搜索结果：Stem cells and development

Objective: To investigate the effects and underlying mechanisms of low-concentration interleukin-6 (IL-6) on the proliferation, migration and osteogenic differentiation of human stem cells from apical papilla (hSCAPs). Methods: hSCAPs were isolated from the apical papilla of third molars extracted in the Department of Oral Maxillofacial Surgery, the First Affiliated Hospital of Zhengzhou University, and identified by flow cytometry and immunofluorescence. Cell counting kit-8 (CCK-8) and Transwell migration assay were used to detect the effects of IL-6 in different concentrations (10, 20, 50 μg/L) on the proliferative and migration ability of hSCAPs. Alizarin red staining, cetyl pyridine chloride quantification, alkaline phosphatase (ALP) staining and ALP activity assay were performed to evaluate the osteogenic differentiation capacity of hSCAPs under inflammatory conditions induced by different concentrations of IL-6, so as to determine the optimal IL-6 concentration to simulate the inflammatory microenvironment of hSCAPs osteogenic differentiation. Real-time fluorescence quantitative PCR (RT-qPCR) and Western blotting were used to detect the osteogenic differentiation ability of hSCAPs treated with low-concentration (10 μg/L) IL-6. Western blotting, Transwell migration assay, ALP staining, ALP activity assay and RT-qPCR were applied to explore the mechanism by which low-concentration IL-6 promotes the migration and osteogenic differentiation of hSCAPs via the Janus kinase 2-signal transducer and activator of transcription 3 (JAK2-STAT3) signaling axis. Results: The isolated and cultured hSCAPs were positive for surface markers CD24 and vimentin, while negative for CD45 and cytokeratin. Different concentrations of IL-6 exerted no significant effect on the proliferation of hSCAPs (P0.05). The number of migrated cells (11 463.33±622.70), cetyl pyridine chloride quantification (2.087±0.121) and ALP activity (94.11±4.06) U/g of hSCAPs in 10 μg/L IL-6 treatment were all significantly increased than those in 0 μg/L IL-6 control group, respectively [8 246.00±674.02, 1.330±0.096, and (53.87±3.70) U/g] (all P0.05). Moreover, the cell migration number and ALP activity in the 10 μg/L IL-6 group were more remarkably increased than in 20 μg/L IL-6 group [9 503.33±275.03, (74.44±3.55) U/g] (both P0.01). After osteogenic mineralization induction of hSCAPs under low-concentration IL-6 intervention, the mRNA expression levels of osteogenic-related factors Osterix (OSX) and Runt-related transcription factor 2 (Runx2) at day 3 (2.033±0.308, 1.433±0.215) and day 7 (3.497±0.153, 3.360±0.061) were significantly upregulated compared to the control group (all P0.05), while the mRNA level of osteocalcin (OCN) was significantly elevated at day 7 (2.950±0.315) (P0.05). The protein expression levels of OCN were significantly increased at day 3 (2.077±0.447) and day 7 (5.023±0.537) than those in the control group (both P0.05), while the protein levels of OSX and Runx2 elevated significantly at day 7 (3.283±0.601, 3.740±0.613) (both P0.05). Compared with the control group, low-concentration IL-6 obviously upregulated the protein expression ratios of p-JAK2/JAK2 and p-STAT3/STAT3, cell migration capacity, ALP activity, as well as the mRNA and protein expression levels of OCN, OSX and Runx2 of hSCAPs (all P0.05). Nevertheless, the JAK2-STAT3 signaling pathway inhibitor AG490 could significantly reverse these promotive effects of IL-6 on cell migration and osteogenic differentiation (all P0.05). Conclusions: IL-6 at concentrations of 10, 20 and 50 μg/L has no obvious effect on the proliferation of hSCAPs. Low-concentration IL-6 promotes the migration and osteogenic differentiation of hSCAPs under inflammatory conditions through activating the JAK2-STAT3 signaling pathway. 目的： 探讨低浓度白细胞介素-6（IL-6）对人根尖牙乳头干细胞（hSCAPs）增殖、迁移与成骨分化的影响及其作用机制。 方法： 从郑州大学第一附属医院口腔颌面外科拔除的第三恒磨牙根尖牙乳头内提取hSCAPs，使用流式细胞术及免疫荧光术进行鉴定；采用细胞计数试剂盒-8（CCK-8）及Transwell迁移实验检测不同质量浓度IL-6（10、20、50 μg/L，以0 μg/L IL-6为对照组）对hSCAPs增殖活性和迁移能力的影响；采用茜素红染色、氯化十六烷基吡啶定量检测、碱性磷酸酶（ALP）染色及ALP活性测定检测不同质量浓度IL-6炎症状态下 hSCAPs 的成骨分化能力，确定最适宜的IL-6浓度模拟hSCAPs成骨分化的炎症环境；采用实时荧光定量PCR（RT-qPCR）和蛋白质印迹法等方法检测低浓度（10 μg/L）IL-6炎症状态下 hSCAPs的成骨分化能力；采用蛋白质印迹法、Transwell迁移实验、ALP染色及ALP活性测定、RT-qPCR实验探究低浓度IL-6炎症状态下hSCAPs通过酪氨酸激酶2-信号转导与转录激活因子3（JAK2-STAT3）信号轴促进迁移和成骨分化的机制。 结果： 分离培养的hSCAPs表面标志物CD24、波形蛋白表达呈阳性，CD45、角蛋白表达呈阴性。不同质量浓度IL-6对hSCAPs的增殖能力均无显著影响（均P0.05）；10 μg/L IL-6炎症状态下 hSCAPs的细胞迁移数量［（11 463.33±622.70）个］、氯化十六烷基吡啶定量结果（2.087±0.121）及ALP活性［（94.11±4.06）U/g］均显著高于对照组［分别为（8 246.00±674.02）个、1.330±0.096、（53.87±3.70）U/g］（均P0.05），且10 μg/L IL-6组细胞迁移数量、ALP活性与20 μg/L IL-6组［分别为（9 503.33±275.03）个、（74.44±3.55）U/g］相比增加更为显著（均P0.01）。在低浓度IL-6炎症状态下hSCAPs成骨矿化诱导后，成骨细胞特异性转录因子Osterix（OSX）、Runt相关转录因子2（Runx2）的基因表达水平在3 d［分别为（2.033±0.308）、（1.433±0.215）］、7 d［分别为（3.497±0.153）、（3.360±0.061）］时均显著高于对照组（均P0.05），7 d时OCN基因表达水平（2.950±0.315）显著高于对照组（P0.05）；在3、7 d时骨钙素（OCN）的蛋白表达水平［分别为（2.077±0.447）、（5.023±0.537）］均显著高于对照组（均P0.05），7 d时OSX、Runx2的蛋白表达水平［分别为（3.283±0.601）、（3.740±0.613）］均显著高于对照组（均P0.05）。低浓度IL-6炎症状态下，与对照组相比，hSCAPs的p-JAK2/JAK2、p-STAT3/STAT3蛋白表达水平、细胞迁移量和ALP活性均显著增加，OCN、OSX、Runx2基因和蛋白表达水平亦显著增加，JAK2-STAT3通道阻断剂AG490能显著逆转这种迁移和成骨分化的促进作用（均P0.05）。 结论： 不同浓度IL-6对hSCAPs的增殖无显著影响，低浓度IL-6炎症状态下hSCAPs通过JAK2-STAT3通路促进细胞迁移和成骨分化。.

The increasing global burden of cancer necessitates innovative therapeutic strategies. Cell therapy represents a major breakthrough in oncology, evolving rapidly from the successful application of chimeric antigen receptor T (CAR-T) cells in hematologic malignancies to a multiplatform landscape characterized by the concurrent development of diverse strategies. Current research focuses on T cell receptor-engineered T (TCR-T) cells, tumor-infiltrating lymphocytes (TILs), gamma delta (γδ) T cells, CAR-natural killer (CAR-NK) cells, CAR-macrophages (CAR-Ms), and various strategies based on dendritic cells (DCs), B cells, and stem cells. The translational paradigm is expanding from the relatively mature field of hematologic malignancies to the more prevalent and mechanistically complex domain of solid tumors. In recent years, this field has exhibited a clear trend toward expansion from autologous therapies to allogeneic "off-the-shelf" platforms. Approaches such as CAR-NK and CAR-natural killer T (CAR-NKT) cell therapies exhibit significant clinical potential because of their low immunogenicity and reduced risk of graft-versus-host disease (GvHD). Concurrently, in vivo engineering technologies that directly deliver CAR genes in situ are emerging as promising approaches to lower costs and simplify manufacturing by bypassing complex ex vivo procedures. This review systematically outlines recent advances in these strategies, focusing on their mechanisms of action, target antigens, and clinical translation. Despite progress, formidable challenges remain, including tumor heterogeneity, the immunosuppressive tumor microenvironment (TME), and therapy-related toxicity. To address these challenges, future research will focus on novel target discovery, enhanced toxicity management, and scalable manufacturing processes. The integration of multidisciplinary technologies, such as multiomics analysis, artificial intelligence, and synthetic biology, will advance cell therapies toward safer, more effective, and widely accessible applications.

Stem cells, with their self-renewal and multi-lineage differentiation potential, hold promise for tissue repair and intractable diseases treatment. Yet clinical translation of stem cell therapies has long been hindered by insufficient scalable stem cell manufacturing, stemness loss and functional decline in 2D expansion, and poor post-transplantation cell retention, unregulated fate control. Programmable microcarriers (MCs) paired with 3D dynamic culture offer an emerging strategy to address these bottlenecks and enable stem cell fate regulation. In this review, we systematically review advanced MC fabrication strategies for stem cell fate regulation, comparing features of emerging technologies (microfluidics, electrospraying, in-air microfluidics, integrated in situ functionalization) and their implications for programmable MC control and scalable manufacturing. We analyze how MCs modulate stem cell behaviors (adhesion, proliferation, stemness maintenance, differentiation) via synergistic static physicochemical cues and dynamic stimuli-responsive properties. We map the latest advances in functionalized MC-mediated stem cell therapy across osteochondral defects, autoimmune, skin, ophthalmic and neurodegenerative diseases. Finally, we pinpoint unresolved challenges for clinical translation of MC-stem cell system and outline key future research directions. This review offers a systematic roadmap for advancing programmable MC fabrication, clinical-grade stem cell biomanufacturing, and precise cell therapy development.

Pancreatic ductal adenocarcinoma (PDAC) remains one of the deadliest cancers due to late diagnosis, early metastasis and resistance to therapy. Cancer stem cells (CSCs) have been implicated in PDAC aggressiveness and treatment failure. MAP kinase-interacting kinase 1 (MNK1) is overexpressed in PDAC and plays a critical role in tumor progression and CSC maintenance. Here, we show the potential of apMNKQ2, a DNA aptamer targeting MNK1, to therapeutically target CSCs and reduce PDAC tumor burden in patient-derived xenografts (PDXs). PDX cell lines and in vivo mouse models were used to assess the effects of apMNKQ2 on cell viability, apoptosis, cell cycle progression, migration, epithelial-to-mesenchymal transition (EMT) and CSC properties. Functional CSC targeting was validated through clonogenic and self-renewal assays as well as extreme limiting dilution analysis. Systemic administration of free apMNKQ2 was tested for biodistribution, pharmacokinetics, toxicity, and antitumor efficacy at escalating doses. apMNKQ2 downregulated MNK1 and anti-apoptotic proteins (MCL1, XIAP), impaired cell proliferation, induced apoptosis, and disrupted cell cycle progression in PDX PDAC cells. Importantly, apMNKQ2 also inhibited migration, mesenchymal properties and angiogenesis in vitro, and lung colonization in vivo. Notably, apMNKQ2 strongly targeted PDAC CSCs, reducing CD24, CD133, CXCR4 and ALDH expression, clonogenicity and in vivo tumor initiation over 600-fold. Free apMNKQ2 (without transfection agents) entered PDAC cells efficiently, retaining anti-CSC activity. Systemic delivery of free apMNKQ2 accumulated in tumors, was well tolerated up to 400 mg/kg and showed no toxicity. Importantly, 10 mg/kg of apMNKQ2 produced strong antitumor effects in PDX models. Increasing the dose 20-fold enhanced tumor uptake but not efficacy, suggesting a therapeutic plateau at 10 mg/kg. MNK1 plays a central role in PDAC progression and CSC maintenance. apMNKQ2 is a potent anti-MNK1 DNA aptamer with robust preclinical activity, including CSC-targeting and anti-invasive effects. Its low toxicity, systemic bioavailability, and efficacy at low doses support further development as a novel therapeutic strategy for PDAC.

Autophagy, an evolutionarily conserved and tightly controlled process in eukaryotic cells, allows them to respond to stress by selectively eliminating dysfunctional or unwanted materials to promote metabolic flexibility and maintain homeostasis. While autophagy is orchestrated by different autophagy (ATG)-related genes, whose regulation varies considerably according to tissue type and developmental stage. In this review, we investigate how regardless of the different players involved in autophagy, ATG5 emerges as a unique, highly conserved, critical molecule that acts as a central rheostat to control the stem cell fate, metabolic adaptability, and govern the immune signature pattern in cells. The journey of ATG5 modulation from physiological developmental variation to pathological scenario brings out the translational impact of ATG5. On the one hand, ATG5 promotes exit from the pluripotent state by c-Myc degradation during differentiation of specific lineages (involved in neurogenesis, adipogenesis, and hematopoiesis), while on the other hand, it has a critical involvement in metabolic circuitry via rewiring autophagy through modulation of lipophagy, mitophagy, and acetyl-CoA epigenetics. Taken together, this work summarizes new findings that centrally place ATG5 as a driver that engineers a coordinated crosstalk between the metabolic state and cell fate decisions and immune responses. These insights position ATG5 as a critical and therapeutic target for developmental disorders, cancer, and immune-metabolic diseases.

To investigate how soluble byproducts derived from a four-species endodontic biofilm model impact the viability, transcriptomic profile, and inflammatory response of human dental pulp stem cells (DPSCs). A sterile-filtered supernatant was extracted from an established interkingdom endodontic biofilm model comprising Streptococcus gordonii, Fusobacterium nucleatum, Porphyromonas gingivalis, and Candida albicans. DPSCs were exposed to the microbial biofilm supernatant (BSN) for 4 and 24 h. Cellular responses were evaluated via MTT, CCK-8, LDH assays, and Annexin V/PI staining. Transcriptomic sequencing was performed to assess gene expression dynamics, with GO and KEGG pathway enrichment analyses. IL6 and IL8 expression was validated by qPCR and ELISA. Data were analyzed using t-tests/ANOVA and RNA-seq differential expression using DESeq. 2 with FDR adjustment. BSN significantly suppressed DPSC metabolic activity without inducing apoptosis or necrosis. RNA-seq revealed 723 significantly differentially expressed genes at 4 h and 1667 at 24 h. Early responses were dominated by upregulation of inflammatory mediators, with enrichment of TNF, NF-κB, and JAK-STAT signaling pathways. At 24 h, the expression profile shifted toward redox regulation and metabolic suppression, including downregulation of glycolytic and purine metabolism pathways. IL6 and IL8 expression was markedly increased at both transcript and protein levels. Soluble factors produced by a biofilm model representative of deep caries and carious pulp exposures induce a time-dependent transcriptional response in DPSCs. This response is characterized by a biphasic pattern of early immune activation followed by later transcriptional metabolic adaptation. These findings highlight the capacity of soluble biofilm-derived products associated with deep caries to modulate DPSC immune-metabolic signaling. They further emphasize the importance of vital pulp therapy strategies that not only target microorganisms but also account for their secreted byproducts.

Androgenetic alopecia (AGA) imposes a significant psychosocial burden, yet current treatments such as minoxidil and finasteride often yield suboptimal responses or adverse effects. Exosomes, nanoscale extracellular vesicles derived from mesenchymal stem cells and dermal papilla cells (DPCs), offer a promising regenerative alternative by modulating key hair-growth pathways. This systematic review evaluates the efficacy, mechanisms, and translational challenges of exosome-based therapies for hair restoration in AGA. A comprehensive search was conducted across Google Scholar, Embase, PubMed, Scopus, and Web of Science for studies published from 2019 to 2025. The search strategy prioritized AGA-related terminology, including androgenetic alopecia, male pattern hair loss, female pattern hair loss, baldness, exosomes, extracellular vesicles, and hair regrowth. Following duplicate removal, 39 studies meeting Population, Intervention, Comparison, Outcome, and Study design criteria were included for qualitative synthesis. Preclinical data demonstrate that exosomes promote hair regeneration through multiple synergistic mechanisms: activation of the Wnt/beta-catenin and Sonic Hedgehog pathways, suppression of transforming growth factor beta (TGF-beta)/SMAD3 signaling, delivery of anti-inflammatory cytokines (e.g., IL-10), and rejuvenation of senescent DPCs via microRNA cargo (e.g., miR-122-5p). Early clinical studies report improvements in hair density (8-20%) and shaft thickness; however, the evidence base remains limited by small sample sizes, retrospective designs, lack of control groups, and inconsistent outcome measures. Emerging delivery systems, such as thermoresponsive hydrogels and microneedle patches, show promise in enhancing follicular penetration but require further validation. Exosome therapy represents a multi-target regenerative approach for AGA with a favorable preliminary safety profile. However, widespread clinical adoption is hindered by critical gaps in manufacturing standardization, scalability, regulatory frameworks, and robust long-term efficacy data. Future research must prioritize large-scale randomized controlled trials with standardized endpoints and Good Manufacturing Practice (GMP)-compliant production protocols to validate these findings and establish exosomes as a mainstream therapeutic option for AGA.

The in vitro culture of immature follicles remains a challenge in reproductive biology, ART, and fertility preservation. This study investigated the impact of α-MEM and SAGE 1-Step media on mouse preantral follicle development and oocyte maturation using two-dimensional (2D) and suspension culture systems. Preantral follicles (∼130 μm) from 14-day-old NMRI mice were cultured for 13 days in either medium under 2D or suspension conditions. On day 13, hCG was added to induce meiotic resumption. We evaluated follicular growth, antrum formation, oocyte maturation, meiotic spindle organization, and expression of genes related to maturation (Bmp15, Gdf9), cumulus expansion (Has2, Ptgs2, Lhr, Adamts1), and apoptosis (Bax, Bcl2). Antral formation was higher in α-MEM in both culture systems (P > 0.05). However, SAGE 1-Step significantly improved oocyte maturation across both systems (P = 0.01 in 2D vs P = 0.006 in suspension compared to α-MEM). Spindle staining demonstrated that suspension culture significantly increased the percentage of oocytes with normal meiotic spindle organization (P = 0.002). Gene expression analysis revealed a significant upregulation of Adamts1 (P = 0.008) and downregulation of Bax (P = 0.01) in suspension culture. Furthermore, the apoptotic ratio of Bax/Bcl2 was significantly decreased in the suspension system (P = 0.01). In conclusion, the SAGE 1-Step medium significantly enhances oocyte maturation, while the suspension system superiorly preserves meiotic spindle architecture and reduces apoptosis. The combination of SAGE 1-Step and suspension culture provides an optimized microenvironment for producing high-quality oocytes. Growing immature egg outside the body, a process known as in vitro follicle culture, is a promising but challenging technique for preserving fertility. This study investigated novel approaches to improve this process, aiming to create healthier, more viable eggs for future use. The researchers found that using a specific medium significantly improved egg development and maturation. Importantly, they found that culturing these immature eggs in suspension, where the follicles remained floating in the medium, rather than in a traditional two-dimensional system where follicles inevitably flatten and partially lose their three-dimensional structure, resulted in significantly better outcomes. Eggs grown in suspension showed more organized cellular structures and reduced cell death compared to those cultured on a flat surface. These findings represent an important step toward reliably producing healthy, developmentally competent eggs outside the body, offering new hope for fertility preservation and assisted reproduction.

Milk oligosaccharides (MO) support intestinal, microbial, and immune development in young pigs. However, modern production practices wean pigs at an early age, removing them from their source of MO prior to intestinal and immune maturation. The purpose of this study was to investigate the effects of dietary supplementation of galacto-oligosaccharides (GOS) and 2'-fucosyllactose (FL) on the jejunal mucosa-associated microbiota, intestinal immune signaling, morphology, and growth performance of nursery pigs. Forty-eight pigs (6.8&#x2009;&#xb1;&#x2009;0.2&#x2009;kg body weight) weaned at 3-weeks-of-age were allotted into six dietary treatments, using a randomized complete block design, with sex and initial body weight as blocks. Dietary treatments were (1) basal diet; (2) basal diet, with supplemental GOS at 1.5% of the diet; (3) basal diet, with supplemental FL at 0.2% of the diet; (4) basal diet, with GOS and FL at 1.5 and 0.2% of the diet, respectively; (5) basal diet, with GOS at 2.3% of the diet; and (6) basal diet, with FL at 0.3% of the diet. These MO were provided alone or in combination at levels mimicking intake at the end of the suckling period, and at 1.5-fold higher, to observe potential dose-dependent responses. Pigs were fed for 21 days in two phases. On d 21, pigs were euthanized for sampling of jejunal mucosa and jejunal tissue. Data were analyzed using the PROC MIXED of SAS 9.4 and contrasts were used to determine the effects of GOS, FL, and their combination (interaction), in addition to the linear effects of increasing dietary GOS or FL. Increasing levels of GOS and FL linearly decreased (P&#x2009;<&#x2009;0.05) Shannon and Simpson alpha diversity of the jejunal mucosa-associated microbiota. Supplementation with FL increased (P&#x2009;<&#x2009;0.05) the absolute abundance of Helicobacter in the jejunal mucosa-associated microbiota, although no dose response was observed. Increasing levels of GOS and FL tended to linearly decrease (P&#x2009;=&#x2009;0.051 and 0.076, respectively) the gene expression of TLR4. Increasing levels of GOS tended to increase (P&#x2009;=&#x2009;0.054) and increasing supplementation of FL increased (P&#x2009;<&#x2009;0.05) the number of Ki-67 proliferative cells in the crypt of the jejunum. Increasing levels of GOS increased (P&#x2009;<&#x2009;0.05) body weight and average daily gain in the early post-weaning period and tended to increase (P&#x2009;=&#x2009;0.093) body weight by the end of the experimental period. Notably, increasing levels of GOS, and GOS in combination with FL, improved growth performance, whereas FL alone did not. Milk oligosaccharides (MO) are indigestible carbohydrates with prebiotic effects for young mammals. Prebiotics support the intestinal microbiota and development of a functional intestine and intestinal immune system. Nursery pigs are weaned at a young age and could benefit from the continued provision of MO beyond the suckling period to mitigate the negative impacts of weaning on intestinal health. The purpose of this study was to evaluate the effects of two MO abundantly present in porcine milk, galacto-oligosaccharides (GOS) and 2&#x2032;-fucosyllactose (FL), on the jejunal mucosa-associated microbiota, immune response, stress status, morphology, and growth performance of nursery pigs. These oligosaccharides were provided alone or in combination at physiologically relevant levels mimicking intake at the end of the suckling period, and at 1.5-fold higher, to observe potential dose-dependent responses. The use of these MO modestly influenced the mucosa-associated microbiota, supplementation with GOS or FL reduced the expression of genes associated with inflammatory signaling and increased intestinal stem cell proliferation. Increasing supplementation of GOS up to 1.5 times the physiologically relevant levels improved growth performance of nursery pigs. The combination of both MO at physiologically relevant levels provided the most consistent benefits related to intestinal function and growth performance of nursery pigs.

Growth factors and cytokines are fundamental regulators of stem cell fate, controlling both self-renewal and differentiation across the embryonic hierarchy. ESCs that originate from the ICM of the blastocyst can self-renew indefinitely and remain pluripotent in response to cytokine-driven signaling cues. In contrast, ASCs, such as HSCs and MSCs, rely on niche-derived growth factors and cytokines for limited self-renewal and lineage commitment. Major extrinsic inputs are key regulators such as LIF, Wnt ligands, FGF2, BMP4, TGF-&#x3b2; family members, and Notch signaling molecules, but their effects are finally controlled by downstream epigenetic pathways. In this review, we highlight that growth factor signals are not understood in isolation but are transduced to stable transcriptional outputs by means of chromatin remodeling, DNA methylation, histone changes, and interactions with transcriptional regulatory networks. These pathways maintain ESC identity, regulate lineage priming, and orchestrate the development of the three germ layers during gastrulation and later tissue-specific differentiation. The same pathways work in specific niches of adult stem cells to balance asymmetric division, quiescence, self-renewal, and differentiation. We also highlight the context-dependent signaling outputs, which are regulated by epigenetic and chromatin changes, in determining stemness, lineage commitment, and tissue regeneration. Insights into the link between extracellular signaling and epigenetic regulation provide a groundwork for tailored techniques in stem cell engineering and regenerative medicine.

The cancer stem cell (CSC) paradigm has evolved from a rigid hierarchical model to a systems-level perspective in which stemness is a reversible and context-dependent phenotype. Evidence from lineage tracing and single-cell/spatial multiomics indicates that tumor cells occupy continuously evolving phenotypic states governed by complex gene regulatory networks. Within this landscape, CSCs can be interpreted as metastable attractors maintained through coupled signaling, epigenetic, metabolic, transcriptional, and microenvironmental interactions. Tumor heterogeneity and therapeutic resistance emerge through phenotypic reprogramming, regulatory network rewiring, and niche-dependent stabilization under environmental and therapeutic stress. This reframes resistance as an emergent property of tumor ecosystems, underscoring the limitations of targeting static CSC populations or single pathways. Therefore, durable therapeutic control will require network-oriented interventions capable of reshaping attractor topology and disrupting stemness-supportive microenvironments.

Lateral root (LR) branching is a vital adaptive strategy for plants to optimize nutrient acquisition, particularly under phosphorus deficiency. While auxin triggers LR formation, the molecular link between inorganic phosphate (Pi) homeostasis and the developmental competence of xylem-pole-pericycle (XPP) cells in roots remains elusive. In this study, we identified CsSPX2 from the suppressor of yeast Gpa1/ phosphate metabolism regulatory protein 81/xenotropic and polytropic retrovirus receptor 1 family as a critical phosphate sensor in cucumber (Cucumis sativus L.) that is strongly induced by Pi starvation. CRISPR/Cas9-generated CsSPX2 knockout lines displayed reduced LR density, shorter LRs, and abnormal root meristems. The loss of CsSPX2 resulted in phosphate overaccumulation and excessive lignin deposition under Pi-sufficient conditions. Concomitantly, we observed abnormal thickening of the XPP cell walls and subsequently impaired auxin responsiveness at LR initiation sites, as evidenced by diminished DR5::GUS activity. Notably, under Pi-deficient conditions, spx2 mutants exhibited obviously decreased lignin content and significantly reduced thickness of the XPP cell wall compared to Pi-sufficient conditions. In addition, the LR developmental defects in spx2 mutants were partially rescued by exogenous application of either a lignin biosynthesis inhibitor or synthetic auxin. Furthermore, CsSPX2 appeared to be important for maintaining the stem cell niche organization and meristematic activity in root tips, as its loss altered quiescent center and columella stem cell behavior and impaired root cap differentiation. Collectively, our findings support a model where CsSPX2 might link phosphate homeostasis to LR development through its involvement in XPP cell wall properties and auxin response. This study deepens our understanding of plant nutrition stress responses and provides a key genetic target for developing phosphate-efficient crops.

Natural killer (NK) cells are innate lymphocytes capable of killing transformed cells without prior sensitization through direct cytolytic activity and, in an antigen-specific manner, by antibody-dependent cellular cytotoxicity (ADCC). Human NK cells mediate ADCC exclusively via the IgG Fc receptor CD16 (Fc&#x3b3;RIIIA), triggering degranulation and cytokine production. Multi-engager complexes, such as BiKEs, TriKEs, TetraKEs, ANKETs, and ICEs, have been developed to link CD16 to tumor antigens. However, CD16 expression can be rapidly downregulated upon NK cell activation by the proteolytic checkpoint, a disintegrin and metalloproteinase-17 (ADAM17). ADAM17 induction in NK cells occurs in response to various stimuli, including the potent activating receptor CD16 and cytokine signaling. CD16 downregulation is further exacerbated within the tumor microenvironment, significantly diminishing ADCC potency by tumor-infiltrating NK cells. Moreover, ADAM17 expression and function are upregulated in various solid tumors, leading to the release of NK cell ligands and tumor-promoting factors. Consequently, developing strategies to inhibit ADAM17 to enhance NK cell function and suppress tumor cell growth is paramount to multi-engager efficacy. This review provides an overview of NK cell biology and the role of ADAM17 in regulating NK cell function and tumor cell growth. We examine the current landscape of NK cell multi-engager complexes in clinical development and discuss emerging strategies for incorporating an ADAM17-blocking component to optimize therapeutic outcomes.

Teeth serve as a powerful model to dissect epithelial-mesenchymal interaction-mediated organogenesis. While most non-mammalian vertebrates exhibit lifelong tooth renewal (polyphyodonty), mammals have largely lost this capacity and become predominantly diphyodont. Elucidating this evolutionary transition is crucial for understanding developmental constraints on regeneration, yet key mechanisms, particularly how the successional dental lamina persists and how dormant stem cell niches are reactivated, remain largely unresolved. Recent advances in lineage tracing, single-cell sequencing, multi-omics profiling and organoid models have uncovered remarkable cellular diversity and lineage potential during tooth development and homeostasis across vertebrates, reshaping our view of dental stem cell identity and its roles in tissue regeneration. In this Review, we summarize recent progress in tooth development, replacement and molecular regulation, and our current understanding of dental stem cells and organoid models, aiming to delineate a roadmap for translational tooth regeneration.

Bronchopulmonary dysplasia (BPD) remains a major complication among extremely low gestational age (ELGA) infants, with long-term respiratory and neurodevelopmental consequences. Despite advances in neonatal care, effective therapies to prevent BPD are lacking. Mesenchymal stromal cells (MSCs), particularly those derived from umbilical cord (UC-MSC), offer promise due to their pleiotropic effects. Preclinical and early-phase clinical studies have demonstrated safety and potential efficacy of MSC in neonatal lung injury. The Helping Underdeveloped Lungs with Cells (HULC)-2 trial aims to evaluate whether multiple intravenous doses of human allogenic UC-MSC can reduce mechanical ventilation duration and improve the respiratory outcome in ELGA infants at high risk of developing BPD. HULC-2 is a multicentre, double-blind, randomised controlled Phase II trial conducted in Canadian Neonatal Intensive Care Units. ELGA infants (gestational age <28 weeks) who remain ventilator-dependent between 4 and 14 days of life will be randomised to receive either 3&#x2009;weekly intravenous doses of UC-MSC (10&#xd7;106&#x2009;cells/kg/dose) or a sham procedure. The primary outcome is ventilation-free days (VFDs) at 120 days postrandomisation, accounting for mortality. Secondary outcomes include cell administration safety, respiratory and neurodevelopmental outcomes and complications of prematurity. A total of 168 participants will be enrolled to detect a clinically meaningful difference in VFDs. The study was approved through Clinical Trials Ontario by the Ottawa Health Science Network Research Ethics Board. Results will be disseminated via peer-reviewed publications, conferences and public engagement platforms. Parent partners are actively involved in study design and dissemination to ensure relevance and transparency. NCT07058025.

Mammary gland development during puberty requires tightly coordinated epithelial proliferation, lineage specification, and branching morphogenesis, processes critically regulated by growth factor signaling. While epidermal growth factor receptor (EGFR) signaling is essential for ductal development, how its activity is quantitatively controlled within mammary epithelial cells (MECs) remains incompletely understood. Here, we identify Rasgrp1, a Ras guanine nucleotide exchange factor, as a key modulator of EGFR signaling in the mammary epithelium. Using Rasgrp1-deficient mice, primary MEC assays, and organoid models, we demonstrate that loss of Rasgrp1 leads to elevated EGFR-Ras-PI3K-AKT and mTORC1-S6 signaling, resulting in enhanced proliferative capacity and aberrant EGF-driven branching. Transcriptomic analysis of organoids reveals that EGF signaling suppresses Wnt/R-spondin-dependent stem-cell gene programs, suggesting that excessive EGFR activity disrupts stem cell maintenance. In vivo, Rasgrp1 deficiency causes impaired ductal elongation, persistent terminal end buds, and increased epithelial proliferation, indicating a breakdown in the spatial and temporal coordination of mammary morphogenesis. Together, our findings establish Rasgrp1 as a signaling rheostat that dampens EGFR pathway activity to support coordinated mammary gland development. These results highlight the importance of precise signaling calibration in epithelial development and suggest broader implications for Ras pathway regulation in tissue homeostasis and disease.

The evolution of the human brain underlies our higher-order cognitive functions. In particular, the cerebral cortex, the outermost layer of the brain, has rapidly evolved to contain a disproportionately large number of neurons relative to the rest of the brain. Much of this expansion is attributed to the enlargement and diversification of the pool of neural precursors, which proliferate and differentiate into the neurons and glia of the brain. How the human cerebral cortex has evolved remains an active area of investigation. With the advent of pluripotent stem cell and brain organoid technologies, comparative genomic studies between humans, mice, and nonhuman primates have identified human-specific genes or pathways during neurodevelopment. The utility of these models relies on the ability of brain organoids to preserve the cytoarchitecture and species-specific developmental trajectories of diverse neural and glial cell types that are observed in&#xa0;vivo. This review will discuss how brain organoids recapitulate aspects of interspecies differences during cortical development, specifically neural precursor expansion, neurogenesis, and gliogenesis, and how these models can be improved to enable a deeper understanding of human brain evolution. Unraveling the cellular and molecular mechanisms underlying cross-species differences in brain expansion could also provide key insights into neurodevelopmental diseases, particularly those where brain size is affected.

Melanoma is an aggressive malignancy with rising global incidence. While early surgical intervention improves survival in localized cases, treatment resistance and recurrence remain a challenge. This underscores the critical need to identify prognostic biomarkers for early diagnosis, personalized treatment, and novel therapeutic development. The GSE126076 and melanoma dataset (Skin Cutaneous Melanoma, TCGA, PanCancer Atlas) were analyzed using an information-theoretical method to identify prognostic factors. Survival analysis was performed via Kaplan-Meier curves and log-rank tests to compare high- and low-mRNA expression groups. In vitro, A375 cells and A2058 cells were treated with the GGT inhibitor 6-diazo-5-oxo-L-norleucin (DON). Cell viability was assessed using the CCK-8 assay and intracellular GSH levels were measured following treatment. Through information-theoretic analysis and survival analysis, we identified GGT6 as a prognostic gene in melanoma. Survival analysis revealed that high GGT6 expression was significantly associated with shorter disease-specific survival across all disease stages. In vitro, 10 &#x3bc;M DON for 72 h reduced A375 cell proliferation by 90.72% versus control (p<0.0001), with an IC50 of 5.14 &#x3bc;M; and reduced A2058 cell proliferation by 60.93% (p<0.0001), with an IC50 of 7.93 &#x3bc;M. Measurement of GSH levels of A375 and A2058 cells revealed that melanoma cells treated with 10 &#x3bc;M DON exhibited lower GSH levels compared with the respective control groups (p<0.0001 and p=0.0042, respectively). Our study is the first to demonstrate the association between GGT6 expression levels and melanoma prognosis, and reveals that GGT inhibition suppresses melanoma cell viability. These findings provide new insights into the mechanisms of melanoma development and progression, and suggest GGT as a potential therapeutic target for clinical treatment.

Dysregulated host-microbe interactions are a hallmark of periodontal disease (PD), a chronic inflammatory condition that causes progressive alveolar bone resorption and tooth loss. MicroRNAs (miRNAs) have become important epigenetic regulators, offering new ways to understand disease pathophysiology and to provide tailored treatments. The current analysis uniquely synthesizes pathogen-specific miRNA signatures, modulation of the Receptor Activator of Nuclear Factor Kappa-B Ligand (RANKL)/Osteoprotegerin (OPG) axis, periodontal ligament stem cells (PDLSCs) osteogenesis, and enhanced delivery platforms into a coherent precision medicine framework, in contrast to other studies that examined these areas independently. We investigate the role of miRNAs in PD, assess their potential as non-invasive diagnostic biomarkers, and review novel therapeutic approaches targeting these molecules. Porphyromonas gingivalis and other periodontal infections cause aberrant miRNA expression that accelerates bone loss by impairing the development of PDLSCs, disrupting the RANKL/OPG axis, and sustaining pro-inflammatory cytokine cascades (IL-1, IL-6, TNF-&#x3b1;). Based on replication across many independent investigations, three of the many identified miRNAs-miR-146a, miR-155, and miR-223-show the most promise for diagnosis and treatment. These miRNAs, found in saliva and gingival crevicular fluid, serve as reliable non-invasive indicators. Preclinical studies show that anti-miR inhibitors and miRNA mimics, administered via hydrogels or nanoparticles, successfully reduce inflammation and promote alveolar bone repair. However, several issues remain unresolved, including miRNA instability, off-target effects, and interpatient variability, as well as contradictory results from multiple studies, such as the opposing functions of miR-21 across various cell types. In summary, targeted host modification by miRNA-based therapies is a paradigm change from traditional symptomatic therapy. To incorporate these strategies into clinical practice and eventually enable regenerative and customized periodontal treatment, it will be crucial to address current delivery and safety constraints through improved nanocarriers and patient-specific profiling.