Phenotypic and spatial heterogeneity of CD8+ tumour infiltrating lymphocytes.

CD8+ T cells are the workhorses executing adaptive anti-tumour response, and targets of various cancer immunotherapies. Latest advances have unearthed the sheer heterogeneity of CD8+ tumour infiltrating lymphocytes, and made it increasingly clear that the bulk of the endogenous and therapeutically induced tumour-suppressive momentum hinges on a particular selection of CD8+ T cells with advantageous attributes, namely the memory and stem-like exhausted subsets. A scrutiny of the contemporary perception of CD8+ T cells in cancer and the subgroups of interest along with the factors arbitrating their infiltration contextures, presented herein, may serve as the groundwork for future endeavours to probe further into the regulatory networks underlying their differentiation and migration, and optimise T cell-based immunotherapies accordingly.

Colorectal polyp segmentation with denoising diffusion probabilistic models.

Early detection of polyps is essential to decrease colorectal cancer(CRC) incidence. Therefore, developing an efficient and accurate polyp segmentation technique is crucial for clinical CRC prevention. In this paper, we propose an end-to-end training approach for polyp segmentation that employs diffusion model. The images are considered as priors, and the segmentation is formulated as a mask generation process. In the sampling process, multiple predictions are generated for each input image using the trained model, and significant performance enhancements are achieved through the use of majority vote strategy. Four public datasets and one in-house dataset are used to train and test the model performance. The proposed method achieves mDice scores of 0.934 and 0.967 for datasets Kvasir-SEG and CVC-ClinicDB respectively. Furthermore, one cross-validation is applied to test the generalization of the proposed model, and the proposed methods outperformed previous state-of-the-art(SOTA) models to the best of our knowledge. The proposed method also significantly improves the segmentation accuracy and has strong generalization capability.

A SAW-Based Programmable Controlled RNA Detecting Device: Rapid In Situ Cytolysis-RNA Capture-RNA Release-PCR in One Mini Chamber.

Viral RNA detection is crucial in preventing and treating early infectious diseases. Traditional methods of RNA detection require a large amount of equipment and technical personnel. In this study, proposed a programmable controlled surface acoustic wave (SAW)-based RNA detecting device has been proposed. The proposed device can perform the entire viral RNA detection process, including cell lysis by cell-microparticle collision through SAW-induced liquid whirling, RNA capture by SAW-suspended magnetic beads, RNA elution through SAW-induced high streaming force, and PCR thermal cycling through SAW-generated heat. The device has completed all RNA detection steps in one mini chamber, requiring only 489 µl reagents for RNA extraction, much smaller than the amount used in manual RNA extraction (2065 µl). The experimental results have shown that PCR results from the device are comparable to those achieved via commercial qPCR instrumental detection. This work has demonstrated the potential of SAW-based lab-on-a-chip devices for point-of-care testing and provided a novel approach for rapidly detecting infectious diseases.

Multi-scale nested UNet with transformer for colorectal polyp segmentation.

BACKGROUND: Polyp detection and localization are essential tasks for colonoscopy. U-shape network based convolutional neural networks have achieved remarkable segmentation performance for biomedical images, but lack of long-range dependencies modeling limits their receptive fields. PURPOSE: Our goal was to develop and test a novel architecture for polyp segmentation, which takes advantage of learning local information with long-range dependencies modeling. METHODS: A novel architecture combining with multi-scale nested UNet structure integrated transformer for polyp segmentation was developed. The proposed network takes advantage of both CNN and transformer to extract distinct feature information. The transformer layer is embedded between the encoder and decoder of a U-shape net to learn explicit global context and long-range semantic information. To address the challenging of variant polyp sizes, a MSFF unit was proposed to fuse features with multiple resolution. RESULTS: Four public datasets and one in-house dataset were used to train and test the model performance. Ablation study was also conducted to verify each component of the model. For dataset Kvasir-SEG and CVC-ClinicDB, the proposed model achieved mean dice score of 0.942 and 0.950 respectively, which were more accurate than the other methods. To show the generalization of different methods, we processed two cross dataset validations, the proposed model achieved the highest mean dice score. The results demonstrate that the proposed network has powerful learning and generalization capability, significantly improving segmentation accuracy and outperforming state-of-the-art methods. CONCLUSIONS: The proposed model produced more accurate polyp segmentation than current methods on four different public and one in-house datasets. Its capability of polyps segmentation in different sizes shows the potential clinical application.

Multiple influence of immune cells in the bone metastatic cancer microenvironment on tumors.

Bone is a common organ for solid tumor metastasis. Malignant bone tumor becomes insensitive to systemic therapy after colonization, followed by poor prognosis and high relapse rate. Immune and bone cells in situ constitute a unique immune microenvironment, which plays a crucial role in the context of bone metastasis. This review firstly focuses on lymphatic cells in bone metastatic cancer, including their function in tumor dissemination, invasion, growth and possible cytotoxicity-induced eradication. Subsequently, we examine myeloid cells, namely macrophages, myeloid-derived suppressor cells, dendritic cells, and megakaryocytes, evaluating their interaction with cytotoxic T lymphocytes and contribution to bone metastasis. As important components of skeletal tissue, osteoclasts and osteoblasts derived from bone marrow stromal cells, engaging in 'vicious cycle' accelerate osteolytic bone metastasis. We also explain the concept tumor dormancy and investigate underlying role of immune microenvironment on it. Additionally, a thorough review of emerging treatments for bone metastatic malignancy in clinical research, especially immunotherapy, is presented, indicating current challenges and opportunities in research and development of bone metastasis therapies.

Osteocyte mitochondria inhibit tumor development via STING-dependent antitumor immunity.

Bone is one of the most common sites of tumor metastases. During the last step of bone metastasis, cancer cells colonize and disrupt the bone matrix, which is maintained mainly by osteocytes, the most abundant cells in the bone microenvironment. However, the role of osteocytes in bone metastasis is still unclear. Here, we demonstrated that osteocytes transfer mitochondria to metastatic cancer cells and trigger the cGAS/STING-mediated antitumor response. Blocking the transfer of mitochondria by specifically knocking out mitochondrial Rho GTPase 1 (Rhot1) or mitochondrial mitofusin 2 (Mfn2) in osteocytes impaired tumor immunogenicity and consequently resulted in the progression of metastatic cancer toward the bone matrix. These findings reveal the protective role of osteocytes against cancer metastasis by transferring mitochondria to cancer cells and potentially offer a valuable therapeutic strategy for preventing bone metastasis.

An On-Demand Collaborative Innate-Adaptive Immune Response to Infection Treatment.

Deep tissue infection is a common clinical issue and therapeutic difficulty caused by the disruption of the host antibacterial immune function, resulting in treatment failure and infection relapse. Intracellular pathogens are refractory to elimination and can manipulate host cell biology even after appropriate treatment, resulting in a locoregional immunosuppressive state that leads to an inadequate response to conventional anti-infective therapies. Here, a novel antibacterial strategy involving autogenous immunity using a biomimetic nanoparticle (NP)-based regulating system is reported to induce in situ collaborative innate-adaptive immune responses. It is observed that a macrophage membrane coating facilitates NP enrichment at the infection site, followed by active NP accumulation in macrophages in a mannose-dependent manner. These NP-armed macrophages exhibit considerably improved innate capabilities, including more efficient intracellular ROS generation and pro-inflammatory factor secretion, M1 phenotype promotion, and effective eradication of invasive bacteria. Furthermore, the reprogrammed macrophages direct T cell activation at infectious sites, resulting in a robust adaptive antimicrobial immune response to ultimately achieve bacterial clearance and prevent infection relapse. Overall, these results provide a conceptual framework for a novel macrophage-based strategy for infection treatment via the regulation of autogenous immunity.

A deep supervised transformer U-shaped full-resolution residual network for the segmentation of breast ultrasound image.

PURPOSE: Breast ultrasound (BUS) is an important breast imaging tool. Automatic BUS image segmentation can measure the breast tumor size objectively and reduce doctors' workload. In this article, we proposed a deep supervised transformer U-shaped full-resolution residual network (DSTransUFRRN) to segment BUS images. METHODS: In the proposed method, a full-resolution residual stream and a deep supervision mechanism were introduced into TransU-Net. The residual stream can keep full resolution features from different levels and enhance features fusion. Then, the deep supervision can suppress gradient dispersion. Moreover, the transformer module can suppress irrelevant features and improve feature extraction process. Two datasets (dataset A and B) were used for training and evaluation. The dataset A included 980 BUS image samples and the dataset B had 163 BUS image samples. RESULTS: Cross-validation was conducted. For the dataset A, the proposed DSTransUFRRN achieved significantly higher Dice (91.04 ± 0.86%) than all compared methods (p < 0.05). For the dataset B, the Dice was lower than that for the dataset A due to the small number of samples, but the Dice of DSTransUFRRN (88.15% ± 2.11%) was significantly higher than that of other compared methods (p < 0.05). CONCLUSIONS: In this study, we proposed DSTransUFRRN for BUS image segmentation. The proposed methods achieved significantly higher accuracy than the compared previous methods.

Survey of open science practices and attitudes in the social sciences.

Open science practices such as posting data or code and pre-registering analyses are increasingly prescribed and debated in the applied sciences, but the actual popularity and lifetime usage of these practices remain unknown. This study provides an assessment of attitudes toward, use of, and perceived norms regarding open science practices from a sample of authors published in top-10 (most-cited) journals and PhD students in top-20 ranked North American departments from four major social science disciplines: economics, political science, psychology, and sociology. We observe largely favorable private attitudes toward widespread lifetime usage (meaning that a researcher has used a particular practice at least once) of open science practices. As of 2020, nearly 90% of scholars had ever used at least one such practice. Support for posting data or code online is higher (88% overall support and nearly at the ceiling in some fields) than support for pre-registration (58% overall). With respect to norms, there is evidence that the scholars in our sample appear to underestimate the use of open science practices in their field. We also document that the reported lifetime prevalence of open science practices increased from 49% in 2010 to 87% a decade later.

Cancer cells reprogram to metastatic state through the acquisition of platelet mitochondria.

Metastasis is the major cause of cancer deaths, and cancer cells evolve to adapt to various tumor microenvironments, which hinders the treatment of tumor metastasis. Platelets play critical roles in tumor development, especially during metastasis. Here, we elucidate the role of platelet mitochondria in tumor metastasis. Cancer cells are reprogrammed to a metastatic state through the acquisition of platelet mitochondria via the PINK1/Parkin-Mfn2 pathway. Furthermore, platelet mitochondria regulate the GSH/GSSG ratio and reactive oxygen species (ROS) in cancer cells to promote lung metastasis of osteosarcoma. Impairing platelet mitochondrial function has proven to be an efficient approach to impair metastasis, providing a direction for osteosarcoma therapy. Our findings demonstrate mitochondrial transfer between platelets and cancer cells and suggest a role for platelet mitochondria in tumor metastasis.

14, 15-EET alleviates neurological impairment through maintaining mitochondrial dynamics equilibrium via AMPK/SIRT1/FoxO1 signal pathways in mice with cerebral ischemia reperfusion.

AIM: To explore whether 14, 15-EET regulates mitochondrial dynamics to exert neuroprotective effects after cerebral ischemia-reperfusion and its underlying mechanisms. METHODS: The mouse middle cerebral artery occlusion reperfusion model was used to observe brain infarct volume and neuronal apoptosis by TTC staining and Tunel assay, modified neurological severity score to detect neurological impairment, HE staining and Nissl staining to observe neuron damage, western blot and immunofluorescence methods to detect the expression of mitochondrial dynamics-related proteins, transmission electron microscopy, and Golgi-Cox staining to detect mitochondrial morphology and neuronal dendritic spines. RESULTS: 14, 15-EET reduced the neuronal apoptosis and cerebral infarction volume induced by middle cerebral artery occlusion reperfusion (MCAO/R), inhibited the degradation of dendritic spines, maintained the structural integrity of neurons, and alleviated neurological impairment. Cerebral ischemia-reperfusion induces mitochondrial dynamics disorders, upregulates the expression of the mitochondrial division protein Fis 1, and inhibits the expression of mitochondrial fusion proteins MFN1, MFN2, and OPA1, while 14, 15-EET treatment reverses this process. Mechanistic studies have shown that 14, 15-EET promotes the phosphorylation of AMPK, upregulates the expression of SIRT1 and phosphorylation of FoxO1, thereby inhibiting mitochondrial division and promoting mitochondrial fusion, preserving mitochondrial dynamics, maintaining neuronal morphological and structural integrity, and alleviating neurological impairment induced by middle cerebral artery occlusion reperfusion. Compound C treatment diminishes the neuroprotective effect of 14, 15-EET following MCAO/R in mice. CONCLUSION: This study elucidates the novel neuroprotective mechanism of 14, 15-EET, providing a novel approach for the development of drugs based on mitochondrial dynamics.

Human γδ T cells induce CD8+ T cell antitumor responses via antigen-presenting effect through HSP90-MyD88-mediated activation of JNK.

Human Vγ9Vδ2 T cells have attracted considerable attention as novel alternative antigen-presenting cells (APCs) with the potential to replace dendritic cells in antitumor immunotherapy owing to their high proliferative capacity and low cost. However, the utility of γδ T cells as APCs to induce CD8+ T cell-mediated antitumor immune response, as well as the mechanism by which they perform APC functions, remains unexplored. In this study, we found that activated Vγ9Vδ2 T cells were capable of inducing robust CD8+ T cell responses in osteosarcoma cells. Activated γδ T cells also effectively suppressed osteosarcoma growth by priming CD8+ T cells in xenograft animal models. Mechanistically, we further revealed that activated γδ T cells exhibited increased HSP90 production, which fed back to upregulate MyD88, followed by JNK activation and a subsequent improvement in CCL5 secretion, leading to enhanced CD8+ T cell cross-priming. Thus, our study suggests that Vγ9Vδ2 T cells represent a promising alternative APC for the development of γδ T cell-based tumor immunotherapy.

Noncontact Manipulation of Intracellular Structure Based on Focused Surface Acoustic Waves.

Cell orientation is essential in many applications in biology, medicine, and chemistry, such as cellular injection, intracellular biopsy, and genetic screening. However, the manual cell orientation technique is a trial-and-error approach, which suffers from low efficiency and low accuracy. Although several techniques have improved these issues to a certain extent, they still face problems deforming or disrupting cell membranes, physical damage to the intracellular structure, and limited particle size. This study proposes a noncontact and noninvasive cell orientation method that rotates a cell using surface acoustic waves (SAWs). To realize the acoustic cell orientation process, we have fabricated a microdevice consisting of two pairs of focused interdigital transducers (FIDTs). Instead of rotating the entire cell, the proposed method rotates the intracellular structure, the cytoplasm, directly through the cell membrane by acoustic force. We have tested the rotational manipulation process on 30 zebrafish embryos. The system was able to orientate a cell to a target orientation with a one-time success rate of 93%. Furthermore, the postoperation survival rate was 100%. Our acoustic rotational manipulation technique is noninvasive and easy to use, which provides a starting point for cell-manipulation-essential tasks, such as single-cell analysis, organism studies, and drug discovery.

Surface-Acoustic-Wave (SAW)-Driven Device for Three-Dimensional Cell Manipulation.

Cell orientation is a necessary step in micromanipulation, which significantly affects the outcomes of cell manipulation. Existing cell orientation techniques include the trial-and-error manual approach that suffers from low efficiency, the mechanical contact approaches that have problems of being invasive, and non-contact approaches that are difficult to set up. OBJECTIVE: This paper proposes a system with a surface-acoustic wave (SAWs) to perform noninvasive cell orientation. METHODS: The system employed a SAW chip with a pair of interdigital transducers (IDTs) to rotate embryos around the X and Y axis using acoustic streaming force. Instead of rotating the entire embryos like other methods, the proposed system rotates the cytoplasm alone through the cell chorion. RESULTS: We evaluated the cellular structure recognition algorithm and the rotation control using 100 embryo images and 30 zebrafish embryos. The system successfully recognized all required cellar structures for visual feedback. Furthermore, it rotated all cells into the desired position, including 26 cases completed within 10s with an orientation angle error of less than 4°. All 30 embryos hatched after manipulation. CONCLUSION: The proposed technique can automatically rotate the cytoplasm through the cell chorion noninvasively. SIGNIFICANCE: The system provides a starting point for noninvasive cell manipulation tasks, such as fast intracellular structure scanning and analysis, and microinjection.

Enhancement of T cell infiltration via tumor-targeted Th9 cell delivery improves the efficacy of antitumor immunotherapy of solid tumors.

Insufficient infiltration of T cells severely compromises the antitumor efficacy of adoptive cell therapy (ACT) against solid tumors. Here, we present a facile immune cell surface engineering strategy aiming to substantially enhance the anti-tumor efficacy of Th9-mediated ACT by rapidly identifying tumor-specific binding ligands and improving the infiltration of infused cells into solid tumors. Non-genetic decoration of Th9 cells with tumor-targeting peptide screened from phage display not only allowed precise targeted ACT against highly heterogeneous solid tumors but also substantially enhanced infiltration of CD8+ T cells, which led to improved antitumor outcomes. Mechanistically, infusion of Th9 cells modified with tumor-specific binding ligands facilitated the enhanced distribution of tumor-killing cells and remodeled the immunosuppressive microenvironment of solid tumors via IL-9 mediated immunomodulation. Overall, we presented a simple, cost-effective, and cell-friendly strategy to enhance the efficacy of ACT against solid tumors with the potential to complement the current ACT.

Metabolic control of CD47 expression through LAT2-mediated amino acid uptake promotes tumor immune evasion.

Chemotherapy elicits tumor immune evasion with poorly characterized mechanisms. Here, we demonstrate that chemotherapy markedly enhances the expression levels of CD47 in osteosarcoma tissues, which are positively associated with patient mortality. We reveal that macrophages in response to chemotherapy secrete interleukin-18, which in turn upregulates expression of L-amino acid transporter 2 (LAT2) in tumor cells for substantially enhanced uptakes of leucine and glutamine, two potent stimulators of mTORC1. The increased levels of leucine and enhanced glutaminolysis activate mTORC1 and subsequent c-Myc-mediated transcription of CD47. Depletion of LAT2 or treatment of tumor cells with a LAT inhibitor downregulates CD47 with enhanced macrophage infiltration and phagocytosis of tumor cells, and sensitizes osteosarcoma to doxorubicin treatment in mice. These findings unveil a mutual regulation between macrophage and tumor cells that plays a critical role in tumor immune evasion and underscore the potential to intervene with the LAT2-mediated amino acid uptake for improving cancer therapies.

Compartmentalized Janus droplets of photoresponsive cholesteric liquid crystals and poly(dimethylsiloxane)-based oligomers.

A new kind of Janus droplet containing photoresponsive cholesteric liquid crystals (CLCs) was fabricated for the first time and their formation, compartment structure, mesophase texture and function were thoroughly investigated. In the droplets, the CLC compartments included a typical nematic LC (4'-pentyl-4-biphenylcarbonitrile) doped with an azobenzene-containing chiral dopant, and the other compartments were formed of a poly(dimethylsiloxane)-based oligomer. Janus droplets were fabricated through microphase separation of the incompatible components in chloroform solution dispersed in an aqueous medium, induced by slow evaporation of chloroform. The mesophase structures of the CLC phase in Janus droplets, both suspended in aqueous medium and spreading on substrates, were controlled by the bulk elastic free energy of the CLC phase, surface anchoring and confining geometries. The helix pitch of the cholesteric phase in the droplets was determined by the doping concentration of the chiral dopant. For the suspended Janus droplets with the helix pitch obviously smaller than the droplet sizes, the CLC compartments mainly possessed a bipolar structure instead of the Frank-Pryce structure typically observed on CLC droplets. After the Janus droplets spread on the substrates, the CLC compartments changed to crescent shapes due to the different wettability characteristics of the two compartments, and the formed stable and metastable CLC configurations were distinctively different from those in the suspensions. Interestingly, when the Janus droplets spreading on substrates were irradiated with a laser beam (λ = 488 nm) of low intensity, the directors in the CLC compartments rearranged to form fingerprint structures with minimum total energy.

Predictive and Prognostic Biomarkers for Lung Cancer Bone Metastasis and Their Therapeutic Value.

Lung cancer is the leading cause of cancer-related death worldwide. Bone metastasis, which usually accompanies severe skeletal-related events, is the most common site for tumor distant dissemination and detected in more than one-third of patients with advanced lung cancer. Biopsy and imaging play critical roles in the diagnosis of bone metastasis; however, these approaches are characterized by evident limitations. Recently, studies regarding potential biomarkers in the serum, urine, and tumor tissue, were performed to predict the bone metastases and prognosis in patients with lung cancer. In this review, we summarize the findings of recent clinical research studies on biomarkers detected in samples obtained from patients with lung cancer bone metastasis. These markers include the following: (1) bone resorption-associated markers, such as N-terminal telopeptide (NTx)/C-terminal telopeptide (CTx), C-terminal telopeptide of type I collagen (CTx-I), tartrate-resistant acid phosphatase isoform 5b (TRACP-5b), pyridinoline (PYD), and parathyroid hormone related peptide (PTHrP); (2) bone formation-associated markers, including total serum alkaline phosphatase (ALP)/bone specific alkaline phosphatase(BAP), osteopontin (OP), osteocalcin (OS), amino-terminal extension propeptide of type I procollagen/carboxy-terminal extension propeptide of type I procollagen (PICP/PINP); (3) signaling markers, including epidermal growth factor receptor/Kirsten rat sarcoma/anaplastic lymphoma kinase (EGFR/KRAS/ALK), receptor activator of nuclear factor κB ligand/receptor activator of nuclear factor κB/osteoprotegerin (RANKL/RANK/OPG), C-X-C motif chemokine ligand 12/C-X-C motif chemokine receptor 4 (CXCL12/CXCR4), complement component 5a receptor (C5AR); and (4) other potential markers, such as calcium sensing receptor (CASR), bone sialoprotein (BSP), bone morphogenetic protein 2 (BMP2), cytokeratin 19 fragment/carcinoembryonic antigen (CYFRA/CEA), tissue factor, cell-free DNA, long non-coding RNA, and microRNA. The prognostic value of these markers is also investigated. Furthermore, we listed some clinical trials targeting hotspot biomarkers in advanced lung cancer referring for their therapeutic effects.

Clinical Features and Serological Markers Risk Model Predicts Overall Survival in Patients Undergoing Breast Cancer and Bone Metastasis Surgeries.

BACKGROUND: Surgical therapy of breast cancer and bone metastasis can effectively improve the prognosis of breast cancer. However, after the first operation, the relationship between preoperative indicators and outcomes in patients who underwent metastatic bone surgery remained to be studied. Purpose 1. Recognize clinical and laboratory prognosis factors available to clinical doctors before the operation for bone metastatic breast cancer patients. 2. Develop a risk prediction model for 3-year postoperative survival in patients with breast cancer bone metastasis. METHODS: From 2014 to 2020, patients who suffered from breast cancer bone metastasis and received therapeutic procedures in our institution were included for analyses (n=145). For patients who underwent both breast cancer radical surgery and bone metastasis surgery, comprehensive datasets of the parameters of interest (clinical features, laboratory factors, and patient prognoses) were collected (n=69). We performed Multivariate Cox regression to identify factors that were associated with postoperative outcome. 3-year survival prediction model and nomograms were established by 100 bootstrapping. Its benefit was evaluated by calibration plot, C-index, and decision curve analysis. The Surveillance, Epidemiology, and End Results database was also used for external validation. RESULTS: Radiotherapy for primary cancer, pathological type of metastatic breast cancer, lymph node metastasis, elevated serum alkaline phosphatase, lactate dehydrogenase were associated with postoperative prognosis. Pathological types of metastatic breast cancer, multiple bone metastasis, organ metastases, and elevated serum lactate dehydrogenase were associated with 3-year survival. Then those significant variables and serum alkaline phosphatase counts were integrated to construct nomograms for 3-year survival. The C-statistic of the established predictive model was 0.83. The calibration plot presents a graphical representation of calibration. In the decision curve analysis, the benefits are higher than those of the extreme curve. The receiver operating characteristic of the external validation of the model was 0.82, indicating a favored fitting degree of the two models. CONCLUSION: Our study suggests that several clinical features and serological markers can predict the overall survival among the patients who are about to receive bone metastasis surgery after breast cancer surgery. The model can guide the preoperative evaluation and clinical decision-making for patients. Level of evidence Level III, prognostic study.