Macrophage hitchhiking for systematic suppression in postablative multifocal HCC.

BACKGROUND AND AIMS: HCC, particularly the multifocal HCC, features aggressive invasion and dismal prognosis. Locoregional treatments were often refractory to eliminate tumor tissue, resulting in residual tumor cells persisting and subsequent progression. Owing to problematic delivery to the tumor tissue, systemic therapies, such as lenvatinib (LEN) therapy, show limited clinical benefit in preventing residual tumor progression. Therefore, more advanced strategies for postablative multifocal HCC are urgently needed. APPROACH AND RESULTS: Motivated by the chemotaxis in tumor penetration of macrophages, we report a strategy named microinvasive ablation-guided macrophage hitchhiking for the targeted therapy toward HCC. In this study, the strategy leverages the natural inflammatory gradient induced by ablation to guide LEN-loaded macrophages toward tumor targeting, which increased by ~10-fold the delivery efficiency of LEN in postablative HCC in vivo. Microinvasive ablation-guided macrophage hitchhiking has demonstrated significant antitumor activity in various HCC models, including the hydrodynamic tail vein injection multifocal HCC mouse model and the orthotopic xenograft HCC rabbit model, systematically inhibiting residual tumor progression after ablation and prolonging the median survival of tumor-bearing mice. The potential antitumor mechanism was explored using techniques such as flow cytometry, ELISA, and immunohistochemistry. We found that the strategy significantly suppressed tumor cell proliferation and neovascularization, and such enhanced delivery of LEN stimulated systemic immune responses and induced durable immune memory. CONCLUSIONS: The macrophage hitchhiking strategy demonstrates exceptional therapeutic efficacy and biosafety across various species, offering promising prospects for clinical translation in controlling residual tumor progression and improving outcomes following HCC ablation.

Development of novel reagents to chicken FLT3, XCR1 and CSF2R for the identification and characterization of avian conventional dendritic cells.

Conventional dendritic cells (cDC) are bone marrow-derived immune cells that play a central role in linking innate and adaptive immunity. cDCs efficiently uptake, process and present antigen to naïve T cells, driving clonal expansion of antigen-specific T-cell responses. In chicken, vital reagents are lacking for the efficient and precise identification of cDCs. In this study, we have developed several novel reagents for the identification and characterization of chicken cDCs. Chicken FLT3 cDNA was cloned and a monoclonal antibody to cell surface FLT3 was generated. This antibody identified a distinct FLT3HI splenic subset which lack expression of signature markers for B cells, T cells or monocyte/macrophages. By combining anti-FLT3 and CSF1R-eGFP transgenic expression, three major populations within the mononuclear phagocyte system were identified in the spleen. The cDC1 subset of mammalian cDCs express the chemokine receptor XCR1. To characterize chicken cDCs, a synthetic chicken chemokine (C motif) ligand (XCL1) peptide conjugated to Alexa Fluor 647 was developed (XCL1AF647 ). Flow cytometry staining of XCL1AF647 on splenocytes showed that all chicken FLT3HI cells exclusively express XCR1, supporting the hypothesis that this population comprises bona fide chicken cDCs. Further analysis revealed that chicken cDCs expressed CSF1R but lacked the expression of CSF2R. Collectively, the cell surface phenotypes of chicken cDCs were partially conserved with mammalian XCR1+ cDC1, with distinct differences in CSF1R and CSF2R expression compared with mammalian orthologues. These original reagents allow the efficient identification of chicken cDCs to investigate their important roles in the chicken immunity and diseases.

Characterization of Subpopulations of Chicken Mononuclear Phagocytes That Express TIM4 and CSF1R.

The phosphatidylserine receptor TIM4, encoded by TIMD4, mediates the phagocytic uptake of apoptotic cells. We applied anti-chicken TIM4 mAbs in combination with CSF1R reporter transgenes to dissect the function of TIM4 in the chick (Gallus gallus). During development in ovo, TIM4 was present on the large majority of macrophages, but expression became more heterogeneous posthatch. Blood monocytes expressed KUL01, class II MHC, and CSF1R-mApple uniformly. Around 50% of monocytes were positive for surface TIM4. They also expressed many other monocyte-specific transcripts at a higher level than TIM4- monocytes. In liver, highly phagocytic TIM4hi cells shared many transcripts with mammalian Kupffer cells and were associated with uptake of apoptotic cells. Although they expressed CSF1R mRNA, Kupffer cells did not express the CSF1R-mApple transgene, suggesting that additional CSF1R transcriptional regulatory elements are required by these cells. By contrast, CSF1R-mApple was detected in liver TIM4lo and TIM4- cells, which were not phagocytic and were more abundant than Kupffer cells. These cells expressed CSF1R alongside high levels of FLT3, MHCII, XCR1, and other markers associated with conventional dendritic cells in mice. In bursa, TIM4 was present on the cell surface of two populations. Like Kupffer cells, bursal TIM4hi phagocytes coexpressed many receptors involved in apoptotic cell recognition. TIM4lo cells appear to be a subpopulation of bursal B cells. In overview, TIM4 is associated with phagocytes that eliminate apoptotic cells in the chick. In the liver, TIM4 and CSF1R reporters distinguished Kupffer cells from an abundant population of dendritic cell-like cells.

Medical micro/nanorobots in complex media.

Medical micro/nanorobots have received tremendous attention over the past decades owing to their potential to be navigated into hard-to-reach tissues for a number of biomedical applications ranging from targeted drug/gene delivery, bio-isolation, detoxification, to nanosurgery. Despite the great promise, the majority of the past demonstrations are primarily under benchtop or in vitro conditions. Many developed micro/nanoscale propulsion mechanisms are based on the assumption of a homogeneous, Newtonian environment, while realistic biological environments are substantially more complex. Moving toward practical medical use, the field of micro/nanorobotics must overcome several major challenges including propulsion through complex media (such as blood, mucus, and vitreous) as well as deep tissue imaging and control in vivo. In this review article, we summarize the recent research efforts on investigating how various complexities in biological environments impact the propulsion of micro/nanoswimmers. We also highlight the emerging technological approaches to enhance the locomotion of micro/nanorobots in complex environments. The recent demonstrations of in vivo imaging, control and therapeutic medical applications of such micro/nanorobots are introduced. We envision that continuing materials and technological innovations through interdisciplinary collaborative efforts can bring us steps closer to the fantasy of "swallowing a surgeon".

Reconfigurable Assembly of Active Liquid Metal Colloidal Cluster.

We report the reconfigurable assembly of rod-shaped eutectic gallium-indium alloy (EGaIn) liquid metal colloidal motors by mimicking the growth behavior of a dandelion. EGaIn nanorods with a diameter of 210 nm and a length of 850 nm were synthesized via an ultrasound-assisted physical dispersion method. The nanorods possess a core-shell structure with a 30 nm GaOOH shell and zero-valent liquid core. The EGaIn motors move autonomously at a speed of 41.2 µm s-1 under an acoustic field. By modulating the frequency of the applied acoustic field, the EGaIn colloidal motors self-organize into various striped and circular patterns, followed by a flower-like cluster. The dandelion-like EGaIn colloidal motor clusters move collectively and redisperse when the applied acoustic frequency is changed. Numerical simulations reveal that the flower-like clusters are created by the acoustic propulsion in combination with steric repulsion and hydrodynamics.

Gold-Nanoshell-Functionalized Polymer Nanoswimmer for Photomechanical Poration of Single-Cell Membrane.

We report an ultrasound-driven gold-nanoshell-functionalized polymer multilayer tubular nanoswimmer that can photomechanically perforate the membrane of a cancer cell by assistance of near-infrared (NIR) light. The nanoswimmers were constructed by a template-assisted layer-by-layer technique and subsequent functionalization of Au nanoshells inside the big opening. The nanoswimmers exhibit efficient and controllable movement toward target cells through the manipulation of the acoustic field. Next, the nanoswimmers with end-on attachment onto the HeLa cells achieve the poration of the cell membrane within 0.1 s under the irradiation of NIR light. The experimental and theoretical results suggest that the instantaneous photothermal effect provides enough photomechanical force to open the cell membrane. Such NIR-light-assisted nanoswimmers-enabled cell membrane poration possesses various advantages including active targeting, short time, and precision in single cells that conventional chemical and physical cell poration techniques could not achieve and, thus, provides considerable promise in a variety of biomedical applications such as gene delivery and artificial insemination.

Macroscale Chemotaxis from a Swarm of Bacteria-Mimicking Nanoswimmers.

Inspired by the dynamics of bacterial swarming, we report a swarm of polymer-brush-grafted, glucose-oxidase-powered Janus gold nanoswimmers with a positive, macroscale chemotactic behavior. These nanoswimmers are prepared through the grafting of polymer brushes onto one side of gold nanoparticles, followed by functionalization with glucose oxidase on the other side. The resulting polymer-brush-functionalized Janus gold nanoswimmers exhibit efficient propulsion with a velocity of up to approximately 120 body lengths s-1 in the presence of glucose. The comparative analysis of their kinematic behavior reveals that the grafted polymer brushes significantly improve the translational diffusion of Janus gold nanoswimmers. Particularly, these bacteria-mimicking Janus gold nanoswimmers display a collectively chemotactic motion along the concentration gradient of a glucose resource, which could be observed at the macroscale.

Phenotypic and genetic variation in the response of chickens to Eimeria tenella induced coccidiosis.

BACKGROUND: Coccidiosis is a major contributor to losses in poultry production. With emerging constraints on the use of in-feed prophylactic anticoccidial drugs and the relatively high costs of effective vaccines, there are commercial incentives to breed chickens with greater resistance to this important production disease. To identify phenotypic biomarkers that are associated with the production impacts of coccidiosis, and to assess their covariance and heritability, 942 Cobb500 commercial broilers were subjected to a defined challenge with Eimeria tenella (Houghton). Three traits were measured: weight gain (WG) during the period of infection, caecal lesion score (CLS) post mortem, and the level of a serum biomarker of intestinal inflammation, i.e. circulating interleukin 10 (IL-10), measured at the height of the infection. RESULTS: Phenotypic analysis of the challenged chicken cohort revealed a significant positive correlation between CLS and IL-10, with significant negative correlations of both these traits with WG. Eigenanalysis of phenotypic covariances between measured traits revealed three distinct eigenvectors. Trait weightings of the first eigenvector, (EV1, eigenvalue = 59%), were biologically interpreted as representing a response of birds that were susceptible to infection, with low WG, high CLS and high IL-10. Similarly, the second eigenvector represented infection resilience/resistance (EV2, 22%; high WG, low CLS and high IL-10), and the third eigenvector tolerance (EV3, 19%; high WG, high CLS and low IL-10), respectively. Genome-wide association studies (GWAS) identified two SNPs that were associated with WG at the suggestive level. CONCLUSIONS: Eigenanalysis separated the phenotypic impact of a defined challenge with E. tenella on WG, caecal inflammation/pathology, and production of IL-10 into three major eigenvectors, indicating that the susceptibility-resistance axis is not a single continuous quantitative trait. The SNPs identified by the GWAS for body weight were located in close proximity to two genes that are involved in innate immunity (FAM96B and RRAD).

Highly Efficient Freestyle Magnetic Nanoswimmer.

The unique swimming strategies of natural microorganisms have inspired recent development of magnetic micro/nanorobots powered by artificial helical or flexible flagella. However, as artificial nanoswimmers with unique geometries are being developed, it is critical to explore new potential modes for kinetic optimization. For example, the freestyle stroke is the most efficient of the competitive swimming strokes for humans. Here we report a new type of magnetic nanorobot, a symmetric multilinked two-arm nanoswimmer, capable of efficient "freestyle" swimming at low Reynolds numbers. Excellent agreement between the experimental observations and theoretical predictions indicates that the powerful "freestyle" propulsion of the two-arm nanorobot is attributed to synchronized oscillatory deformations of the nanorobot under the combined action of magnetic field and viscous forces. It is demonstrated for the first time that the nonplanar propulsion gait due to the cooperative "freestyle" stroke of the two magnetic arms can be powered by a plane oscillatory magnetic field. These two-arm nanorobots are capable of a powerful propulsion up to 12 body lengths per second, along with on-demand speed regulation and remote navigation. Furthermore, the nonplanar propulsion gait powered by the consecutive swinging of the achiral magnetic arms is more efficient than that of common chiral nanohelical swimmers. This new swimming mechanism and its attractive performance opens new possibilities in designing remotely actuated nanorobots for biomedical operation at the nanoscale.

Histone deacetylase 5 promotes the migration and invasion of hepatocellular carcinoma via increasing the transcription of hypoxia-inducible factor-1α under hypoxia condition.

Hypoxia plays a critical role in the progression and metastasis of hepatocellular carcinoma by activating the key transcription factor, hypoxia-inducible factor-1. This study aims to identify the novel mechanisms underlying the dysregulation of hypoxia-inducible factor-1α in hepatocellular carcinoma. We found that histone deacetylase 5, a highly expressed histone deacetylase in hepatocellular carcinoma, strengthened the migration and invasion of hepatocellular carcinoma cells under hypoxia but not normoxia condition. Furthermore, histone deacetylase 5 induced the transcription of hypoxia-inducible factor-1α by silencing homeodomain-interacting protein kinase-2 expression, which was also dependent on hypoxia. And then knockdown of hypoxia-inducible factor-1α decreased the expressions of mesenchymal markers, N-cadherin, and Vimentin, as well as matrix metalloproteinases, MMP7 and MMP9; however, the epithelial marker, E-cadherin, increased. Phenotype experiments showed that the migration and invasion of hepatocellular carcinoma cells were impaired by knockdown of histone deacetylase 5 or hypoxia-inducible factor-1α but rescued when eliminating homeodomain-interacting protein kinase-2 in hepatocellular carcinoma cells, which suggested the critical role of histone deacetylase 5-homeodomain-interacting protein kinase-2-hypoxia-inducible factor-1α pathway in hypoxia-induced metastasis. Finally, clinical analysis confirmed the positive correlation between histone deacetylase 5 and hypoxia-inducible factor-1α in hepatocellular carcinoma specimens and a relatively poor prognosis for the patients with high levels of histone deacetylase 5 and hypoxia-inducible factor-1α. Taken together, our findings demonstrated a novel mechanism underlying the crosstalk between histone deacetylase 5 and hypoxia-inducible factor-1 in hepatocellular carcinoma.

Self-thermophoretic motion of controlled assembled micro-/nanomotors.

As artificial active colloids, micro-/nanomotors (MNMs) can convert energy from the environment into mechanical motion in different fluids, showing potential applications in diverse fields such as targeted drug delivery and photothermal therapy. However, chemical fuels for typical catalytic MNMs, e.g., hydrogen peroxide, are highly toxic to organisms, and thus fuel-free MNMs are required. Recently, we have developed near-infrared light (NIR) propelled MNMs through integrating plasmonic gold nanoshells into nanoparticles or layer-by-layer assemblies in an asymmetric manner. In this perspective, we give an account of self-thermophoresis motion of these NIR-powered MNMs. The design of the motor architectures, as well as the theoretical study on the propulsion mechanism, is highlighted. We believe that the insights into self-thermophoretic motion would pave the way to access powerful MNMs for future applications and to explore interesting collective behaviors of active matter.

Sequence of a complete chicken BG haplotype shows dynamic expansion and contraction of two gene lineages with particular expression patterns.

Many genes important in immunity are found as multigene families. The butyrophilin genes are members of the B7 family, playing diverse roles in co-regulation and perhaps in antigen presentation. In humans, a fixed number of butyrophilin genes are found in and around the major histocompatibility complex (MHC), and show striking association with particular autoimmune diseases. In chickens, BG genes encode homologues with somewhat different domain organisation. Only a few BG genes have been characterised, one involved in actin-myosin interaction in the intestinal brush border, and another implicated in resistance to viral diseases. We characterise all BG genes in B12 chickens, finding a multigene family organised as tandem repeats in the BG region outside the MHC, a single gene in the MHC (the BF-BL region), and another single gene on a different chromosome. There is a precise cell and tissue expression for each gene, but overall there are two kinds, those expressed by haemopoietic cells and those expressed in tissues (presumably non-haemopoietic cells), correlating with two different kinds of promoters and 5' untranslated regions (5'UTR). However, the multigene family in the BG region contains many hybrid genes, suggesting recombination and/or deletion as major evolutionary forces. We identify BG genes in the chicken whole genome shotgun sequence, as well as by comparison to other haplotypes by fibre fluorescence in situ hybridisation, confirming dynamic expansion and contraction within the BG region. Thus, the BG genes in chickens are undergoing much more rapid evolution compared to their homologues in mammals, for reasons yet to be understood.

Chemotaxis-Guided Hybrid Neutrophil Micromotors for Targeted Drug Transport.

Engineering self-propelled micromotors with good biocompatibility and biodegradability for actively seeking disease sites and targeted drug transport remains a huge challenge. In this study, neutrophils with intrinsic chemotaxis capability were transformed into self-guided hybrid micromotors by integrating mesoporous silica nanoparticles (MSNs) with high loading capability. To ensure the compatibility of neutrophil cells with drug-loaded MSNs, bacteria membranes derived from E. coli were coated on MSNs in advance by a camouflaging strategy. The resulting biohybrid micromotors inherited the characteristic chemotaxis capability of native neutrophils and could effectively move along the chemoattractant gradients produced by E. coli. Our studies suggest that this camouflaging approach, which favors the uptake of MSNs into neutrophils without loss of cellular activity and motility, could be used to construct synthetic nanoparticle-loaded biohybrid micromotors for advanced biomedical applications.

Light-Activated Active Colloid Ribbons.

We report a dynamic self-organization of self-propelled peanut-shaped hematite motors from non-equilibrium driving forces where the propulsion can be triggered by blue light. They result in one-dimensional, active colloid ribbons with a positive phototactic characteristic. The motion of colloid motors is ascribed to the diffusion-osmotic flow in a chemical gradient by the photocatalytic decomposition of hydrogen peroxide fuel. We show that self-propelled peanut-shaped colloids readily form one-dimensional, slithering ribbon structures under the out-of-equilibrium collisions. This self-organization intrinsically results from the competition among the osmotically driven motion, the phoretic attraction and the inherent magnetic moments. The giant size number fluctuation in colloid ribbons is observed above a critical point 4.1 % of the surface density of colloid motors. Such phototactic colloid ribbons may provide a model system to understand the emergence of function in biological systems and have potential to construct bioinspired active materials based on different active building blocks.

Functional annotation of the T-cell immunoglobulin mucin family in birds.

T-cell immunoglobulin and mucin (TIM) family molecules are cell membrane proteins, preferentially expressed on various immune cells and implicated in recognition and clearance of apoptotic cells. Little is known of their function outside human and mouse, and nothing outside mammals. We identified only two TIM genes (chTIM) in the chicken genome, putative orthologues of mammalian TIM1 and TIM4, and cloned the respective cDNAs. Like mammalian TIM1, chTIM1 expression was restricted to lymphoid tissues and immune cells. The gene chTIM4 encodes at least five splice variants with distinct expression profiles that also varied between strains of chicken. Expression of chTIM4 was detected in myeloid antigen-presenting cells, and in γδ T cells, whereas mammalian TIM4 is not expressed in T cells. Like the mammalian proteins, chTIM1 and chTIM4 fusion proteins bind to phosphatidylserine, and are thereby implicated in recognition of apoptotic cells. The chTIM4-immunoglobulin fusion protein also had co-stimulatory activity on chicken T cells, suggesting a function in antigen presentation.

Near Infrared Light-Powered Janus Mesoporous Silica Nanoparticle Motors.

We describe fuel-free, near-infrared (NIR)-driven Janus mesoporous silica nanoparticle motors (JMSNMs) with diameters of 50, 80, and 120 nm. The Janus structure of the JMSNMs is generated by vacuum sputtering of a 10 nm Au layer on one side of the MSNMs. Upon exposure to an NIR laser, a localized photothermal effect on the Au half-shells results in the formation of thermal gradients across the JMSNMs; thus, the generated self-thermophoresis can actively drive the nanomotors to move at an ultrafast speed, for instance, up to 950 body lengths/s for 50 nm JMSNMs under an NIR laser power of 70.3 W/cm(2). The reversible "on/off" motion of the JMSNMs and their directed movement along the light gradient can be conveniently modulated by a remote NIR laser. Moreover, dynamic light scattering measurements are performed to investigate the coexisting translational and rotational motion of the JMSNMs in the presence of both self-thermophoretic forces and strong Brownian forces. These NIR-powered nanomotors demonstrate a novel strategy for overcoming the necessity of chemical fuels and exhibit a significant improvement in the maneuverability of nanomotors while providing potential cargo transportation in a biofriendly manner.

Recent Progress on Bioinspired Self-Propelled Micro/Nanomotors via Controlled Molecular Self-Assembly.

The combination of bottom-up controllable self-assembly technique with bioinspired design has opened new horizons in the development of self-propelled synthetic micro/nanomotors. Over the past five years, a significant advances toward the construction of bioinspired self-propelled micro/nanomotors has been witnessed based on the controlled self-assembly technique. Such a strategy permits the realization of autonomously synthetic motors with engineering features, such as sizes, shapes, composition, propulsion mechanism, and function. The construction, propulsion mechanism, and movement control of synthetic micro/nanomotors in connection with controlled self-assembly in recent research activities are summarized. These assembled nanomotors are expected to have a tremendous impact on current artificial nanomachines in future and hold potential promise for biomedical applications including drug targeted delivery, photothermal cancer therapy, biodetoxification, treatment of atherosclerosis, artificial insemination, crushing kidney stones, cleaning wounds, and removing blood clots and parasites.

Superfast Near-Infrared Light-Driven Polymer Multilayer Rockets.

A gold nanoshell-functionalized polymer multilayer nanorocket performs self-propulsion upon the irradiation with NIR light in the absence of chemical fuel. Theoretical simulations reveal that the NIR light-triggered self-thermophoresis drives the propulsion of the nanorocket. The nanorocket also displays -efficient NIR light-triggered propulsion in -biofluids and thus holds considerable promise for various potential biomedical applications.

Stem Cell Membrane-Coated Nanogels for Highly Efficient In Vivo Tumor Targeted Drug Delivery.

Stem cell membrane-coated nanogels can effectively evade clearance of the immune system, enhance the tumor targeting properties and antitumor chemotherapy efficacy of gelatin nanogels loaded doxorubicin in mice.

Catalytic Polymer Multilayer Shell Motors for Separation of Organics.

A catalytic polymer multilayer shell motor has been developed, which effects fast motion-based separation of charged organics in water. The shell motors are fabricated by sputtering platinum onto the exposed surface of silica templates embedded in Parafilm, followed by layer-by-layer assembly of polyelectrolyte multilayers to the templates. The catalytic shell motors display high bubble propulsion with speeds of up to 260 µm s(-1) (13 body lengths per second). Moreover, the polyelectrolyte multilayers assembled at high pH (pH>9.0) adsorb approximately 89% of dye molecules from water, owing to the electrostatic interaction between the positively charged polymers and the anionic dye molecules, and subsequently release them at neutral pH in a microfluidic device. The efficient propulsion coupled with the effective adsorption behavior of the catalytic shell motors in a microfluidic device results in accelerated separation of organics in water and thus holds considerable promise for water analysis.