Autism spectrum disorder (ASD) is a complex neurodevelopmental disorder that significantly impacts children's physical and mental health, yet effective pharmacological treatments remain limited. The primary objective of this study was to investigate the therapeutic effects of human umbilical cord blood mesenchymal stem cells (hUC-MSCs) on ASD, evaluate the safety profile of hUC-MSCs, and elucidate their underlying mechanisms and functional roles.

Large intestinal crypts are tightly packed tubular indentations of the epithelium that act both as an absorptive surface and as a protective barrier. The crypt base consists of stem cells involved in the continuous regeneration of the intestinal surface. Thus, these crypts serve as the etiology of 95% of colon cancer and other colon-related pathologies such as inflammatory bowel disease. However, we still need a better understanding of these important structures in the human intestine. We collected and scanned four different regions of the large intestine (cecum, descending colon, sigmoid colon, and rectum) from 15 cadavers to study the surface characteristics of the crypts such as crypt density, opening area, depth, and volume. The cecum had a significantly larger crypt opening area when compared to the rectum. Further analysis showed that this difference is age dependent and only observed in older cadavers. The rectum also showed age-dependent changes in crypt depth. The varying morphology of crypts between the proximal/distal regions and individuals could be attributed to functional differences and naturally occurring adaptations to protect areas of the colon that are more susceptible to colon-related pathologies.

Autologous peripheral blood stem cell (PBSC) collections are routinely performed in the outpatient setting and typically involve placement of an apheresis-compatible central venous catheter (CVC). There is reluctance to discharge outpatients with non-tunneled CVCs due to safety concerns. We characterized our center's practice of multiday non-tunneled CVC use in vetted adult outpatients undergoing autologous PBSC collections. No patient with social, cognitive, hygienic, caregiver, or bleeding concerns had non-tunneled CVC placement. All patients and caregivers were counseled about potential risks of non-tunneled CVCs and were provided with basic CVC care education. In total, 117 autologous PBSC collections were performed in 65 adult outpatients via non-tunneled CVCs. Apheresis nursing staff removed CVCs from all patients on their final PBSC collection day. There was one minor CVC site bleed that was successfully controlled with manual compression. In our small population, multiday non-tunneled CVC use in vetted adult outpatients undergoing autologous PBSC collections was feasible and well tolerated.

CD90 (THY1) is a cell surface glycoprotein that plays a crucial role in the occurrence and development of various malignant tumors. It affects tumor cell proliferation, metastasis, angiogenesis, stem cell characteristics, and energy metabolism. CD90 is an important biomarker for many malignant tumors and is closely related to tumor prognosis. However, the role and specific mechanisms of CD90 in glutamine metabolism in gastric cancer (GC) cells remain incompletely understood. In this study, we report that CD90 affects glutamine metabolism in GC cells through SLC1A5, ultimately promoting GC progression. Mechanistically, CD90 promotes the binding of YY1 to the SLC1A5 promoter, enhances the transcriptional activity of the SLC1A5 promoter, and increases its transcriptional level. This, in turn, affects SLC1A5-mediated glutamine metabolism and ferroptosis in GC cells, ultimately facilitating GC progression. Our results suggest that CD90 plays an important role in regulating glutamine metabolism in GC cells, providing important experimental evidence for elucidating the pathogenesis of GC.

Current single-cell RNA atlases largely capture polyadenylated transcripts while missing critical regulatory layers from noncoding RNA. To address this, we develop a generalizable framework that adapts total RNA profiling for use in standard droplet-based platforms and captures a broad complement of coding and noncoding RNAs using a unified pipeline. Applying this approach to the developing human brain, we generate a dataset mapping diverse RNA biotypes across all neuronal and non-neuronal lineages, revealing biotype-specific expression programs with cell-type and temporal specificity. Tracking microRNA dynamics in Cajal-Retzius neurons, transient and early-born neurons in the cortex, we show the enrichment and target anticorrelation of MIR137, associated with schizophrenia and intellectual disability, suggesting tight regulatory control. We apply TotalX to human peripheral blood mononuclear cells and identify transcriptional modules combining coding and noncoding RNAs and tRNA dynamics. In addition, we analyze dengue-infected hepatocytes and capture non-adenylated viral transcripts that distinguish infection states. This expanded coverage helps with understanding cellular identity and gene regulation at the atlas scale.

GD2, a tumor-associated carbohydrate antigen, has been validated as a promising immunotherapeutic target for the treatment of high-risk neuroblastoma. Historically, cancer therapies have largely focused on protein targets. However, the FDA approval of dinutuximab, a GD2-specific monoclonal antibody, in 2015 marked a significant shift by establishing carbohydrates as viable therapeutic targets. Since then, anti-GD2 has become a key component of the standard treatment regimen for high-risk neuroblastoma. Beyond neuroblastoma, GD2 is also expressed in several other malignancies, including melanoma, glioma, small cell lung cancer, and sarcoma. A wide array of therapeutic strategies targeting GD2 has been developed, encompassing murine, chimeric, and humanized antibodies; bispecific antibodies; immunocytokines; chimeric antigen receptor (CAR)-T and CAR natural killer T (NKT) cells; and vaccine-based approaches. A thorough understanding of the strengths and limitations of each of these modalities is essential to inform the design of more effective combination therapies and to improve outcomes for patients with GD2-expressing tumors.

Pluripotent stem cells (PSCs) are a unique population of cells, capable of infinite self-renewal and differentiation into many cell types, making them essential for regenerative medicine, tissue engineering, and drug discovery. With the increasing use of PSCs in clinical trials, it is crucial to accurately assess their differentiation status and understand the diversity within PSC populations. Glycan markers have emerged as key tools for evaluating the undifferentiated state of PSCs, as well as for live and fixed imaging, and for isolating particular cell populations by applying approaches such as fluorescence-activated cell sorting (FACS). This article summarizes the glycan markers used to assess the undifferentiated states of both mouse and human PSCs, which include embryonal carcinoma cells (ECCs), embryonic stem cells (ESCs), and induced pluripotent stem cells (iPSCs). By offering a summary of these markers, we aim to advance our understanding of their role in PSC biology, facilitating further developments in stem cell research and broadening medical applications.

Although cancer immunotherapy has been flourishing, the number of targets for Food and Drug Administration (FDA)-approved cancer immunotherapeutics has remained small and mostly limited to proteins. The approval of dinutuximab, an anti-GD2 for treating high-risk neuroblastoma in 2015, marks the first new agent targeting glycosphingolipids (GSLs). Recently, our research group demonstrated that another GSL, Globo H ceramide (GHCer), which was previously reported to be the most prevalent cancer-associated antigen in many epithelial cancers, is an independent poor prognostic factor for breast cancer, hepatoma, cholangiocarcinoma, and gallbladder cancer. We had further shown that GHCer is shed from tumor cells to extracellular vesicles (EVs), which are then incorporated into endothelial cells in the tumor microenvironment to promote angiogenesis. Molecular dynamics simulation and other studies also revealed fucose-dependent changes in the glycan conformation of GHCer conducive for a complex formation between translin-associated factor X (TRAX) protein and GHCer, thus highlighting the molecular mechanism of how GHCer facilitated dissociation of phospholipase C beta 1 (PLCβ1) from TRAX, thereby enhancing angiogenesis. Thus, GHCer is not only a tumor antigen associated with adverse prognostic impact but also acts as an immune checkpoint and an angiogenic factor to shape the tumor microenvironment. These findings provide strong rationales for developing Globo H-targeted immunotherapy.

Serendipity is to come across a nice coincidence or to discover something unexpected. Also, it is to look for something and happen to find something of value other than what you are looking for. In simple, it is to seize upon good fortune by a random chance. I described examples of serendipity in scientific discoveries in the glycoscience field and their background in the 2011 publication of MICC-3 under the title "Serendipity in scientific discoveries: some examples in glycosciences." We have learned that the progress in the scientific field has been remarkable, and many discoveries have been made, including some discoveries by serendipity. In this chapter, I filled the gaps in serendipitous discoveries in the biomedical sciences left by the MICC-3 as follows: (i) the discovery of intramolecular lactonization of sialic acids, (ii) the discovery of ancient Chinese script (), (iii) the discovery of oriental beauty oolong tea, and (iv) Yamanaka's Nobel Prize-winning discovery of induced pluripotent stem (iPS) cells. Finally, the planning for serendipity was discussed.

Human trophoblast stem (TS) cells provide a powerful in vitro model to study trophoblast lineage specification, placental development, and pregnancy-related disorders. This chapter presents detailed methodologies for the maintenance, derivation, and differentiation of TS cells, with a focus on reproducibility. We describe optimized culture conditions that preserve stem cell identity, including culture medium composition, extracellular matrix requirements, and passaging techniques. Step-by-step protocols for inducing differentiation into extravillous trophoblast and syncytiotrophoblast are presented, along with morphological, molecular, and functional assays to assess lineage commitment. Additionally, we describe a procedure to generate organoids from TS cells. Practical guidance is provided for maintaining reproducibility across experiments. These protocols can be implemented to investigate the role of diverse proteins, including members of the ADAMTS family, in human placental development. By enabling reproducible and controlled TS cell expansion and trophoblast cell differentiation, this methodology supports both basic research and disease modeling.

Isocitrate dehydrogenase 1 (IDH1) mutations confer distinct biological properties to gliomas, including the reshaping of the tumor immune microenvironment. While T cell dysfunction in glioblastoma has been extensively characterized, the role of innate lymphoid cells (ILCs)-critical regulators of tissue homeostasis and early immune responses- remains poorly understood.

Induced pluripotent stem cells (iPSCs) have emerged as a powerful tool in biomedical research, enabling the study of cellular function and early disease mechanisms within patient-specific genetic contexts. Traditionally, iPSCs have been used to model monogenic diseases, where highly penetrant variants produce robust cellular phenotypes detectable in few cell lines. Recent advances in scalability and standardisation now enable systematic comparisons across many donors. This development is particularly relevant for complex diseases, which are driven by numerous genetic variants with small individual effects and therefore require population-scale designs to resolve genotype-phenotype relationships. However, several limitations of iPSC technology continue to challenge the reliability and reproducibility of such studies, constraining their translational relevance. Here, we review the challenges and opportunities of using iPSCs to model complex diseases, structured around three key themes: detecting subtle effects, modelling environmental context, and expanding genetic diversity.

Alternative lengthening of telomeres (ALT) is a telomere maintenance mechanism deployed in embryonic stem cells and cancer cells. High levels of the heterochromatin mark H3 lysine 9 trimethylation (H3K9me3) at telomeres are critical for ALT, but how this is achieved remains unclear. Telomeric association of orphan nuclear receptors (NRs)-such as COUP-TF1, COUP-TF2, TR2, and TR4-has been shown previously to facilitate ALT activation. Here, we show that orphan NRs regulate telomeric H3K9me3 through TRIM28, a corepressor of ZNF transcription factors, to promote ALT. We report that H3K9me3 is induced by telomeric association of orphan NRs in cultured human fibroblast and ALT cancer cell lines. Moreover, TRIM28 is required for the orphan-NR-induced H3K9 methylation and ALT phenotypes. Importantly, physical interaction of TRIM28 with orphan NRs induces telomeric localization of TRIM28. A TRIM28 variant defective in orphan-NR interaction fails to localize to telomeres and is unable to promote H3K9me3 and ALT phenotypes. These findings indicate that telomeric orphan NRs recruit TRIM28 for telomeric H3K9me3 and ALT activation, emphasizing the role of chromatin structure in ALT activation.

Animal study.

Parkinson's disease (PD), characterized by α-Synuclein aggregation and dopaminergic neuronal loss, has no current cure. Autophagy is critical for α-Synuclein clearance, yet its real-time dynamics remain challenging to assess in human-relevant systems. Here, we used live-cell imaging to assess autophagy within human neuronal cultures and midbrain organoids (hMOs) derived from induced pluripotent stem cells (iPSCs) of PD patients carrying a triplication of the α-Synuclein gene (3xSNCA). Using the LC3-Rosella dual-fluorescent reporter, we quantified autolysosomes dynamics in real time. In 3xSNCA neuronal cultures, we detected early autophagy defects. In 3xSNCA hMOs, reduced autolysosome area, increased total and phosphorylated α-Synuclein (pS129), and decreased electrophysiological activity were observed at 50 days of differentiation (DoD). By 70 DoD, autophagy impairment became more pronounced, overlapping with dopaminergic neuron dysfunction. These findings support the use of human iPSCs-derived models to study autophagy dysfunction in PD and demonstrate a temporal correlation between impaired autophagy, α-Synuclein pathology and neuronal degeneration.

The epigenetic silencing or remarkably diminished expression of STING in cancer cells, along with the structural and functional impairment of the endoplasmic reticulum (ER) and Golgi apparatus, represents a unique mechanism of tumor immune escape and poses an important challenge for STING-targeted therapies. Here, we develop a cell membrane-derived nanodisc system (ND-cGAMP-HP; HP, heparin), which is capable of presenting activated STING proteins in their native form by means of cell membrane-directed display and biological self-assembly techniques. It can directly introduce activated STING protein to tumor cells and circumvent the translocation process between the ER and Golgi apparatus, selectively activating the IFN-I signaling pathway without initiating the inflammation-related pathway NF-κB. ND-cGAMP-HP triggers potent cellular immune responses and remodels the tumor immune microenvironment. Moreover, it augments immune memory by promoting the differentiation of TCF1+ stem cell-like T cells. We thus manifest a strategy based on STING therapy that does not depend on the ER and Golgi apparatus pathways to activate the IFN-I pathway, for cancer immunotherapy.

Chimeric antigen receptor (CAR)-T cell therapy has transformed treatment of relapsed/refractory DLBCL, yet resistance driven by regulatory T cells (Tregs) limits its efficacy. Here we identify Timosaponin AIII (TAIII), a clinical-stage natural product, as an effective modulator of CAR-T function that depletes CAR-Tregs while enhancing effector activity. Mechanistically, TAIII acts as an allosteric A2AR inhibitor by competing with cholesterol, suppressing CREB-dependent FoxP3 transcription and disrupting the A2AR-Treg axis. Ablation of A2AR or Tregs in vitro and in vivo abolishes TAIII activity, confirming specificity. Furthermore, TAIII reduces intratumoral Tregs, increases CD8⁺ T cells infiltration, and potentiates PD-1 blockade in solid tumor models. Importantly, TAIII promotes central memory T-cell formation and enhances CAR-T cytotoxic cytokine secretion. Combining or pretreating CAR-T cells with TAIII markedly improves antitumor efficacy and prevents late relapse across preclinical models. These findings establish TAIII as a combinatorial strategy to deplete CAR-Tregs, enhance CAR-T activity, and extend therapeutic durability.

Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) hold tremendous promise for in vitro modeling to assess native myocardial function and disease mechanisms, as well as testing drug safety and efficacy. However, current hiPSC-CMs are functionally immature, resembling in vivo CMs of fetal or neonatal developmental states. The use of targeted culture media and organoid formats have been identified as potential high-yield contributors to improve CM maturation. This study presents an hiPSC-CM maturation medium formulation, designed using a differential evolutionary approach targeting metabolic functionality for iterative optimization. Relative to existing high-performing reference formulations, our medium significantly matured morphology, Ca2+ handling, electrophysiology, and metabolism, which was further validated by multi-omic screening, for cells in either pure or co-cultured microtissue formats. Together, these findings not only provide a reliable workflow for highly functional hiPSC-CMs for downstream use, but also demonstrate the power of high-dimensional optimization processes in evoking advanced biological function in vitro.

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive and often fatal cancer with limited treatment options. Small nucleolar RNA host gene 10 (SNHG10) has emerged as a key regulator in the progression and metastasis of human cancers. However, the potential of SNHG10 in PDAC tumorigenesis, gemcitabine resistance, and the underlying molecular mechanisms remains poorly understood. Our data analysis revealed significant upregulation of the SNHG10 transcript in 179 PDAC cases compared with 171 normal pancreatic specimens, with a positive association with clinical stages of PDAC. Further, we confirmed the significant overexpression of the SNHG10 transcript in several PDAC cell lines compared to normal pancreatic cells. Our results revealed that the downregulation of SNHG10 significantly decreased the cellular proliferation, clonogenic ability, cell migration, and the epithelial to mesenchymal transition, leading to the induction of cell cycle arrest and apoptosis of PDAC cells. Mechanistically, the downregulation of SNHG10 significantly inhibited the expression of vimentin, N-cadherin, survivin, CDK4, CDK6, cyclin B1, cyclin D1, aurora kinase A, and aurora kinase B, with an increased expression of E-cadherin and p21. The bioinformatics analysis, RNA Immunoprecipitation, and qRT-PCR results showed the physical interaction among SNHG10, miR-150-5p, and VEGF-A, which are the integral parts of the ternary complex in PDAC cell lines. Interestingly, silencing of SNHG10 led to the significant induction of miR-150-5p, which repressed the expression of VEGF-A in PDAC cells. Moreover, the miR-150-5p rescued the VEGF-A expression in PDAC cells even during the silencing of SNHG10. Interestingly, the downregulation of SNHG10 enhanced gemcitabine sensitivity in gemcitabine-resistant PDAC cells. The depletion of SNHG10 in the PDAC xenograft model significantly reduced tumor growth, volume, and weight. Importantly, downregulation of SNHG10 suppressed the phosphorylation of EGFR, AKT, ERK1/2, mTOR, and c-MET signaling pathways in both in vitro and xenograft models of PDAC. Our study unveils the oncogenic potential of SNHG10 in tumorigenesis through modulation of the EGFR/AKT/ERK/mTOR, and miR-150-5p/VEGF-A axis, as well as gemcitabine resistance of PDAC as a prospective therapeutic strategy.The graphical abstract represented the potential role of SNHG10 and its regulated molecular mechanisms in the tumorigenesis and gemcitabine resistance in PDAC. Silencing of SNHG10 decreases cell survival, proliferation, clonogenicity, EMT tumor growth through the EGFR/AKT/ERK/mTOR axis, and restores the expression of miR-150-5p, which eventually downregulates VEGF-A. SNHG10 downregulation enhanced the gemcitabine sensitivity in gemcitabine-resistant PDAC cells.

Extracellular vesicles (EVs) derived from human induced pluripotent stem cells (hiPSC-EVs) hold great therapeutic promise, yet challenges in scalable production and the impact of 3D cellular architecture on EV content and function continue to hinder clinical translation. Here, we demonstrate a perfusion-based stirred-tank bioreactor (BR) system to scale up hiPSC-EV production, eliminating labor-intensive static cultures while preserving physiologically relevant cellular characteristics. The system was successfully scaled from 0.2 l BRs to 2 l BRs, yielding clinically relevant EV doses (1011 EVs per 2 l BR). Notably, only EVs secreted under BR conditions elicited significant proangiogenic effects in vitro, promoting endothelial cell survival, proliferation, migration, and sprouting. Small RNA and proteomic profiling revealed enrichment of proangiogenic miRNA and proteins regulating endothelial function in BR-derived EVs, indicating a shift driven by the culture environment. These findings underscore the role of BR dynamics in shaping EV properties and supporting the scalable production of therapeutically potent hiPSC-EVs.