation modifications that have been found to be associated with various human diseases and cancer development. Recent studies have revealed that protein ubiquitination is involved in the progression of ovarian cancer and plays an important role in the tumor immune process. Moreover, the combination of ubiquitinase/deubiquitinase inhibitors and cancer immunotherapy approaches can effectively reduce treatment resistance and improve treatment efficacy, which provides new ideas for cancer treatment. Herein, we review the role of protein ubiquitination in relation to ovarian cancer immunotherapy and recent advances in the use of ubiquitinase/deubiquitinase inhibitors in combination with cancer immunotherapy.

Cardiovascular diseases (CVDs) impose a significant burden worldwide. Despite the elucidation of the etiology and underlying molecular mechanisms of CVDs by numerous studies and recent discovery of effective drugs, their morbidity, disability, and mortality are still high. Therefore, precise risk stratification and effective targeted therapies for CVDs are warranted. Recent improvements in single-cell RNA sequencing and spatial transcriptomics have improved our understanding of the mechanisms and cells involved in cardiovascular phylogeny and CVDs. Single-cell RNA sequencing can facilitate the study of the human heart at remarkably high resolution and cellular and molecular heterogeneity. However, this technique does not provide spatial information, which is essential for understanding homeostasis and disease. Spatial transcriptomics can elucidate intracellular interactions, transcription factor distribution, cell spatial localization, and molecular profiles of mRNA and identify cell populations causing the disease and their underlying mechanisms, including cell crosstalk. Herein, we introduce the main methods of RNA-seq and spatial transcriptomics analysis and highlight the latest advances in cardiovascular research. We conclude that single-cell RNA sequencing interprets disease progression in multiple dimensions, levels, perspectives, and dynamics by combining spatial and temporal characterization of the clinical phenome with multidisciplinary techniques such as spatial transcriptomics. This aligns with the dynamic evolution of CVDs (e.g., “angina–myocardial infarction–heart failure” in coronary artery disease). The study of pathways for disease onset and mechanisms (e.g., age, sex, comorbidities) in different patient subgroups should improve disease diagnosis and risk stratification. This can facilitate precise individualized treatment of CVDs.

The immune responses play a profound role in the progression of lung lesions in both infectious and non-infectious diseases. Dendritic cells, as the “frontline” immune cells responsible for antigen presentation, set up a bridge between innate and adaptive immunity in the course of these diseases. Among the receptors equipped in dendritic cells, Toll-like receptors are a group of specialized receptors as one type of pattern recognition receptors, capable of sensing environmental signals including invading pathogens and self-antigens. Toll-like receptor 4, a pivotal member of the Toll-like receptor family, was formerly recognized as a receptor sensitive to the outer membrane component lipopolysaccharide derived from Gram-negative bacteria, triggering the subsequent response. Moreover, its other essential roles in immune responses have drawn significant attention in the past decade. A better understanding of the implication of Toll-like receptor 4 in dendritic cells could contribute to the management of pulmonary diseases including pneumonia, pulmonary tuberculosis, asthma, acute lung injury, and lung cancer.

Pneumonia is an inflammatory condition of the lung with symptoms that include productive dry cough, fever, chest pain, and difficulty breathing, and it is usually caused by viruses and bacteria, but also other microorganisms (such as fungi and parasites). Community-acquired pneumonia (CAP) is a major cause of infectious diseases, hospitalization, and mortality, especially in the elderly population.1 We have undertaken a case–control study on CAP patients by way of sequencing the complete exome of 300 patients and 438 healthy controls (Table S1 for demographic and clinical characteristics of patients). This study is by far the largest exome sequencing project to date aimed at exploring the potential genetic causes behind CAP.

Hepatocellular carcinoma (HCC), a highly malignant tumor, faces a major challenge with its high post-hepatectomy recurrence impacting patient survival. Mechanisms and preventive strategies remain to be resolved. Zrt-/Irt-like protein 4 (ZIP4, also called SLC39A4/solute carrier family 39 member 4) belongs to the zinc transporter ZIP superfamily of which the members play an important role in maintaining zinc steady state, and its high expression in various tumors is associated with the prognosis of cancer patients.1 Recently, it has been reported that ZIP4 promoted the development of pancreatic cancer through the zinc finger E-box binding homeobox 1 (ZEB1)/Integrin α3β1/equilibrative nucleoside transporter (ENT1) 1,2 and zona occludens 1 (ZO-1)/claudin-1/ZEB1 pathways.3 In nasopharyngeal carcinoma, ZIP4 induces epithelial–mesenchymal transition (EMT) through the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway and promotes migration and invasion.4 In our previous study, ZIP4 was found to be highly expressed in HCC and promoted HCC invasion and metastasis.5 But so far, the direct regulation mechanism for ZIP4 promotes liver cancer is still unclear. In this study, we found that ZIP4 binds to Ephrin-B1 to inhibit the ubiquitination of Ephrin-B1, regulating the Wnt family member 5A (Wnt5A)/Jun N-terminal kinase (JNK)/ZEB1 signaling pathway and promoting EMT, thereby inducing invasion and metastasis of liver cancer cells.

Fetal hydrops is a rare but serious fetal developmental abnormality characterized by the abnormal accumulation of large amounts of fluid in the fetus resulting in generalized edema, and clinically manifested by abnormal functioning of multiple organs and systems.1 The ryanodine receptor 1 (RYR1) gene encodes the ryanodine receptor found in skeletal muscle and is expressed predominantly in cardiac and skeletal muscle.2 The encoded protein functions as a calcium release channel in the sarcoplasmic reticulum, but is also used to connect the sarcoplasmic reticulum to the transverse tubules. Mutations in this gene have been associated with malignant hyperthermia susceptibility, central core disease, and micro-nucleated cardiomyopathy with extraocular muscle paralysis.3 Existing studies have also found that mutations in the RYR1 gene, as well as mutation-induced purebred shear variants, can be associated with fetal hydrops.4,5 Here, we report a novel heterozygote intronic variant affecting RYR1 gene splicing may cause fetal hydrops.

The treatment of prostate cancer (PCa) needs to be improved.1 Micro-RNAs (miRNAs) are a subtype of non-coding, single-stranded RNAs that influence cellular survival and death by modulating mRNAs. Among these miRNAs, MIR375 has a critical role in the regulation of tumorigenesis2 and holds promise as a novel therapeutic target for future PCa treatment. Recombinant adeno-associated virus (rAAV) exhibits non-pathogenicity, low-grade inflammation, and robust and long-lasting expressions of target genes. We have previously described the rAAV9 as a valid vector to transfer target miRNAs and genes,3 so rAAV9 could be used as a vector to deliver MIR375 into the mouse prostate and PCa cells.

Neuroblastoma (NB) is a common pediatric extracranial solid tumor that exhibits varied characteristics, clinical features, and prognosis.1 Totally 1%–2% of cases show familial history with genetic links like ALK, PHOX2B mutations, and 1p36 or 11q14-23 locus deletions. Gastrointestinal stromal tumors (GISTs) are rare mesenchymal neoplasms of the gastrointestinal tract. Approximately 85% of pediatric patients with GIST lack oncogenic mutations in cKIT or platelet-derived growth factor receptor alpha (PDGFRA), and a majority of these are characterized by molecular alterations in the succinate dehydrogenase (SDH) family of genes.2,3 Genetic changes, germline mutations, and variant-phenotype links in NB and GIST remain largely unexplored. Here, we reported a case of biallelic SDHA variant and copy number deletion causing pediatric recurrent NB with GIST to enhance the understanding of this rare clinical scenario.

The prevalence of sleep disorders is increasing worldwide, prompting greater efforts to understand the biological mechanisms underlying sleep. Despite the growing interest in this area, a continuous transcriptome analysis for studying sleep quality is still lacking. This study performed a regression analysis of the Pittsburgh sleep quality index (PSQI) from the blood transcriptome to identify marker genes associated with sleep quality. In this cohort study, sleep quality scores were defined for 100 participants, and molecular properties associated with the scores were analyzed using blood transcriptome analysis. Gene expression and pathway enrichment analyses revealed that immune-related dysregulation was prominently associated with sleep quality. To understand the genetic influences on our observations, a genome-wide association study (GWAS) was conducted to examine the relationship between sleep quality and immune indicators. The results revealed a relationship between sleep and the immune system and confirmed a genetic correlation. Our study provides valuable insights into the complex interplay between the genetic and environmental factors that determine sleep quality and their impact on overall health.

Infertility affects around 8%–12% of couples globally, and in about 50% of these cases, male factors are either the primary cause or contribute significantly to infertility. Any defects during spermiogenesis may result in male subfertility or complete infertility in mammals.1 Previously, we found that CFAP53 is localized in the manchette and sperm tail, and it plays an essential role in sperm flagellum biogenesis. CFAP53 knockout leads to male infertility due to multiple morphological abnormalities of the flagella.2 To investigate the mechanism of CFAP53 during spermiogenesis, we searched for potential CFAP53-interacting proteins and identified a new CFAP53-interacting protein, CCDC178 (Fig. 1A). Western blotting of this protein revealed a significant decrease of CCDC178 in the testes of Cfap53 knockout mice (Fig. S1A–C). We further showed that CCDC178 colocalized with the centrosome marker γ-tubulin and CFAP53 in either HeLa cells or HEK 293T cells (Fig. S1D). Furthermore, we found that CCDC178 is an evolutionarily conserved protein and is predominantly expressed in the testis (Fig. 1B; Fig. S1E–H), suggesting that this protein might also participate in spermiogenesis.

Constitutive androstane receptor (CAR) is a nuclear receptor that is encoded by the gene NR1I3 (nuclear receptor subfamily 1 group I member 3) and is almost exclusively expressed in the liver.1 As reported in our recent publication, CAR is well established as a xenosensor for drugs and energy metabolism with newer implications in the regulation of normal liver physiology and liver regeneration.2 However, many controversies exist regarding the biological roles of CAR in human liver cancer2 and the species difference between the role of CAR in animal and human liver cancers are evident in the existing experimental and epidemiological data. Activation of CAR in animal models facilitates pro-carcinogenic pathways, eventually leading to the development of hepatocellular carcinoma (HCC). The same phenomenon is not seen following the activation of CAR in human liver cancer models. Recently, a few studies have highlighted the possible tumor-suppressive role of CAR in human cancer including liver cancer.3,4 Here, we aimed to unveil the clinical implications of CAR in human HCC patients.

N6-methyladenosine (m6A) RNA methylation exerts significant functions in regulating various cardiovascular diseases. Following the recognition of the first m6A demethylase, namely fat and obesity-associated protein (FTO), mounting evidence has shown that polymorphism of the FTO gene leads to an increased occurrence of cardiovascular-related risk factors including obesity, diabetes, and inflammation.1 As shown previously, FTO is directly involved in the physiological processes of hypertension, ischemic cardiomyopathy, and heart failure, and it could be a pathogenic factor and potential therapeutic target for various cardiovascular diseases.2 It is experimentally confirmed that m6A deficiency leads to abnormal cardiac function in mice; m6A-deficient mice are more inclined to develop cardiac dysfunction stimulated by serum-induced myocardial hypertrophy.3 Therefore, m6A modification is required to maintain cardiac homeostasis. FTO ameliorates ischemia-induced cardiac systolic dysfunction, which is induced by the demethylating effect of FTO, leading to an inhibition of degrading cardiac contractile transcripts and an enhanced expression of transcripts in the ischemic state.4 These researches showed that FTO might be pivotal in various cardiovascular diseases, particularly in cardiac hypertrophy, by demethylation. However, the specific mechanism of action remains unelucidated. Here, we investigated whether FTO regulates cardiac hypertrophy and the underlying mechanism of its regulatory effect.

Craniopharyngioma (CP) is a rare, histologically benign tumor located in the sellar region which is defined as a grade I tumor by the World Health Organization (WHO) classification.1 There are mainly two different clinicopathological subtypes of CP, the adamantinomatous CP (ACP) and the papillary CP (PCP).1 Although both variations have distinct histomorphological characteristics, an accurate diagnosis might be difficult to make, especially in tiny and/or fragmented specimens. Furthermore, there is a continuous scientific dispute about the occurrence of mixed forms and the cell of origin of these tumors.2 The CTNNB1 gene mutation has been demonstrated to play a significant role in the tumorigenesis of ACP, and a growing body of research supported a high prevalence of CTNNB1 mutation in ACP, while many studies have failed to detect the CTNNB1 mutation in some ACP samples, thus resulting in a CTNNB1 mutation rate of 16%–100%.3,4 Previously, work by Apps et al (2020) has demonstrated the high prevalence of CTNNB1 mutations in ACP (100% in this study) when using a more sensitive TAm-seq sequencing method rather than Sanger sequencing.5 All the 22 ACP samples analyzed were found to carry the CTNNB1 mutation by TAm-seq. A low mutant allelic frequency was found to correlate with the failure to detect the CTNNB1 mutation by Sanger sequencing. Here, to figure out why the mutation rate of CTNNB1 in ACP was inconsistent in the literature and to highlight the importance of the mutation for CP subtyping, Sanger sequencing was used to detect CTNNB1 mutation in fresh-frozen tissues, formalin-fixed paraffin-embedded (FFPE) tissues, and primary ACP cells. Briefly, we observed that the CTNNB1 mutation detection was influenced by the wet keratin/calcification, diaphragma sellae, and reactive glial tissue in ACP tissues. Hematoxylin and eosin (H&E) staining can be used to guide mutation identification to enhance the rate of CTNNB1 mutation detection. An alternative for improving the mutation detection rate is to use primary ACP cells. Finally, The CTNNB1 mutation is critical for CP subtyping, particularly for atypical CP.

Female breast cancer (BCa) has overtaken lung cancer as the most diagnosed cancer worldwide.1 The overall survival for BCa patients without metastasis for five years is over 80%.2 However, relapse and metastasis remain the primary challenge for BCa. In various carcinomas, Holliday junction recognition protein (HJURP) has shown up-regulated patterns and belongs to the CENP-A pre-nucleosomal complex.3, 4, 5 HJURP's expression pattern, roles, and prognosis remain largely unknown in BCa. Nevertheless, we investigated HJURP's expression in BCa and explored its possible role in cancer development. We found that HJURP was significantly higher in BCa tissues and associated with pathological characteristics, and down-regulation of HJURP in BCa cells could suppress the proliferation and migration of MDA-MB-231 cells through the PI3K-AKT pathway. A significant role for HJURP in BCa progression has been identified, and it could provide a valuable biomarker for the diagnosis and prognosis of BCa. Also, considering HJURP as the novel diagnosis and treatment target will provide new strategies for early diagnosis, precise treatment, and effective prevention of BCa.

Adenomatous polyposis coli (APC) is a key tumor suppressor gene playing a central role in the Wnt signaling pathway through β-catenin down-regulation.1 APC germline pathogenic variants are associated with familial adenomatous polyposis (FAP), an autosomal dominant colorectal cancer (CRC) predisposition syndrome characterized by the development of hundreds to thousands of colorectal adenomatous polyps.1 Depending on the number of polyps and age of onset, FAP can be classified into four forms: profuse FAP (>1000 adenomas), intermediate FAP (100-1000 adenomas), attenuated FAP (10–100 adenomas), and gastric polyposis and desmoid FAP (<50 polyps).1 Disease severity seems to be correlated with the location of APC pathogenic variants, most of which occur over the 5′ half of the gene, leading to the elimination of domains involved in β-catenin level regulation and Axin binding.1

Diabetic nephropathy (DN) is now the most common cause of end-stage renal disease in the world. Inflammatory and immune responses play an important role in the pathogenesis of DN, but the specific mechanisms responsible for the development of this disease have yet to be fully elucidated. Numerous epidemiological and preclinical studies have shown that inflammatory response and immune response play an important role in the early stages of DN pathogenesis.1 Still, the exact mechanism for the development of this disease has not been fully elucidated.

Malignant pleural mesothelioma (MPM) is a rare and aggressive cancer with low survival probability as it is generally diagnosed at later stages.1 Using a combination of immune-checkpoint inhibitors (ICIs) ipilimumab (IPI) and nivolumab (NIVO) as a first-line treatment for unresectable MPM, the CheckMate 743 trial reported higher overall survival and prolonged duration of response compared with traditional chemotherapy.1 This combination has recently been approved as a new first-line standard of care for patients with advanced MPM, although the incidence of immune-related adverse events reached 80 %, with 31% of patients experiencing grade 3-4 immune-related adverse events (irAEs).1 The unpredictable nature and severity of immune-related adverse events emphasize a critical need to conduct in-depth translational studies to characterize the tumor microenvironment and systemic immune response that contribute to the efficacy and safety outcomes of ICI therapy.

Endoplasmic reticulum (ER) stress is a procedure that results from increased protein release or improper ER protein folding, which is emerging as a possible driver of pathological conditions such as cancer, cardiometabolic diseases, rheumatic disease, and neurodegenerative diseases.1,2 Activating transcription factor 4 (ATF4), which is considered the primary controller of the cellular reaction when subjected to external stress,3 plays a vital role in amino acid metabolism, differentiation, metastasis, angiogenesis, and stress-related oxidative resistance.4 However, the prognosis value and immune signature of ATF4 activating genes in tumors are still unclear. Therefore, a better understanding of the role of ATF4 activating genes will promote new approaches to tumor treatment. In this work, we set up a model based on the expression level of ATF4 signaling-related genes, and ATF4 signaling score, which reflects the level of ER stress. Then we explored the expression profile of genes relevant to ATF4 signaling and evaluated the association between ATF4 signaling score and the prognosis of cancer patients. Additionally, our study explored the connection between ATF4 signaling score and tumor immunologic characteristics. Therefore, the current work provides a thorough analysis of the expression of 27 ATF4 signaling-related genes in 33 different kinds of cancers. Our results further demonstrate the potential of ATF4 signaling in tumor development and immunotherapy.

Friedreich's ataxia (FRDA) is a rare genetic disorder characterized by motor discoordination and cerebellar involvement due to mutations in the frataxin (FXN) gene, which encodes a mitochondrial protein involved in iron-sulfur cluster biogenesis and iron handling.1 While progress has been made in understanding FRDA's pathophysiology and cerebellar degeneration caused by frataxin deficiency, the role of central nervous system (CNS)-resident non-neuronal cells, as microglia, necessitates further investigation. Microglia play crucial roles in CNS development, neurogenesis, apoptosis, and synaptic remodeling, acting as sentinels with homeostatic functions. In neurodegenerative diseases, microglia respond rapidly to injury, potentially leading to sustained neuroinflammation and therefore, contributing to foster neuron damage.2 Similarly, in FRDA, cerebellar neuron degeneration may be influenced by glial cells in a non-cell-autonomous process. Indeed, in FRDA, microglia contribute to reactive oxygen species accumulation in the CNS, particularly in cerebellar regions, possibly participating in cerebellar susceptibility in ataxias. Circumstantial evidence supports the involvement of neuroinflammatory mechanisms in FRDA pathogenesis,3 as indicated by the presence of hypertrophic and reactive microglia in brain regions of FRDA mouse models and patients, with increased neuroimmune activity correlating with earlier symptom onset and shorter disease duration.4 However, the extent to which microglia dysfunction contributes to FRDA remains uncertain.

PDZ and LIM domain protein 1 (PDLIM1) is a cytoplasmic LIM protein that regulates cytoskeleton organization and functions as a platform to coordinate various cellular processes.1 Accumulated evidence suggests that PDLIM1 plays important roles in tumorigenesis, tumor progression, and response to cancer therapy. Consistently, our results demonstrated that silencing PDLIM1 significantly inhibited the migration and the invasion of HeLa cells compared with control cells (Fig. S1). However, the feasibility of targeting PDLIM1 as an effective strategy for radiotherapy remains to be determined. Here, we knocked down PDLIM1 while irradiating with 60Co γ-rays (IR), and impaired clonogenic survival was observed in PDLIM1-depleted HeLa cells (Fig. 1A, B). Furthermore, the heightened radiosensitivity prompted by PDLIM1 knockdown was effectively counteracted through the expression of siRNA-resistant PDLIM1 protein (Fig. S2A, B). To determine whether PDLIM1 affects DNA damage repair following IR exposure, we performed a neutral comet assay. PDLIM1 knockdown cells have significantly more unrepaired IR-induced DNA double strands breaks (DSBs) compared with control cells, as monitored by comet tail moment (Fig. 1C). There are two primary mechanisms for the restoration of DNA DSBs: homologous recombination (HR) and non-homologous end joining (NHEJ). Therefore, the quantitative assessment of NHEJ and HR efficiencies was conducted in vivo using a CRISPR/Cas9-mediated oligodeoxynuceotide (ODN) detection system.2 The loss of PDLIM1 significantly reduces both NHEJ and HR activity within HeLa cells (Fig. 1D). Subsequently, HeLa cells were subjected to IR, followed by immunofluorescence staining targeting γH2AX, which represents a prominent chromatin modification trigger by several phosphatidylinositol 3-kinase-related family members after DNA DSBs. Interestingly, the depletion of PDLIM1 substantially abrogated the formation of γH2AX after IR exposure (Fig. 1E). The recruitment of mediator of DNA damage checkpoint protein 1 (MDC1) by γH2AX is recognized as the initial step in which the site of a DSB undergoes preparation for the activation of DNA damage response (DDR). Immunofluorescence staining also revealed that IR-induced MDC1 foci formation was attenuated in PDLIM1-deficient cells at the early time points post-IR compared with control cells (Fig. 1F). Furthermore, the hypothesis proposing that the recruitment of DDR factors to DNA DSBs necessitates the presence of PDLIM1 was further substantiated by the observation that the localization of 53BP1 (Fig. 1G) and BRCA1 (Fig. S2C) — both critical factors in NHEJ and HR pathway, respectively — at DSBs sites was diminished in cells depleted of PDLIM1, as compared with the control cells.

Cell typing is an important step in the single-cell RNA sequencing (scRNA-seq) analysis. Although some cell marker databases and cell typing tools have been proposed, limited roles are in prostate cell typing. Through literature review, we found prostate cell typing relied much on researchers' knowledge and experience, thus different markers were used to label the same cell type, leading to the divergences between studies, emphasizing the importance of a sound epistemological foundation for prostate cell typing in single-cell data. Therefore, we designed this study to provide performance analysis for prostate cell markers using eight integrated human prostate scRNA-seq datasets of 170,438 cells from 41 peoples (methods were described in Supplementary Data 2 in detail). Using unsupervised learning, information entropy, F1-score, and local outlier factor score, an objective performance analysis report was obtained, based on which, stable and specific human prostate main and fine cell markers were proposed. Our findings will help decide to select suitable markers for human prostate cell typing in single-cell data.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder causing memory loss, cognitive decline, language impairment, and disorientation, which impose an enormous burden on caregivers and the public health sector. A673T as a protective mutation has great therapeutic potential in AD.1,2 Therefore, a combination of stem cell therapy and A673T mutation existing in natural people based on gene targeting techniques such as CRISPR-Cas9 have been suggested as promising and exciting new developments. Homology-directed repair (HDR), relatively rare in mammalian cells, is necessary to generate a specific sequence change such as point mutations.3 Efficient HDR and inefficient non-homologous DNA end joining are needed.4,5 Moreover, the HDR pathway repairs double-strand breaks via homologous recombination; a synthesized single-stranded oligodeoxynucleotide (ssODN) with a homologous sequence as donor templates is simultaneously introduced, thereby enabling intended sequence changes.5 However, obstacles remain. HDR pathway disrupted by indels arises from subsequent re-editing in previously edited loci. The undesirable re-editing will recut the target loci until they are sufficiently modified to prevent further detection (Fig. 1A).

Properdin (FP) is a soluble glycoprotein with a key role in the regulation of the alternative pathway (AP) of the complement system.1 Neutrophils are the main source of FP, although monocytes, bone marrow progenitors, and T cells also synthesize this complement regulator. FP is codified by the gene CFP, located at Xp11.23–p11.3, and formed by 10 exons, of which exons 2 to 10 are translated conforming six thrombospondin type I repeats (TSR1-6).1 FP deficiency (MIM # 312060) is a rare X-linked disorder that contributes to increased susceptibility to infectious diseases, mainly caused by rare strains of Neisseria meningitidis such as serogroups W-135 and Y. Since its initial description in 1982,2 more than 100 cases have been reported worldwide demonstrating susceptibility to meningococcal infections and sepsis.1 FP deficiency can be divided into three subtypes: total deficiency (type I), partial deficiency (type II), and deficiency due to a dysfunctional protein (type III). In the present study, we investigated the members of a non-consanguineous Spanish family with a novel CFP mutation causing type I FP deficiency.

RNA binding protein is a highly conserved protein family with the RNA binding domain to bind RNA and alter RNA metabolism and function.1 Abnormal expression of RNA binding protein is common in malignant tumors and connected to migration, invasion, death, and proliferation.2 RBM43, a member of the RNA binding protein family, was verified to be considerably down-regulated in hepatocellular carcinoma (HCC), indicating poor prognosis in HCC patients. RBM43 acted as a tumor suppressor that modulated CCNB1 expression to regulate the cell cycle, providing sufficient evidence for the significance of RBM43 in HCC.3 However, it is unknown if RBM43 has an impact on the process of HCC metastasis. Here, we discovered that Rbm43-deficient mice treated with diethylnitrosamine and carbon tetrachloride developed more intrahepatic and lung metastasis tumors. Consistently, the ability of migration and invasion was dramatically promoted in RBM43-deficient HCC cells. In terms of mechanisms, RBM43 suppressed HCC cell migration through the regulation of Slug mRNA stability. Additionally, in contrast to primary HCC and non-tumorous tissues, RBM43 expression was markedly decreased in metastatic HCC. In summary, our findings implied that RBM43 played a pivotal role in regulating the metastatic process of HCC, and highlighted the RBM43-Slug axis as a prospective HCC therapeutic target.

The BCR-ABL1 fusion gene is a driver and hallmark of leukemia and a classic structural variant.1 Single nucleotide variations (SNVs) occurring in the ABL1 gene kinase domain (KD)2 and aberrant DNA methylation modifications, specifically 5-methylcytosine (5mC) of the BCR gene promoter,3 have strong clinical implications, such as tyrosine kinase inhibitor resistance, therapeutic responsiveness, and disease progression. Therefore, detecting the presence of KD region mutations and/or promoter 5mC modifications in the BCR-ABL1 fusion gene may help clinicians formulate individualized treatment regimens for patients with leukemia. Currently, the clinical detection of structural variants, SNVs, and 5mC modifications relies on a variety of independent techniques, and comprehensive techniques that can simultaneously detect all three events in one assay are urgently needed. Here, we report the direct detection of all three events using nanopore Cas9-targeted sequencing (nCATS),4 which combines Cas9-mediated target enrichment and the advantages of long-read length and direct sequencing of the Oxford Nanopore Technologies platform.

Breast cancer is the most common cancer and the leading cause of cancer death in women worldwide.1 Tumor-infiltrating myeloid cells (TIMs), key components of tumor microenvironment, are considered to be potential therapeutic targets for cancer recently,2,3 however, their heterogeneity remains insufficiently characterized in different breast cancer subtypes. A more detailed TIM transcriptional atlas across breast cancer subtypes at the single-cell level is required to better understand the underlying mechanisms influencing the prognosis of breast cancer patients. Here, we painted the landscapes of TIMs and investigated the diversity of TIM subsets across breast cancer subtypes. Particularly, we identified a novel subset, CXCL10+ conventional dendritic cells (cDCs), which are abundant in triple-negative breast cancer (TNBC) and have the ability to recruit tumor-associated macrophages (TAMs). We further discovered the developmental origins of CXCL10+ cDCs. Moreover, we identified novel subsets of macrophages and mast cells and systematically analyzed their characteristics associated with clinical outcomes in different breast cancer subtypes. Our findings provide valuable information for the identification of potential targets and biomarkers to better direct breast cancer immunotherapies.

Mutations in CYBB, encoding gp91phox subunit of NADPH oxidase in phagocytes, impair the respiratory burst of neutrophils and result in X-linked chronic granulomatous disease (CGD). While inflammatory response and NETosis are important modalities employed by neutrophils for pathogen clearance, variants in these cell functions in CGD neutrophils (CGD-PMN) could possibly explain the insufficient defense and accumulation of phagocytes in the sites of infection. To decipher the intrinsic features of CGD-PMN, neutrophils from X-linked CGD patients with CYBB mutations and age-matched healthy donors (HD-PMN) were compared. Our study found an enhanced spontaneous neutrophil extracellular trap (NET) formation with histone hypercitrullination in resting CGD-PMN. RNA sequencing (RNA-seq) analysis and qPCR validation further revealed a prominent activated type I interferon (IFN) gene signature. This suggested that in CGD patients an inefficient clearance of pathogen caused a chronic stimulation to provoke a spontaneous type I IFN-mediated neutrophil activation and NET formation which were never seen in resting neutrophils of healthy controls.

The BRAF (V600E) mutation is common in malignant melanoma patients and drives constitutive activation of the MEK/ERK signaling pathway and cancer progression.1 To date, precision therapies targeting the BRAF/MEK/ERK pathway, including BRAF inhibitor (BRAFi) monotherapy using vemurafenib (VMF) and dabrafenib and combination therapy using dabrafenib and the MEKi trametinib, have been developed and have led to great advances in malignant melanoma therapy.2 Unfortunately, many tumors initially responsive to these drugs develop resistance, which leads to patient death. Therefore, the exploration of alternative targets is important and urgently needed to improve the treatment of malignant melanoma patients. Studies have shown that abnormally high expression levels of TMEM16A (also called ANO1), a calcium-activated chloride channel (CaCC), contribute to the occurrence and proliferation of many kinds of tumor cells.3 Here, we report that TMEM16A/ANO1 is significantly up-regulated in A375 human malignant melanoma cells, which carry the BRAF(V600E) mutation. In vitro, inhibition of TMEM16A by two CaCC inhibitors (T16Ainh-A01 and CaCCinh-A01) and shRNA-mediated knockdown of TMEM16A suppressed the proliferation of A375 cells. In vivo, the growth of engrafted A375 tumors in nude mice was mitigated by TMEM16A knockdown. Mechanistically, the activation of the MEK/ERK and AKT signaling pathways was inhibited.

Depression is the leading global cause of disability, affecting about 300 million people worldwide.1,2 Depending on the number and severity of symptoms, depressive episodes can be classified as mild, moderate, and severe. Previous studies have typically focused on the treatment of severe refractory depression, while there have been few studies on the treatment of mild-to-moderate depression. However, patients with mild-to-moderate depression may develop severe episodes if not properly treated. Antidepressant dietary supplements have been suggested as an alternative treatment for depression, but the effectiveness of these supplements varies widely, and their efficacy has not been rigorously tested in clinical trials. NeuroWell contains all plant extracted ingredients and affects multiple pathways to provide a multi-targeted treatment. Preliminary results indicated that NeuroWell is safe and meets the U.S. FDA standards for dietary supplements. NeuroWell is currently in the U.S. market to relieve depression and anxiety and has exhibited remarkable clinical effects. However, no systematic clinical trials have been performed to investigate the effects of NeuroWell on mild-to-moderate depression and anxiety. Deanxit is an antidepressant widely used in the treatment of depression. It is a mixture of flupentixol and melitracine, which increases the concentration of neurotransmitters in the intracerebral synaptic space. It is mainly used in patients with mild-to-moderate anxiety and depression with major somatization symptoms.3, 4, 5 Thus, Deanxit was selected as the positive control for NeuroWell in this clinical trial.

Mucolipidosis (ML) II (OMIM 252500) and III α/β (OMIM 252600) are a group of rare lysosomal storage disorders caused by mis-sorting of lysosomal hydrolases and the subsequent accumulation of nondegraded macromolecules. These disorders manifest as multiple-systemic abnormalities throughout the body, mainly including severe dysplasia, short stature, scoliosis, joint stiffness, joint contractures, claw-hand deformities, craniofacial deformities, retinal degeneration, mental retardation, cognitive impairment, and internal organ dysfunction. ML II has an early onset, and patients usually die in early childhood. For ML III α/β, symptoms are less severe due to residual lysosomal enzymes, with a late onset and slow progression; these patients usually die in adulthood.1 Both ML II and ML III α/β are inherited in an autosomal recessive manner and caused by mutations in GNPTAB.

RE1 silencing transcription factor (REST) plays a key role in embryonic development and fetal cardiac gene reactivation.1 However, understanding of the role of REST in cardiac remodeling is very limited. A recent study has shown that cardiac-specific REST knockout increases Gαo expression, and impairs Ca2+ processing in ventricular myocytes, leading to cardiac dysfunction.2 Moreover, REST could bind to the neuron-restrictive silencer element region of the UCHL1 (ubiquitin carboxy-terminal hydrolase L1) gene promoter and regulate the expression of UCHL1.3 A recent study has reported that UCHL1 is significantly up-regulated in hypertrophic hearts, and positively regulates cardiac hypertrophy through stabilizing epidermal growth factor receptors.4 Here, we found that cardiac-specific REST knockout (REST cKO) mice showed more severe fibrosis and inflammation following pressure overload conditions. REST deficiency up-regulated UCHL1 expression, which then exacerbated cardiac hypertrophy. Whatmore, the application of UCHL1 inhibitor LDN-57444 in the REST cKO mice could alleviate fibrosis and inflammation in hypertrophic hearts via the NF-κB (nuclear factor-κB) and STAT3 (signal transducer and activator of transcription 3) signaling pathways.