Cinnamoylquinic acid derivatives, namely, 3,4,5-tri-caffeoylquinic acid (TCQA) and its structurally modified analogue, 3,4,5-tri-feruloylquinic acid (TFQA), have demonstrated promising neuroprotective and pro-neurogenic activities. However, it remains unclear how the substitution of caffeoyl groups with feruloyl groups influences their molecular activity. Therefore, we performed an integrated multi-omics analysis, combining post hoc transcriptomic profiling of TCQA- and TFQA-treated neural stem cells (NSCs) isolated from 6 to 8-week-old adult male ICR mouse brains with targeted metabolomic analysis in SH-SY5Y neuroblastoma cells. We applied an interaction-only nested linear model, which revealed gene sets uniquely responsive to TFQA compared with TCQA. TFQA elicited broader pathway enrichment involving cell signaling, immune modulation, and metabolic regulation, whereas TCQA produced narrower transcriptional shifts. Nested comparative modeling identified 709 genes differentially regulated between TFQA and TCQA, with TFQA uniquely enriching pathways such as glutamatergic synapse, long-term potentiation, TRP channel regulation, and apelin signaling. Downregulation of cell cycle-related pathways and cytokine secretion suggested that TFQA promotes neuronal differentiation while reducing inflammatory activity. Metabolomic profiling further demonstrated TFQA-specific alterations in central energy metabolism, redox balance, amino acid utilization, and neurotransmitter-related metabolites. TCQA showed carbohydrate metabolism and glycan turnover. Integrated pathway analysis and gene-metabolite network modeling revealed coordinated TFQA-associated molecular reprogramming, with metabolites such as ATP, GTP, glutamate, GABA, and L-arginine emerging as central hubs linking transcriptomic and metabolic responses. Collectively, this study provides a multi-layered, systems-level comparison of TCQA and TFQA across a cross-species, cross-cell-type framework, revealing both conserved and divergent molecular responses and offering insights into the structure-activity principles shaped by feruloyl substitution.

Friedreich's ataxia (FRDA) is a multisystem, autosomal recessive disease caused by biallelic expansion of GAA repeats in intron 1 of the frataxin gene (FXN). While ∼96% of FRDA patients carry expanded GAA repeats on both FXN alleles, ∼4% are compound heterozygous with expanded GAA repeats on one allele and another mutation on the second allele. We generated induced pluripotent stem cells from blood lymphocytes from a FRDA patient carrying the FXN c.165 + 5G > C point mutation, which interferes with canonical splicing of intron 1 of the FXN gene. These cells allow for development of therapeutic approaches that target splicing defect in FRDA.

CD8αα+TCRαβ+ intraepithelial lymphocytes (IELs) play crucial roles in maintaining intestinal homeostasis and host protection. However, the functional regulation of these cells remains unclear. Here, we have discovered and characterized two distinct developmental stages within intestinal CD8αα+ αβ IELs: a stem-like, defined by BCL6, TCF1, and CD160 expression, and an effector-like, with granzyme B expression and Prdm1 transcription. The differentiation from stem-like to effector-like CD8αα+ αβ IELs is promoted by T cell receptor (TCR) and IL-12 signaling and is controlled by the opposing actions of BCL6 and BLIMP1. Loss of BCL6 promotes the development of effector-like CD8αα+ αβ IELs, leading to increased effector molecule expression and heightened inflammation under dextran sodium sulfate (DSS)-induced colitis. In contrast, BLIMP1 deficiency perturbs the effector differentiation and reduces the susceptibility to gut inflammation. Our study thus reveals a critical antagonistic function between BCL6 and BLIMP1 in governing the fate decisions of CD8αα+ αβ IEL subsets.

HC-HA/PTX3 (a complex formed by high molecular weight hyaluronan covalently linked to heavy chain 1 of inter-α-trypsin inhibitor and tightly bound to pentraxin 3) is a unique extracellular matrix from human amniotic membrane that exerts an anti-scarring action and reprograms human corneal fibroblasts (HCF) and myofibroblasts to corneal stromal keratocytes in the absence of transforming growth factor β1 (TGFβ1) by downregulating canonical Smad-mediated signaling and upregulating bone morphogenetic protein (BMP) signaling. It remains unclear whether HC-HA/PTX3 can further reprogram HCF into neural crest (NC) progenitors in the presence of TGFβ1.

Hyperglycemia, a hallmark of diabetes mellitus, is a metabolic condition that highly affects the nervous system. While evidence from epidemiological and animal studies links diabetes to dopaminergic dysfunction and an increased risk of Parkinson's disease, the underlying mechanisms remain unclear. Here, we examined the effects of high glucose on human iPSC-derived dopaminergic neurons and glial cells to better understand the pathogenic alterations that lead to neurotoxicity. Previous implication of neurotrophins in the neurological manifestations of diabetes prompted us to focus on the role of p75NTR neurotrophin receptor (p75NTR) in dopaminergic neurodegeneration under hyperglycemic conditions.

Cancer stem cells are critically involved in the initiation and progression of hepatocellular carcinoma (HCC). Understanding their biological properties and underlying mechanisms is essential for the development of effective targeted therapies.

Current therapeutic models for ischemic stroke (IS) are shifting from a narrow focus on neuroprotection to a broader concept of cytoprotection. This new paradigm emphasizes rescuing damaged brain cells and maintaining their structural and functional integrity through organelle transfer between healthy and damaged cells. Mounting evidence have supported that intracellular mitochondrial transfer is an intrinsic response to IS, playing a critical role in mitigating neural damage. Consequently, mitochondrial transplantation from stem cell is emerging as a therapeutic avenue for IS.

Post-acute sequelae of COVID-19 (PASC) affect millions of people worldwide and are increasingly recognized as a disorder of failed innate immune resolution rather than a persistent viral infection. Emerging evidence shows that residual SARS-CoV-2 antigens, host-derived alarmins, reactivated latent viruses, and mucosal microbiome-derived products from oral-nasopharyngeal and gut reservoirs sustain the chronic activation of pattern-recognition receptors, inflammasomes, and complement pathways. In parallel, deficits in specialized pro-resolving mediators, impaired efferocytosis, and persistent tissue injury prevent physiological termination of inflammation. These unresolved cues drive long-lasting epigenetic and metabolic reprogramming of hematopoietic stem cells and myeloid lineages, creating maladaptive trained immunity states characterized by hyper-responsiveness or exhaustion of these cells. Thromboinflammatory processes, including aberrant NETosis and sustained interface signalingling, further reinforce self-perpetuating inflammatory circuits. Together, these pathways give rise to reproducible molecular endotypes, including thromboinflammatory, interferon-driven, and neuroinflammatory phenotypes, which explain clinical heterogeneity. Framing PASC as a disorder of impaired immune resolution within a mucosal microbial viral context provides a unifying mechanistic scaffold for biomarker identification and host-directed therapies. This review proposes that restoring active resolution programs, rebalancing metabolic-epigenetic networks, and dismantling pathogenic innate feedback loops are promising strategies for reversing the chronic immune imprint of PASC.

Human induced pluripotent stem cell (hiPSC)-derived neural models combined with microelectrode array (MEA)-based readouts are increasingly used in next-generation neurotoxicity assessment. However, neural induction of hiPSCs into multipotent neural progenitor cells (hiNPCs) remains highly variable, and current quality control (QC) efforts focus largely on the pluripotent starting material. As a result, failed neural inductions are often recognized only after weeks of differentiation during functional network analysis, causing substantial resource loss. Here, we present a tiered QC framework spanning the entire workflow from hiPSC banking, neural induction into proliferative hiNPCs, the differentiation of hiNPCs into 3D neuron-glia BrainSpheres, and their subsequent organization into functional 2D networks on MEAs, with emphasis on early detection of induction failures. Using eleven independent dual-SMAD inductions derived from a single hiPSC line, we show that hiPSCore, a machine-learning-based classifier for early cell-fate decisions, reliably distinguishes successful from unsuccessful neuroectodermal inductions at early stages (days 6-12). Successful induction depends on timely neuroectodermal commitment, reflected in hiPSCore trajectories and PAX6 expression, as well as preserved hiNPC proliferation. Inductions passing these early QC checkpoints generated BrainSphere-derived networks with reproducible MEA maturation trajectories, balanced neurotransmitter-responsive unit distributions, and conserved subtype-specific pharmacological responses across GABAergic, glutamatergic, dopaminergic, and serotonergic modalities. In contrast, signaling pathway-related markers associated with dual SMAD inhibition (BMP, TGFβ, and MAPK) were not predictive of downstream QC performance. Together, these findings demonstrate that early fate-level QC is predictive of the functional performance of hiPSC-derived neural networks. Our adaptable QC framework allows early termination of failed inductions, reduces resource burden, and strengthens confidence in hiPSC-based neural network assays for neurotoxicity testing.

Propranolol is currently the first-line therapy for infantile hemangioma (IH); however, its use is limited by notable adverse effects. The non-β-blocker enantiomer of propranolol, R-propranolol, represents a promising alternative for IH treatment. In this study, the efficacy of R-propranolol and its underlying mechanism in IH involution were investigated. The protein kinase RNA-like endoplasmic reticulum kinase (PERK) signaling pathway was required for the adipogenic potential of hemangioma stem cells (HemSCs), whereas prolonged PERK activation led to apoptosis. R-propranolol and racemic propranolol induce comparable HemSC adipogenesis. R-propranolol showed a stronger ability to disrupt protein homeostasis in HemSCs than racemic propranolol, as indicated by increased protein misfolding and enhanced activation of the PERK signaling pathway. Notably, R-propranolol induced HemSC apoptosis more effectively than racemic propranolol. This effect was attenuated by the PERK inhibitor GSK2606414. Immunostaining of clinical specimens further showed that PERK expression was higher in involuting IH samples than in proliferating IH samples. Together, these findings demonstrate that R-propranolol promotes adipogenesis and subsequent apoptosis of HemSCs through activation of the PERK signaling pathway.

Some mammalian tissues can replace lost cells within one lineage, but organ-level regeneration-restoring diverse cell types across lineages-remains rare. Here, we show that late embryonic full-thickness skin injuries heal by regenerating epithelial, mesenchymal, neuronal, and vascular tissues with proper connectivity. However, this ability is lost soon after birth, resulting in failure to restore most cell types and hyperinnervation within the wound bed. Single-cell sequencing identified a postnatal wound-specific fibroblast (PWF) population absent after embryonic wounding. Through an in vivo screen, we discovered that three PWF-enriched genes-Timp1, Cxcl12, and Ccl7-inhibit organ-level regeneration and cause hyperinnervation when overexpressed in embryonic wounds. Reducing hyperinnervation in postnatal wounds through the depletion of Cxcl12 in fibroblasts or nerve ablation enables regeneration of diverse lineages after injury. Our study identifies mechanisms that transition an organ from regenerative to non-regenerative, discovers fibroblast-driven hyperinnervation as a key barrier, and demonstrates that removing this barrier unlocks organ-level regeneration.

T cell-based immunotherapies represent one of the most promising areas in cancer treatment, offering the dual advantages of precise molecular targeting and durable therapeutic effects. Notch signaling platforms facilitate the ex vivo generation of T lineage cells from hematopoietic stem and progenitor cells sourced from bone marrow, peripheral blood, cord blood, or pluripotent stem cells. These platforms can be tailored to produce either T cell progenitors or fully differentiated T cells. The use of T cell precursors in adoptive cell therapy enables off-the-shelf immunotherapy, as even fully HLA-mismatched allogeneic T cell precursors undergo thymic selection and are tolerized during maturation in the recipient's thymus. Furthermore, T lineage cells produced via Notch-based systems can be engineered to express chimeric antigen receptors, resulting in potent, tumor-specific therapeutic products. Notch-based in vitro T cell development platforms range from stromal cell-dependent co-cultures to clinically suitable, stromal cell-free systems using immobilized Notch ligands, ligand-coated microbeads or soluble Notch agonists. Enhancements such as incorporating VCAM-1 alongside Notch signaling have been introduced to improve the efficiency of T cell production. This review highlights the various Notch signaling platforms that have contributed - and continue to contribute - to the advancement of adoptive T cell immunotherapy.

Obesity is the most common chronic metabolic disorder, that exacerbates low-grade systemic inflammation and insulin resistance. Insulin resistance promoted by chronic caloric excess, results in systemic inflammation, ectopic lipid deposition and adipose hypertrophy. Type 2 diabetes mellitus (T2DM) is fuelled by persistent adiposity, and approximately half of T2DM patients develop diabetic neuropathies. Diabetic neuropathy causes severe morbidity, yet current treatments are only based on symptomatic and not able to regenerate damaged nerve. Due to these limitations, regenerative cell-free therapies, such as adipose stem cell derived-exosomes (ADSC-EXOs) has gained more attention. ADSC-EXOs have emerged as a promising platform for nerve regeneration. ADSC-EXOs carry a rich cargo of regulatory miRNAs reflective of the parent cell phenotype as well as angiogenic and neurotrophic factors. ADSC-EXOs avoid immune rejection and tumorigenicity. Additionally, engineered ADSC-EXOs loaded with therapeutic miRNAs such as neurotrophin-3 significantly improves axonal regrowth and remyelination. ADSC-EXOs enhance insulin signalling and glucose metabolism via modulating phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) pathway and exosomal miRNAs (e.g., miR-155) that enhances neuronal insulin response. They also downregulate neuroinflammation by inhibiting nuclear factor kappa-light-chain-enhancer of activated B cells/ mitogen-activated protein kinase (NF-κB/MAPK) signalling and polarizing macrophages/microglia toward anti-inflammatory phenotypes. ADSC-EXOs restore mitochondrial function and ATP production in damaged neurons. ADSC-EXOs also encourage Schwann cells to release neurotrophic factors, divide and survive to help in nerve regeneration. Pro-regenerative environment was created through these combined effects in diabetic neuropathy that encourages axonal regeneration and the functional repair of injured neurons. ADSC-EXOs show promise as a potentially ground-breaking medicinal development, particularly for diabetic neuropathy.

Retinitis pigmentosa (RP) is a group of inherited retinal degenerative disorders characterized by progressive visual dysfunction. In this study, we generated an induced pluripotent stem cell (iPSC) line, CSUASOi014-A, from peripheral blood cells of a 40-year-old Chinese female patient clinically diagnosed with RP. Peripheral blood mononuclear cells were isolated and reprogrammed using an integration-free Sendai virus system. The established iPSC line harbors a heterozygous PRPF31 c.528-39_531del mutation. CSUASOi014-A provides a patient-specific cellular resource for studying the pathogenic mechanisms underlying PRPF31-associated RP.

Phosphatase and tensin homolog (PTEN) is a critical regulator of cell proliferation, differentiation, and inflammatory balance. However, its downstream proteomic effects in periodontal ligament stem cells (PDLSCs) remain poorly understood. This study aimed to elucidate the proteomic alterations induced by PTEN inhibition and identify potential molecular pathways underlying periodontal regeneration.

Neural organoids can exhibit variability in both tissue shape and tissue identity. Here, we present a pipeline for rapid, protocol-agnostic quality control screening of brain organoids based on their overall gross morphology. We describe a semi-automated image analysis of organoid size, shape, and texture from 2D bright-field imaging. We provide a reference dataset of brain organoids with complex morphology. We show how to integrate input and reference organoids and perform the unbiased sample selection by k-means clustering. For complete details on the use and execution of this protocol, please refer to Chiaradia et al.1.

Mutations in the presenilin (PS/PSEN) genes cause early-onset familial Alzheimer's disease (AD) by enhancing cerebral accumulation of amyloid-β (Aβ) peptides and microtubule-associated protein tau (MAPT). How PS mutations affect Aβ generation is well characterized, but the precise cellular mechanisms by which PS dysfunction drives neuronal tau pathology are not fully understood. Here, we investigated the mechanisms linking PS/γ-secretase-dependent tau pathology and autophagy/proteasome by employing pathological, imaging and molecular approaches in human brains, fibroblasts and induced pluripotent stem cells (iPSC)-derived neurons from PSEN1-linked familial AD carriers, and in a novel neuronal PS-deficient tauopathy transgenic mouse. We found enhanced levels and colocalization of pathological phosphorylated tau (pTau) and ubiquitin factor p62 in the hippocampus of dementia patients with familial AD-linked PSEN1 mutations, corticobasal degeneration and Pick's disease, suggesting disrupted proteasomal degradation in tauopathies. Human primary fibroblasts from PSEN1 G206D and/or L286P carriers showed elevated LC3-I and autolysosomes indicating autophagy flux alterations. Human iPSC-derived neurons harboring the familial-AD linked PSEN1 G206D mutation showed increased aggregated tau and reduced secreted tau, whereas pharmacological proteasome inhibition reduced significantly total and pTau (Ser396/404) while increasing its release. Consistently, proteasomal inhibition decreased intracellular tau and pTau and promoted tau release in human tau-expressing neurons through a mechanism that partially depends on PS. In the hippocampus of neuronal PS-deficient mice, Akt activation and GSK3β inhibition were associated with elevated levels of phosphorylated and aggregated tau and the ubiquitin-binding protein p62. In conclusion, PS function is required for autophagy/proteasome-mediated tau elimination in neurons, whereas that FAD-linked PSEN1 mutations cause progressive tau pathology by disrupting the proteasome and autophagy/lysosomal pathways.

Periodontitis can impair the osteogenic function of periodontal ligament stem cells (PDLSCs), thereby compromising their capacity for periodontal tissue regeneration. In this study, we explored the impact of a synthetic small molecule, DS96432529 (DS), on the osteogenic differentiation potential of PDLSCs and its underlying mechanism.

Mesenchymal stromal cells (MSCs) are widely used in regenerative medicine, but their clinical utility is limited by replicative senescence. Strategies that reverse aging while maintaining MSC identity are urgently needed.