The xenotransplantation of human cells into porcine hosts holds immense potential in the fields of regenerative medicine and organ transplantation. However, the low survival rate of human-derived cells within porcine remains a critical bottleneck constraining the application of xenotransplantation. Whether porcine cells exert negative effect on human cell growth is not studied. Here, we established an in vitro direct co-culture model of human and porcine mesenchymal stem cells (hMSCs and pMSCs) to investigate the competitive relationship between human and porcine-derived cells. The results demonstrated that the proliferation capability of hMSCs in the co-culture system was significantly suppressed compared to those cultured in isolation. Moreover, an increasing number of pMSCs exhibited enhanced inhibition of hMSC proliferation. Notably, results from transwell assays and treatment with porcine-conditioned medium indicated that the inhibition of hMSCs by pMSCs was not mediated through soluble cytokines. To elucidate the underlying molecular mechanisms, RNA sequencing analysis was performed and the result revealed that direct co-culture significantly downregulated the expression of proliferation-related genes in hMSCs, including CYP1B1, SLC7A11, TFAP2C, and PSAT1. Concurrently, the co-culture paradigm disrupted endoplasmic reticulum function and multiple amino acid transport processes within hMSCs, while activating the NF-κB signaling pathway, thereby achieving negative regulation of hMSC proliferation. Collectively, our primary study characterized the competitive interactions between hMSCs and pMSCs and uncovered possible underlying mechanisms which provided new experimental foundations for improving human cell survival in porcine hosts to advance xenotransplantation application.

A substantial association has been established between diabetes and an elevated risk of lung cancer. This study aimed to elucidate the prognosis and characterize alterations in the immune microenvironment linked to the protein kinase C substrate 80K-H (PRKCSH) gene in the context of diabetic lung cancer.

[This retracts the article DOI: 10.3892/etm.2012.707.].

Hybridization chain reaction (HCR), which typically consists of two hairpins for signal amplification, has emerged as a versatile tool in bioanalytical applications. Here, a novel HCR nanowire based on a single DNA hairpin structure is reported. The hairpin stem is rationally engineered with a palindromic sequence, which enables a self-hybridization chain reaction (SHCR) upon the introduction of the initiator DNA strand. Compared to the conventional two hairpin-based HCR nanowire, the single DNA hairpin-based SHCR nanowire achieves nearly a two-fold improvement in the signal-to-noise ratio and exhibits better selectivity for single-base mismatch. By integrating the initiator DNA strand with adenosine triphosphate (ATP) aptamer, the single DNA hairpin-based nanowire has been applied for sensitive ATP detection, capable of monitoring ATP both in living cells and that released from dead cancer cells post-radiotherapy. The SHCR nanowire we proposed here has significantly simplified the sequence design of HCR and holds promise as a potential alternative to the conventional HCR nanowire.

Breast cancer remains the most commonly diagnosed cancer among women worldwide, and multiple studies now link its development and progression to disturbances in metabolic and immune regulation. Among these factors, the gut microbiota is increasingly recognized as a modulator of host physiology through its metabolism of dietary tryptophan (Trp). Here we focus on the current understanding of the microbial metabolism of Trp, which primarily generates bioactive metabolites through the kynurenine (Kyn) pathway and the indole pathway. These metabolites can serve as endogenous ligands, activating the aryl hydrocarbon receptor (AhR) signaling pathway. They can also promote tumor stem cell characteristics, epithelial-mesenchymal transition (EMT), and metastasis via serotonin receptors (such as HTR1B/1D, HTR2B). The activation of such pathways contributes to the remodeling of the tumor immune microenvironment, alters the functions of immune cells, and directly influences the proliferation, invasion, and metastatic behavior of breast cancer cells. By integrating findings from preclinical and clinical studies, this review organizes current evidence around the "gut microbiota-Trp metabolism-breast cancer" axis and discusses clinical implications and current limitations. Targeting this metabolic network may provide new opportunities for breast cancer prevention and therapeutic intervention.

Leukemia is a malignant clonal disease of hematopoietic stem cells. Currently, primary treatments include chemotherapy, targeted drugs, hematopoietic stem cell transplantation, and immunotherapy. Although advances in treatment have improved survival for leukemia patients, treatment failure still occurs in some individuals for various reasons. This necessitates the discovery of new pathogenic mechanisms and the exploration of novel biological targets. Tumor Necrosis Factor Receptor-Associated Factors (TRAFs) are a family of cytoplasmic adapter proteins, typically comprising seven members. The TRAF protein family is widely involved in cell proliferation, differentiation, survival, and apoptosis, and also regulates immune and inflammatory responses. TRAFs perform dual roles in a broad range of biological activities-as adapter proteins and as E3 ubiquitin ligases-both essential for activating receptor-mediated signaling. In recent years, growing evidence has highlighted the significant role of TRAFs in leukemia, linking them to leukemic stem cell activity, drug resistance, apoptosis, and autophagy. This review introduces the functions and characteristics of TRAFs and summarizes research progress on their involvement in leukemia, underscoring their potential as novel therapeutic targets for the disease.

The study aimed to compare the incidence and course of febrile neutropenia (FN) and factors affecting mortality in hematologic patients undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT) with either fresh or cryopreserved grafts.

Myelodysplastic syndrome (MDS) is a malignant clonal disorder originating from hematopoietic stem and progenitor cells and is characterized by ineffective hematopoiesis and a high propensity for transformation into acute leukemia. Research has indicated that the pathogenesis of MDS is closely linked to genetic mutations and that its progression may encompass multiple stages, including cytogenetic and molecular alterations, leading to the acquisition of oncogenic mutations. The widespread application of next-generation sequencing (NGS) technologies, including whole-genome sequencing, whole-exome sequencing, and RNA sequencing, has significantly improved our understanding of the genetic alterations and transcriptomic modifications underlying MDS. These technologies not only deepen our understanding of molecular mechanisms but also facilitate the identification of potential therapeutic targets and prognostic biomarkers. Consequently, the 2022 World Health Organization Classification and the International Consensus Classification have incorporated molecular features into the MDS classification system, and the Molecular International Prognostic Scoring System (IPSS-M) was introduced. This review aims to summarize the role of NGS in the precise diagnosis, classification, risk assessment, treatment selection, and evaluation of therapeutic effectiveness in MDS, with implications for advancing precision medicine in hematological malignancies.

Gamma delta (γδ) T cells are defined by their unique ability to recognize a limited repertoire of non-peptide, non-MHC-associated antigens on transformed and pathogen-infected cells. In addition to their inability to mediate GvHD, γδ T cells exhibit properties distinct from other lymphocyte subsets, prompting significant interest in their development as an off-the-shelf cellular immunotherapeutic. However, their low abundance in circulation, heterogeneity, limited methods for ex vivo expansion, and under-developed methodologies for genetic modification have hindered basic study and clinical application of γδ T cells. Here, we implement a feeder-free, scalable approach for ex vivo manufacture of polyclonal, non-virally modified, gene edited chimeric antigen receptor (CAR)-γδ T cells for therapeutic application. Engineered CAR-γδ T cells demonstrate robust functionality in vitro and in vivo. Longitudinal in vivo pharmacokinetic profiling of adoptively transferred polyclonal CAR-γδ T cells uncover subset-specific responses to IL-15 cytokine armoring and multiplex base editing. Our results present a robust platform for genetic modification of polyclonal CAR-γδ T cells and present unique opportunities to further define synergy and the contribution of discrete, engineered CAR-γδ T cell subsets to therapeutic efficacy in vivo.

Analyze the predictive value of body mass index (BMI), prognostic nutritional index (PNI), C-reactive protein (CRP), and CD3+CD8+T for the prognosis of lymphoma patients undergoing autologous hematopoietic stem cell transplantation.

Paediatric high-grade gliomas (pHGGs) are the most lethal brain tumours in children, characterised by profound epigenetic dysregulation and limited treatment options. The 2021 WHO Classification has established a molecular framework that distinguishes pHGGs as biologically distinct from adult glioblastoma, recognising four major subtypes: H3K27-altered diffuse midline glioma, H3G34-mutant diffuse hemispheric glioma, infant-type hemispheric glioma, and the rare IDH-mutant gliomas. Each subtype exhibits unique epigenetic landscapes, metabolic dependencies, and therapeutic vulnerabilities, necessitating subtype-specific treatment strategies. This review explores the molecular classification of pHGGs and examines the critical role of the tumour microenvironment in disease progression. We focus on glioma stem cells as central drivers of tumour initiation, maintenance, and therapeutic resistance, highlighting their remarkable cellular plasticity and ability to dynamically transition between different states. Particular attention is given to metabolic reprogramming in pHGGs, including alterations in glucose and lipid metabolism, and the exceptional metabolic flexibility of glioma stem cells that enables adaptation to microenvironmental pressures. Importantly, we discuss the intimate crosstalk between metabolism and epigenetic regulation, whereby metabolites serve as essential cofactors for chromatin-modifying enzymes-exemplified by α-ketoglutarate maintaining low H3K27me3 in H3K27M tumours and 2-hydroxyglutarate driving hypermethylation in IDH-mutant gliomas. We review preclinical models that have advanced pHGG research and discuss emerging immunotherapeutic approaches, including CAR T-cell therapies and oncolytic viruses. By synthesising current understanding of pHGG biology, this review aims to identify promising therapeutic avenues that exploit the unique metabolic and epigenetic vulnerabilities of each molecular subtype.

Naive human pluripotent stem cells (hPSCs) represent a pre-implantation epiblast state able to efficiently differentiate into embryonic and extraembryonic pre-implantation lineages and to self-organise in vitro into blastocyst-like structures called blastoids. Naive hPSC maintenance routinely relies on co-culture with mouse embryonic fibroblast (MEFs) as feeder cells, a method prone to variability and analytical confounders. Here, we describe a feeder-free culture system based on serum coating that supports long-term maintenance of naive hPSCs. Across five laboratories, 30 serum batches were evaluated for the expansion of eight naive hPSCs lines for up to 25 passages. Mass spectrometry analysis identified fibronectin and collagens as extracellular matrix proteins consistently present in serum coating. Cells cultured on serum coating displayed growth kinetics, clonogenic capacity, mutation rates, and global gene expression profiles comparable to MEF-based cultures. Importantly, serum-cultured naive hPSCs efficiently underwent germ layer specification, retained trophectoderm competence, and generated blastoids with efficiency similar to MEF-based cultures. Collectively, serum coating provides a scalable, cost-effective, and robust alternative to feeder-based systems, preserving genomic stability and developmental potential while eliminating MEF-associated disadvantages and variability. This platform facilitates large-scale applications of naive hPSCs and enables more reproducible mechanistic studies.

Mosaic variegated aneuploidy (MVA), a rare human congenital disorder that causes microcephaly, is characterized by extensive abnormalities in chromosome number and results from mutations in genes involved in accurate mitotic chromosome segregation. To characterize the cellular mechanisms underlying this disease, here we generated a Drosophila model of microcephaly caused by the depletion of a single spindle assembly checkpoint (SAC) gene in the neural stem cell (NSC) compartment. We present evidence that loss of stemness - compromised identity and proliferative capacity of NSCs- plays an important role in MVA and results in a reduced number of neurons and glial cells. We show that loss of stemness arises from the accumulation over time of an unbalanced number of gains and losses of more than one chromosome, rather than a direct consequence of chromosomal instability-induced DNA damage or the production of simple aneuploidies. We unravel a contribution of proteostasis failure and mitochondrial dysfunction to the negative impact of complex aneuploidies on stemness, a highly energy demanding cellular state. We identify overexpression of Radical Oxygen Species scavengers, mitochondria chaperones and apoptosis inhibition as genetic interventions capable of dampening the deleterious effects of aneuploidy on brain size.

The maintenance of human embryonic stem cell (hESC) self-renewal and pluripotency is governed by distinct signaling pathways, yet endogenous pluripotency-supporting pathways remain understudied despite extensive exogenous signaling research. Here, we identify a previously unrecognized role of endogenous VEGF signaling in sustaining primed hESC pluripotency. VEGF signaling is robustly activated in primed hESCs, quiescent in naïve cells, and inactivated upon differentiation. Strikingly, targeted VEGFR inhibition (pharmacological, soluble decoy receptors [sFLT1/sKDR], or CRISPR-mediated VEGFR1/2 knockout) in primed hESCs disrupts self-renewal and induces trophoblast-like differentiation. Mechanistically, VEGFR inhibition activates the BMP pathway and down-regulates NANOG, which directly binds and represses select BMP components and trophoblast lineage-specific genes. Functionally, BMP inhibition partially and NANOG overexpression substantially rescue the phenotype induced by VEGF signaling ablation. Collectively, our work uncovers a pivotal VEGF-dependent network maintaining primed pluripotency, providing valuable insights into integrated pluripotency and lineage regulation by signaling cascades and transcription factors.

Retrotransposons are DNA elements capable of inserting themselves into various regions of the genome, potentially leading to genomic changes. Their expression is tightly regulated by epigenetic and environmental factors, such as DNA methylation and radiation. For a long time, the effects of retrotransposons were poorly understood and largely overlooked. However, recent studies have demonstrated the significant role retrotransposons play in hematopoiesis under both healthy and stress conditions, such as during pregnancy or infection. Additionally, the expression of retrotransposons has been strongly correlated with various types of cancer. Depending on the cancer type, retrotransposons appear to exert both protumorigenic and antitumorigenic effects. This review summarizes current findings on the role of retrotransposons in hematopoiesis and cancer and highlights their potential as valuable tools for diagnosis and treatment.

The rapid development of allogeneic engineered therapeutic cells has intensified the challenge of host immune-mediated rejection. Advances in molecular immunology, genetic engineering, and induced pluripotent stem cell-based multigene editing have enabled the creation of 'stealth' allogeneic cells designed to evade immune detection while maintaining function. Key strategies include the deletion of human leukocyte antigen class I and class II molecules to limit T cell recognition, the expression of natural killer (NK) cell inhibitory ligands to prevent NK cell-mediated killing, and the upregulation of CD47 to suppress phagocytosis. An expanding repertoire of immune-modulatory molecules, receptor-ligand interactions, and experimental assays is refining these approaches. Together, stealth designs are accelerating the translation of allogeneic cell therapies toward more durable and broadly applicable clinical use.

The motor symptoms of Parkinson's disease are linked to age-dependent degeneration of dopamine neurons. Traditionally, the loss of these neurons has been thought to stem mainly from cell-autonomous cellular dysfunctions. However, there is growing evidence suggesting that it could result from complex interactions between cell-autonomous and noncell-autonomous mechanisms. This review article examines evidence for cell-autonomous mechanisms linked to mitochondrial, lysosomal, or proteasomal perturbations and for noncell-autonomous processes arising from glial and peripheral immune cells. We discuss how these pathways can converge to create chronic cellular stress and trigger cell death mechanisms. Beyond highlighting apoptosis as a key mode of degeneration, we consider the contribution of other mechanisms, including pyroptosis and ferroptosis. Understanding the relative dominance of these mechanisms across disease stages and patient subgroups could help guide the development of mechanism-based neuroprotective strategies.

Human induced pluripotent stem cell (hiPSC) line FLENIi004-A was generated from peripheral blood mononuclear cells (PBMCs) using the lentiviral-hSTEMCCA-loxP vector. FLENIi004-A was derived from a 41-year-old Argentinean Caucasian male patient with Autosomal-Dominant Alzheimer's Disease, who carried the heterozygous PSEN1 p.M146L mutation. The hiPSC line retained the PSEN1 mutation, and its pluripotency and trilineage differentiation potential were confirmed.