Aging is driven by interconnected molecular and cellular processes (senescence, inflammation, proteostasis decline, mitochondrial dysfunction) that reduce tissue repair and organismal resilience, resulting in an increased prevalence of chronic degenerative conditions. Accordingly, the development of strategies to prevent, delay, or mitigate multiple age-related diseases is imperative. Embryonic stem cells (ESCs) and other pluripotent stem cells (PSCs) offer substantial therapeutic potential owing to their self-renewal and cellular plasticity, but their direct clinical use is limited by concerns of immunogenicity, tumorigenicity, and ethical constraints. ESC- and PSC-derived extracellular vesicles (ESC-EVs/PSC-EVs) have emerged as promising alternatives that may capture many of the regenerative and modulatory benefits of their parent cells while reducing associated risks. Cumulative evidence shows that EVs from young or stem-like cells mainly produce rejuvenating effects across tissues and molecular pathways. Here we summarize key biological features of stem cells and their EVs, outline mechanisms by which EVs impact aging hallmarks, and assess preclinical and early clinical findings on stem cell and EVs interventions for aging and age-related diseases. Careful optimization of EVs production, standardized characterization, and thorough safety and efficacy testing in diverse disease models and clinical contexts are essential to translate PSC-EVs approaches into viable therapies.

Extracellular vesicles derived from mesenchymal stem cells (MSC-EVs) have biocompatibility and low immunogenicity. They can effectively mediate intercellular communication by transmitting various information molecules, including polypeptides, lipids, and nucleic acids, thereby demonstrating unique potential in the field of drug delivery. Given their crucial roles in immune regulation and regenerative medicine, an increasing number of studies are focusing on developing MSC-EVs as drug carriers. This paper presents a systematic review of the future development directions of MSC-EVs as drug carriers, with a particular emphasis on their origins and engineering modification or pretreatment strategies. By employing engineering techniques to modify MSC-EVs, it is possible to precisely regulate key properties such as drug-loading capacity and targeting ability, thereby providing a novel approach for efficient drug delivery. Meanwhile, this paper delves into the safety evaluation of MSC-EVs in clinical applications across different diseases, comprehensively assessing their safety and efficacy as drug carriers based on clinical studies in the fields of lungs, skin, nervous system, eyes, and gastrointestinal tract, among others. Furthermore, we provide an outlook on the potential value and challenges associated with the clinical application of MSC-EVs as drug carriers in various diseases in the future, aiming to offer theoretical support and research directions for promoting MSC-EVs as safe and effective drug carriers.

BACKGROUND:Premature ovarian insufficiency (POI) causes irreversible follicular depletion and infertility, and current treatments do not reliably preserve fertility after gonadotoxic chemotherapy. Recent work suggests extracellular vesicles from embryonic stem cells (ESC-EVs) can promote tissue regeneration, but it is unknown whether ESC-EVs can prevent or reverse POI, and which cellular programs they engage to restore ovarian function. METHODS:Here we tested ESC-EVs in a cyclophosphamide (Cy)-induced POI mouse model. We evaluated ovarian function by monitoring estrous cyclicity, follicle quantification, fertility, histology, fibrosis, and cellular senescence. We isolated granulosa cells for transcriptomic profiling to define ESC-EV-driven molecular changes. RESULTS:ESC-EV treatment restored estrous cyclicity and improved fertility in Cy-induced mice, recovered follicle numbers, reduced stromal fibrosis and senescence marker expression, and activated estrogen‑responsive and gametogenesis gene programs. Granulosa cell transcriptomes showed reversal of Cy‑induced dysregulation of POI‑related genes (Figla, Nobox, Gdf9, Bmp15, Bcl2, Nfe2l2) and modulation of Nrf2 and DNA damage-response pathways. CONCLUSIONS:ESC‑EVs rejuvenate the granulosa-oocyte niche by attenuating fibrosis and senescence and reactivating folliculogenic and estrogen‑responsive programs, thereby restoring ovarian function in a chemotherapy‑induced POI model. These results identify ESC‑EVs as a promising cell‑free regenerative approach for fertility preservation and ovarian rejuvenation.

Non-small cell lung cancer (NSCLC) is a leading cause of cancer-related morbidity and mortality worldwide. Therefore, there is an urgent need to discover precise diagnostic markers and novel therapeutic strategies. A genome-wide association study (GWAS) was conducted using the Kunshan Elderly Cohort to identify genetic susceptibility factors underlying NSCLC. GWAS results identified that seven single-nucleotide polymorphisms (SNPs) were associated with NSCLC risk, including SPOCK1 (SPARC/osteonectin, CWCV, and Kazal-like domains proteoglycan (1), which emerged as a key candidate oncogene driving NSCLC progression. High expression of SPOCK1 in NSCLC tissues was associated with poor prognosis. Subsequently, using NSCLC cell lines with shRNA-mediated SPOCK1 knockdown (loss of function) and plasmid-mediated SPOCK1 overexpression (gain of function), we demonstrated that SPOCK1 significantly promotes in vitro NSCLC cell proliferation, migration, invasion, and cell cycle progression. Furthermore, in vivo mouse studies demonstrated that xenografted SPOCK1-knockdown NSCLC cells exhibited significantly reduced tumor growth and metastasis, whereas xenografted SPOCK1-overexpressing NSCLC cells enhanced tumor progression and metastasis. Mechanistically, SPOCK1 activates the JAK2/STAT3 signaling pathway by interacting with integrin alpha5 beta1 (α5β1) and subsequently upregulating SLC3A2 expression. This regulatory cascade promotes glutathione (GSH) biosynthesis and mitigates the accumulation of reactive oxygen species (ROS) and lipid peroxidation LPO), thereby suppressing ferroptosis in the NSCLC cells. Furthermore, Heat Shock Factor 1 (HSF1) is a critical upstream transcriptional regulator of SPOCK1. Notably, SPOCK1 acts as a pivotal mediator of the immunosuppressive microenvironment in the NSCLC tumors by inducing T-cell exhaustion and promoting infiltration of regulatory T cells (Tregs). In summary, SPOCK1 drives NSCLC progression by inhibiting ferroptosis and inducing an immunosuppressive microenvironment. Therefore, SPOCK1 is a promising therapeutic target in NSCLC.

Understanding how skeletal muscle influences bone formation is essential for uncovering the mechanisms of muscle-bone communication and developing therapies for osteoporosis. Here, we demonstrate that extracellular vesicles (EVs) derived from atrophic skeletal muscle (Aged-SKM-EVs) inhibit bone formation during aging. Utilizing a muscle-specific EV tracking transgenic mouse model, we found that Aged-SKM-EVs were significantly increased and taken up by osteoblasts in bone during aging. Notably, pharmacological blockade of muscle EV generation via a skeletal muscle-targeted delivery of GW4869 significantly restored osteoblast activity and alleviated bone loss in aged mice. Functional studies revealed that Aged-SKM-EVs suppressed bone formation and inhibited osteogenic differentiation both in vivo and in vitro. Mechanistically, we identified miR-125a-5p as a key cargo enriched in EVs from sarcopenic patients and aged mice. Muscle-specific overexpression of miR-125a-5p inhibited osteogenesis and exacerbated muscle atrophy and bone loss, whereas silencing miR-125a-5p in skeletal muscle effectively reversed these effects. Further investigation demonstrated that miR-125a-5p inhibits osteogenic differentiation by directly targeting Sirt7 in preosteoblasts, thereby disrupting SIRT7-mediated histone deacetylation at the Sp7 promoter and suppressing Sp7 transcription. Our findings reveal a novel endocrine pathway from muscle to bone mediated by EV-associated miRNA and highlight miR-125a-5p as a promising therapeutic target for sarcopenia-related osteoporosis.

KRAB zinc finger proteins (KRAB-ZFPs) are the most abundant transcriptional regulators and epigenetic repressors in mammals and contribute to the silencing of transposable elements (TEs) during embryonic development. However, the early effectors of the KRAB-ZFP family during embryonic development and cell state transition remain largely unexplored. Here, we identify that zinc finger protein 560 (ZFP560) is involved in the regulation of early embryonic development and the transition from totipotency in mice. ZFP560 safeguards heterochromatin structure by recruiting KAP1 to facilitate the exit of totipotency. The deficiency of ZFP560 disrupted heterochromatin formation, significantly halting the transition from totipotency to pluripotency. Similarly, the overexpression of ZFP560 promotes exit from the MERVL-positive state and suppresses expression of 2-cell (2C) transcription factors by recruiting KAP1 via its KRAB domain, thereby reducing chromatin accessibility. Taken together, these findings reveal that ZFP560 is a highly expressed 2C-specific KRAB-ZFP transcription factor and a mediator that facilitates the exit from totipotency, providing insights into the epigenetic regulation of early embryonic development.

Identifying novel therapeutic targets for pancreatic cancer (PC) is crucial for improving patient outcomes. This study identified the functions, expression, and associated mechanisms of adhesion G protein-coupled receptor G6 (ADGRG6/ GPR126) in PC. Bioinformatics analyses revealed substantial upregulation of ADGRG6 in human PC, correlating with poor survival rates and advanced tumor stages. Elevated ADGRG6 expression has been observed in human PC tissues and cell lines. Targeted depletion of ADGRG6 via the CRISPR/Cas9 knockout (KO) or lentiviral shRNA technology in established and primary PC cells (priPC-1) resulted in a substantial decrease in cell cycle progression, cell proliferation, viability, as well as reduced migratory and invasive capabilities. Conversely, ADGRG6 overexpression further enhanced the malignant behavior of PC cells. Mechanistically, ADGRG6 is crucial for Akt-mTOR cascade activation. ADGRG6 depletion markedly decreased Akt, S6, and 4E-BP1 phosphorylation. Constitutively active mutant Akt1 (S473D, caAkt1) reversed the anti-proliferative and anti-migratory effects of ADGRG65 shRNA and restored Akt-mTOR phosphorylation. Further analysis revealed that ADGRG6-driven Akt-mTOR activation is mediated by G protein inhibitory subunit 3 (Gαi3). ADGRG6 shRNA significantly inhibited subcutaneous PC xenograft growth in mice, accompanied by reduced Akt-mTOR signaling activation. In contrast, ADGRG6 overexpression promotes xenograft growth. Together, these findings establish ADGRG6 as a critical mediator of PC progression via Gαi3-dependent activation of the Akt-mTOR axis. Targeting ADGRG6 is a promising therapeutic strategy for combating PC.

Transposable elements, long considered genomic intruders, have been found to play significant and intriguing roles during early embryonic development based on the paradigm shift that has undergone in recent years. Long interspersed element-1 (LINE-1) is the predominant class of retrotransposons with autonomous retrotransposition capabilities in mammals and has emerged as a crucial element of preimplantation development. In this review, we elucidate the expression dynamics of key transposable elements throughout preimplantation development and their contribution to the regulation of developmental progression and totipotency. We also explore the critical function of LINE-1 activation and its rich functional reservoir, which is exploited by the host to provide cis-regulatory elements and functional proteins. Particular highlights of the widespread activities in preimplantation development of LINE-1 during multiple epigenetic modifications such as DNA methylation, histone methylation, ubiquitination, and RNA methylation. The silencing complex and RNA exosome also coordinate with LINE-1 across distinct developmental stages. Accordingly, the up-regulated expression of LINE-1 retrotransposons and their protein products plays a key role in various processes, including the opening of chromatin architecture, zygotic genome activation, aging, and age-related disorders. It may reflect an effect on totipotency and pluripotency of mammalian development. Underscoring its pivotal significance, the nuanced regulation of LINE-1 illuminates its indispensable role in orchestrating the precise coordination essential for the regulation of cellular pluripotency and the intricate mechanisms of zygotic genome activation.

The transcription factor c-Maf, a member of the Maf family characterized by its basic domain and b-Zip DNA-binding motif, is a pivotal regulator of immune cells development and function. It governs immune cells growth, differentiation, function, and immune responses. This review explores the mechanistic role of c-Maf and its associated signaling networks in modulating autoimmunity and inflammation. We highlight its dual function as an immune checkpoint that suppresses pathological inflammation while promoting protective immunity, underscoring its therapeutic potential in autoimmune diseases.

Here, we report a new method that employs a simple and readily available small molecule, 2-cyano-6-hydroxybenzothiazole (CBT), to selectively quench the nucleophilicity of an N-terminal Cys-containing leaving group, thereby shifting the equilibrium of peptidyl asparaginyl ligase (PAL)-mediated ligation toward the product side. This method can be applied not only to the N- and C-terminal modifications of proteins but also to efficiently mediate protein-protein ligation.

The role of 5-HT in maintaining glucose homeostasis during metabolic stress and inhibiting glucagon secretion is well documented, however, its effect on α cell identity remained unclear. In this study, we demonstrated that 5-HT suppressed the expression of α cell markers, such as Arx and Gcg, while enhancing the expression of β cell markers in mouse pancreatic α cell lines. We further found that treatment with 5-HT significantly increased the percentage of Gcg+Ins+ and Gcg+Nkx6.1+ cells in isolated human and mouse islets. Using pancreatic α cell lineage-tracing Gcgcre+; tdTomato/tdTomato mice, we observed that 5-HT treatment significantly reduced random blood glucose levels and increased tdTomato+Ins+, Gcg+Ins+ and Gcg+Nkx6.1+ cells in a high fat diet and streptozotocin (HFD + STZ) induced diabetes model. Additionally, in situ detection of 5-HT production in the human pancreas revealed a reduction of 5-HT expression in β cells of human T2D patients. These findings suggest that 5-HT treatment induces the transdifferentiation of α to β cells, potentially contributing to the recovery of β cell mass in T2D.

Osteoarthritis is a common degenerative joint disease, in which mechanical overloading disrupts subchondral bone remodeling before cartilage degeneration and the osteocytes in the subchondral bone are mainly responsible for mechanosensing. However, their functional role in the early osteoarthritis is still unclear. Here we show that mechanical stress induces osteocytes in subchondral bone to secrete extracellular vesicles that accelerate cartilage metabolic dysregulation in patients with both sexes and male mice. The miR-23b-3p in extracellular vesicles promotes cartilage catabolism and inhibits anabolism by targeting OTUD4, disrupting mitophagy in chondrocytes. Inhibiting miR-23b-3p in osteocytes or chondrocytes reduces cartilage degeneration and osteoarthritis progression in male mice. Together, our findings highlight that osteocyte-derived extracellular vesicles mediate communication with chondrocytes and suggest miR-23b-3p as a potential therapeutic target for osteoarthritis.

Mammalian development commences with the zygote, which can differentiate into both embryonic and extraembryonic tissues, a capability known as totipotency. Only the zygote and embryos around zygotic genome activation (ZGA) (two-cell embryo stage in mice and eight-cell embryo in humans) are totipotent cells. Epigenetic modifications undergo extremely extensive changes during the acquisition of totipotency and subsequent development of differentiation. However, the underlying molecular mechanisms remain elusive. Recently, the discovery of mouse two-cell embryo-like cells, human eight-cell embryo-like cells, extended pluripotent stem cells and totipotent-like stem cells with extra-embryonic developmental potential has greatly expanded our understanding of totipotency. Experiments with these in vitro models have led to insights into epigenetic changes in the reprogramming of pluri-to-totipotency, which have informed the exploration of preimplantation development. In this review, we highlight the recent findings in understanding the mechanisms of epigenetic remodeling during totipotency capture, including RNA splicing, DNA methylation, chromatin configuration, histone modifications, and nuclear organization.

The potassium channel Kv1.3 plays an important role in regulating immune cell functions in many inflammatory diseases whereas rarely in osteoarthritis (OA). Here, it is demonstrated that the Kv1.3 of macrophages is upregulated in response to LPS stimulation, as well as in human OA synovium samples than non-OA. Administration of Stichodactyla toxin (ShK), a Kv1.3 blocker, significantly inhibited cartilage degeneration and synovial inflammation in animal models of OA in vivo by inhibiting M1 macrophage polarization and reducing the production of inflammatory factors. In this study, a transgenically engineered human umbilical cord mesenchymal stem cell (UCMSC) delivery system is developed that secreted a peptide ShK, a Kv1.3 potassium blocker, into the knee articular cavity. Collectively, the results identified Kv1.3 as a potential therapeutic target for OA and demonstrated the efficacy of using ShK transgenic engineered UCMSCs as a delivery for the peptide in OA treatment.

Transposable elements (TEs) are essential components of eukaryotic genomes and subject to stringent regulatory mechanisms to avoid their potentially deleterious effects. However, numerous studies have verified the resurrection of TEs, particularly long interspersed nuclear element-1 (LINE-1), during preimplantation development, aging, cancer, and other age-related diseases. The LINE-1 family has also been implicated in several aging-related processes, including genomic instability, loss of heterochromatin, DNA methylation, and the senescence-associated secretory phenotype (SASP). Additionally, the role of the LINE-1 family in cancer development has also been substantiated. Research in this field has offered valuable insights into the functional mechanisms underlying LINE-1 activity, enhancing our understanding of aging regulation. This review provides a comprehensive summary of current findings on LINE-1 and their roles in aging and age-related diseases.

BACKGROUND AND PURPOSE:Reducing hypertensive myocardial fibrosis is the fundamental approach to preventing hypertensive ventricular remodelling. C1q/TNF-related protein-3 (CTRP3) is closely associated with hypertension. However, the role and mechanism of CTRP3 in hypertensive myocardial fibrosis are unclear. In this study, we aimed to explore the effect of CTRP3 on hypertensive myocardial fibrosis and the potential mechanism. METHODS AND RESULTS:WKY and SHR rats were employed, blood pressure, body weight, heart weight, H/BW were measured, and fibrotic-related proteins, CTRP3 and Collagen I were tested in myocardium at 12 and 20 weeks by immunohistochemical staining and Western blotting, respectively. The results showed that compared with the WKY, SBP, DBP, mean arterial pressure and heart rate (HR) were all significantly increased in SHR at 12 and 20 weeks, while heart weight and H/BW were only increased at 20 weeks. Meanwhile, CTRP3 decreased, while Collagen I increased significantly in the SHR rat myocardium at 20 weeks, which compared to the WKY. Moreover, the expression of α-SMA increased from 12 weeks, Collagen I/III and MMP2/9 increased and TIMP-2 decreased until 20 weeks. In order to explore the function and mechanism of CTRP3 in hypertensive fibrosis, Angiotensin II (Ang II) was used to induce hypertension in primary neonatal rat cardiac fibroblasts in vitro . CTRP3 significantly inhibited the Ang II induced activation of fibrotic proteins, purinergic 2X7 receptor (P2X7R)-NLRP3 inflammasome pathway. The P2X7R agonist BzATP significantly exacerbated Ang II-induced NLRP3 inflammasome activation, which was decreased by the P2X7R antagonists A43079, CTRP3 and MCC950. CONCLUSION:CTRP3 expression was decreased in the myocardium of SHR rats, and exogenous CTRP3 inhibited Ang II-induced fibrosis in cardiac fibroblasts by regulating the P2X7R-NLRP3 inflammasome pathway, suggesting that CTRP3 is a potential drug for alleviating myocardial fibrosis in hypertensive conditions.