Post-translational modifications (PTMs) of proteins represent central regulatory mechanisms within cells, enabling rapid and targeted modulation of protein functions. PTMs can directly influence protein activity, stability, or protein-protein interactions, while histone modifications also play a crucial role in gene expression regulation. To date, over 400 types of PTMs have been identified, with new types continually emerging. Among these, lysine lactylation (Kla), crotonylation (Kcr), and lysine 2-hydroxyisobutyrylation (Khib) have recently been recognized as novel PTMs with critical roles in plant growth, development, and stress responses. Unlike previous reviews that broadly summarize plant PTMs, this article provides a focused synthesis on these three newly characterized acylations, highlighting their distinct biochemical features, regulatory mechanisms, and functional relevance in plant growth, development, and stress adaptation. We summarize recent proteomic and functional studies that uncover (i) the role of histone and non-histone lactylation in metabolic reprogramming and stress resilience; (ii) the contribution of crotonylation to transcriptional regulation, enzyme activity, and abiotic stress tolerance; and (iii) the emerging function of 2-hydroxyisobutyrylation in photosynthesis, stem development, and plant pathogen interactions. Furthermore, we discuss cross-talk among Khib, Kcr, and Kac (lysine acetylation), revealing a coordinated acylation network that fine-tunes chromatin dynamics and metabolic homeostasis. By integrating findings across multiple species including rice, wheat, maize, pepper, and fungi this review proposes a comparative and mechanistic framework for understanding how these acylations bridge cellular metabolism with epigenetic and physiological regulation.

We report on the generation of human induced pluripotent stem cell (iPSC) lines, BIHi005-A-86 and BIHi005-A-87, carrying the KIT D816V mutation associated with Indolent Systemic Mastocytosis (ISM). To overcome the confounding genetic backgrounds of existing leukemic models, we introduced this gain-of-function mutation into the healthy BIHi005-A line using CRISPR/Cas9 editing. The resulting clones were validated via whole-genome sequencing (WGS) to confirm specific on-target editing and lack of predicted or disease-relevant off-target effects, while maintaining genomic stability. Together with the parental line, this resource provides an isogenic controlled platform for investigating KIT D816V-driven pathogenesis.

Oral mucosal ageing represents a fundamental reprogramming of the tissue microenvironment, a dynamic process that underlies the functional decline and heightened disease susceptibility observed in the elderly. This review synthesises current evidence to reconceptualise oral mucosal ageing as an active reprogramming of the tissue microenvironment, delineating the interplay between structural, molecular, and immunological changes, and exploring how these alterations drive functional decline and increase susceptibility to age-related oral diseases. Through a comprehensive analysis of experimental and clinical studies from human and animal models, we demonstrate that the ageing process fundamentally transforms the oral mucosa. Key findings include structural changes such as epithelial atrophy, extracellular matrix remodelling, and salivary gland degeneration, driven molecularly by genomic instability, accumulation of proinflammatory senescent cells, stem cell exhaustion, and dysregulated stress responses. These are compounded by an immunological state of 'inflammaging' and functional decline in innate and adaptive immunity, further exacerbated by shifts in the oral microbiome. Collectively, these deficits lead to impaired regeneration, diminished sensory function, and reduced salivary secretion, creating a permissive landscape for chronic oral diseases. In conclusion, oral mucosal ageing is a dynamic process of microenvironmental reprogramming driven by cellular senescence, immunosenescence, and structural decay. This actively underpins the heightened vulnerability to oral disease in the elderly, providing a mechanistic foundation for developing targeted interventions to preserve oral health in ageing populations.

Magnesium alloys have significant advantages as biodegradable orthopedic implants. However, their rapid degradation in physiological environments and lack of efficient osteointegration severely limit clinical applications. To effectively regulate the degradation rate of ZK60 magnesium alloy and enhance its bone defect regeneration ability, this study employed micro-arc oxidation (MAO) and hydrothermal treatment to create a microporous coating, followed by a layered double hydroxide (LDH) coating to seal the micropores and enhance corrosion resistance. Meanwhile, polydopamine (PDA) modification and BMP2 loading were applied to construct a functional composite coating with both corrosion resistance and osteogenic activity. The corrosion resistance and bone defect regeneration performance were systematically evaluated in vitro and in vivo. Results showed that the LDH coating exhibited a typical layered structure and effectively sealed the MAO coating micropores, forming a dense protective layer. Electrochemical results showed that compared with pristine ZK60, the corrosion current density of BMP2@PDA-LDH/ZK60 decreased from 6.18 × 10-5 A/cm² to 8.15 × 10-6 A/cm², indicating the coating significantly retarded the corrosion rate. In vitro and in vivo biological assessments demonstrated that the coating effectively promoted rat bone marrow mesenchymal stem cells (BMSCs) adhesion and bone defect regeneration; specifically, the bone mineral density (BMD) value of the BMP2@PDA-LDH/ZK60 group was 454.25 ± 31.41 mg/cm³ , which was 1.44 times higher than that of the LDH/ZK60 group.This study provides a novel strategy for biodegradable magnesium alloys with excellent corrosion resistance and osteogenic potential.

Neuroblastomas are heterogeneous pediatric tumors of the sympathetic nervous system for which treatments are still limited. Fundamental and applied approaches have been enabled thanks to the generation of patient-derived tumoroids (PDT), ex vivo 3D structures used as avatars of the original tumor. We generated neuroblastoma PDT and quantified the spatial distribution of CD133+ cancer stem cells using immunohistochemistry. We observed that those cells tend to aggregate in the PDT. In order to better understand the set of rules needed for generating such structures, we implemented a multiscale agent-based neuroblastoma tumoroid model. Model rules specify single cell's fate based on their intracellular content, which dynamically evolves according to a stochastic gene regulatory network. The state of this network can be modulated by cell-to-cell signaling through neighbor cell fate decisions and, possibly, spatial location. We first observed that in the absence of any spatial rules for inter-cellular interactions, no spatial structure emerged. The addition of simple rules (signaling by cell-to-cell contact or differential cell adhesion) only marginally improved the quantitative agreement to the experimental dataset. In sharp contrast, the addition of short-range pro-stem cell diffusive signaling among stem cells produced very realistic 3D PDT-like structures. This works highlights the power of our multiscale approach to discard too simplistic rules and to propose a minimal set of hypotheses required to reproduce qualitatively and quantitatively experimentally observed spatial structures. In the case of neuroblastoma-derived PDT, short-range spatial diffusion of stem-to-stem cell signaling proved to play a key role in successfully reconstructing the spatial structure.

There is currently no biomarker to predict the maximum morphine dose (MMD) for newborns experiencing withdrawal from chronic prenatal opioid exposure (POE). This is due, in part, to a lack of understanding about how the developing brain is altered by chronic opioid exposure and withdrawal on a molecular level. We previously developed a human induced pluripotent stem cell-derived model of POE and withdrawal to examine the impact on neural progenitor cell fates. Here, we leveraged our model to investigate the role of two microRNAs implicated in both neural stem cell differentiation and opioid signaling: miR-149 and miR-23b. Further, we asked if these microRNAs were related to the need for morphine treatment and MMD in the saliva of infants with POE. Levels of miR-149 (One-way ANOVA, F = 34.18, p < 0.0001), but not miR-23b (One-way ANOVA, p = 0.14), were significantly decreased in human neural progenitors after chronic morphine exposure (Tukey's, adj. p = 0.004), and decreased further in those that underwent withdrawal compared to vehicle exposed controls (Tukey's, adj. p < 0.0001). The relevance of miR-149 to neonates experiencing withdrawal after POE was confirmed by decreased salivary levels of miR-149 compared to levels in healthy infants 24-96 hours after birth (n = 56, 28 unexposed and 28 infants with POE) (Mann-Whitney U, p < 0.0001). Stratifying infants with POE by need for pharmacotherapy revealed a further decrease in levels of miR-149 in infants that required treatment (One-way ANOVA, p < 0.0001). In a hierarchical linear regression model utilizing infant demographic factors, addition of miR-149 levels in neonatal saliva improved performance for predicting the MMD necessary for symptom control (R = 0.673, p = 0.002). These results indicate the potential relevance of miR-149 levels in infants with prenatal opioid exposure. Validation in larger cohorts is necessary.

Human induced pluripotent stem cell (hiPSC)-derived microglia (hiMG) constitute a powerful platform to study human microglial biology in health and disease. Here, we present a protocol for microglial differentiation and efficient gene delivery to these cells using adeno-associated virus (AAV). We describe steps for differentiation, viral transduction, and validation using flow cytometry and fluorescence imaging. Finally, to demonstrate the utility of this approach, we outline procedures to record and analyze calcium activity in hiMG using the genetically encoded sensor GCaMP8s. For complete details on the use and execution of this protocol, please refer to Fruhwürth et al.1 and Heiss et al.2.

Various therapeutic approaches have been developed for lung cancer, including chemotherapy, radiation therapy, and immune checkpoint inhibitors. However, these approaches, including chimeric antigen receptor (CAR)-T cell therapy, have shown limited efficacy against solid tumors, especially in advanced disease.To enhance the therapeutic effect, we focused on the multiple effects of a new modality of cell therapy and created engineered natural killer (eNK) cells, which are gene-engineered induced pluripotent stem cell (iPSC)-derived NK cells armed with CC motif ligand 19 (CCL19), CC chemokine receptor type 2B (CCR2B), high-affinity cluster of differentiation 16 (CD16), interleukin (IL)-15, and natural killer group 2, member D (NKG2D)-DNAX-activating protein 10 (DAP10) complex. In vitro studies showed that eNK cells exhibit significant long-lasting cytotoxicity and antibody-dependent cell-mediated cytotoxicity (ADCC) against human lung cancer cell lines. In vivo, eNK cells achieved near-complete tumor regression in orthotopic and subcutaneous cell line-derived xenograft (CDX) models. In contrast, in patient-derived xenograft (PDX) models, eNK cells demonstrated modest tumor growth inhibition (28% reduction) as monotherapy and significantly enhanced efficacy (53% inhibition) in combination with cetuximab via antibody-dependent cellular cytotoxicity. In treated PDX tumors, human CD45-positive cells were detected within the tumor parenchyma, supporting intratumoral presence of administered human cells.These findings support the potential contribution of the five-gene modifications in enhancing tumor homing, persistence, and cytotoxicity in solid tumor treatment. This study underscores the potential of eNK cells as a novel, "off-the-shelf" allogeneic therapy for refractory solid tumors, including lung cancer.

Acute myeloid leukemia (AML) is an aggressive disease with suboptimal overall survival, especially in relapsed/refractory patients. The primary goal of salvage therapy in this patient is to achieve optimal disease control, thereby allowing the transition to hematopoietic stem cell transplantation (HSCT), which remains the only curative option for a subset of these patients. Allogeneic KIR ligand-mismatched CD56+ NK/NKT-like cells have demonstrated antileukemic activity and represent a promising platform for the development of novel cellular therapies.

Immunotherapy has revolutionized cancer treatment, offering durable responses across various malignancies. However, blood cancers present unique challenges due to significant heterogeneity, an immunosuppressive tumor microenvironment (TME), and potent mechanisms of immune evasion. This review synthesizes the current landscape of immune checkpoint blockade (ICB), evaluating both established agents and emerging targets (e.g., LAG-3, TIM-3, CD47) in hematologic malignancies. We critically examine the role of cancer stem cells (CSCs) in driving therapeutic resistance and recurrence, highlighting how aberrant signaling pathways (Wnt, Notch, m6A) fuel immune avoidance. Furthermore, we assess the transformative potential of cellular immunotherapies, specifically CAR-T and CAR-NK cells, proposing strategies to enhance their persistence and safety. By integrating insights from recent preclinical and clinical studies, this report provides a roadmap for developing rational, multi-pronged combination strategies that disrupt the “resistance ecosystem” of blood cancers, paving the way for personalized curative regimens.

The most robust functional synaptic refinement in the dorsal lateral geniculate nucleus (dLGN) occurs around the time of eye opening. This study aimed to identify the major glial phagocyte responsible for eliminating excess synapses during this critical window, and to elucidate the molecular mechanisms underlying its regulation.

Hedgehog (Hh) signaling is essential for embryonic development and tissue homeostasis in organisms. However, Caenorhabditis elegans lacks canonical Hh signaling due to the absence of key components, such as smoothened (SMO) and obvious Hh ligands. Despite this, C. elegans retains Patched homologs, ptc-1 and ptc-3, which have specialized independent functions. Although ptc-1 is predominantly expressed in the germ line and ptc-3 in somatic tissues, we demonstrate that both genes are required to maintain germ cell populations and proper actin cytoskeletal architecture in the progenitor zone of the germ line. Disruption of actin-encoding genes impairs germ cell S-phase and reduces the number of cells in the progenitor zone, indicating that cytoskeletal integrity is critical for maintaining the germ line. Furthermore, defects observed upon loss of Patched function are linked to disruptions in cholesterol metabolism. We show that the phenotypes observed in the gonads due to dietary cholesterol changes can be modulated through Patched receptors. Together, our findings reveal a role for Patched receptors in regulating gonad architecture and germ cell development through cholesterol-sensitive functions, offering insights into how metabolic cues influence the organization of complex tissues.

Despite advances in the development of mesenchymal stem cells (MSCs), the ultimate benefits of MSCs against current cell-based therapies are still limited. This study aimed to assess the safety, feasibility, and efficacy of Cytopeutics® umbilical cord-derived MSCs (Chondrocell-EX) in patients with knee cartilage injury.

Musashi RNA-binding proteins are important post-transcriptional regulators of stem cell homeostasis and are known to be involved in viral infections. However, their role in SARS-CoV-2 infection remains largely unknown. Using computational studies, in vivo RNA immunoprecipitation, and biochemical assays, here, we establish that Musashi 1 (Msi1) interacts with viral genomic RNA through direct binding to the SARS-CoV-2 3'UTR. Importantly, binding of Msi1 to the viral 3'UTR results in translational repression that could be mediated by inhibition of poly(A) binding protein. Conversely, Msi1 knockout promotes robust viral replication and increased viral protein expression. Using 2D cell cultures, stem cells, and 3D organoids, we show that depletion of Msi1 in intestinal cells augments infection. This finding explains why the human intestine serves as a reservoir for SARS-CoV-2, in which differentiated enterocytes with negligible Msi1 levels are particularly affected. Contrarily, stem cells, which are enriched for Msi1 expression, are known to be less permissive to SARS-CoV-2 infection despite expressing the entry receptors. Our findings show how translational repression of SARS-CoV-2 by stem cell RNA-binding proteins, such as Msi1, could help evade infection.

Anticancer therapeutics have evolved from small-molecule drugs to monoclonal antibodies and, more recently, to cell and gene therapies (CGTs). This progress has been driven by the pursuit of greater drug specificity, potency, and safety. Recent breakthroughs in chimeric antigen receptor T-cell (CAR-T) therapy for B-cell hematologic malignancies have accelerated the development of CAR-X CGTs, including CAR-T, CAR-natural killer (CAR-NK), and CAR-macrophage approaches. In this article, we compare candidate CAR-X platforms for T-cell-related diseases, such as T-cell hematologic malignancies, and propose the most suitable modality. Therefore, we analyzed the advantages and limitations of CAR-T, CAR-NK, and CAR-macrophage therapies. In T-cell-related diseases, CAR-T therapy faces multiple challenges, including fratricide, T-cell aplasia, and substantial barriers to the generation of allogeneic CAR-T products. CAR-macrophage therapies, in contrast, are constrained by relatively limited efficacy. In contrast, CAR-NK cells do not cause fratricide or T-cell aplasia and can be manufactured efficiently as allogeneic, "off-the-shelf" products. Collectively, to sustain and extend the advances in CGT initiated by CAR-T cells in B-cell malignancies, prioritizing CAR-NK research infrastructure for T-cell-related diseases represents a rational and strategic approach.

Transcription factor acetylation is a critical yet often overlooked regulator of cell fate. Although traditionally studied in the context of histone modifications, many acetyltransferases and deacetylases also modify transcription factors directly, thereby controlling lineage-specific transcriptional programs. At the molecular level, acetylation fine-tunes transcription factor activity by modulating DNA binding, protein stability, cofactor interactions, and nucleo-cytoplasmic trafficking. These molecular effects frequently intersect with other post-translational modifications, establishing acetylation as a versatile molecular switch of transcriptional output. These molecular effects scale into cellular outcomes that determine identity and plasticity. In pluripotent stem cells, defined acetylation events on core regulators stabilize the pluripotency network and prime lineage-specific enhancers. In hematopoiesis, transcription factor acetylation modulates transitions from stem and progenitor states to committed lineages, while in myogenesis, it governs progenitor differentiation and regenerative capacity. Importantly, differential acetylation of distinct lysine residues can yield context-dependent outcomes, underscoring the precision and adaptability of this modification in controlling cell identity. Recognizing transcription factor acetylation as a central axis of epigenetic regulation reframes our understanding of lineage specification and cellular plasticity. Beyond developmental biology, it provides a mechanistic rationale for therapeutic strategies that target acetylation dynamics, not only altering chromatin states but also reprogramming transcription factor function. This review synthesizes current knowledge of transcription factor acetylation in hematopoietic and myogenic contexts, highlighting its significance as a bridge between molecular mechanisms and cellular identity, and as a promising target in disease intervention.

Male infertility is a rising problem globally with male factors contributing up to 50% of all couple infertility cases. The sperm quality decline raises serious concerns regarding future population sustainability and male reproductive health. Adipose-derived mesenchymal stem cell (AdMSC) secretome, defined as a cell-free product comprising paracrine factors secreted by these cells, has emerged as a promising cell-free regenerative therapy for testicular injury, offering advantages of accessibility and therapeutic potential. This systematic review aimed to evaluate the effectiveness of AdMSC secretome in chemically induced testicular injury models.

Human cytomegalovirus (HCMV) reactivation is a common and significant complication after allogeneic haematopoietic stem cell transplantation (allo-HSCT), causing potentially life-threatening disease. Antiviral therapy approaches for HCMV are often limited by toxicities and the risk of developing antiviral drug resistance. This review article (Part 2) provides an overview of current antiviral pharmacotherapies for HCMV and the application of virus-specific adoptive T cell therapies for the prevention or treatment of HCMV reactivation in allo-HSCT recipients. The number of available antiviral drugs for HCMV is expanding, and letermovir primary prophylaxis is increasingly being adopted due to its favourable safety profile. Treatment resistant/refractory infections, end-organ disease, and late HCMV reactivations after antiviral therapy withdrawal continue to pose challenges. Adoptive HCMV-specific T cell therapies are a promising strategy for promoting immune-mediated control of HCMV reactivation in allo-HSCT recipients. The administration of HCMV-specific T cell products, generated through ex vivo expansion of donor-derived or partially HLA matched, third party HCMV-specific T cells, have demonstrated efficacy in combatting clinically significant HCMV infection in clinical trials. Adoptive HCMV-specific T cell therapies represent a powerful alternative approach for managing drug resistant HCMV infections in allo-HSCT recipients and reducing the reliance on antiviral pharmacotherapies.

Deuterium is unevenly distributed in natural waters, while the same applies to the content of deuterium in ice on Mars. Moreover, changes in the deuterium content of drinking water are known to affect the bodies of mammals. Thus, since plans are in place to send people to Mars in the coming years, understanding the effects of water with a Martian isotopic composition is necessary. Therefore, this study aimed to evaluate the impact of water with an increased deuterium content of 1200 ppm on the dynamics of indicators in the body of mammals.

Human neurons derived from stem cells show increased structural complexity and stronger synaptic connections after exposure to psilocin, the active metabolite of the psychedelic psilocybin.