The color patterns of mammalian fur are determined by melanocytes' ability to respond to and adapt to microenvironmental cues. However, these patterns can be lost following injury or under pathological conditions, and the underlying biological mechanisms remain poorly understood. In this study, reconstituted hair-bearing skin is generated using skin organoids derived from dissociated epidermal cells, dermal cells, and melanocyte progenitors. The reconstituted skin exhibited pigmented hair patterns. By investigating the molecular cues involved in re-establishing pigment patterns, it is demonstrated that this process is regulated through a two-step mechanism. First, during skin organoid culture, signaling from dermal fibroblasts to melanocytes via the COL6A3-CD44 pathway promotes the early maintenance of organotypic melanocytes. Subsequently, during hair follicle morphogenesis after skin organoid transplantation, signaling from the bulge to melanocytes via the SEMA3C-NRP1 pathway regulates microtubule stability. This regulation guides melanocytes to migrate to their bulge stem cell niche, thereby enhancing hair pigmentation by promoting the adaptive patterning of melanocytes within the hair follicle. The study reveals two novel signaling mechanisms that shape melanocyte adaptive patterning and highlight the hair follicle as a regulatory hub for melanocyte physiological behaviors. These findings may inspire new clinical strategies for preventing hair greying.

Neuroblastoma (NB) is the most common extracranial solid tumor in children characterized by poor immune infiltration and resistance to adaptive immunity, contributing to its limited response to immunotherapy. A key mechanism underlying immune evasion in cancer is autophagy, a cellular process that plays many roles in cancer by supporting tumor survival and regulating immune interactions. In this study, we investigate the impact of autophagy inhibition on NB tumor growth, immune modulation, and the efficacy of immunotherapy. Using both murine and human NB cell lines, we demonstrate that genetic and pharmacological inhibition of autophagy significantly reduces 3D spheroid growth and upregulates major histocompatibility complex class I (MHC-I) expression. In vivo studies further confirm that targeting autophagy suppresses tumor progression and promotes immune infiltration into the tumor. Notably, we observe a significant increase in CD8+ T cell recruitment and activation, suggesting that autophagy inhibition reshapes the immune landscape of NB, rendering it more susceptible to immune-mediated clearance. Crucially, autophagy inhibition also sensitizes NB cells to T cell-mediated cytotoxicity and enhances the therapeutic efficacy of GD2.CAR T-cell therapy. In vitro co-culture assays reveal increased CAR T cell-mediated tumor killing upon autophagy blockade, while in vivo models show prolonged tumor control and improved survival in treated mice compared to CAR T-cell therapy alone. These findings highlight autophagy as a key regulator of immune evasion in NB and suggest that its inhibition could serve as a promising therapeutic strategy to enhance immune recognition and improve the efficacy of immunotherapy.

Dyslipidemia has been extensively documented as a key driver of cardiovascular pathology. Regulating lipid homeostasis holds promise for treating atherosclerosis (AS). Although the protein phosphatase 2 catalytic subunit beta (PPP2CB) is involved in post-transcriptional gene regulation, its role in AS-associated dyslipidemia is not well understood.

Multiplexing samples from distinct individuals prior to sequencing is a promising step towards achieving population-scale single-cell RNA sequencing by reducing the restrictive costs of the technology. Individual genetic demultiplexing tools resolve the donor-of-origin identity of pooled cells using natural genetic variation but present diminished accuracy on highly multiplexed experiments, impeding the analytic potential of the dataset. In response, we introduce Ensemblex: an accuracy-weighted, ensemble genetic demultiplexing framework that integrates four distinct algorithms to identify the most probable subject labels. Using computationally and experimentally pooled samples, we demonstrate Ensemblex's superior accuracy and illustrate the implications of robust demultiplexing on biological analyses.

Aging is linked to various dysfunctions of the immune system, including the decline of its primary developmental source: the hematopoietic stem cell (HSC) niche. This decline leads to chronic inflammation, increased vulnerability to infections, cancer, autoimmune diseases, and reduced vaccine efficacy. As individuals age, the HSC niche undergoes significant changes, including greater adipocyte accumulation and alterations in the molecular microenvironment, which may influence the development and function of immune cells. Among these cells, the impact of the aging HSC niche on dendritic cell (DC) function is less understood. Heterochronic autologous HSC transplantation is a promising intervention to prevent age-related disorders, contributing to the extension of healthspan and longevity, however, several murine experiments failed to produce the expected results, which led us to presume that the problem lies within the old HSC niche. Therefore, we created in vitro models of young and old HSC niches and examined how these microenvironments affect the differentiation and maturation and functionality of BM-derived DCs (BMDCs).

The aim to this study was to explore the effect of p75NTR on the mineralization and development of maxillofacial processes. Moreover, we tried to elaborate the potential mechanism of p75NTR's effect on the odontogenic or osteogenic differentiation ability of neural crest cells (s).

The maintenance of the haematopoietic stem cells and the production of platelets thereof is acutely dependent upon the thrombopoietin receptor (TpoR). TpoR dimerizes in the presence of its ligand thrombopoietin leading to the activation of downstream JAK2-STAT signalling. Alternatively, the receptor dimerizes in the presence of activating mutations of JAK2 (JAK2 V617F), calreticulin or TpoR leading to myeloproliferation. These effects are dependent upon the surface expression of the receptor. TpoR is secreted through the Golgi-dependent pathway. Although, an unconventional autophagosome-lysosome mediated traffic has been postulated; questions remained whether the observed lysosomal localization was indicative of degradation or secretion. We fused the pH-sensitive FRET pair of TOLLES-YPet with TpoR (SRAI-TpoR) that showed FRET quenching at low pH environment of lysosomes. Using this construct, we demonstrated the presence of quenched SRAI-TpoR on the cell surface indicating that TpoR could indeed be secreted through low pH compartments. We further demonstrated that the lysosome-autophagosome-mediated secretion of TpoR was promoted by JAK2 V617F mutation, partially utilized by TpoR W515L but abrogated by calreticulin mutations. Finally, we showed that the lysosome-autophagosome mediated secretion was dependent upon Rab1A. Our study conclusively showed the unconventional traffic of TpoR and its modulation by the driver mutations causing myeloproliferation.

Microtubules and microtubule-associated proteins are critical regulators of cerebral cortex development, and their defects can lead to severe cortical malformations. EML1/Eml1 (Echinoderm microtubule-associated protein-like 1) is a microtubule-binding protein whose mutations cause subcortical heterotopia in both humans and mice. While perturbations of Eml1 in neural progenitor cells have been associated with defects in cilia and progenitor cell detachment from the ventricular zone, the regulatory role of Eml1 at the protein level remains understudied. To reveal global changes in the absence of Eml1, we perform a comparative proteomic analysis of the cortices and neural progenitor cells of Eml1 conditional-knockout mice during cerebral cortex development. Our comprehensive analyses reveal that Eml1 depletion causes significant downregulation of multiple centrosomal and spindle proteins in neural progenitor cells. The absence of Eml1 significantly reduces microtubule polymerization and stability. Several microtubule-associated proteins, including Eml4 and Septins, lose their affinity with microtubules in the absence of Eml1. Our findings support the central role of Eml1 in the regulation of microtubules and provide a valuable resource for the investigation of the underlying mechanisms of heterotopia-based pathophysiological conditions.

HUWE1, a member of HECT E3 ubiquitin ligase family, is implicated in a variety of cellular processes. Recent studies find that HUWE1 also plays critical roles in germ cell development and inactivation of HUWE1 causes germ cell loss in both male and female mice. In this study, we found that Huwe1 was also highly expressed in testicular Sertoli cells. Inactivation of Huwe1 in Sertoli cells resulted in loss of cell polarity, which in turn caused germ cells loss and male infertility. Further study revealed that dysregulation in the expression of cytoskeletal and adhesion-related molecules, as well as a significant increase in EMT-related trans-factors SNAI1&2 in Huwe1-deficient Sertoli cells. Intriguingly, the protein level of WT1 was significantly increased in Huwe1-deficient Sertoli cells, and overexpression of Wt1 in Sertoli cells also caused the defects in spermatogenesis which was consistent with Huwe1 CKO mouse model. Furthermore, the defect of spermatogenesis in Huwe1 CKO mice was partially rescued by deleting one allele of Wt1 gene. Mechanistic studies revealed that WT1 interacts with HUWE1 protein and it could be ubiquitinated by HUWE1. Our study demonstrates that HUWE1 is involved in the establishment of Sertoli cell polarity mainly by regulating the protein level of WT1 gene.

As DNA variants accumulate in somatic stem cells, become selected or evolve neutrally, they may ultimately alter tissue function. When, and how, selection occurs in homeostatic tissues is incompletely understood. Here, we introduce SCIFER, a scalable method that identifies selection in an individual tissue, without requiring knowledge of the driver event. SCIFER also infers self-renewal and mutation dynamics of the tissue's stem cells, and the size and age of selected clones. Probing bulk whole-genome sequencing data of nonmalignant human bone marrow and brain, we detected pervasive selection in both tissues. Selected clones in hematopoiesis, with or without known drivers, were initiated uniformly across life. In the brain, we found pre-malignant clones with glioma-initiating mutations and clones without known drivers. In contrast to hematopoiesis, selected clones in the brain originated preferentially from childhood to young adulthood. SCIFER is broadly applicable to renewing somatic tissues to detect and quantify selection.

Tumor immune microenvironment plays a crucial role in determining the prognosis of lung adenocarcinoma (LUAD), with the interaction of immune cells within this microenvironment contributing to a poorer prognosis. We sought to investigate the causal relationship and underlying biological mechanisms between immune cell characteristics and LUAD to offer new insights for enhancing treatment strategies. We evaluated the association between immune cell characteristics and LUAD using Mendelian randomization (MR) analysis based on genome-wide association studies summary statistics. Sensitivity analysis was performed to verify the robustness of MR results. Immune cell infiltration analysis and machine learning on bulk RNA-sequencing data were conducted to further identify immune cells associated with LUAD. Prognostic genes of LUAD were identified using single-cell RNA-sequencing data and high dimensional weighted gene co-expression network analysis. MR analysis identified three immune cell characteristics associated with LUAD, including CCR2 on granulocyte, CD25 on CD45RA + CD4 not Treg, and plasmacytoid dendritic cell, and sensitivity analysis confirmed the robustness of the associations. Machine learning identified neutrophils, hematopoietic stem cells, B cells, and myeloid progenitor cells as key immune cell characteristics related to LUAD. Neutrophil was identified as the target cell of LUAD based on the MR analysis and machine learning. Subcequently, single-cell RNA-sequencing mapped the immune microenvironment of LUAD and identified down-regulated neutrophil. In addition, robust neutrophil-macrophage communication in LUAD was revealed using the CellChat package. Finally, nine neutrophil-related prognostic genes of LUAD were identified, three of which potentially regulated neutrophil-macrophage communication. This study showed a significant correlation between neutrophil and LUAD, particularly highlighting the neutrophil-macrophage communication. This finding may enhance our comprehension of LUAD immune microenvironment and hopefully promote the discovery of new immunotherapeutic targets for LUAD and the development of related drugs.

A novel LMNA p.(Glu105Leu) variant was identified in five families with dilated cardiomyopathy (DCM), revealed as a local founder variant originating approximately 650 years ago. Genetic testing and clinical analysis of 795 DCM patients demonstrated that probands with this variant typically present with severe DCM in their sixties, characterized by high prevalence of late gadolinium enhancement, arrhythmias, and conduction disorders. Time-to-event analysis suggested a later onset of clinical symptoms compared to other LMNA variants, with a trend towards longer event-free survival. Microscopic imaging of patient fibroblasts, induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs), and heart tissue confirmed structural nuclear LMNA-associated abnormalities. Patient iPSC-CMs exhibited distinct sarcomeric disorganization, increased glucose uptake and glycogen content, reduced mitochondrial function and biogenesis, and delayed contractile function. These findings support the pathogenicity of the variant and demonstrate its profound impact on structural and metabolic functions in cardiomyocytes.

To mitigate the necessity for multiple invasive procedures in treating malignant osteosarcoma, an innovative therapeutic approach is imperative to achieve controllable tumor-killing effects and subsequent bone repair. Here, we propose the de novo design of sono-activable and biocatalytic nanoparticles-modified 3D-printed hydroxyapatite (HA) scaffold (HS-ICTO) for intelligently sequential therapies in osteosarcoma eradication and bone defect regeneration. The engineered HS-ICTO scaffold displays superior, spatiotemporally controllable H2O2-catalytic performances, which promptly generate massive reactive oxygen species via multienzyme-like mechanisms coupled with sono-activation, thus augmenting tumor cell apoptosis. Furthermore, HS-ICTO can intelligently switch to catalyze H2O2 to O2 within the inflammatory bone defect microenvironment, effectively blocking endogenous H2O2-mediated oxidative stress, which positively modulates the osteogenic differentiation of stem cells and ultimately facilitates defect regeneration. We validate that this multifaceted HS-ICTO scaffold possesses robust and on-demand abilities to prevent neoplastic recurrence and promote anti-inflammatory osseous tissue repair, representing a promising platform for precision oncological intervention and regenerative medicine.

Meniscus-derived stem cells (MeSCs) hold great promise for cytotherapy of the meniscus. Large-scale cell expansion in vitro is usually accompanied by decreased proliferation and migration and increased senescence, leading to a decrease in treatment efficiency. Therefore, the present study aimed to compare the effects of different concentrations of glucose on the proliferation, migration, senescence and protein expression of MeSCs. In this study, human MeSCs were cultured in two types of expansion media: low-glucose (LG) DMEM (5.5 mM glucose) and high-glucose (HG) DMEM (25 mM glucose). At specific passage number, the proliferation rate was evaluated by cell counting, the migration rate was evaluated by a scratch-wound assay, cell senescence was evaluated by β-GAL assays, and protein expression/phosphorylation was evaluated by ELISA. The results showed that a lower concentration of glucose promoted proliferation and migration, reduced cell senescence, and activated PI3K/Akt-related signaling pathways. This study provides a preliminary basis for the use of this medium composition for MeSC expansion.

Chimeric antigen receptor T (CAR-T) cell therapies have transformed the treatment of relapsed/refractory (R/R) B-cell malignancies and multiple myeloma by redirecting activated T cells to CD19- or BCMA-expressing tumor cells. However, this approach has yet to be approved for acute myeloid leukemia (AML), the most common acute leukemia in adults and the elderly. Simultaneously, CAR-T cell therapies continue to face significant challenges in the treatment of solid tumors. The primary challenge in developing CAR-T cell therapies for AML is the absence of an ideal target antigen that is both effective and safe, as AML cells share most surface antigens with healthy hematopoietic stem and progenitor cells (HSPCs). Simultaneously targeting antigen expression on both AML cells and HSPCs may result in life-threatening on-target/off-tumor toxicities such as prolonged myeloablation. In addition, the immunosuppressive nature of the AML tumor microenvironment has a detrimental effect on the immune response. This review begins with a comprehensive overview of CAR-T cell therapy for cancer, covering the structure of CAR-T cells and the history of their clinical application. It then explores the current landscape of CAR-T cell therapy in both hematologic malignancies and solid tumors. Finally, the review delves into the specific challenges of applying CAR-T cell therapy to AML, highlights ongoing global clinical trials, and outlines potential future directions for developing effective CAR-T cell-based treatments for relapsed/refractory AML.

Transplantation of neural stem cells (NSCs) holds promise for repairing traumatic brain injury (TBI) but their therapeutic performance is hindered due to the low efficient differentiation into neurons. Direct injection of differentiation modulators to the lesion site has limited improvement to neuronal differentiation as they tend to diffuse or be degraded. In the present study, we report a simple and versatile strategy to engineer the NSCs with a micropatch to improve their therapeutic performance in TBI treatment. The micropatches are fabricated through microcontact printing technique and can adhere to the membrane with negligible detachment or internalization within 14 days after surface moficiation. The micropatches on the cell membrane can move together with stem cells and sustainedly release retinoic acid, a neuronal differentiation modulator, to regulate the surrounding microenvironment of NSCs, improving their neuronal differentiation rate from 28.0% to 54.2%. The micropatches-engineered NSCs can be implanted to the injured brain tissue through a minimally invasive microinjection approach and show outperformance in repairing damaged neural tissue of TBI mice compared to normal stem cells. Overall, this work highlights a new pathway to engineer stem cells and holds great potential in nerve regeneration and neurodegenerative diseases treatment.

Endothelial cells (ECs), which line the inner surface of blood vessels, continuously respond to biomechanical forces from blood flow, extracellular matrix, and intracellular tension. Recent advances have highlighted the pivotal role of these forces in regulating cellular plasticity during endothelial-to-hematopoietic transition (EHT) and endothelial-to-mesenchymal transition (EndMT), two processes essential for embryogenesis, tissue repair, and disease progression. EHT contributes to hematopoietic stem cell formation, and EndMT to valve formation and vessel sprouting. When misregulated, both processes cause vascular pathologies such as fibrosis, cancer metastasis, and atherosclerosis. This review provides an overview of how biomechanical cues influence EC fate decisions and behavioral transitions. We explore how external biomechanical forces are sensed at the endothelial cell surface, transmitted through intracellular adaptors, and affect changes at the transcriptional level. Understanding these mechanotransduction pathways during cell fate transition not only deepens our knowledge of endothelial cell plasticity but also provides insight into potential root causes of and treatments for vascular diseases.

Natural killer-cell therapies are limited by donor cell sourcing and dose-to-dose variability. FT516 is an induced pluripotent stem cell (iPSC)-derived natural killer-cell therapy expressing high-affinity, non-cleavable CD16 to optimise antibody-dependent cellular cytotoxicity in combination with therapeutic monoclonal antibody. We aimed to assess the safety of FT516 in patients with relapsed or refractory B-cell lymphoma.

Micro/Nano-plastics (MNPs), including microplastics (MPs; <5mm) and nanoplastics (NPs; <100nm), have become pervasive environmental pollutants due to extensive plastic production and insufficient recycling practices. These particles originate from the degradation of larger plastic materials through processes such as photo-oxidation, thermo-oxidation, and incomplete biodegradation, resulting in chemically reactive fragments that persist in air, water, and food. Once released, MNPs enter the human body primarily via ingestion, inhalation, and dermal absorption, ultimately accumulating in various tissues, including reproductive organs. This review provides a comprehensive summary of current knowledge regarding the toxicological effects of MNPs on male and female reproductive health, with a focus on mammalian models and relevance to human exposure. In males, MNPs have been associated with testicular damage, impaired spermatogenesis, reduced sperm count and motility, and disruptions in the hypothalamic-pituitary-gonadal axis. In females, exposure has been linked to altered folliculogenesis, disrupted ovarian hormone levels, impaired oocyte quality, and placental dysfunction. These effects are largely driven by mechanisms involving oxidative stress, inflammation, endocrine disruption, mitochondrial dysfunction, and apoptosis. Furthermore, MNPs have been shown to disrupt gut microbiota composition, contributing to systemic inflammation and reproductive dysfunction through emerging pathways such as the gut-testis axis. Given their widespread presence and multifaceted modes of action, MNPs pose a serious threat to human reproductive health. Therefore, there is an urgent need for stricter environmental regulations, improved waste management, and further research to understand the long-term and transgenerational consequences of MNP exposure.

The thymidine phosphorylase (TYMP) gene encodes a pivotal enzyme involved in nucleoside metabolism, playing a critical role in the processing of chemotherapeutic agents. Despite its recognized importance, the influence of TYMP on tumor immunology and clinical outcomes remains largely unexplored. This study sought to elucidate the roles and regulatory pathways of TYMP across a spectrum of cancers and whether TYMP is related to DNA damage repair, thereby promoting stem cell maintenance, chemotherapy resistance and immunosuppression. Utilizing pan-cancer bulk sequencing and digital platforms, the study investigated the impact of TYMP on patient outcomes, genomic instability, cancer stemness, DNA repair, and immune cell dynamics. Moreover, single-cell datasets of TISCH, spatial transcriptomic data of SpatialDB, and multiple fluorescence staining were used to validate the association between TYMP expression and macrophages. In vitro viability (trypan blue), wound healing, Transwell, Cell cycle and M1 macrophage infiltration assays were performed to determine the biological functions of TYMP in Kidney renal clear cell carcinoma (KIRC) cells. Tools such as ROCplotter and cMap were employed to evaluate treatment responses and identify compounds targeting TYMP, while PDB and Autodock Vina facilitated structural modeling and binding simulations. TYMP was an oncogene in many cancer types. High TYMP was associated with lower genome stability and high expression of mismatch repair genes, stemness, homologous repair gene signature. Also, immune checkpoint CD276 was positively relevant to TYMP. Subsequently, we validated TYMP as the macrophage marker and showed its connection with other immunosuppressive cells and CD8+ T-cell depression. In vitro experiments showed that TYMP promoted migration, and invasion of KIRC cells and recruited the M2 macrophage to KIRC cells. Finally, potential TYMP -targeted drugs were screened out and docked to TYMP protein. TYMP was a novel oncogene, and it correlated with immunosuppression and DNA repair. TYMP was highly associated with immune checkpoint CD276 and was an macrophage biomarker in many cancers. This study will reveal TYMP's roles in pan-cancer and its potential as a novel therapeutic target, especially KIRC.