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Context: Good health and longevity depend on dynamic interactions between biological, social, psychological, and environmental factors. Aging is globally a big challenge, particularly with the demographic transition, including population growth, and an emerging burden to society. Knowledge, behavior, diet, and consumption of animal source food were related to aging and emerged as the key factors modulating healthy aging.Objective: The study was designed to understand the main healthy aging factors, such as knowledge, social network, and diet of elders, and to derive mutual learning from it for healthy aging.Methods: A qualitative approach has been applied to explore health-related knowledge, attitude, and diet of elders from Ebetsu (Japan) and Tiassalé (Côte d’Ivoire) health districts, using focus group discussions and comparative context analysis between high- and low-income countries.Results: The study shows that living longer is a common feature of people in Japan compared to Côte d’Ivoire, where the life expectancy is still low. Both groups of elders have social networks that support them, and both offer their gained experience to society. While Japanese elders depend on pension and insurance for income and medical treatments, Ivorians depend mostly on their children and social network in old age. The worries of elders differ between the two regions. In Ebetsu, elder members of the community are concerned about the future burden they pose for the younger generation if they develop ill-health, making them more resilient to aging. In Taabo, elders are considered to be culturally and socially useful to the society. Elders in Ebetsu pointed out that for healthy aging, education on diet at a younger age, physical activities, and access to basic social services are the key aspects. This was not observed in Taabo’s context. Being inactive and dependent on others were described as the most worrying situations for elders in Ebetsu, as it is perceived to increase the risk of non-communicable diseases and anxiety. Elders in Ebetsu have good knowledge on what constitutes a healthy diet, and they believe that diversifying their diet, reducing portions, and substituting red meat with good animal and vegetable proteins are best eating practices to maintain good health. In Côte d’Ivoire, the diet is imbalanced and the whole family consumes the same meal made mainly with high-energy staples and little protein. However, it is observed in both societies that adopting a good diet is very expensive.Conclusion: The consciousness of aging is universal, but healthy aging varies according to the social systems, education, and knowledge on diet transition. Physical activities, protein–energy balance in diet, and social networks are the key for healthy aging in both contexts. The challenge is to find ways to increase knowledge regarding healthy aging and to strengthen the support system so that healthy aging becomes affordable.

CO2 observations in the free troposphere can be useful for constraining CO2 source and sink estimates at the surface since they represent CO2 concentrations away from point source emissions. The thermal infrared (TIR) band of the Thermal and Near Infrared Sensor for Carbon Observation (TANSO) Fourier transform spectrometer (FTS) on board the Greenhouse Gases Observing Satellite (GOSAT) has been observing global CO2 concentrations in the free troposphere for about 8 years and thus could provide a dataset with which to evaluate the vertical transport of CO2 from the surface to the upper atmosphere. This study evaluated biases in the TIR version 1 (V1) CO2 product in the lower troposphere (LT) and the middle troposphere (MT) (736–287 hPa), on the basis of comparisons with CO2 profiles obtained over airports using Continuous CO2 Measuring Equipment (CME) in the Comprehensive Observation Network for Trace gases by AIrLiner (CONTRAIL) project. Bias-correction values are presented for TIR CO2 data for each pressure layer in the LT and MT regions during each season and in each latitude band: 40–20° S, 20° S–20° N, 20–40° N, and 40–60° N. TIR V1 CO2 data had consistent negative biases of 1–1.5 % compared with CME CO2 data in the LT and MT regions, with the largest negative biases at 541–398 hPa, partly due to the use of 10 µm CO2 absorption band in conjunction with 15 and 9 µm absorption bands in the V1 retrieval algorithm. Global comparisons between TIR CO2 data to which the bias-correction values were applied and CO2 data simulated by a transport model based on the Nonhydrostatic ICosahedral Atmospheric Model (NICAM-TM) confirmed the validity of the bias-correction values evaluated over airports in limited areas. In low latitudes in the upper MT region (398–287 hPa), however, TIR CO2 data in northern summer were overcorrected by these bias-correction values; this is because the bias-correction values were determined using comparisons mainly over airports in Southeast Asia, where CO2 concentrations in the upper atmosphere display relatively large variations due to strong updrafts.

The Thermal and Near Infrared Sensor for Carbon Observation (TANSO)–Fourier Transform Spectrometer (FTS) on board the Greenhouse Gases Observing Satellite (GOSAT) has been observing carbon dioxide (CO2) concentrations in several atmospheric layers in the thermal infrared (TIR) band since its launch. This study compared TANSO-FTS TIR version 1 (V1) CO2 data and CO2 data obtained in the Comprehensive Observation Network for TRace gases by AIrLiner (CONTRAIL) project in the upper troposphere and lower stratosphere (UTLS), where the TIR band of TANSO-FTS is most sensitive to CO2 concentrations, to validate the quality of the TIR V1 UTLS CO2 data from 287 to 162 hPa. We first evaluated the impact of considering TIR CO2 averaging kernel functions on CO2 concentrations using CO2 profile data obtained by the CONTRAIL Continuous CO2 Measuring Equipment (CME), and found that the impact at around the CME level flight altitudes (∼ 11 km) was on average less than 0.5 ppm at low latitudes and less than 1 ppm at middle and high latitudes. From a comparison made during flights between Tokyo and Sydney, the averages of the TIR upper-atmospheric CO2 data were within 0.1 % of the averages of the CONTRAIL CME CO2 data with and without TIR CO2 averaging kernels for all seasons in the Southern Hemisphere. The results of comparisons for all of the eight airline routes showed that the agreements of TIR and CME CO2 data were worse in spring and summer than in fall and winter in the Northern Hemisphere in the upper troposphere. While the differences between TIR and CME CO2 data were on average within 1 ppm in fall and winter, TIR CO2 data had a negative bias up to 2.4 ppm against CME CO2 data with TIR CO2 averaging kernels at the northern low and middle latitudes in spring and summer. The negative bias at the northern middle latitudes resulted in the maximum of TIR CO2 concentrations being lower than that of CME CO2 concentrations, which led to an underestimate of the amplitude of CO2 seasonal variation.

Recent advancements in spatial transcriptomics and spatial proteomics enabled the high-throughput profiling of single or multi-cell types and cell states with spatial information. They transformed our understanding of the higher-order architectures and paired cell-cell interactions within a tumor microenvironment (TME). Within less than a decade, this rapidly emerging field has discovered much crucial fundamental knowledge and significantly improved clinical diagnosis in the field of immuno-oncology. This review summarizes the conceptual frameworks to understand spatial omics data and highlights the updated knowledge of spatial immuno-oncology., Competing Interests: Declaration of Competing Interest All authors have nothing to declare, about any financial and personal relationships with other people or organizations that could inappropriately influence or bias their work. Examples of potential competing interests include:, (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Recently, injectable hydrogels have garnered significant attention in tissue engineering due to their controlled flowability, strong plasticity, adaptability, and good biocompatibility. However, research on readily injectable in situ -forming hydrogels capable of forming functional three-dimensional (3D) tissue condensations remains limited. This study explores the development and evaluation of a carboxymethyl chitosan (CMCTS)/oxidized hyaluronic acid (oHA) hydrogel incorporated with silver sulfadiazine (AgSD) for tissue engineering applications with inherent antibacterial activity. Through physicochemical analysis, the optimal formulation of CMCTS/oHA hydrogels was established. The hydrogel demonstrated excellent injectability, enabling minimally invasive in situ delivery. In vitro cytotoxicity assays identified 0.1 % AgSD as the optimal concentration, supporting cell proliferation while exhibiting antimicrobial efficacy against S. mutans and E. faecalis . In vivo studies revealed complete hydrogel degradation and good biocompatibility, with no adverse tissue reactions. The hydrogel's ability to form 3D cell aggregates and support tissue regeneration underscores its potential for future 3D tissue engineering applications. Consequently, the injectable CMCTS/oHA/AgSD hydrogel developed in this study holds significant potential for application in a wide range of bioengineering fields, including antibacterial substance delivery systems and 3D tissue engineering, indicating potential for future clinical application., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2025 The Authors.)

Cell therapies such as CAR-T have demonstrated significant clinical successes, driving the investigation of immune cell surface engineering using natural and synthetic materials to enhance their therapeutic performance. However, many of these materials do not fully replicate the dynamic nature of the extracellular matrix (ECM). This study presents a cell surface engineering strategy that utilizes phase-separated peptide coacervates to decorate the surface of immune cells. We meticulously designed a tripeptide, Fmoc-Lys-Gly-Dopa-OH (KGdelta; Fmoc=fluorenylmethyloxycarbonyl; delta=Dopa, dihydroxyphenylalanine), that forms coacervates in aqueous solution via phase separation. These coacervates, mirroring the phase separation properties of ECM proteins, coat the natural killer (NK) cell surface with the assistance of Fe 3+ ions and create an outer layer capable of encapsulating monoclonal antibodies (mAb), such as Trastuzumab. The antibody-embedded coacervate layer equips the NK cells with the ability to recognize cancer cells and eliminate them through enhanced antibody-dependent cellular cytotoxicity (ADCC). This work thus presents a unique strategy of cell surface functionalization and demonstrates its use in displaying cancer-targeting mAb for cancer therapies, highlighting its potential application in the field of cancer therapy., (© 2024 The Author(s). Angewandte Chemie International Edition published by Wiley-VCH GmbH.)

Background: Chimeric antigen receptor (CAR)-NK cell therapy has shown remarkable clinical efficacy and safety in the treatment of hematological malignancies. However, this efficacy was limited in solid tumors owing to hostile tumor microenvironment (TME). Radiotherapy is commonly used for solid tumors and proved to improve the TME. Therefore, the combination with radiotherapy would be a potential strategy to improve therapeutic efficacy of CAR-NK cells for solid tumors., Methods: Glypican-3 (GPC3) was used as a target antigen of CAR-NK cell for hepatocellular carcinoma (HCC). To promote migration towards HCC, CXCR2-armed CAR-NK92 cells targeting GPC3 were first developed, and their cytotoxic and migration activities towards HCC cells were evaluated. Next, the effects of irradiation on the anti-tumor activity of CAR-NK92 cells were assessed in vitro and in HCC-bearing NCG mice. Lastly, to demonstrate the potential mechanism mediating the sensitized effect of irradiation on CAR-NK cells, the differential gene expression profiles induced by irradiation were analyzed and the expression of some important ligands for the NK-cell activating receptors were further determined by qRT-PCR and flow cytometry., Results: In this study, we developed CXCR2-armed GPC3-targeting CAR-NK92 cells that exhibited specific and potent killing activity against HCC cells and the enhanced migration towards HCC cells. Pretreating HCC cells with irradiation enhanced in vitro anti-HCC effect and migration activity of CXCR2-armed CAR-NK92 cells. We further found that only high-dose (8 Gy) but not low-dose (2 Gy) irradiation in one fraction could significantly enhanced in vivo anti-HCC activity of CXCR2-armed CAR-NK92 cells. Irradiation with 8 Gy significantly up-regulated the expression of NK cell-activating ligands on HCC cells., Conclusions: Our results indicate the evidence that irradiation could efficiently enhance the anti-tumor effect of CAR-NK cells in solid tumor model. The combination with radiotherapy would be an attractive strategy to improve therapeutic efficacy of CAR-NK cells for solid tumors., (© 2024. The Author(s).)

Objective: The color matching of single-shade resin composites after in-office whitening was investigated., Materials and Methods: Four single-shade resin composites were used. A total of 35% hydrogen peroxide was used as the whitening agent. The resin composite was placed in a cavity of an artificially discolored bovine tooth. The color differences between the restoration and surrounding enamel before and after whitening were determined based on ΔE* ab , ΔE 00 , and ΔWI D . The color stability, surface roughness (Sa), and surface gloss (GU) of the resin composite alone were also evaluated. Statistical analyses were performed using repeated-measures analysis of variance with the Tukey-Kramer test., Results: Based on the 50:50% of perceptibility and acceptability thresholds of ΔE* ab and ΔE 00 , none of the resin composite restorations were clinically acceptable before or after whitening. Regarding ΔWI D , although all resin composites showed "acceptable match" in the baseline, they showed "mismatch" after the third session of whitening. Most of the resin composites alone were stable in color against whitening., Conclusions: Although the single-shade resin composites failed to achieve the expected color matching on discolored teeth either before or after the whitening, the impact of the whitening on the color of the resin composite alone may be negligible., (© 2024 Wiley Periodicals LLC.)

Microfluidic chips have emerged as significant tools in cell culture due to their capacity for supporting cells to adopt more physiologically relevant morphologies in 3D compared with traditional cell culture in 2D. Currently, irreversible bonding methods, where chips cannot be detached from their substrates without destroying the structure, are commonly used in fabrication, making it challenging to conduct further analysis on cells that have been cultured on-chip. Although some reversible bonding techniques have been developed, they are either restricted to certain materials such as glass, or require complex processing procedures. Here, we demonstrate a simple and reversible polydimethylsiloxane (PDMS)-polystyrene (PS) bonding technique that allows devices to withstand extended operations while pressurized, and supports long-term stable cell cultures. More importantly, it allows rapid and gentle live cell extraction for downstream manipulation and characterization after long-term on-chip culturing, and even further subculturing. Our new approach could greatly facilitate microfluidic chip-based cell and tissue cultures, overcoming current analytical limitations and opening up new avenues for downstream uses of on-chip cultures, including 3D-engineered tissue structures for biomedical applications.

SARS-CoV-2 provokes devastating tissue damage by cytokine release syndrome and leads to multi-organ failure. Modeling the process of immune cell activation and subsequent tissue damage is a significant task. Organoids from human tissues advanced our understanding of SARS-CoV-2 infection mechanisms though, they are missing crucial components: immune cells and endothelial cells. This study aims to generate organoids with these components. We established vascular immune organoids from human pluripotent stem cells and examined the effect of SARS-CoV-2 infection. We demonstrated that infections activated inflammatory macrophages. Notably, the upregulation of interferon signaling supports macrophages' role in cytokine release syndrome. We propose vascular immune organoids are a useful platform to model and discover factors that ameliorate SARS-CoV-2-mediated cytokine release syndrome., (© 2024. The Author(s).)

COVID-19 emerged in September 2020 as a disease caused by the virus SARS-CoV-2. The disease presented as pneumonia at first but later was shown to cause multisystem infections and long-term complications. Many efforts have been put into discovering the exact pathogenesis of the disease. In this review, we aim to discuss an emerging tool in disease modeling, organoids, in the investigation of COVID-19. This review will introduce some methods and breakthroughs achieved by organoids and the limitations of this system., Competing Interests: Conflict of interest statement. None declared., (© The Author(s) 2023. Published by Oxford University Press on behalf of Society for Leukocyte Biology.)

This commentary investigates the findings presented in the article by Yang et al. in 2023, published in the Journal of Leukocyte Biology. This commentary first summarizes the spatial-temporal dynamics of regulatory T cells derived from mice (Tabula Muris Senis) of different ages (3, 18, and 24 mo) at different anatomic niches like lymph nodes and bone marrow. We also reported possible combinations of receptor-ligand interactions among T follicular regulatory cells, T follicular helper cells, and germinal center B cells, such as the calmodulin/Fas axis and PSGL-1/L-selectin axis. Then, we have elaborated on the significance of understanding aging regulatory T cells and offered some possible future research directions for Yang et al., contributing to a critical analysis of their recent study. Building on these foundations, further investigations and studies can be conducted to delve deeper into the mechanisms by which regulatory T cells influence health upon aging, potentially unveiling novel therapeutic targets to ameliorate age-related pathogenicity., Competing Interests: Conflict of interest statement. The authors declare no financial and personal conflicts of interest., (© The Author(s) 2023. Published by Oxford University Press on behalf of Society for Leukocyte Biology.)

Hematopoiesis is a highly orchestrated biological process sustaining the supply of leukocytes involved in the maintenance of immunity, O2 and CO2 exchange, and wound healing throughout the lifetime of an animal, including humans. During early hematopoietic cell development, several waves of hematopoiesis require the precise regulation of hematopoietic ontogeny as well as the maintenance of hematopoietic stem and progenitor cells in the hematopoietic tissues, such as the fetal liver and bone marrow. Recently, emerging evidence has suggested the critical role of m6A messenger RNA (mRNA) modification, an epigenetic modification dynamically regulated by its effector proteins, in the generation and maintenance of hematopoietic cells during embryogenesis. In the adulthood, m6A has also been demonstrated to be involved in the functional maintenance of hematopoietic stem and progenitor cells in the bone marrow and umbilical cord blood, as well as the progression of malignant hematopoiesis. In this review, we focus on recent progress in identifying the biological functions of m6A mRNA modification, its regulators, and downstream gene targets during normal and pathological hematopoiesis. We propose that targeting m6A mRNA modification could offer novel insights into therapeutic development against abnormal and malignant hematopoietic cell development in the future., Competing Interests: Conflict of interest statement: The authors declare no conflict of insterest., (© The Author(s) 2023. Published by Oxford University Press on behalf of Society for Leukocyte Biology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Chimeric antigen receptors (CAR) are synthetic receptors engineered to target a user-defined antigen. They comprise an extracellular single-chain variable fragment for target recognition and intracellular signalling domains commonly derived from immune cells. CAR-T cells have proven to be successful in therapy of some cancers. CAR-T cells are activated upon antigen-priming and subsequent intracellular signalling. However, tonic signalling in CAR-T cells remains a challenge in developing CAR-T therapeutics of high efficacy as it causes early T-cell exhaustion, limiting therapeutic persistence. Moreover, a poor choice of target antigen leads to off-target cytotoxicity, often hampering the host's survival. In addition, conventional methods of delivering CAR gene circuits utilise viral vectors, such as lentiviruses and retroviruses, which insert the CAR gene circuits into transcriptionally active sites in the genome. This increases the risks of malignant transformation due to improper genome integration. Optimisation in CAR-T engineering, from the architecture of CAR gene circuits to the structure of CAR and the behaviour of CAR-T cells, is paramount to ensure high efficacy, persistence, and precision in CAR-T therapy. This review provides insights into engineering CAR-T cells for precision in cancer therapy by highlighting the key strategies recently developed to optimise the function and efficiency of CARs. The delivery method of CAR gene circuits, circuit and structural modification of CAR, T-cell phenotype manipulation and T-cell arming will be discussed to accentuate their interplay in regulating CAR-T therapy's safety, precision, and efficacy., (Copyright © 2024. Published by Elsevier Inc.)

We report the case of a 41-year-old man who presented with gross hematuria and a bladder tumor on ultrasonography. Magnetic resonance imaging indicated a possible muscle-invasive bladder cancer or urachal carcinoma. Following transurethral resection of the bladder tumor, histopathological findings revealed an adenocarcinoma similar to colorectal cancer. The patient was diagnosed with an urachal carcinoma in the urinary bladder dome. Since multiple lung metastases were observed on computed tomography, and his serum carcinoembryonic antigen level was 116 ng/dL, his final diagnosis was a stage IVb urachal carcinoma. He received 11 courses of mFOLOX6, and underwent a laparoscopy-assisted partial cystectomy and pelvic lymph node dissection. Pathological examination confirmed negative surgical margins, but remained tumor cells were confirmed. The patient continued mFOLFOX6 treatment for 12 months postoperatively, with no disease progression observed.

Transfection of proteins, mRNA, and chimeric antigen receptor (CAR) transgenes into immune cells remains a critical bottleneck in cell manufacturing. Current methods, such as viruses and bulk electroporation, are hampered by low transfection efficiency, unintended transgene integration, and significant cell perturbation. The Nanostraw Electro-actuated Transfection (NExT) technology offers a solution by using high aspect-ratio nanostraws and localized electric fields to precisely deliver biomolecules into cells with minimal disruption. We demonstrate that NExT can deliver proteins, polysaccharides, and mRNA into primary human CD8 + and CD4 + T cells, and achieve CRISPR/Cas9 gene knockout of CXCR4 and TRAC in CD8 + T cells. We showcase NExT's versatility across a range of primary human immune cells, including CD4 + T cells, γδ-T cells, dendritic cells, NK cells, T reg cells, macrophages, and neutrophils. Finally, we developed a scalable, high-throughput multiwell NExT system capable of transfecting over 14 million cells and delivering diverse cargoes into multiple cell types from various donors simultaneously. This technology holds promise for streamlining high-throughput screening of allogeneic donors and reducing optimization costs for large-scale CAR-immune cell transfection., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 Elsevier Ltd. All rights reserved.)

The rapid emergence of spatial transcriptomics (ST) technologies is revolutionizing our understanding of tissue spatial architecture and biology. Although current ST methods, whether based on next-generation sequencing (seq-based approaches) or fluorescence in situ hybridization (image-based approaches), offer valuable insights, they face limitations either in cellular resolution or transcriptome-wide profiling. To address these limitations, we present SpatialScope, a unified approach integrating scRNA-seq reference data and ST data using deep generative models. With innovation in model and algorithm designs, SpatialScope not only enhances seq-based ST data to achieve single-cell resolution, but also accurately infers transcriptome-wide expression levels for image-based ST data. We demonstrate SpatialScope's utility through simulation studies and real data analysis from both seq-based and image-based ST approaches. SpatialScope provides spatial characterization of tissue structures at transcriptome-wide single-cell resolution, facilitating downstream analysis, including detecting cellular communication through ligand-receptor interactions, localizing cellular subtypes, and identifying spatially differentially expressed genes., (© 2023. The Author(s).)

Programmed death ligand 1 (PD-L1) serves as a pivotal immune checkpoint in both the innate and adaptive immune systems. PD-L1 is expressed in macrophages in response to IFNγ. We examined whether PD-L1 might regulate macrophage development. We established PD-L1 KO ( CD274 -/- ) human pluripotent stem cells and differentiated them into macrophages and observed a 60% reduction in CD11B + CD45 + macrophages in CD274 -/- ; this was orthogonally verified, with the PD-L1 inhibitor BMS-1166 reducing macrophages to the same fold. Single-cell RNA sequencing further confirmed the down-regulation of the macrophage-defining transcription factors SPI1 and MAFB Furthermore, CD274 -/- macrophages reduced the level of inflammatory signals such as NF-κB and TNF, and chemokine secretion of the CXCL and CCL families. Anti-inflammatory TGF-β was up-regulated. Finally, we identified that CD274 -/- macrophages significantly down-regulated interferon-stimulated genes despite the presence of IFNγ in the differentiation media. These data suggest that PD-L1 regulates inflammatory programs of macrophages from human pluripotent stem cells., (© 2023 Cao et al.)

Haematological malignancies comprise a diverse set of lymphoid and myeloid neoplasms which can arise during any stage of haematopoiesis in the bone marrow. Accumulating evidence suggests that chronic inflammation generated by inflammatory cytokines secreted by tumour and the tumour-associated cells within the bone marrow microenvironment initiates signalling pathways in malignant cells, resulting in activation of master transcription factors including Smads, STAT3, and NF-κB which confer cancer stem cell phenotypes and drive disease progression. Deciphering the molecular mechanisms for how immune cells interact with malignant cells to induce such epigenetic modifications, specifically DNA methylation, histone modification, expression of miRNAs and lnRNAs to perturbate haematopoiesis could provide new avenues for developing novel targeted therapies for haematological malignancies. Here, the complex positive and negative feedback loops involved in inflammatory cytokine-induced cancer stem cell generation and drug resistance are reviewed to highlight the clinical importance of immune-epigenetic crosstalk in haematological malignancies., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Wong and Sugimura.)

Cell differentiation is achieved by acquiring a cell type-specific transcriptional program and epigenetic landscape. While the cell type-specific patterning of enhancers has been shown to precede cell fate decisions, it remains unclear how regulators of these enhancers are induced to initiate cell specification and how they appropriately restrict cells that differentiate. Here, using embryonic stem cell-derived hematopoietic cell differentiation cultures, we show the activation of some hematopoietic enhancers during arterialization of hemogenic endothelium, a prerequisite for hematopoiesis. We further reveal that ZEB2, a factor involved in the transcriptional regulation of arterial endothelial cells, and a hematopoietic regulator MEIS1 are independently required for activating these enhancers. Concomitantly, ZEB2 or MEIS1 deficiency impaired hematopoietic cell development. These results suggest that multiple regulators expressed from an earlier developmental stage non-redundantly contribute to the establishment of hematopoietic enhancer landscape, thereby restricting cell differentiation despite the unrestricted expression of these regulators to hematopoietic cells., Competing Interests: The authors declare no conflict of interest., (© 2023 The Authors.)

Adoptive cell therapies (ACT) based on chimeric antigen receptor (CAR)-modified immune cells have made great progress with six CAR-T cell products approved by the U.S. FDA for hematological malignancies. Compared with CAR-T cells, CAR-NK cells have attracted increasing attention owing to their multiple killing mechanisms, higher safety profile, and broad sources. Induced pluripotent stem cell (iPSC)-derived NK (iPSC-NK) cells possess a mature phenotype and potent cytolytic activity, and can provide a homogeneous population of CAR-NK cells that can be expanded to clinical scale. Thus, iPSC-derived CAR-NK (CAR-iNK) cells could be used as a standardized and "off-the-shelf" product for cancer immunotherapy. In this review, we summarize the current status of the manufacturing techniques, genetic modification strategies, preclinical and clinical evidence of CAR-iNK cells, and discuss the challenges and future prospects of CAR-iNK cell therapy as a novel cellular immunotherapy in cancer., Competing Interests: Declaration of Competing Interest We declare no competing interests., (Copyright © 2023 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Hematopoietic stem cells (HSCs) are multipotent progenitor cells that can differentiate into various mature blood cells and immune cells, thus reconstituting hematopoiesis. By taking advantage of the tremendous potential of HSCs, varied hereditary and hematologic diseases are promised to be alleviated or cured. To solve the contradiction between the growing demand for HSCs in disease treatment and the low population of HSCs in both cord blood and bone marrow, ex vivo HSC expansion along with multiple protocols has been investigated for harvesting adequate HSCs over the past two decades. This review surveys the state-of-the-art techniques for ex vivo HSC self-renewal and provides a concise summary of the effects of diverse intrinsic and extrinsic factors on the expansion of HSCs. The remaining challenges and emerging opportunities in the field of HSC expansion are also presented., Competing Interests: Declaration of competing interest Authors declare no conflict of interest., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Adaptive cellular immunotherapy, especially chimeric antigen receptor-T (CAR-T) cell therapy, has advanced the treatment of hematological malignancy. However, major limitations still remain in the source of cells comes from the patients themselves. The use of human pluripotent stem cells to differentiate into immune cells, such as T cells, NK cells, and macrophages, then arm with chimeric antigen receptor (CAR) to enhance tumor killing has gained major attention. It is expected to solve the low number of immune cells recovery from patients, long waiting periods, and ethical issues(reprogramming somatic cells to produce induced pluripotent stem cells (iPS cells) avoids the ethical issues unique to embryonic stem cells (Lo and Parham, 2009). However, there are still major challenges to be further solved. This review summarizes the progress, challenges, and future direction in human pluripotent stem cell-based immunotherapy., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

The differentiation of natural killer (NK) cells from human pluripotent stem cells allows for research on and the manufacture of clinical-grade cellular products for immunotherapy. Described here is a two-phase protocol that uses a serum-free commercial medium and a cocktail of cytokines (interleukin [IL]-3, IL-7, IL-15, stem cell factor [SCF], and FMS-like tyrosine kinase 3 ligand [Ftl3L]) to differentiate human expanded potential stem cells (hEPSCs) into cells that possess NK cell properties in vitro with both 3-dimensional (3D) and 2-dimensional (2D) culture technology. Following this protocol, CD3-CD56+ or CD45+CD56+ NK cells are consistently generated. When cocultured with tumor targets for 3 h, the differentiated products display mild cytotoxicity as compared to an IL-2-independent permanent cell line, NK92mi cells. The protocol preserves the complexity of the differentiation microenvironment by the generation of 3D structures, thus facilitating the study of the spatial relationships between immune cells and their niches. Meanwhile, the 2D culture system enables the routine phenotypical validation of cell differentiation without harming the delicate differentiation niche.

Valid and relevant models are critical for research to have biological relevance or to proceed in the right path. As well-established two-dimensional cell cultures lack niches and cues and rodent models differ in species, three-dimensional organoids emerged as a powerful platform for research. Cultured in vitro from stem cells, organoids are heterogeneous in cells and closely resemble the in vivo settings. Organoids also recapitulate the unique human features if cultured from a human source and are subjected to genetic modification. However, one type of organoid possesses only a limited selection of cells. In particular, the absence of vasculature and immune cells restricts the organoids from nutrition, cues, or critical interactions, undermining the validity of organoids as physiological or pathological models. To fill the current gap, there is an urgent need to provide organoids with vasculature and immune cells. In this paper, we review the methods to generate physiological and pathological organoid models and summarize ways to vascularize or immunize them. Our discussion continues with some advantages and disadvantages of each method and some emerging solutions to current problems., (© 2023 The Author(s). Published by S. Karger AG, Basel.)

The use of natural killer (NK) cells in cancer immunotherapy demonstrates promising potential, yet its efficacy is often limited due to the loss of tumor-killing capacity and lack of specificity in vivo. Here, we review current approaches to confer enhanced tumor-killing capacity and specificity by genetic engineering. Increasing sensitivity to cytokines and protecting NK cells from the immune checkpoint endowed sustainability of NK cells in the tumor microenvironment. Transducing chimeric antigen receptor (CAR) in NK cells successfully targeted both hematologic and solid tumors in preclinical models. The use of human pluripotent stem cells as an expandable and genetically amenable platform offers a stable source of engineered NK cells for cancer immunotherapy. We highlight that CAR-NK cells from human pluripotent stem cells are a promising approach for cancer immunotherapy., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2022. Published by The Company of Biologists Ltd.)

Tumor tissue comprises a highly complex network of diverse cell types. The tumor microenvironment (TME) can be mainly subdivided into cancer cells and stromal cell compartments, the latter include different types of immune cells, fibroblasts, endothelial cells, and pericytes. Tumor cells reprogram immune cells and other stromal cells in the TME to constrain their antitumor capacity by creating an immunosuppressive milieu and metabolism competition. Moreover, the reprogramming effect on immune cells is localized not only in the tumor but also at the systemic level. With wide application of single-cell sequencing technology, tumor-specific characteristics of immune cells and other stromal cells in the TME have been dissected. In this review, we mainly focus on how tumor cells reprogram immune cells both within the TME and peripheral blood. This information can further help us to improve the efficiency of current immunotherapy as well as bring up new ideas to combat cancer.

Chimeric antigen receptors (CAR) T cells (CAR-T) mark a significant step towards producing safe and effective personal anticancer treatments. CAR-T strategies engineers the T cells from the patients to allow specific binding to a tumour-specific antigen. CAR-Ts are a second-wave offensive strategy to clear out remaining chemotherapy-resistant tumour cells. Though showing practical antitumor abilities in multiple haematological malignancies and solid tumour cancers, the issues of antigen escape, tumour infiltration/penetration, and toxicity side effects limit the usage of prolonged CAR-T therapies. However, engineering immunology has exploited human stem cell-based CAR-T therapies and the development of CAR-M (macrophage) therapies to combat the disadvantages of conventional CAR-T therapies. In this review, we will highlight the challenges of CAR-T therapies and combat them with engineering immunology for cancer immunotherapy., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Wang, Ting Cheung and Sugimura.)

MicroRNA-124a (miR-124a) is one of the most abundantly expressed microRNAs in the central nervous system and is encoded in mammals by the three genomic loci miR-124a-1/2/3; however, its in vivo roles in neuronal development and function remain ambiguous. In the present study, we investigated the effect of miR-124a loss on neuronal differentiation in mice and in embryonic stem (ES) cells. Since miR-124a-3 exhibits only background expression levels in the brain and we were unable to obtain miR-124a-1/2/3 triple knockout (TKO) mice by mating, we generated and analyzed miR-124a-1/2 double knockout (DKO) mice. We found that these DKO mice exhibit perinatal lethality. RNA-seq analysis demonstrated that the expression levels of proneural and neuronal marker genes were almost unchanged between the control and miR-124a-1/2 DKO brains; however, genes related to neuronal synaptic formation and function were enriched among downregulated genes in the miR-124a-1/2 DKO brain. In addition, we found the transcription regulator Tardbp/TDP-43, loss of which leads to defects in neuronal maturation and function, was inactivated in the miR-124a-1/2 DKO brain. Furthermore, Tardbp knockdown suppressed neurite extension in cultured neuronal cells. We also generated miR-124a-1/2/3 TKO ES cells using CRISPR-Cas9 as an alternative to TKO mice. Phase-contrast microscopic, immunocytochemical, and gene expression analyses showed that miR-124a-1/2/3 TKO ES cell lines were able to differentiate into neurons. Collectively, these results suggest that miR-124a plays a role in neuronal maturation rather than neurogenesis in vivo and advance our understanding of the functional roles of microRNAs in central nervous system development., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

The purpose of this study was to investigate the protective effect of a glass ionomer cement (GIC) consisting of fluoro-zinc-silicate glass on the demineralization of bovine dentin using the ultrasonic pulse-echo method. The findings were compared with those obtained using a conventional GIC. Slabs of dentin from bovine teeth were sliced, shaped into a rectangular form, and immersed in 0.5 M ethylenediaminetetraacetic acid solution at 25ºC for 6 days, followed by storage in distilled water for 3 days. After demineralization, they were immersed in artificial saliva with and without the GIC specimens. The propagation times of the longitudinal ultrasonic waves in the samples were measured. The ultrasonic velocities of the fluoro-zinc-silicate glass-containing GICs were significantly increased 2-3 days after the start of the experiment and showed an upward trend thereafter. These findings indicate that the GICs containing fluoro-zinc-silicate glass might exhibit the potential to promote remineralization in the dentin.

R-loops are prevalent in mammalian genomes and involved in many fundamental cellular processes. Depletion of BRCA2 leads to aberrant R-loop accumulation, contributing to genome instability. Here, we show that ZFP281 cooperates with BRCA2 in preventing R-loop accumulation to facilitate DNA replication in embryonic stem cells. ZFP281 depletion reduces PCNA levels on chromatin and impairs DNA replication. Mechanistically, we demonstrate that ZFP281 can interact with BRCA2, and that BRCA2 is enriched at G/C-rich promoters and requires both ZFP281 and PRC2 for its proper recruitment to the bivalent chromatin at the genome-wide scale. Furthermore, depletion of ZFP281 or BRCA2 leads to accumulation of R-loops over the bivalent regions, and compromises activation of the developmental genes by retinoic acid during stem cell differentiation. In summary, our results reveal that ZFP281 recruits BRCA2 to the bivalent chromatin regions to ensure proper progression of DNA replication through preventing persistent R-loops., (© 2022. The Author(s).)

The tumor microenvironment encompasses various innate immune cells which regulate tumor progression. Exploiting innate immune cells is a new frontier of cancer immunotherapy. However, the classical surface markers for cell-type classification cannot always well-conclude the phenotype, which will further hinge our understanding. The innate immune cells include dendritic cells, monocytes/macrophages, natural killer cells, and innate lymphoid cells. They play important roles in tumor growth and survival, in some cases promoting cancer, in other cases negating cancer. The precise characterization of innate immune cells at the single-cell level will boost the potential of cancer immunotherapy. With the development of single-cell RNA sequencing technology, the transcriptome of each cell in the tumor microenvironment can be dissected at a single-cell level, which paves a way for a better understanding of the cell type and its functions. Here, we summarize the subtypes and functions of innate immune cells in the tumor microenvironment based on recent literature on single-cell technology. We provide updates on recent achievements and prospects for how to exploit novel functions of tumor-associated innate immune cells and target them for cancer immunotherapy., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Sugimura and Chao.)

Innate Lymphoid Cells (ILCs) are an elusive type of innate immune cell that was only discovered recently. Their tissue residency and dependency makes them a niche group of cells that bridge the adaptive and innate immune system. The nomenclature and classification of ILCs have been challenging due to their heterogeneity. The currently agreed ILC classification splits the cells into two categories including cytotoxic and helper ILCs. The tumour microenvironment is often hostile for immune cells. Remodeling the microenvironment and regulating other immune cells-achieved by ILCs-can enhance anti-tumor effects. How ILCs regulate other immune cells in the tumor microenvironment remains to be understood. Here we review current understanding of the role of ILCs in the tumor microenvironment. ILCs recruit CD8 positive T and memory T cells in PDAC, ILCs are also able to help CD108 positive B cells migrate toward tumour locations. In NSCLC, ILC3s are seen helping resident macrophages enhancing the mucus immunity to cancer cells. We then highlight the roles of cytokines and immune checkpoint pathways in ILCs and its implication in immunotherapy., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Sugimura and Wang.)

The tumor microenvironment (TME) is a complex milieu consisting of lymphoid cells, myeloid cells, fibroblasts, and multiple molecules, which play a key role in tumor progression and immunotherapy. TME is characterized by immune-suppressive features, which release anti-inflammatory cytokines such as IL-4 and TGFβ to skew the T cells to a Th2 state as well to polarize tumor-associated macrophages (TAMs) to an anti-inflammatory phenotype to curb the immunotherapy. Considering the heterogeneity of the TME and its role in determining response to chimeric antigen receptor (CAR)-T cells, delineating TME at a single-cell level will provide useful information for cancer treatment. First, we discuss cellular and molecular features that curb the response to CAR-T cells, for example, high expression of immune checkpoint molecules (PD-1, LAG3) and anti-inflammatory cytokines (IL-4, TGFb) that block CAR-T cell function. Then, we summarize how newly invented single-cell technologies such as spatial multi-omics would benefit the understanding of cancer immunotherapy. Finally, we will further describe recent attempts of CAR-T to remodel TME by arming the CAR-T with anti-PD-1 single-chain variants or Th1 triggering cytokines (such as IL-7, IL-12) to remodel TME into a pro-inflammatory state. Herein, we review the single-cell-level signatures of TME and the strategies of CAR-T to remodel TME., (© 2022. The Author(s), under exclusive license to Springer Nature Switzerland AG.)

Autologous chimeric antigen receptor (CAR) T cells have expanded the scope and therapeutic potential of anti-cancer therapy. Nevertheless, autologous CAR-T therapy has been challenging due to labor some manufacturing processes for every patient, and the cost due to the complexity of the process. Moreover, T cell dysfunction results from the immunosuppressive tumor microenvironment in certain patients. Considering technical challenges in autologous donors, the development of safe and efficient allogeneic CAR-T therapy will address these issues. Since the advent of the generation of immune cells from pluripotent stem cells (PSCs), numerous studies focus on the off-the-shelf generation of CAR-immune cells derived from the universal donor PSCs, which simplifies the manufacturing process and standardizes CAR-T products. In this review, we will discuss advances in the generation of immune cells from PSCs, together with the potential and perspectives of CAR-T, CAR-macrophages, and CAR-natural killer (NK) cells in cancer treatment. The combination of PSC-derived immune cells and CAR engineering will pave the way for developing next-generation cancer immunotherapy., (© 2022. The Author(s), under exclusive license to Springer Nature Switzerland AG.)

A 71-year-old man with a history of hoarseness and right upper extremity numbness was referred to our department for evaluation of an intrathoracic mass that was detected on chest radiography and a right kidney tumor observed on computed tomography (CT). Histopathological examination of percutaneous kidney biopsy and bronchoscopic lung biopsy specimens revealed renal clear cell carcinoma with multiple lung metastases. The patient showed a poor risk based on the International Metastatic renal cell carcinoma Database Consortium score, and nivolumab plus ipilimumab were initiated as first-line therapy. His symptoms gradually improved, following four courses of nivolumab plus ipilimumab treatment, and CT revealed shrinkage of all lesions. However, he developed diarrhea, rash, anemia, and elevated serum C-reactive protein levels (CRP) following this therapy. Diarrhea and rash were considered immune-related adverse events, and he was treated with oral prednisolone and topical corticosteroid. Nivolumab administration was discontinued because anemia worsened together with elevated serum CRP levels despite improvement in diarrhea. He subsequently developed constipation and abdominal bloating, following further treatment for 4 months. CT revealed intestinal tumor-induced intussusception, necessitating partial resection of the small intestinal tumor, which was histopathologically diagnosed as metastases. Both anemia and elevated CRP improved postoperatively. Currently, all metastatic lesions other than the resected intestine have continued to respond to treatment over 12 months after initiation of nivolumab plus ipilimumab therapy.

Hematopoietic stem cells (HSCs) play a core role in blood development. The ability to efficiently produce HSCs from various pluripotent stem cell sources is the Holy Grail in the hematology field. However, in vitro or in vivo HSC production remains low, which may be attributable to the lack of understanding of hematopoiesis. Here, we review the recent progress in this area and introduce advanced technologies, such as single-cell RNA-seq, spatial transcriptomics, and molecular barcoding, which may help to acquire missing information about HSC generation. We finally discuss unresolved questions, the answers to which may be conducive to HSC production, providing a promising path toward HSC-based immunotherapies.

Purpose: To examine the effects of an ion-releasing filler-containing gel on the remineralization of dentin using optical coherence tomography (OCT)., Methods: Dentin slabs of bovine teeth were sliced and shaped into a rectangular form. Specimens were treated with undersaturated 0.1 M lactic acid buffer solution (pH 4.75) for 10 minutes and then placed in artificial saliva (pH 7.0). This procedure was repeated three times a day for 28 days. The dentin remineralization effects of a fluoride/S-PRG filler-containing gel (PRG) and a 38% SDF solution (SDF) on dentin slabs of bovine teeth were compared. After treatment, the dentin slabs were immersed in a 0.1 M lactic acid buffer solution and then placed in artificial saliva. This procedure was repeated three times a day for 28 days. OCT imaging was conducted on the selected location of the dentin surface. The peak intensity and width at 1/e² were recorded in each of the six areas on the sample and averaged. Each group had a sample size of 10. Knoop hardness number (KHN) measurements and scanning electron microscopy (SEM) observations were also conducted. The data for each group were subjected to a one-way repeated-measures ANOVA and Tukey tests (α= 0.05). The samples were also observed using SEM., Results: The peak signal intensities of SDF and PRG decreased on day 7 and then slightly increased during the experimental period for the one-off application groups and then decreased for frequent-time application groups. Although the width at 1/e² in the untreated specimens decreased over the test period, SDF and PRG for the one-off application groups exhibited an increase in widths on day 7 followed by a slight decrease, whereas it increased for the frequent-time application groups. The average KHN of the dentin samples exhibited the same tendency as the width at 1/e². Closure of the dentin tubules and crystal precipitation were detected on the surface of both SDF and PRG groups., Clinical Significance: S-PRG filler-containing gel might have the ability to prevent dentin demineralization and could be useful for the prevention of hard-to-access lesions. This material achieved remineralization of the demineralized root dentin and had the same remineralization ability as SDF in vitro., Competing Interests: The authors certify that they have no proprietary, financial, or other personal interest of any nature or kind in any product, service, and/or company that is presented in this article. This work was supported, in part, by Nakao Foundation, Grants-in-Aid for Scientific Research (C) 19K10158, 19K10159, from the Japan Society for the Promotion of Science. This project was also supported, in part, by the Sato Fund, Uemura Yasuo-Haruko Fund, and by a grant from the Dental Research Center of Nihon University School of Dentistry, Japan., (Copyright©American Journal of Dentistry.)